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Whole blood can be used as an alternative to isolated peripheral blood mononuclear cells to measure in vitro specific T-cell responses in human samples J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-23 Philippe Moris; Aurélie Bellanger; Opokua Ofori-Anyinam; Erik Jongert; Juan-Pablo Yarzabal Rodriguez; Michel Janssens
Vaccinology is confronted with diseases for which the control of T-cell responses by the vaccine is essential. