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  • sgp120 and the contact system in hereditary angioedema: A diagnostic tool in HAE with normal C1 inhibitor
    Mol. Immunol. (IF 3.064) Pub Date : 2020-01-16
    Blas Larrauri; C. Garren Hester; Haixiang Jiang; Vojislav D. Miletic; Alejandro Malbran; Konrad Bork; Allen Kaplan; Michael Frank

    Mutations in Factor XII, plasminogen gene, angiopoietin-1 gene and kininogen 1 gene have been found in some patients with hereditary angioedema with normal C1 inhibitor (HAE-nl-C1inh), but the underlying disease mechanisms remain unclear. Additionally, there are no accepted biomarkers for this disease. Because the contact system has been implicated in hereditary angioedema with C1 inhibitor deficiency (HAE-C1inh), we studied the fragmentation patterns of serum glycoprotein 120 (sgp120), a protein that is highly susceptible to cleavage by kallikrein, in 31 HAE-C1inh and 13 HAE-nl-C1inh patient plasma samples. Compared to normal controls, the majority of plasma samples from patients with HAE-C1inh contained fragmented sgp120. These samples also showed increased kallikrein amidolytic activity indicating spontaneous contact system activation. In contrast, most samples from HAE-nl-C1inh patients exhibited intact sgp120. However, if these samples were incubated at 4 °C in plastic, significant sgp120 fragmentation and spontaneous contact system activation were observed. Concurrently, there was C1 inhibitor fragmentation that generated the nonfunctional 94 kD fragment and a reduction in C1 inhibitor function. Normal samples did not show sgp120 or C1 inhibitor fragmentation after incubation. We sequenced sgp120 and found it to be identical to inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4). These results suggest that sgp120 or ITIH4 is cleaved when the contact system is activated and that this cleavage could be used as a biomarker in patients with HAE-nl-C1inh.

    更新日期:2020-01-17
  • Function characterization and expression regulation of two different-sized 3’ untranslated region-containing interferon genes from clone F of gibel carp Carassius auratus gibelio
    Mol. Immunol. (IF 3.064) Pub Date : 2020-01-15
    Zhao-Xing Zang; Cheng Dan; Li Zhou; Qi-Ya Zhang; Jian-Fang Gui; Yi-Bing Zhang

    Fish interferon (IFN)-mediated antiviral innate immunity is the first line of defense against virus invasion. In the present study, we identify two fish IFN genes (here tentatively named IFNa and IFNc) with different-sized 3′ UTRs from clone F strain of gibel carp Carassius auratus gibelio. Carp IFNa has a relatively short 3′UTR without AU-rich elements (AREs) but IFNc has a long one with 9 AREs. Functionally, carp IFNa and IFNc display significantly antiviral potential to viral infection, likely through induction of downstream IFN-stimulated genes (ISGs). Both carp IFN genes are induced by viral infection, poly(I:C) treatment and IRF3/7, which are ascribed to the IFN-sensitive response elements (ISRE) within their promoters. Carp IFN genes are also induced by each other and by themselves, indicating existence of a positive feedback loop in fish IFN-mediated antiviral immune response. Comparative analyses of 3′UTR-mediated expression regulation at mRNA and protein levels show that the ARE-containing 3′UTR of carp IFNc rather than the short 3′UTR of carp IFNa promotes mRNA decay but instead results in high-level protein expression, indicating that 3′UTR of fish IFN mRNAs might be a potential factor for regulation of IFN-mediated antiviral immune response. Considering a fact that a given protein function is largely related to its protein level, these results suggest that both promoter and 3′UTR contribute to the transcription and translation of fish IFN genes, thus shaping their eventually antiviral potential.

    更新日期:2020-01-15
  • Molecular and functional characterization of ovis aries IFN-epsilon
    Mol. Immunol. (IF 3.064) Pub Date : 2020-01-08
    Yongli Guo; Zhifeng Song; Xintong Cheng; Yujiao Wang; Xiuxin Luo; Ran An; Junwei Wang; Mingchun Gao

    Interferon-epsilon (IFN-ε) is a type I IFN playing an essential role in innate and adaptive immunity against viral infection. Ovis aries IFN-ε (OvIFN-ε), consisting of 582 bp and which encodes a protein of 193 amino acids containing a signal peptide of 21 amino acids, was cloned and characterized. OvIFN-ε shares 51.6∼ 86.5% similarity to other species of IFN-ε, and evolves from the IFN-ε branch but not the other types of IFN. Additionally, OvIFN-ε gene is well conserved during evolution, and is highly transcribed in the liver, lung, brain, skin, ovary and uterus. Recombinant protein of OvIFN-ε was expressed in Escherichia coli and purified with nickel chelated column, which exhibited broad antiviral activity in vitro, sensitivity of trypsin, and stability of pH and temperature to some extent. Furthermore, OvIFN-ε could induce the transcription of ISG15, Mx1 and OAS in a time-dependent manner, as well as inhibit the VSV and BVDV replication in Ovis aries peripheral blood lymphocyte cells and MDBK cells. This study revealed OvIFN-ε has the typical characterization of type I IFNs and exerts antiviral activity against VSV and BVDV, and induces the expression of ISGs, which not only enriches the understanding of IFN-ε, but also facilitates further research on the antiviral defense responses of Ovis aries.

    更新日期:2020-01-09
  • Molecular characterization of a MOSPD2 homolog in the barbel steed (Hemibarbus labeo) and its involvement in monocyte/macrophage and neutrophil migration
    Mol. Immunol. (IF 3.064) Pub Date : 2020-01-09
    Wei Jiang; Jie Chen; Zhi-Ping Guo; Le Zhang; Guang-Ping Chen

    Motile sperm domain containing 2 (MOSPD2) is a single-pass membrane protein to which until recently little function had been ascribed. Although its mammalian homologs have been identified, the status of the mospd2 gene in lower vertebrates is still unknown. In the present study, cDNA of the mospd2 gene of barbel steed (Hemibarbus labeo) was cloned and sequenced to characterize its potential involvement in the innate immune system of this fish. Sequence analysis revealed that the predicted barbel steed MOSPD2 protein contained an N-terminal extracellular portion composed of a CRAL-TRIO domain, a motile sperm domain, and a transmembrane domain, as well as a short C-terminal intracellular domain. Phylogenetic tree analysis indicated that barbel steed MOSPD2 is closely related to that of zebrafish. Barbel steed mospd2 transcripts were detected in a wide range of tissues, with the highest level being found in the gill. In response to lipopolysaccharide (LPS) treatment or Aeromonas hydrophila infection, mospd2 gene expression was significantly altered in the head kidney, spleen, and mid-intestine. The expression of mospd2 gene was detected in monocytes/macrophages (MO/MФ), neutrophils, and lymphocytes, and was found to be mainly expressed in MO/MФ. At the same time, using flow cytometry, we also confirmed that MOSPD2 protein is located on MO/MФ, neutrophil, and lymphocyte membranes. Following treatment with LPS or A. hydrophila, MOSPD2 protein expression was induced in these immune cells. The migration of MO/MФ and neutrophils decreased significantly upon MOSPD2 blockade with anti-MOSPD2 IgG in a dose-dependent manner, whereas this treatment had no significant effect on lymphocytes migration. To the best of our knowledge, our study, for the first time, provides evidence that MOSPD2 mediates the migration of MO/MФ and neutrophils in a fish species.

    更新日期:2020-01-09
  • Identification, cloning, and immunological studies on a major eggplant (Solanum melongena L.) allergen Sola m 1: A new member of profilin allergen family
    Mol. Immunol. (IF 3.064) Pub Date : 2020-01-02
    Sukhendu Maity; Subham Bhakta; Moumita Bhowmik; Gaurab Sircar; Swati Gupta Bhattacharya
    更新日期:2020-01-02
  • Up-regulation of PTEN via LPS/AP-1/NF-κB pathway inhibits trophoblast invasion contributing to preeclampsia
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-30
    Pingping Xue; Wenqiang Fan; Zhenyu Diao; Yujing Li; Chengcai Kong; Xiuliang Dai; Yue Peng; Li Chen; Huiyan Wang; Yali Hu; Zhibin Hu

    Preeclampsia, a pregnancy-specific disorder, is characterized by abnormal vascular remodeling of the spiral arteries due to deficient trophoblast invasion. Lipopolysaccharide (LPS) administration to pregnant rats on day 5 of pregnancy could induce excessive immune response at the maternal-fetal interface contributing to poor early placentation that culminate in the preeclampsia-like syndrome. Furthermore, the expression of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), a critical tumor suppressor, is markedly increased in the placentas of patients with preeclampsia. Our goal was to investigate the association of PTEN with preeclampsia and the pathways involved using human-trophoblast-derived cell line (HTR-8/SVneo) stimulated with LPS. We found that the expression of PTEN was significantly increased in the placentas of patients with severe preeclampsia and preeclamptic rat model induced by LPS. In vitro trophoblasts results showed that significantly differential expression of PTEN with corresponding changes in JunB/FosB (subunits of AP-1) and NF-κB activity after LPS stimulation. We further demonstrated that LPS-induced PTEN expression was dependent on AP-1 and NF-κB in trophoblasts. The trophoblasts with enforced expression of PTEN showed a reduced ability to invasion. Taken together, LPS may undermine remodelling of the human-trophoblast-derived HTR-8/SVneo cells by increasing PTEN, acting in part through the AP-1 and NF-κB pathways.

    更新日期:2019-12-31
  • BMSC-EVs regulate Th17 cell differentiation in UC via H3K27me3
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-30
    Qianyun Chen; Xueyun Duan; Meng Xu; Heng Fan; Yalan Dong; Hui Wu; Man Zhang; Yujin Liu; Zhen Nan; Shuangjiao Deng; Xingxing Liu

    In our previous studies, we found that extracellular vesicles in mesenchymal stem cells can alleviate ulcerative colitis. In view of the fact that extracellular vesicles have the same immunomodulatory effects as their maternal cells and considering the important role of Th17 cells in the pathogenesis of ulcerative colitis, we aimed to investigate whether extracellular vesicles from mesenchymal stem cells can affect the differentiation of Th17 cells in ulcerative colitis. Histone H3K27me3 can regulate the expression of Th17 cell-related genes. We focused on determining whether the effect of extracellular vesicles on Th17 cells in ulcerative colitis is related to H3K27me3. For our experiments, we used low, medium and high doses of extracellular vesicles from mesenchymal stem cells to interfere with TNBS-induced colitis in rats and then evaluated the alleviation of inflammation and observed the impact of the extracellular vesicles on the differentiation of Th17 cells in ulcerative colitis. In addition, we detected the levels of histone H3K27me3 and the expression of its upstream methyltransferase and demethylase in the colon tissues of each group. Our data showed that extracellular vesicles from bone marrow mesenchymal stem cells can inhibit the abnormal differentiation of Th17 cells in ulcerative colitis, and the content of histone H3K27me3 was also changed accordingly. Our study suggests that extracellular vesicles from mesenchymal stem cells could inhibit the differentiation of Th17 cells in ulcerative colitis by regulating H3K27me3. This study reveals that H3K27me3 is an important target for inflammatory immune diseases associated with abnormal Th17 cell differentiation and indicates that mesenchymal stem cell extracellular vesicles are promising agents for the treatment of ulcerative colitis.

    更新日期:2019-12-31
  • Trigonelline: An alkaloid with anti-degranulation properties
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-30
    Arita Dewi Nugrahini; Momoko Ishida; Toshifumi Nakagawa; Kosuke Nishi; Takuya Sugahara

    Trigonelline, one of the alkaloids contained in coffee, is important not only as one of the constituents of aroma and flavor in coffee but also as a useful source of nutrition. Its anti-microbial, anti-carcinogenic, and anti-hyperglycemic effects have been investigated in previous studies. However, there have not been any studies examining the anti-degranulation effect of trigonelline. In this study, the anti-degranulation effect of trigonelline was evaluated in in vitro and in vivo models using a rat basophilic leukemia cell line, RBL-2H3 cells, and a passive cutaneous anaphylaxis (PCA) reaction in mice, respectively. In the β-hexosaminidase release assay, trigonelline effectively suppressed antigen-induced degranulation of RBL-2H3 cells in a dose-dependent manner without cytotoxicity. Trigonelline also inhibited FcεRI-mediated intracellular signaling pathways, such as phosphorylation of PLCγ1, PI3 K, and Akt, in antigen-stimulated RBL-2H3 cells and suppressed the PCA response in mice. Moreover, trigonelline also inhibited the microtubule formation in RBL-2H3 cells, indicating that trigonelline could inhibit IgE-sensitized mast cell degranulation by attenuating both the intracellular calcium-dependent and independent pathways. These results revealed that trigonelline possesses the anti-degranulation effect against the development of allergic diseases.

    更新日期:2019-12-31
  • Expressing the immunodominant projection domain of infectious bursal disease virus fused to the fragment crystallizable of chicken IgY in yellow maize for a prospective edible vaccine
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-24
    Reda Salem; Shireen K. Assem; Omar A. Omar; Ahmed A. Khalil; Mahmoud A. Basry; Fatma R. Waly; Noha Samir; Alaa A. El-Kholy

    Control of Infectious bursal disease virus (IBDV) in endemic countries has been based on early immunization of chicks using conventional live or inactivated vaccines that became not fully effectual and have biosafety concerns. This endeavor seeks generating a recombinant chimeric protein merging the projection domain (PD) of IBDV VP2 capsid with the fragment crystallizable (Fc) of avian IgY (FcIgY), in maize as a prospective poultry edible vaccine. The PD sequence was built on the basis of very virulent IBDV isolates circulating in Egypt. After optimization of codon-usage in maize, sequences of PD and FcIgY were effectively expressed in two elites of yellow maize via bombardment transformation in immature embryos. Chimeric protein amount in stable transgenic samples ranged from1.36% to 3.03% of the total soluble protein based on tissue age and maize cultivar. IBDV VP2 coding sequence was amplified from viral RNA, cloned, and expressed in E. coli. A group of Balb/C mice were hyper-immunized with purified recombinant VP2 protein for raising anti- recombinant VP2 antibodies (anti-rVP2 Ab). Proper expression in maize and immunoreactivity of the chimeric protein (PD-FcIgY) to chicken anti- IBDV and anti-rVP2 Ab were confirmed by both direct and indirect double antibody sandwich (DAS)-ELISAs as well as western blotting. Seeds of regenerated transgenic maize will be validated for chickens as edible vaccination in further studies.

    更新日期:2019-12-27
  • Myeloid-driven mechanisms as barriers to antitumor CD8+ T cell activity
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-26
    Sean H. Colligan; Stephanie L. Tzetzo; Scott I. Abrams

    The adaptive immune system is essential for host defense against pathogenic challenges, and a major constituent is the CD8+ cytotoxic T cell. Ordinarily, CD8+ T cells are endowed with a unique ability to specifically recognize and destroy their targets. However, in cases where disease emerges, especially in cancer, the efficacy of the CD8+ T cell response is frequently counterbalanced in a ‘tug-of-war’ by networks of tumor-driven mechanisms of immune suppression. As a result, antitumor CD8+ T cell activity is hampered, which contributes to clinical manifestations of disease. It is now well-recognized that prominent elements of that network include myeloid-derived suppressor cells (MDSC) and macrophages which assume tumor-supportive phenotypes. Both myeloid populations are thought to arise as consequences of chronic inflammatory cues produced during the neoplastic process. Numerous preclinical studies have now shown that inhibiting the production, trafficking and/or function of these immune suppressive myeloid populations restore antitumor CD8+ T cell responses during both immune surveillance or in response to immune-targeted interventions. Correlative studies in cancer patients support these preclinical findings and, thus, have laid the foundation for ongoing clinical trials in patients receiving novel agents that target such myeloid elements alone or in combination with immunotherapy to potentially improve cancer patient outcomes. Accordingly, this review focuses on how and why it is important to study the myeloid-T cell interplay as an innovative strategy to boost or reinvigorate the CD8+ T cell response as a critical weapon in the battle against malignancy.

    更新日期:2019-12-27
  • Reduced activated regulatory T cells and imbalance of Th17/activated Treg cells marks renal involvement in ANCA-associated vasculitis
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-26
    Yanlin Wang; Shulan Zhang; Na Zhang; Min Feng; Zhaojun Liang; Xiangcong Zhao; Chong Gao; Yan Qin; Yanyao Wu; Guangying Liu; Jinfang Zhao; Hui Guo; Jing Luo

    The role of naturally occurring regulatory T cells (Treg) in the control of the immune tolerance of ANCA-associated vasculitis (AAV) has not been well defined. Therefore, we separate the phenotypically heterogeneous Treg cells into different subsets based on the expression of FOXP3 and CD45RA during AAV pathogenesis. Fifty-four AAV patients (38 patients with renal involvement) and 19 healthy controls (HCs) were enrolled in this study. Levels of CD4+T cell subsets and cytokines were detected by flow cytometry. Treg immunesuppression capacity was measured in co-culture experiments. The diagnostic value for Treg subsets was evaluated by the areas under the receiver operating characteristic curves (AUC). Patients with AAV had lower percentages and numbers of activated Treg cells (aTreg, P = 0.044, P = 0.002), while higher levels of total Treg cells (P = 0.001, P = 0.026) with diminished immunosuppression capacity. The proportions of effector memory T-cell subpopulation (P < 0.001) were increased in AAV patients. Interestingly, the AUC of the aTreg improved significantly the diagnostic potential of AAV. Furthermore, the ratio of Th17/aTreg was significantly increased in active and renal vasculitis patient and positive correlation between Th17/Treg subset ratio and creatinine or BUN. In addition, we found that cytokine IL-2 and IL-4 exhibited a downward while IL-6, IL-10, TNF-α, IFN-γ and IL-17A trend upward in AAV patients. Increase in total Treg levels, along with functional deficiency, and decrease in aTreg cells constitute potential novel biomarkers for AAV. And the ratio of Th17/aTreg might serve as an important tool to recognize and monitor AAV patients with renal involvement and disease remission.

    更新日期:2019-12-27
  • Exosomal long non-coding RNA GAS5 suppresses Th1 differentiation and promotes Th2 differentiation via downregulating EZH2 and T-bet in allergic rhinitis
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-13
    Xiaoyuan Zhu; Xueping Wang; Ying Wang; Yulin Zhao

    The imbalance of helper T cell (Th) 1/Th2 differentiation is involved in the development of allergic rhinitis (AR). Recent studies reveal the regulatory function of exosomes on Th1/Th2 differentiation. However, the key mediator in exosomes that modulate such response remains unclear. In this study, the expression of long-noncoding RNA GAS5 (LncGAS5) was detected in exosomes which were isolated from AR patient nasal mucus (AR-EXO) and ovalbumin (OVA)-stimulated nasal epithelial cells (OVA-EXO). Th1/Th2 differentiation was induced in naïve CD4+ T cells, and the percentage of IFN-γ expressing cells (Th1 cells) and IL-4 expressing cells (Th2 cells) was detected using flow cytometry. The result showed that LncGAS5 was upregulated in AR epithelial samples, AR-EXO, and OVA-EXO. The coincubation of AR-EXO and CD4+ T cells suppressed Th1 differentiation and promoted Th2 differentiation, which is mediated by LncGAS5 in AR-EXO. The LncGAS5 in AR-EXO inhibited transcription and expression of EZH2, and it also inhibited T-bet expression at mRNA and protein levels. The gain-of-function and loss-of-function experiments suggested that LncGAS5 mediates Th1/Th2 differentiation partly through downregulating T-bet and EZH2. In summary, our findings demonstrated that LncGAS5 in AR epithelium-derived exosomes is the key mediator in Th1/Th2 differentiation, providing a possible therapeutic target of AR.

    更新日期:2019-12-27
  • T and B lymphocytes immune responses in flounder (Paralichthys olivaceus) induced by two forms of outer membrane protein K from Vibrio anguillarum: Subunit vaccine and DNA vaccine
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-13
    Jing Xing; Zhiqi Zhang; Keke Luo; Xiaoqian Tang; Xiuzhen Sheng; Wenbin Zhan

    To further elucidate the roles of T and B lymphocytes in fish subunit and DNA candidate vaccines for immunisation, the immune responses of T and B lymphocytes to recombinant protein (rOmpK) and plasmid OmpK (pOmpK) from Vibrio anguillarum plus cyclosporine A (CsA) were investigated in flounder (Paralichthys olivaceus). The results showed that in the rOmpK-immunised groups, the percentages of CD4-1+ and CD4-2+ T (PCD4-1+ and PCD4-2+ T) lymphocytes significantly increased to a peak on days 5 or 7. The percentages of IgM+ B (PIgM+ B) lymphocytes and specific antibodies markedly increased to a peak at weeks 4 or 5. The nine immune-related genes were significantly up-regulated and the expression levels of CD4-1, CD4-2 and MHC II genes were higher than that of CD8α, CD8β and MHC I genes. The CD4+ T lymphocytes, IgM+ B lymphocytes, and specific antibodies were significantly inhibited by CsA. Therefore, the responses of CD4+ T lymphocytes influenced the responses of the B lymphocytes and antibodies. In the pOmpK-immunised groups, the PCD4-1+, PCD4-2+, and PCD8β+ T lymphocytes significantly increased to a peak on days 11 or 14, days 9 or 11, and days 7 or 9, respectively. The PIgM+ B lymphocytes and specific antibodies significantly increased to a peak at weeks 5 or 6. Immune related genes upregulated, and CD4+ and CD8+ T lymphocytes, IgM+ B lymphocytes and specific antibodies all suppressed by CsA, suggesting that the responses of T lymphocytes subpopulations influenced B lymphocytes and antibodies responses. Therefore, the subpopulations of T lymphocytes played an important role in the immune responses induced by subunit and DNA candidate vaccines of OmpK and regulated the immune responses of B lymphocytes in flounder.

    更新日期:2019-12-27
  • Interleukin-22 regulates interferon lambda expression in a mice model of pseudomonas aeruginosa pneumonia
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-16
    Alexis Broquet; Anissa Besbes; Jérôme Martin; Cédric Jacqueline; Mickaël Vourc’h; Antoine Roquilly; Jocelyne Caillon; Régis Josien; Karim Asehnoune

    Background Interleukin (IL)-22 is a cytokine involved in tissue protection and repair following lung pathologies. Interferon (IFN)-λ cytokines displayed similar properties during viral infection and a synergy of action between these two players has been documented in the intestine. We hypothesize that during Pseudomonas aeruginosa challenge, IL-22 up-regulates IFN-λ and that IFN-λ exhibits protective functions during Pseudomonas aeruginosa acute pneumonia model in mice. Methods Using an in vitro human alveolar epithelial cell line A549, we assessed the ability of IL-22 to enhance IFN-λ expression during infection. IFN-λ protective function was evaluated in an acute mouse pneumonia model. Results We first demonstrated in murine lungs that only type-II alveolar cells express IL-22 receptor and that IL-22 treatment of A549 cell line up-regulates IFN-λ expression. In a murine acute pneumonia model, IL-22 administration maintained significant IFN-λ levels in the broncho-alveolar fluids whereas IL-22 neutralization abolished IFN-λ up-regulation. In vivo administration of IFN-λ during Pseudomonas aeruginosa pneumonia improves mice outcome by dampening neutrophil recruitment and decreasing epithelium damages. Discussion We show here that IL-22 regulates IFN-λ levels during Pseudomonas aeruginosa pneumonia.

    更新日期:2019-12-27
  • Effect of dehydroepiandrosterone on the immune response and gut microbiota in dextran sulfate sodium-induced colitis mice
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-17
    Ji Cao; Huihui Zhang; Zhongmiao Yang; Jinlong Zhao; Haitian Ma

    Dehydroepiandrosterone (DHEA) possess anti-inflammatory, anti-oxidant and immune-regulating function in animals and humans, but there is not enough information about the mechanisms underlying its beneficial effects. The present study investigated the effect and mechanism of DHEA in dextran sulfate sodium (DSS)-induced colitis mice. The findings showed that DHEA relieved the decreasing of body weight, the increasing of disease activity index, the enhancing of spleen weight, the shortening of colon length and the rising of myeloperoxidase activity; meanwhile, histopathological analysis showed that DHEA maintained a relatively intact structure of colon in DSS-induced colitis mice. DHEA decreased the malondialdehyde content, superoxide dismutase activity and inducible nitric oxide synthase protein level; meanwhile, DHEA also inhibited the secretion of tumor necrosis factor-α, interleukin-1β and interleukin-6 in DSS-induced colitis mice. Importantly, our results showed that DHEA blocked the activation of nuclear factor-kappa B (NF-κB) and p38 mitogen-activated protein kinase (MAPK) pathways; and it inhibited the Nod-like receptor protein 3 inflammasome activation in DSS-induced colitis mice. Furthermore, DHEA markedly promoted the intestinal barrier function by up-regulation zonula occludens-1 expression level. The 16S rDNA gene sequencing demonstrated that DHEA decreased the Pseudomonas abundance in DSS-induced colitis mice. In conclusion, our data demonstrated that DHEA reduces oxidative damage through regulating antioxidant enzyme activity; inhibits pro-inflammatory cytokines production by blocking the activation of p38 MAPK and NF-κB signal pathway; protects colon barrier integrity via increasing tight junction protein expression and modulating gut microbiota taxa; all that finally alleviates DSS-induced experimental colitis in mice.

    更新日期:2019-12-27
  • 2′-5′-Oligoadenylate synthetase-like protein inhibits intracellular M. tuberculosis replication and promotes proinflammatory cytokine secretion
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-17
    G. Leisching; A. Ali; V. Cole; B. Baker

    Host cytoplasmic surveillance pathways are known to elicit type I interferon (IFN) responses which are crucial to antimicrobial defense mechanisms. Oligoadenylate synthetase-like (OASL) protein has been extensively characterized as a part of the anti-viral mechanism, however a number of transcriptomic studies reveal its upregulation in response to infection with a wide variety of intracellular bacterial pathogens. To date, there is no evidence documenting the role (if any) of OASL during mycobacterium tuberculosis infection. Using two pathogenic strains differing in virulence only, as well as the non-pathogenic M. bovis BCG strain, we observed that pathogenicity and virulence strongly induced OASL expression after 24 h of infection. Further, we observed that OASL knock down led to a significant increase in M. tb CFU counts 96 h post-infection in comparison to the respective controls. Luminex revealed that OASL silencing significantly decreased IL-1β, TNF-α and MCP-1 secretion in THP-1 cells and had no effect on IL-10 secretion. We therefore postulate that OASL regulates pro-inflammatory mediators such as cytokines and chemokines which suppress intracellular mycobacterial growth and survival.

    更新日期:2019-12-27
  • p38 mitogen-activated protein kinases (MAPKs) are involved in intestinal immune response to bacterial muramyl dipeptide challenge in Ctenopharyngodon idella
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-17
    Yuandong Sun; Wenqian Xu; Die Li; Hui Zhou; Fufa Qu; Shenping Cao; Jianzhou Tang; Yonghua Zhou; Zhimin He; Hang Li; Zhigang Zhou; Zhen Liu

    The p38 mitogen-activated protein kinases (MAPKs) are essential cytoplasmic signal molecules of innate immune pathways that play a vital role in host immune defense responses to pathogenic challenges. In this study, two fish p38 genes (Cip38α and Cip38β) were characterized for the first time from the grass carp Ctenopharyngodon idella. Similar to other reported p38MAPKs, both Cip38α and Cip38β contained a conserved phosphorylation motif (Thr-Gly-Tyr, TGY) and a substrate binding site (Ala-Thr-Arg-Trp, ATRW) in the serine/threonine protein kinase (S_TKc) domain. Expression profile analysis showed that Cip38α and Cip38β mRNAs were broadly expressed in all of the examined tissues and developmental stages of C. idella. In addition, in vivo injection experiments directly revealed that Cip38α and Cip38β showed strong responsiveness to Aeromonas hydrophila and muramyl dipeptide (MDP) challenges, and their expression levels were significantly upregulated in the intestine of grass carp. Additionally, the MDP-induced expression levels of intestinal inflammatory cytokines (TNF-α and IL-15) and an antimicrobial peptide (β-defensin) were significantly inhibited by the p38MAPK-specific inhibitor SB203580. Moreover, the nutritional dipeptide carnosine and Ala-Gln were found to significantly suppress the bacterial MDP-induced expression of p38MAPK pathway genes and inflammatory cytokines in the intestine of grass carp. Finally, overexpression analysis demonstrated that Cip38α and Cip38β could act as efficient activators in the regulation of AP-1 signaling pathways through interaction with CiMKK6. Altogether, this study provided experimental evidence of the presence of a functional p38 pathway in grass carp, which revealed its involvement in the intestinal immune response to bacterial challenges in bony fish.

    更新日期:2019-12-27
  • Complement Receptor 1 (CR1/CD35)-expressing retinal pigment epithelial cells as a potential therapy for age-related macular degeneration
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-26
    Ken T. Simmons; John L. Mazzilli; Stacey L. Mueller-Ortiz; Aleksey Y. Domozhirov; Charles A. Garcia; Eva M. Zsigmond; Rick A. Wetsel

    The purpose of this study was to identify a membrane-bound complement inhibitor that could be overexpressed on retinal pigment epithelial cells (RPE) providing a potential therapy for age-related macular degeneration (AMD). This type of therapy may allow replacement of damaged RPE with cells that are able to limit complement activation in the retina. Complement Receptor 1 (CR1) is a membrane-bound complement inhibitor commonly found on erythrocytes and immune cells. In this study, QPCR and flow cytometry data demonstrated that CR1 is not well-expressed by RPE, indicating that its overexpression may provide extra protection from complement activation. To screen CR1 for this ability, a stable CR1-expressing ARPE19 line was created using a combination of antibiotic selection and FACS. Cell-based assays were used to demonstrate that addition of CR1 inhibited deposition of complement proteins C3b and C6 on the transfected line. In the end, this study identifies CR1 as a complement inhibitor that may be overexpressed on stem cell-derived RPE to create a potential “enhanced” cell therapy for AMD. A combination cell/complement therapy may create transplantable RPE better suited to avoid complement-mediated lysis and limit chronic inflammation in the retina.

    更新日期:2019-12-27
  • Carboplatin chemoresistance is associated with CD11b+/Ly6C+ myeloid release and upregulation of TIGIT and LAG3/CD160 exhausted T cells
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-18
    Doxakis Anestakis; Savvas Petanidis; Kalliopi Domvri; Drosos Tsavlis; Paul Zarogoulidis; Theodora Katopodi
    更新日期:2019-12-27
  • Single-domain antibodies for functional targeting of the signaling scaffold Shoc2
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-20
    HyeIn Jang; Patricia G. Wilson; Mary Sau; Udeep Chawla; David W. Rodgers; Emilia Galperin

    The accurate transmission of signals by the canonical ERK1/2 kinase pathway critically relies on the proper assembly of an intricate multiprotein complex by the scaffold protein Shoc2. However, the details of the mechanism by which Shoc2 guides ERK1/2 signals are not clear, in part, due to the lack of research tools targeting specific protein binding moieties of Shoc2. We report generation and characterization of single domain antibodies against human Shoc2 using a universal synthetic library of humanized nanobodies. Our results identify eight synthetic single-domain antibodies and show that two evaluated antibodies have binding affinities to Shoc2 in the nanomolar range. High affinity antibodies were uniquely suited for the analysis of the Shoc2 complex assembly. Selected single-domain antibodies were also functional in intracellular assays. This study illustrates that Shoc2 single-domain antibodies can be used to understand functional mechanisms governing complex multiprotein signaling modules and have promise in application for therapies that require modulation of the ERK1/2-associated diseases.

    更新日期:2019-12-27
  • miRNA-20a suppressed lipopolysaccharide‐induced HK‐2 cells injury via NFκB and ERK1/2 signaling by targeting CXCL12
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-23
    Li zhang; Shuai He; Yun Wang; Xinyu Zhu; Wenying Shao; Qian Xu; Zhangke Cui

    Acute kidney injury (AKI) has one of the highest mortalities in terms of inflammatory sepsis. MiR-20a is involved in a variety of inflammatory reactions, but its role in AKI remains unknown. The purpose of this study was to investigate specific in vitro function and mechanisms of miR‐20a in AKI. We used, lipopolysaccharide (LPS) against human proximal tubular epithelial (HK-2) cells to induce an in vitro model of AKI. Then, our data showed that miR-20a expression levels were down-regulated in LPS-treated HK-2 cells. Overexpression of miR-20a promoted cell viability, inhibited apoptosis rate and inhibited the expression of apoptotic factors and inflammatory cytokines in HK-2 cells after LPS stimulation. In addition, CXCL12 was identified as a direct target of miR-20a by luciferase reporter gene assay, and CXCL12 expression was negatively regulated by miR-20a. Moreover, CXCR4 attenuated the suppression of miR-20a on inflammation and apoptosis in LPS-stimulated HK-2 cells, and further data indicated that miR-20a deactivated CXCL12/CXCR-4, NFκB and ERK1/2 signaling by targeting CXCL12. Therefore, our data revealed that miR-20a may play an anti-inflammatory and antiapoptotic roles in LPS-induced HK-2 cells via deactivation of CXCL12/CXCR-4, NFκB and ERK1/2 signaling.

    更新日期:2019-12-27
  • B cells in Crohn’s patients presented reduced IL-35 expression capacity
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-23
    Mingning Zhao; Jun Gu; Zhongchuan Wang

    Interleukin (IL)-35 has capacity to ameliorate experimental colitis and is upregulated in immune cells from active Crohn’s disease (CD) patients. Nonetheless, CD continues to be an incurable disease with characteristics of chronic relapsing-remitting inflammation. In this study, we investigated the production of IL-35 by B cells from active CD patients and non-CD controls. Immediately ex vivo, the mRNA transcription of IL-12/IL-35 subunit IL12A was significantly higher in CD B cells than in control B cells, and the mRNA transcription of IL-27/IL-35 subunit EBI3 was significantly higher in mild CD and moderate CD B cells than in control B cells. However, we also found that CpG-activated B cells and BCR + CD40-activated B cells from CD subjects presented significantly lower IL12A and lower EBI3 transcription than their counterparts from control subjects. We further evaluated IL-35 protein secretion and confirmed that B cell-mediated IL-35 protein secretion was lower in CD patients than in controls. However, IL-35-Fc preconditioning was able to significantly increase IL-35 production from B cells and to eliminate the difference in IL-35 production capacity between controls and CD patients. Furthermore, these IL-35-Fc-preconditioned B cells were able to suppress IFN-γ and IL-17 production from CD4+CD25− T cells more potently than Fc control-preconditioned B cells. Rh IL-27Rα-Fc, a soluble form of B cell-specific IL-35 receptor, significantly increased IFN-γ and IL-17 production. Together, these data supported a role of B cell-mediated IL-35 in suppressing IFN-γ and IL-17 inflammation. However, despite the fact that CD B cells presented higher transcription of IL-35 subunits directly ex vivo, CD B cells also presented reduced capacity for further IL-35 production upon activation.

    更新日期:2019-12-27
  • Complement fragments are biomarkers of antibody-mediated endothelial injury
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-26
    Erik Stites; Brandon Renner; Jennifer Laskowski; Moglie Le Quintrec; Zhiying You; Brian Freed; James Cooper; Diana Jalal; Joshua M. Thurman

    Antibody-mediated rejection (AbMR) adversely affects long-term graft survival in kidney transplantation. Currently, the diagnosis of AbMR requires a kidney biopsy, and detection of complement C4d deposition in the allograft is one of the diagnostic criteria. Complement activation also generates several soluble fragments which could potentially provide non-invasive biomarkers of the process. Furthermore, microvesicles released into the plasma from injured cells can serve as biomarkers of vascular injury. To explore whether soluble complement fragments or complement fragments bound to endothelial microvesicles can be used to non-invasively detect AbMR, we developed an in vitro model in which human endothelial cells were exposed to anti-HLA antibodies and complement sufficient serum. We found that complement fragments C4a and sC5b-9 were increased in the supernatants of cells exposed to complement-sufficient serum compared to cells treated complement-deficient serum. Furthermore, complement activation on the cell surface was associated with the release of microvesicles bearing C4 and C3 fragments. We next measured these analytes in plasma from kidney transplant recipients with biopsy-proven acute AbMR (n = 9) and compared the results with those from transplant recipients who also had impaired allograft function but who did not have AbMR (n = 30). Consistent with the in vitro results, complement fragments C4a and Ba were increased in plasma from patients with AbMR compared to control subjects (P < 0.001 and P < 0.01, respectively). Endothelial microvesicle counts were not increased in patients with AbMR, however, and the number of microvesicles with C4 and C3 bound to the surface was actually lower compared to control subjects (both P < 0.05). Our results suggest that plasma complement activation fragments may be useful as non-invasive biomarkers of antibody-mediated complement activation within the allograft. Complement-opsonized endothelial microvesicles are decreased in patients with AbMR, possibly due to enhanced clearance of microvesicles opsonized with C3 and C4 fragments.

    更新日期:2019-12-27
  • TLR4 induced Wnt3a-Dvl3 restrains the intensity of inflammation and protects against endotoxin-driven organ failure through GSK3β/β-catenin signaling
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-26
    Dongqiang Yang; ShuJian Li; Xiaoxian Duan; Junling Ren; Shuang Liang; Lan Yakoumatos; Yi Kang; Silvia M. Uriarte; Jia Shang; Wei Li; Huizhi Wang

    Background Accumulating evidence suggests a regulatory role of Wnt proteins in innate immune responses. However, the effects of Wnt3a signaling on TLR4-mediated inflammatory responses are controversial and the signaling crosstalk between TLR4 and Wnt3a remains uncertain. Methods Gain- and Loss- of function approaches were utilized to determine the function of Wnt3a signaling in TLR4-mediated inflammatory responses. Cytokine production at protein and mRNA levels and phosphorylation of signaling molecules were measured by ELISA, qRT-PCR, and Western Blot, respectively. Endotoxemia mouse model was employed to assess the effect of Wnt3a on systemic inflammatory cytokine levels and neutrophil infiltration. Results LPS stimulation leads to an increase of Wnt3a expression and its downstream molecule, Dvl3, in primary monocytes. Inhibition or silence of Wnt3a or Dvl3 significantly increases the production of pro-inflammatory cytokines (IL-12, IL-6, TNFα), robustly reduces β-catenin accumulation, and enhances the phosphorylation of NF-κB P65 and its DNA binding activity. These results were confirmed by multiple gain- and loss- of function approaches including specific siRNA and ectopic expression of Dvl3, GSK3β, and β-catenin in monocytes. Moreover, in vivo relevance was established in a murine endotoxin model, in which Wnt3a inhibition enhances the inflammatory responses by augmenting the systemic pro-inflammatory cytokine levels and neutrophil infiltration. Conclusions TLR4 activation promotes Wnt3a-Dvl3 signaling, which acts as rheostats to restrain the intensity of inflammation through regulating GSK3β-β-catenin signaling and NF-κB activity. General significance Wnt3a-Dvl3-β-catenin signaling axis could be a potential interventional target for manipulating the direction and intensity of inflammatory responses.

    更新日期:2019-12-27
  • Evaluation the potential of recombinant anti-CD3 nanobody on immunomodulatory function
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-26
    Shima Moradi-Kalbolandi; Azadeh Sharifi-K; Behrad Darvishi; Keivan Majidzadeh-A; Neda jalili; Solmaz Sadeghi; Marjan Mosayebzadeh; Hassan Sanati; Malihe Salehi; Leila Farahmand

    T cells are the most predominant effector cells in immune-mediated elimination of cancer and circumventing tumor progression. Among various approaches, T cells activation by specific antibodies independently of their TCR specificity, is considered as an effective approach to circumvent tumor progression. The most common surface marker for all T cells which is crucial for T cell activation is regarded as CD3. Therefore, the goal of our study was to evaluate the preclinical efficacy of recombinant anti-CD3 nanobody. To this end, anti-CD3 sequence, was PCR amplified, following cloning and expression in E.coli and purification, the purified nanobody with a molecular weight of ∼17 kDa was confirmed by western blot. Furthermore, flow cytometry analysis demonstrated that purified nanobody could bind to CD3 on Jurkat cell line. Subsequently, results from inoculation of 3 μg/g of nanobody to tumor bearing balb/c mice indicate inhibition of tumor growth. Furthermore, circulating levels of tumoricidal cytokines such as IL-2 and IFNγ were raised whereas tolerogenic cytokines such as IL-4, 6 and 10 were decreased at the end of the treatment. Moreover, IHC analysis confirmed the presence and also the percentage of TILs in tumor sites in response to anti-CD3 therapy. Hence, our results suggest that the purified anti-CD3 nanobody may become a promising candidate for targeting and activating CTLs to induce anti-tumor responses and may provide groundwork for future studies involving other kind of cancers.

    更新日期:2019-12-27
  • Inflammasome in HIV infection: Lights and shadows
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-10
    Vinicius Nunes Cordeiro Leal, Edione Cristina Reis, Alessandra Pontillo

    The importance of inflammasome, and related cytokines IL-1ß and IL-18, in host defense against pathogens is well documented, however, at the same time, dysregulation of inflammasome has been associated to multifactorial diseases characterized by chronic inflammation (i.e.: metabolic disorders, cardiovascular diseases, neurodegenerative diseases, autoimmunity, cancer). Inflammasome activation has been described in response to HIV-1 and possibly contributes to the resistance against virus establishment, however, on the other hand, when viral infection becomes chronic, independently from antiretroviral therapy, the increase constitutive activation of inflammasome has been eventually associated to a worse prognosis, raising the question about the role played by inflammasome and/or some specific receptors in this context. Due to the chance to imply targeted therapies that inhibit inflammasome activation and/or cytokines release, it will be important to define the impact of the complex in the pathogenesis of HIV. The purpose of this review is to depict the double-faced inflammasome role in HIV-1 infection, trying to unveil whether besides its role in first line defense against the virus, it exerts a harmful effect during the chronic phase of infection.

    更新日期:2019-12-11
  • Knockdown PEG10 deteriorates H2O2-injury of PC-12 cells by targeting miR-34a-5p/TLX
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-10
    Shizhen Niu, Yong Ni, Tong Niu, Jianwen Gao

    Objective Spinal cord injury (SCI) is a neurological disease with high incidence and disability. In this paper, we made a foundational study in long non-coding RNA paternally expressed gene 10 (lncRNA PEG10) at PC-12 cells. Methods We used CCK8, flow cytometry, migration and invasion assay to detect changes at the cellular level. PEG10, microRNA-34a-5p (miR-34a-5p) and TLX transfected by transfection assay and amplified by real-time quantitative PCR (qRT-PCR). TLX expression and proteins about apoptosis and pathways were investigated by Western blot. Additionally, the relationship between PEG10 and miR-34a-5p, miR-34a-5p and TLX were examined through luciferase assay. Results H2O2-injury model was established. Knockdown PEG10 deteriorated H2O2-injury. miR-34a-5p was confirmed as a target of PEG10 and miR-34a-5p inhibitor remitted PEG10-induced H2O2-injury. Moreover, TLX was confirmed as a target miR-34a-5p and TLX remitted H2O2-injury. At last, TLX worked in H2O2-injury via Smad and Wnt/β-catenin pathways. Conclusion Knockdown PEG10 remitted H2O2-injury of PC-12 cells by targeting miR-34a-5p/TLX, and the behavior may be involved in Smad and Wnt/β-catenin pathways.

    更新日期:2019-12-11
  • Control of memory CD8+ T cell longevity and effector functions by IL-15
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-06
    Jeffrey C. Nolz, Martin J. Richer

    IL-15 is a member of the common gamma chain family of cytokines and plays important roles in regulating several aspects of innate and adaptive immunity. Besides its established role in controlling homeostatic proliferation and survival of memory CD8+ T cells and natural killer cells, recent findings demonstrate that inflammatory IL-15 can also stimulate a variety of effector functions, such as enhanced cytotoxicity, entry into the cell cycle, and trafficking into non-lymphoid tissues. Here, we discuss how IL-15 is critical in regulating many functions of memory CD8+ T cells and how these processes act collectively to ensure optimal protective cellular immunity against re-infections.

    更新日期:2019-12-07
  • Early secreted antigenic target of 6-kDa of Mycobacterium tuberculosis induces transition of macrophages into epithelioid macrophages by downregulating iNOS / NO-mediated H3K27 trimethylation in macrophages
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-06
    Jiahui Lin, Yuyin Jiang, Dan Liu, Xueting Dai, Min Wang, Yalei Dai

    Background Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (Mtb). Granuloma is a pathological feature of tuberculosis and is a tight immune cell aggregation caused by Mtb. The main constituent cells are macrophages and their derivative cells including epithelioid macrophages. However, the molecular mechanism of the transition has not been reported. The purpose of this study was to investigate whether early secreted antigenic target of 6-kDa (ESAT6) can induce the transition of bone marrow-derived macrophages (BMDMs) into epithelioid macrophages and its possible molecular mechanism. Methods The recombinant ESAT6 protein was obtained from E.coli carrying esat6 gene after isopropyl β-d-thiogalactopyranoside (IPTG) induction. BMDMs were isolated from bone marrow of mice hind legs. Cells viability was detected by Cell Counting Kit 8 (CCK8) assays. The expression levels of mRNA and proteins were detected by qPCR and Western blot, or evaluated by flow cytometry. The expression level of nitric oxide (NO) was measured with a nitric oxide indicator. Results ESAT6 could significantly induce mRNA and protein expression levels of a group of epithelioid macrophages marker molecules (EMMMs), including E-cadherin, junction plakoglobin, ZO1, desmoplakin, desmoglein3 and catenin porteins, in BMDMs. These events could be abrogated in macrophage from TLR2 deficiency mice. ESAT6 could also markedly induce iNOS/NO production that could significantly inhibit trimethylation of H3K27 in the cells. ESAT6-induced expressions of epithelioid macrophages marker molecules were significantly inhibited in the presence of H3K27 histone demethylase inhibitor GSK J1. Furthermore, ROS scavenging agent N,N'-Dimethylthiourea (DMTU) could markedly inhibit the transition induced by ESAT6 in macrophages. Conclusion This study demonstrates that ESAT6 bound with TLR2 can activate iNOS/NO and ROS signalings to reduce the trimethylation of H3K27 resulting in the increment of EMMMs expression that is beneficial to the transition of macrophages into epithelioid macrophages. However, hypoxia can inhibit this transition event. This study has provided new evidence of pathogenesis of granuloma caused by Mtb and also proposed new ideas for the treatment of TB.

    更新日期:2019-12-07
  • LncRNA NEAT1 reversed the hindering effects of miR-495-3p/STAT3 axis and miR-211/PI3K/AKT axis on sepsis-relevant inflammation
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-05
    Demeng Xia, Renqi Yao, Panyu Zhou, Chen Wang, Yan Xia, Shuogui Xu

    Objective This investigation was intended to elucidate lncRNA-miRNA networks that could explain inflammation underlying sepsis progression. Methods In the first place, four kinds of mice models were established, namely, SHAM group (n = 30), trauma (TH) group (n = 30), lipopolysaccharide (LPS) group (n = 30) and TH + LPS group (n = 30). Their lung, spleen and liver tissues were gathered for determination of TNF-α, IL-6, IL-10 and MCP-1 levels. Furthermore, mouse mononuclear macrophage leukemia cell line (RAW264.7) was stimulated by LPS to establish inflammation cell models. Then si-NEAT1s, pcDNA3.1-NEAT1, miR-495-3p mimic, miR-495-3p inhibitor, miR-NC, miR-211 mimic and miR-211 inhibitor were, respectively, transfected into the cells, so as to observe the impacts of NEAT1, miR-495-3p and miR-211 on cytokine levels released by the cells. Results The survival condition of mice in the TH + LPS group was undesirable, in relative to mice in the LPS group and SHAM group (both P < 0.05). High-level NEAT1 and low-level miR-495-3p/miR-211 were associated with poor survival of mice in the TH+LPS group (P < 0.05). Additionally, the correlation between NEAT1/miR-495-3p/miR-211 level and cytokine level was the strongest among TH+LPS-treated mice, in comparison to mice treated by TH or LPS. Furthermore, up-regulation of NEAT1 level and down-regulation of miR-495-3p/miR-211 level could aggravate inflammation in LPS-treated RAW264.7 cells. The miR-495-3p and miR-211 herein, were both subjected to sponging of NEAT1, possibly affected inflammation responses in RAW264.7 cells, respectively, through modulating STAT3 and PI3K/AKT signaling. Conclusion LncRNA NEAT1 exhibited great potential sepsis diagnosis and treatment, considering its modifying miR-495-3p/STAT3 axis and miR-211/PI3K/AKT axis in inflammation cell models.

    更新日期:2019-12-05
  • Enhancement of adaptive immune responses of aged mice by dietary intake of β-glucans, with special emphasis on anti-aging activity
    Mol. Immunol. (IF 3.064) Pub Date : 2019-12-01
    Lili Song, Jiangshui Yuan, Shousheng Ni, Yang Zhou, Xia Wang, Yan Chen, Shicui Zhang

    The naturally occurring polysaccharide, β-1,3-glucans, a well-known immunostimulant, has been highly valued for many years for their health-promoting and anti-aging properties, but its mode of action is poorly understood. In this study, we first showed that oral administration of β-1,3-glucans did not affect the general condition and physiology of male mice throughout the trial period. We then showed that dietary intake of β-1,3-glucans induced a significant increase in T helper cells (CD4+) in young, middle-aged and aged male mice. We also showed that β-1,3-glucans supplementation considerably increased the delayed-type hypersensitivity (DTH) response, a T cell-mediated immune response, in young and aged mice. In addition, we found that β-1,3-glucans supplementation remarkably promoted the production of total anti-keyhole limpet hemocyanin (KLH) immunoglobulin G (IgG), anti-KLH IgG1, and anti-KLH IgG2a in young and aged mice without disturbing immune homeostasis. These data together indicate that oral administration of β-1,3-glucans enhanced the adaptive immune responses of aged mice without disturbing their general condition and physiology, supporting the idea that β-1,3-glucans are capable of counteracting the immunosenescence in mice. They also suggest that β-1,3-glucans can be clinically useful to help the elderly generate an improved response to vaccine with stronger humoral and cell-mediated immune responses.

    更新日期:2019-12-02
  • Comparative transcriptomic analysis provides insights into immune responses of lamprey larvae under three pathogens infections
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-29
    Meng Gou, Lie Ma, Jiali Lu, Xinya Wang, Yue Pang, Qingwei Li

    Based on the large number of deaths of lamprey larvae infected by pathogens in the culture process, the infection response of lamprey larvae to different pathogens was studied. Thus, high-throughput sequencing is firstly used to analyze the lamprey larvae infected by Aeromonas hydrophila, Vibrosplendidus, and Saprolegnia parasitica. A high quality of 338,628,426 Reads were obtained and 203,829 transcripts were assembled. A total of 158,001 Unigenes were annotated by Blast comparison, and a relatively comprehensive transcriptome database of lamprey larvae was established. Based on the transcriptome of lamprey larvae infected by three aquatic pathogens, classification results of COG and GO show that the genes classified into signal transduction mechanism accounted for the largest proportion, and 2316 differentially expressed genes were identified and screened out. After infection by the three pathogens, 16 genes and 19 genes expression were up-regulated and down-regulated, respectively. KEGG pathway analysis of differentially expressed genes showed that lamprey larvae were mainly affected their metabolism and oxidative phosphorylation pathways after infection by two gram-negative bacteria, Vibrosplendidus and Aeromonas hydrophila. As a fungus, Saprolegnia parasitica mainly affected ribosome synthesis in lamprey larvae. The results were validated by detecting the expression levels of 13 DEGs at 24 h after pathogens treatment using Q-PCR. The results provide valuable information for further research into the development of immunity and the innate immune response against pathogen invasion during the artificial propagation of lamprey larvae.

    更新日期:2019-11-30
  • From structure to function for the characterization of ERAP1 active site in Behçet syndrome. A novel polymorphism associated with known gene variations
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-29
    Maria Carmela Padula, Pietro Leccese, Nancy Lascaro, Teresa Carbone, Antonina Rita Limongi, Rosa Paola Radice, Angela Anna Padula, Salvatore D'Angelo, Giuseppe Martelli

    Introduction ERAP1 has been recently proposed as risk marker of Behçet syndrome (BS). Gene single nucleotide polymorphisms (SNPs) could affect the enzymatic activity and the conserved active site is pivotal for the aminopeptidase function. This study aims to characterize the ERAP1 active site in a cohort of BS patients vs healthy controls (HC) integrating genomics, transcriptomics and bioinformatics approach. Materials and methods We recruited 109 consecutive Italian BS patients (63M:46 F; mean age: 45.07 ± 12.28 years) and 106 matched HC (55M:51 F; mean age: 42.57 ± 12.29 years). DNA was isolated and amplified using PCR with home made-primer pairs. PCR products were directly sequenced and computational analyses were performed to search active site SNPs (NCBI-BlastN tool), to predict SNPs functional effect (PolyPhen-2 software) and to obtain protein 3D modelling (Protean3D software). In a second phase of analysis, RNA was extracted and reverse transcribed. Quantitative Real-Time PCR (qPCR) was performed to assess ERAP1 mRNA level in presence (target) and in absence (control) of gene polymorphisms. The Fold change was calculated for the relative quantification of gene expression. Results A novel coding variation (NG_027839.1:g.25637 T > G; NP_057526.3:p.Phe360Cys, HGSV nomenclature) was found in heterozygosity state in 5/109 BS patients (4.59 % of cases) and none of HC. It was recognized in association with rs2287987, rs30187, rs17482078, and rs27044 BS-related polymorphisms for 4 out of 5 patients. All patients carrying the novel SNP were HLA-B*51-positive. The novel SNP was released in GenBank database with MK140632.1 ID. The SNP was predicted to be damaging and resides within the Zn-binding HEXXH(X)18E region of the active site, changing the structurally conserved region for the amminopeptidase function. In fact, the change in energy (ΔE) score between wild-type and SNP-containing protein showed a less stable protein in presence of p.Cys360 (ΔE:3.584) (Protean3D prediction). Preliminary qPCR results underlined a significant difference in fold change value when target and control values were compared (p < 0.05), suggesting a reduced expression of ERAP1 mRNA in presence of the novel SNP. Conclusions Our study strengthens the association between ERAP1 and BS. The most significant point was the localization of the novel p.Phe360Cys SNP within the Zn-binding region of protein active site that was predicted to affect its function, causing protein destabilization. Our findings need to be tested in larger genetic studies.

    更新日期:2019-11-30
  • High-fat diet triggers obesity-related early infiltration of macrophages into adipose tissue and transient reduction of blood monocyte count
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-25
    Yanxia Liu, Xinping Lu, Xialian Li, Peijie Du, Guijun Qin

    Infiltration of adipose tissue macrophages (ATMs) is a typical feature of obesity, and circulating immune cells may indicate immune cell accumulation. However, it remains unclear whether this is true in the early stages of obesity. This study aimed to define the role of blood monocytes in obesity and the relationship between blood monocytes and ATMs in early-stage obesity. Two groups of male C57BL/6 J mice were fed on a 60 % high-fat diet (HFD) or a 10 % fat normal diet (ND), respectively, and monitored at 1, 2, 3, 7, and 12 weeks. Populations of circulating blood monocytes (CD11b + CD115+), ATMs (F4/80+CD11b+), and their subtypes were collected and analyzed using flow cytometry and immunofluorescence. Some cytokines (TNF-a, IL-1β) and chemokines (CCL2, CCL7) were also analyzed by real-time PCR. HFD induced obesity, dramatic fat expansion, and accumulation of ATMs in mice after 12 weeks. However, an acute and transient reduction of circulating monocyte count, elevated expression of CD11c in ly6clow monocytes, and concurrent infiltration of ATMs into visceral adipose tissues (VAT) were observed as early as 1 week after initiating HFD. Further, HFD-induced changes in VAT, but not blood monocyte count, were partially reversed upon reverting to ND for 6 weeks. An acute but transient reduction of blood monocyte count was observed at the early stages of HFD feeding, which might be related to early infiltration of macrophages into adipose tissues. We believe that blood monocytes could be targeted as a new obesity treatment following additional studies.

    更新日期:2019-11-26
  • Analyses of RAG1 and RAG2 genes suggest different evolutionary rates in the Cetacea lineage
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-25
    Bruna C. Dias, Mariana F. Nery

    V(D)J recombination is a process of somatic recombination catalyzed by proteins encoded by RAG1 and RAG2 genes, both restricted to the genome of jawed vertebrates. Their proteins constitute the enzymatic core of V(D)J recombination machinery and are crucial for jawed vertebrate adaptive immunity. Mammals possess great ecological diversity, and their complex evolutionary history associated with radiation to different environments presented many distinct pathogenic challenges from these different habitats. Cetaceans comprise a mammalian order of fully aquatic mammals that have arisen from a complete terrestrial ancestor and, accordingly, was confronted with challenges from changing environmental pathogens while they transitioned from land to sea. In this study we undertook molecular evolutionary analyses of RAG1 and RAG2 genes, exploring the possible role of natural selection acting on these genes focusing on the cetacean lineage. We performed phylogenetic reconstructions on IQ-TREE, together with selection analyses in the codeml program of the PAML package, and in the FITMODEL program for codon evolution and switching on both the RAG1 and RAG2 genes. Our findings demonstrate that RAG1 and RAG2 remained fairly conserved among tetrapods, with purifying selection acting on both genes, with evidence for a few punctuated shifts in nucleotide substitution rates of both genes along tetrapod evolution. We demonstrate differential evolution in the closely linked genes RAG1 and RAG2 specifically in cetaceans.

    更新日期:2019-11-26
  • Granulocyte colony stimulating factor (G-CSF) regulates neutrophils infiltration and periodontal tissue destruction in an experimental periodontitis
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-22
    Zheng Zhang, Wei Yuan, Junjie Deng, Danyang Wang, Tianyi Zhang, Li Peng, Huan Tian, Zuomin Wang, Jie Ma

    Although granulocyte colony-stimulating factor(G-CSF) has pathogenic roles in several immune inflammatory diseases, its role in periodontitis has not been investigated. Here we detected local expression of G-CSF using public datasets in the Gene Expression Omnibus (GEO) database, and immune cell infiltration into gingival tissue was estimated based on single-sample gene set enrichment analysis (ssGSEA). G-CSF expression and neutrophil infiltration were also confirmed by human gingival biopsies analysis. Moreover, anti-G-CSF neutralizing antibody was locally administrated to investigate the effects of G-CSF neutralization on neutrophils infiltration and periodontal tissue destruction in periodontitis mice model. Two public datasets (GSE10334 and GSE16134), which included 424 patients with periodontitis and 133 health controls, were used in the analysis. Markedly increased immune cell infiltration and G-CSF expression in gingival tissues were found in the periodontitis group as compared to the control group. The higher expression of G-CSF was correlated with higher infiltration of immune cells, especially with neutrophil infiltration. Analysis of gingival biopsies further confirmed high neutrophil infiltration and G-CSF expression. In addition, anti-G-CSF antibody-treated mice with periodontitis showed significantly reduced alveolar bone resorption and neutrophil infiltration when compared with periodontitis mice treated with isotype control antibody. Also, anti-G-CSF antibody treatment significantly reduced mRNA expression of CXC chemokines (CXCL1, CXCL2 and CXCL3), interleukin 1β (IL-1β), IL-6, matrix metalloproteinases 9, receptor activator of nuclear factor κB ligand/osteoprotegerin (RANKL/OPG) ratio and osteoclasts number in periodontal tissues. In summary, neutrophil infiltration and G-CSF expression levels were significantly increased in inflamed gingival tissues. G-CSF neutralization in periodontal inflammation could alleviate neutrophil infiltration and periodontal tissue destruction in experimental periodontitis.

    更新日期:2019-11-22
  • Monomeric C-reactive protein regulates fibronectin mediated monocyte adhesion
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-22
    Naeem Ullah, Fu-Rong Ma, Jin Han, Xiao-Ling Liu, Yu Fu, Yu-Tong Liu, Yu-Lin Liang, Hanyue Ouyang, Hai-Yun Li
    更新日期:2019-11-22
  • rhBMP2 alone does not induce macrophage polarization towards an increased inflammatory response
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-20
    Emily L. Durham, Rajiv Kishinchand, Zachary J. Grey, James J. Cray

    Once thought to have revolutionized therapeutic intervention in surgery, Recombinant Human Bone Morphogenic Protein 2 (rhBMP2) is now in its second decade of sustained controversy over the side effects associated with its use. Side effects associated with clinical use of rhBMP2 (Infuse, Medtronic Inc) include a marked inflammatory response, pain, therapeutic failures, ectopic bone, tissue degradation, and death. What is missing, despite the depth of literature on the subject, is a direct interrogation of rhBMP2, specifically for inflammation. Here we set out to determine if rhBMP2 alters traditional macrophage markers associated with pro-inflammatory responses, and pro-reparative responses to injury. Based on our previous work, we hypothesized there would be no direct effect of the peptide on macrophage polarization. Here we utilized commercially available murine macrophages, RAW 264.7, and treated these cells with rhBMP2 in standard growth media or macrophage polarizing media (M1 and M2) at several doses of the peptide. Our readouts were cell viability, apoptosis, gene expression of M1 and M2 markers, and ELISA for M1 marker iNOS, and M2 marker Arg1. Our data give very little evidence to support an alteration in macrophage phenotype by rhBMP2 alone, or alteration of the phenotype when cultured in enriched M1 or M2 media. These results further suggest that other factors associated with the clinical use of Infuse, likely supraphysiological rhBMP2 doses and off label usage, are more likely the culprit for poor outcomes. This further reinforces the utility of rhBMP2 and other peptides in tissue engineering therapies when conditions are tightly controlled.

    更新日期:2019-11-21
  • IFNγ potentiates TNFα/TNFR1 signaling to induce FAT10 expression in macrophages
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-20
    Michal Kandel-Kfir, Rolando Garcia-Milan, Itai Gueta, Irit Lubitz, Ilan Ben-Zvi, Aviv Shaish, Lidar Shir, Dror Harats, Milind Mahajan, Allon Canaan, Yehuda Kamari

    Introduction The tight regulation of the cytokine network during macrophage activation is of prime importance to enable a fast and potent innate immune response against exogenous pathogens. The inflammation mediating ubiquitin-like protein HLA-F adjacent transcript number 10 (FAT10) was shown to be transcriptionally regulated by and also regulate the nuclear factor-κB (NFκB) signaling pathway. However, very little is known about the regulation of FAT10 gene expression during macrophage activation. Results RNA sequencing of interferon (IFN)γ-stimulated mouse peritoneal macrophages analyzed by ingenuity pathway analysis revealed significant involvement of tumor necrosis factor receptor 1 (TNFR1) signaling in addition to IFNγ signaling. Subsequently, IFNγ robustly upregulated FAT10 expression compared to a milder induction seen with TNFα or lipopolysaccharide (LPS) stimulation. While low dose IFNγ with TNFα synergistically elevated FAT10 expression, preincubation of macrophages with IFNγ strongly augmented TNFα-induced FAT10 expression. Moreover, a short preincubation with IFNγ, which did not elevate FAT10, was sufficient to potentiate the induction of FAT10 by TNFα. A double augmentation mechanism of TNFα signaling was demonstrated, where IFNγ rapidly induced the expression of TNFα and TNFR1, which further augmented the induction of TNFα and TNFR1 expression by TNFα. Importantly, the induction of FAT10 by IFNγ in macrophages from TNFα-deficient or TNFR1-deficient mice was completely inhibited compared to macrophages from wild type (WT) mice. Finally, we show that TNFα-induced FAT10 expression is dependent on NFκB signaling. Conclusion IFNγ potentiates the TNFα/TNFR1 signaling pathway to induce FAT10 expression in mouse macrophages, mediated through NFκB network.

    更新日期:2019-11-21
  • Recombinant C1q variants modulate macrophage responses but do not activate the classical complement pathway
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-15
    Victoria Espericueta, Ayla O. Manughian-Peter, Isabelle Bally, Nicole M. Thielens, Deborah A. Fraser
    更新日期:2019-11-18
  • MicroRNA-223-3p modulates dendritic cell function and ameliorates experimental autoimmune myocarditis by targeting the NLRP3 inflammasome
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-16
    Liangqi Chen, Xinyu Hou, Maomao Zhang, Yang Zheng, Xianghui Zheng, Qingyuan Yang, Jing Li, Nan Gu, Min Zhang, Yong Sun, Jian Wu, Bo Yu

    Autoimmune myocarditis is a cause of dilated cardiomyopathy and heart failure. MicroRNAs regulate many immune processes, but their role in aberrant inflammation during autoimmune myocarditis remains unclear. In this study, we investigated the role of miR-223-3p in experimental autoimmune myocarditis (EAM). We found that miR-223-3p expression was significantly lower in EAM mice than that in normal mice. miR-223-3p inhibited NLRP3 inflammasome expression, promoting the polarization of dendritic cells (DCs) towards a tolerogenic DC phenotype. miR-223-3p effectively induced regulatory T cell (Treg) generation by inhibiting the function of antigen-presenting DCs. Transfer of miR-223-3p-overexpressing DCs protected mice against the development of EAM. Our findings suggest that miR-223-3p is involved in the induction of the tolerogenic DC phenotype and regulates tolerance in autoimmune myocarditis.

    更新日期:2019-11-18
  • FGL2 regulates IKK/NF-κB signaling in intestinal epithelial cells and lamina propria dendritic cells to attenuate dextran sulfate sodium-induced colitis
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-16
    Tang Li, Ru-ru Chen, Hong-peng Gong, Bin-feng Wang, Xi-xi Wu, Yue-qiu Chen, Zhi-ming Huang

    Inflammatory bowel disease (IBD) is an autoimmune disease characterized by an abnormal immune response. Fibrinogen-like protein 2 (FGL2) is known to have immunoregulatory and anti-inflammatory activity. The level of FGL2 is elevated in patients with IBD; however, its comprehensive function in IBD is almost unknown. In our study, we explored the effect of FGL2 on dextran sulfate sodium (DSS)-induced colitis in mice and on NF-κB signaling in intestinal epithelial cells (IECs) and lamina propria dendritic cells (LPDCs). We founded that FGL2−/− mice in the colitis model showed more severe colitis manifestations than WT mice did, including weight loss, disease activity index (DAI), and colon histological scores. FGL2−/− mice treated with DSS produced more proinflammatory cytokines (IL-1β, IL-6, TNF-α) in serum than WT mice did and demonstrated upregulated expression of TNF-α and inflammatory marker enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (Cox-2) in the colon tissue. Our data suggested that DSS-treated FGL2−/− mice showed stronger activation of NF-κB signaling, especially in IECs. Next, we demonstrated that recombinant FGL2 (rFGL2) inhibited the production of proinflammatory cytokines and the expression of inflammatory marker enzymes by downregulating the NF-κB signaling in HT-29 cells. Finally, we discovered that LPDCs from the colon of DSS-treated FGL2−/− mice showed significantly upregulated expression of surface maturation co-stimulatory molecules, including CD80, CD86, CD40, and MHC class II molecules compared with that in WT mice. In addition, LPDCs in FGL2−/− treated with DSS exhibited excessive NF-κB activity and the administration of rFGL2 to FGL2−/− mice could rescue the aggravated results of FGL2−/− mice. Taken together, our findings demonstrated that FGL2 might be a target for further therapy of IBD.

    更新日期:2019-11-18
  • JAK/STAT proteins and their biological impact on NK cell development and function
    Mol. Immunol. (IF 3.064) Pub Date : 2019-01-28
    Alexander Vargas-Hernández, Lisa R. Forbes

    NK cells are important early effectors in the innate immune response to a variety of viral infections and for elimination of tumor cells. The JAK/STAT signaling cascade is critical for NK cell development, maturation, survival, and proliferation, therefore, it is important to understand the role of this pathway in NK cell biology. Many cytokines can activate multiple JAK/STAT protein family members, creating a severe phenotype when mutations impair their function or expression. Here we discuss the impact of defective JAK/STAT signaling pathways on NK cell development, activation and cytotoxicity.

    更新日期:2019-11-18
  • KIR2DL4-HLAG interaction at human NK cell-oligodendrocyte interfaces regulates IFN-γ-mediated effects
    Mol. Immunol. (IF 3.064) Pub Date : 2018-11-24
    P.P. Banerjee, L. Pang, S.S. Soldan, S.M. Miah, A. Eisenberg, S. Maru, A. Waldman, E.A. Smith, Y. Rosenberg-Hasson, D. Hirschberg, A. Smith, D.V. Ablashi, K.S. Campbell, J.S. Orange

    Interactions between germline-encoded natural killer (NK) cell receptors and their respective ligands on tumorigenic or virus-infected cells determine NK cell cytotoxic activity and/or cytokine secretion. NK cell cytokine responses can be augmented in and can potentially contribute to multiple sclerosis (MS), an inflammatory disease of the central nervous system focused upon the oligodendrocytes (OLs). To investigate mechanisms by which NK cells may contribute to MS pathogenesis, we developed an in vitro human model of OL-NK cell interaction. We found that activated, but not resting human NK cells form conjugates with, and mediate cytotoxicity against, human oligodendrocytes. NK cells, when in conjugate with OLs, rapidly synthesize and polarize IFN-γ toward the OLs. IFN-γ is capable of reducing myelin oligodendrocyte and myelin associated glycoproteins (MOG and MAG) content. This activity is independent of MHC class-I mediated inhibition via KIR2DL1, but dependent upon the interaction between NK cell-expressed KIR2DL4 and its oligodendrocyte-expressed ligand, HLA-G. NK cells from patients with MS express higher levels of IFN-γ following conjugation to OLs, more actively promote in vitro reduction of MOG and MAG and have higher frequencies of the KIR2DL4 positive population. These data collectively suggest a mechanism by which NK cells can promote pathogenic effects upon OLs.

    更新日期:2019-11-18
  • Protective effect of ginsenoside Rg1 on LPS-induced apoptosis of lung epithelial cells
    Mol. Immunol. (IF 3.064) Pub Date : 2018-11-22
    Qijian Ji, Zhaorui Sun, Zhizhou Yang, Wei Zhang, Yi Ren, Weijun Chen, Mengya Yao, Shinan Nie

    Sepsis-induced acute lung injury (ALI) is a life-threatening medical condition with high mortality and morbidity in the critical care units. Though, it was commonly accepted that inflammation and apoptosis of lung epithelial cells played an essential role in the pathogenesis of ALI, the underlying mechanism remain unknown. In our study, we found that LPS-induced cell apoptosis could be counteracted by elevated cell autophagy. In LPS-treated MLE-12 cells, suppression of autophagy via 3-MA could aggravate LPS-induced apoptosis, while activation of autophagy via Rapamycin could effectively impair the apoptosis of MLE-12 cells induced by LPS. In order to further discover the molecular regulation mechanism between apoptosis and autophagy in LPS-treated MLE-12 cells, we demonstrated that autophagy could induced the expression of Nrf2, followed with the decrease of p-p65. Targeted inhibition of Nrf2 could induce enlarged cell apoptosis via increasing the level of p-p65. In addition, we demonstrated that ginsenoside Rg1 protected MLE-12 cells from LPS-induced apoptosis via augmenting autophagy and inducing the expression of Nrf2. Our data implicates that activation of autophagy and Nrf2 by ginsenoside Rg1 may provide a preventive and therapeutic strategy for ALI.

    更新日期:2019-11-18
  • Generation of novel Id2 and E2-2, E2A and HEB antibodies reveals novel Id2 binding partners and species-specific expression of E-proteins in NK cells
    Mol. Immunol. (IF 3.064) Pub Date : 2018-08-23
    Jai Rautela, Laura F. Dagley, Tobias Kratina, Angaleena Anthony, Wilford Goh, Elliot Surgenor, Rebecca B. Delconte, Andrew I. Webb, Ngaire Elwood, Joanna R. Groom, Fernando Souza-Fonseca-Guimaraes, Lynn Corcoran, Nicholas D. Huntington
    更新日期:2019-11-18
  • MicroRNA regulation of natural killer cell development and function in leukemia
    Mol. Immunol. (IF 3.064) Pub Date : 2018-08-09
    Jennifer N. Saultz, Aharon G. Freud, Bethany L. Mundy-Bosse
    更新日期:2019-11-18
  • Genome-wide analyses and functional profiling of human NK cell lines
    Mol. Immunol. (IF 3.064) Pub Date : 2018-07-24
    Justin T. Gunesch, Laura S. Angelo, Sanjana Mahapatra, Raquel P. Deering, Johanna E. Kowalko, Patrick Sleiman, John W. Tobias, Linda Monaco-Shawver, Jordan S. Orange, Emily M. Mace

    Natural killer (NK) cell lines, including YTS, NK92, NK3.3, and NKL, represent excellent models for the study of human natural killer cells. While phenotypic and functional differences between these cell lines have been reported, a multi-parametric study, encompassing genomic, phenotypic, and functional assays, has not been performed. Here, using a combination of techniques including microarray and copy number analyses, flow cytometry, and functional assays, we provide in-depth genetic, functional, and phenotypic comparison of YTS, NK92, NK3.3, and NKL cell lines. Specifically, we found that while the cell lines shared similarities in enrichment of growth and survival pathways, they had differential expression of 557 genes, including genes related to NK cell development, survival, and function. In addition, we provide genetic and phenotypic analyses that demonstrate distinct developmental origins of NK92, YTS, and NKL cell lines. Specifically, NK92 has a phenotype associated with the CD56bright NK cell subset, while both YTS and NKL appear more CD56dim-like. Finally, by classifying cell lines based on their lytic potential, we identified genes differentially expressed between NK cell lines with high and low lytic function. Taken together, these data provide the first comprehensive genetic, phenotypic, and functional analyses of these commonly used NK cell lines and provides deeper understanding into their origins and function. This will ultimately improve their use as models for human NK cell biology.

    更新日期:2019-11-18
  • The expanding role of murine class Ib MHC in the development and activation of Natural Killer cells
    Mol. Immunol. (IF 3.064) Pub Date : 2018-05-20
    Katharine J. Goodall, Angela Nguyen, Lucy C. Sullivan, Daniel M. Andrews

    Major Histocompatibility Complex-I (MHC-I) molecules can be divided into class Ia and class Ib, with three distinct class Ib families found in the mouse. These families are designated as Q, T and M and are largely unexplored in terms of their immunological function. Among the class Ib MHC, H2-T23 (Qa-1b) has been a significant target for Natural Killer (NK) cell research, owing to its homology with the human class Ib human leukocyte antigen (HLA)-E. However, recent data has indicated that members of the Q and M family of class Ib MHC also play a critical role in the development and regulation NK cells. Here we discuss the recent advances in the control of NK cells by murine class Ib MHC as a means to stimulate further exploration of these molecules.

    更新日期:2019-11-18
  • Natural killer cell metabolism
    Mol. Immunol. (IF 3.064) Pub Date : 2017-11-24
    Takumi Kobayashi, Stephen R. Mattarollo

    Natural killer (NK) cells are a critical component in the innate immune response against disease. NK cell function is tightly regulated by specific cytokine and activation/inhibitory receptor signalling, leading to diverse effector responses. Like all living cells, energy metabolism is a fundamental requirement for NK cell activation and survival. There is growing evidence that distinct functional profiles of NK cells are determined by alterations to cellular metabolic pathways. In this review, we summarise current literature that has explored NK cell metabolism to provide insight into how metabolic regulation controls NK cell function. We focus on metabolism pathways induced by different NK cell stimuli, metabolic regulatory proteins, and nutrient and hormonal levels in health and disease which impact on NK cell metabolic and functional activity.

    更新日期:2019-11-18
  • Enhancement of immune response against Mycobacterium tuberculosis HspX antigen by incorporation of combined molecular adjuvant (CASAC)
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-15
    Min Han Lew, Mohd Nor Norazmi, Gee Jun Tye

    Tuberculosis (TB) is one of the deadliest human diseases worldwide caused by mycobacterial infection in the lung. Bacillus Calmette-Guerin (BCG) vaccine protects against disseminated TB in children, but its effectiveness is still questionable due to highly variable protections in adolescence and elderly individuals. Targeting the latency M.tb antigen is a recent therapeutic approach to eradicate dormant pathogen that could possibly lead to disease activation. In this study, we aimed to potentiate immune responses elicited against 16 kDa α-crystalline (HspX) tuberculosis latency antigen by incorporation of Combined Adjuvant for Synergistic Activation of Cellular immunity (CASAC). Histidine-tagged recombinant HspX protein was initially produced in Escherichia coli and purified using Ni-NTA chromatography. To evaluate its adjuvanticity, C57BL/6 mice (n = 5) were initially primed and intradermally immunised in 2-weeks interval for 4 rounds with recombinant HspX, formulated with and without CASAC. Humoral and cell-mediated immune responses elicited against HspX antigen were evaluated using ELISA and Flow Cytometry. Our findings showed that CASAC improved humoral immunity with increased antigen-specific IgG1 and IgG2a antibody response. Stronger CD8+ and Th1-driven immunity was induced by CASAC formulation as supported by elevated level of IFN-γ, TNF-α, IL-12 and IL-17A; and with low IL-10 secretion. Interestingly, adjuvanted HspX vaccine triggered a higher percentage of effector memory T-cell population than those immunised with unadjuvanted vaccine. In conclusion, CASAC adjuvant has great potential to enhance immunogenicity elicited against HspX antigen, which could be an alternative regimen to improve the efficacy of future therapeutic vaccine against Mycobacterium tuberculosis.

    更新日期:2019-11-15
  • Molecular cloning and functional characterization of duck Janus kinase 1
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-13
    Dejian Liu, Huijun Zheng, Yaqian Li, Peng Zhou, Hui Jin, Rui Luo

    Janus kinase 1 (JAK1) is a member of JAK family of non-receptor protein tyrosine kinases that plays critical roles in transducing cytokine signals via JAK-signal transducer and activator of transcription (STAT) signaling pathway. The importance of JAK1 in innate immunity has been well-studied in mammals and fish, yet in avian remains largely unknown. Here, we cloned the full-length of the duck JAK1 (duJAK1) gene for the first time. DuJAK1 encoded a protein of 1152 amino acids and possessed high amino acid identity with goose and budgerigar JAK1s. The duJAK1 was expressed in all detected tissues, especially high in the thymus and bursa of Fabricius. Overexpression of duJAK1 significantly activated ISRE promoter activity and induced duck viperin, 2′, 5′-OAS, MX, PKR and ZAP expression. Knockdown of duJAK1 by small interfering RNA significantly inhibited duck Tembusu virus (DTMUV)-, duck Enteritis virus (DEV)-, poly (I:C)-, poly (dA:dT)- or Sendai virus (SeV)-induced ISRE promoter activation. Furthermore, duJAK1 exhibited antiviral activity against DTMUV infection. These results will help us understand the function of JAK family proteins in duck antiviral immunity.

    更新日期:2019-11-13
  • Deiminated proteins in extracellular vesicles and serum of llama (Lama glama)—Novel insights into camelid immunity
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-13
    Michael F. Criscitiello, Igor Kraev, Sigrun Lange

    Peptidylarginine deiminases (PADs) are phylogenetically conserved calcium-dependent enzymes which post-translationally convert arginine into citrulline in target proteins in an irreversible manner, causing functional and structural changes in target proteins. Protein deimination causes generation of neo-epitopes, affects gene regulation and also allows for protein moonlighting. Furthermore, PADs have been found to be a phylogenetically conserved regulator for extracellular vesicle (EVs) release. EVs are found in most body fluids and participate in cellular communication via transfer of cargo proteins and genetic material. In this study, post-translationally deiminated proteins in serum and serum-EVs are described for the first time in camelids, using the llama (Lama glama L. 1758) as a model animal. We report a poly-dispersed population of llama serum EVs, positive for phylogenetically conserved EV-specific markers and characterised by TEM. In serum, 103 deiminated proteins were overall identified, including key immune and metabolic mediators including complement components, immunoglobulin-based nanobodies, adiponectin and heat shock proteins. In serum, 60 deiminated proteins were identified that were not in EVs, and 25 deiminated proteins were found to be unique to EVs, with 43 shared deiminated protein hits between both serum and EVs. Deiminated histone H3, a marker of neutrophil extracellular trap formation, was also detected in llama serum. PAD homologues were identified in llama serum by Western blotting, via cross reaction with human PAD antibodies, and detected at an expected 70 kDa size. This is the first report of deiminated proteins in serum and EVs of a camelid species, highlighting a hitherto unrecognized post-translational modification in key immune and metabolic proteins in camelids, which may be translatable to and inform a range of human metabolic and inflammatory pathologies.

    更新日期:2019-11-13
  • Immunogenetics of marsupial B-cells
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-11
    Andrea L. Schraven, Hayley J. Stannard, Oselyne T.W. Ong, Julie M. Old

    Marsupials and eutherians are mammals that differ in their physiological traits, predominately their reproductive and developmental strategies; eutherians give birth to well-developed young, while marsupials are born highly altricial after a much shorter gestation. These developmental traits also result in differences in the development of the immune system of eutherian and marsupial species. In eutherians, B-cells are the key to humoral immunity as they are found in multiple lymphoid organs and have the unique ability to mediate the production of antigen-specific antibodies in the presence of extracellular pathogens. The development of B-cells in marsupials has been reported and hypothesised to be similar to that of eutherians, except that haematopoiesis occurs in the liver, postpartum, until the bone marrow fully matures. In eutherians, specific genes are linked to specific stages in B-cell development, maturation, and differentiation processes, and have been identified including immunoglobulins (heavy and light chains), cluster of differentiation markers (CD10, 19, 34 and CD79α/β), signal transduction molecules (BTK, Lyn and Syk) and transcriptional regulators (EBF1, E2A, and Pax5). This review aims to discuss the known similarities and differences between marsupial and eutherian B-cells, in regards to their genetic presence, homology, and developmental stages, as well as to highlight the areas requiring further investigation. By enhancing our understanding of the genes that are involved with B-cells in the marsupial lineage, it will, in turn, aid our understanding of the marsupial immune system and support the development of specific immunological reagents for research and wildlife conservation purposes.

    更新日期:2019-11-13
  • Toll-like receptor 4 knockout protects against diabetic-induced imbalance of bone metabolism via autophagic suppression
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-12
    Yongze Zhang, Ximei Shen, Ling Cheng, Ran Chen, Fengying Zhao, Shuai Zhong, Chao Lan, Sunjie Yan

    Objective This study aimed to elucidate the mechanism of autophagy in bone metabolism in high-glucose environments and its relationship with Toll-like receptor 4 (TLR4). Methods A TLR4 knockout diabetic rat model and MC3T3-E1 with TLR4 silencing by small interfering RNA were used to observe the protective mechanism of TLR4 knockdown or silencing in hyperglycemia-induced bone injury. Results The inhibition of TLR4 expression improved bone metabolism and bone structure; promoted alkaline phosphatase (ALP) and osteocalcin (OCN) secretion, enhanced bone morphogenic protein (BMP)-2 expression, promoted bone mineralization, and reduced hyperglycemia-induced osteoblast apoptosis. TLR4 knockdown or silencing reduced the levels of inflammatory factors interleukin (IL)-1, IL-6, and tumor necrosis factor-alpha at the animal and cell levels, inhibited the expression of inflammatory pathway proteins, and downregulated the expression of Beclin 1 and LC3II/LC-1 proteins. The inhibition of autophagic activity enhanced the osteoprotective effect of TLR4 knockdown, improved cell viability, reduced the apoptosis rate of osteoblasts, and promoted the BMP-2 protein level and ALP and OCN secretion. Conversely, the activation of autophagy significantly aggravated osteoblast apoptosis, reduced BMP-2 protein levels, and inhibited ALP and OCN secretion. Conclusion Taken together, the experimental results supported the hypothesis that TLR4 deficiency might alleviate hyperglycemia-induced apoptosis and differentiation suppression in osteoblasts and exert osteoprotective effects via autophagic inhibition.

    更新日期:2019-11-13
  • LncRNA NEAT1 sponges miR-214 to regulate M2 macrophage polarization by regulation of B7-H3 in multiple myeloma
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-12
    Yin Gao, Peng Fang, Wen-Jin Li, Jian Zhang, Guang-Ping Wang, Duan-Feng Jiang, Fang-Ping Chen

    Background LncRNA NEAT1 was associated with the tumorigenesis of multiple myeloma (MM). However, the mechanisms of M2 macrophage polarization involved with NEAT1 in MM are still unknown. Methods Bone marrow samples, multiple myeloma cells RPMI 8226 and monocyte cell line THP-1 were used in this study. The expression of NEAT1 and miR-214 was modified by transfection with the shNEAT1 or miR-214 inhibitor. The expression of NEAT1, miR-214 and B7-H3 in MM patient tissues and cells was analyzed by RT-qPCR. ELISA assay was used to determine the release of B7-H3 in the supernatant of cell culture. The patient survival curve was analyzed using Kaplan-Meier method. The macrophage polarization markers were examined by RT-qPCR and western blotting. The interaction between NEAT1, miR-214 and B7-H3 was analyzed by Dual-Luciferase reporter and RIP assays. AG490 was used to block the JAK2/STAT3 signaling. Co-culture of THP-1 and RPMI 8226 cells was used for macrophage polarization. Results NEAT1 and B7-H3 were up-regulated, but miR-214 was obviously down-regulated in MM patients. B7-H3, NEAT1 and miR-214 were associated with overall survival time of MM patients. NEAT1 silencing induced miR-214 and inhibited the expression and release of B7-H3 and then suppressed M2 macrophage polarization via inhibiting the JAK2/STAT3 signaling. NEAT1 directly targeted miR-214, and miR-214 directly bound to B7-H3. MiR-214 inhibitor reversed the down-regulation and release of B7-H3 and M2 macrophage polarization caused by shNEAT1. The specific JAK2/STAT3 signaling inhibitor AG490 abrogated M2 macrophage polarization. Conclusion NEAT1 promoted M2 macrophage polarization by sponging miR-214 and then regulating B7-H3, thus accelerating MM progression via the JAK2/STAT3 signaling pathway. Our study revealed novel mechanisms of M2 macrophage polarization and provided new potential clinical therapeutic targets for MM.

    更新日期:2019-11-13
  • Contribution of endothelial cell and macrophage activation in the alterations induced by the venom of Micrurus tener tener in C57BL/6 mice
    Mol. Immunol. (IF 3.064) Pub Date : 2019-10-07
    Emelyn Salazar, Ana María Salazar, Peter Taylor, Izaskun Urdanibia, Karin Pérez, Alexis Rodríguez-Acosta, Elda E. Sánchez, Belsy Guerrero

    An acute inflammatory response, cellular infiltrates, anemia, hemorrhage and endogenous fibrinolysis activation were previously described in C57BL/6 mice injected with M. tener tener venom (Mtt). As the endothelium and innate immunity may participate in these disturbances and due to our poor understanding of the alterations produced by these venoms when the neurotoxic component is not predominant, we evaluated the effects in an in vitro model. At 24 h, the release of pro-inflammatory mediators was detected in peritoneal macrophages. At different times, the release of pro-inflammatory (TNF-α, IL-6, NO and E-Selectin), pro-coagulant (vWF and TF) and pro-fibrinolytic (uPA) mediators were seen in liver sinusoidal endothelial cells (LSECs). These results suggest that Mtt venom activates macrophages and endothelium, thus inducing the release of mediators, such as TNF-α, that orchestrate the acute inflammatory response and the later infiltration of mononuclear cells into liver in C57BL/6 mice. In addition, endothelium activation promotes TF expression, which may in turn modulate the inflammatory and hemostatic response. These findings suggest crosstalk between inflammation and hemostasis in the alterations observed in Micrurus envenomation, where the neurotoxic manifestations do not predominate.

    更新日期:2019-11-11
  • Curcumin inhibits proliferation and soluble collagen synthesis of NIH/3T3 cell line by modulation of miR-29a and via ERK1/2 and β-catenin pathways
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-08
    Jianhua Zhang, Mei Song, Weifeng Li, Feng Zhao, Yuguo Li

    Background Scars affects the appearance and results in tissue damage. In this research, we preliminarily studied the function and mechanism of curcumin (CUR) on cell proliferation and soluble collagen synthesis in NIH-3T3 cells. Methods CCK-8 was used to detect the IC50 of CUR. Moreover, Western blot was used to measure the expression of cell proliferation-related, soluble collagen synthesis and pathway-related proteins. Sircol assay was determined the expression of soluble collagen. Furthermore, reverse transcription quantitative PCR (RT-qPCR) was used to determined miR-29a, α-smooth muscle aorta (α-SMA), soluble collagen 1 (Col 1) and Col 3 expression. Results CUR inhibited cell viability and proliferation-related proteins expression. Transforming growth factor-β (TGFβ1)-induced heightened the expression of proliferation-related proteins and soluble collagen synthesis-related proteins. CUR inhibited TGFβ1-induced proliferation and soluble collagen synthesis. Furthermore, CUR positively related miR-29a and miR-29a mimic inhibited TGFβ1-induced proliferation and soluble collagen synthesis. Besides, transfection with miR-29a inhibitor could partly reverse the effects of CUR. CUR inhibited the ERK1/2 and β-catenin pathways and the miR-29a inhibitor reversed the above results. Otherwise, soluble collagen 1 (Col 1) partly reversed the effects of CUR on proliferation and soluble collagen synthesis and silenced Col 1/3 could inhibit ERK1/2 and β-catenin signaling pathways. Conclusion CUR restrained TGFβ1-induced proliferation and soluble collagen synthesis in NIH-3T3 cells by up-regulation of miR-29a via ERK1/2 and β-catenin signaling pathways.

    更新日期:2019-11-11
  • TRAV gene segments further away from the TRAJ gene segment cluster appear more commonly in human tumor and blood samples
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-05
    Nagehan Pakasticali, Tommy Gill, Boris I. Chobrutskiy, Wei Lue Tong, Michael Ramsamooj, George Blanck

    We considered the possibility that the greater the distance between an immune receptor V and J, the more likely the V usage. Such a hypothesis is supported by results from mouse experiments. And, such a hypothesis is consistent with the fundamental nature of recombination and genomic distance: the further the distance, the greater the chance of a DNA break. Thus, we exploited the vast dataset of V and J recombination reads available for the human TRA gene, particularly from cancer and blood specimens, to assess the frequency of TRAV usage with respect to distance from the TRAJ cluster. Results indicated that, indeed, over the entire TRAV cluster, there is a greater chance of V usage the further the distance from the J cluster. These results do not address causation, and are not consistent for certain individual V gene segments, but the results do indicate that overall, the larger the distance between the V and J gene segment cluster, the more likely the appearance of at least a subset of TRAV segments, particularly among tumor infiltrating lymphocytes. With a similar approach, the distal TRAV gene segments were also found to be more commonly associated with a subset of distal TRAJ segments. These results have implications for restrictions on the apparent TRA repertoire in disease settings.

    更新日期:2019-11-06
  • A chimeric recombinant infectious hematopoietic necrosis virus induces protective immune responses against infectious hematopoietic necrosis and infectious pancreatic necrosis in rainbow trout
    Mol. Immunol. (IF 3.064) Pub Date : 2019-11-05
    Jing-Zhuang Zhao, Miao Liu, Li-Ming Xu, Zhen-Yu Zhang, Yong-Sheng Cao, Yi-Zhi Shao, Jia-Sheng Yin, Hong-Bai Liu, Tong-Yan Lu

    Infectious pancreatic necrosis virus (IPNV) and infectious hematopoietic necrosis virus (IHNV) are two common viral pathogens that cause severe economic losses in all salmonid species in culture, but especially in rainbow trout. Although vaccines against both diseases have been commercialized in some countries, no such vaccines are available for them in China. In this study, a recombinant virus was constructed using the IHNV U genogroup Blk94 virus as a backbone vector to express the antigenic gene, VP2, from IPNV via the reverse genetics system. The resulting recombinant virus (rBlk94-VP2) showed stable biological characteristics as confirmed by virus growth kinetic analyses, pathogenicity analyses, indirect immunofluorescence assays and western blotting. Rainbow trout were immunized with rBlk94-VP2 and then challenged with the IPNV ChRtm213 strain and the IHNV Sn1203 strain on day 45 post-vaccination. A significantly higher survival rate against IHNV was obtained in the rBlk94-VP2 group on day 45 post-vaccination (86%) compared with the PBS mock immunized group (2%). Additionally, IPNV loads decreased significantly in the rBlk94-VP2 immunized group in the liver (28.6-fold to 36.5-fold), anterior kidney (21.7-fold to 44.2-fold), and spleen (14.9-fold to 22.7-fold), as compared with the PBS mock control group. The mRNA transcripts for several innate and adaptive immune-related proteins (IFN-γ, IFN-1, Mx-1, CD4, CD8, IgM, and IgT) were also significantly upregulated after rBlk94-VP2 vaccination, and neutralizing antibodies against both IHNV and IPNV were induced on day 45 post-vaccination. Collectively, our results suggest that this recombinant virus could be developed as a vaccine vector to protect rainbow trout against two or more diseases, and our approach lays the foundations for developing live vaccines for rainbow trout.

    更新日期:2019-11-06
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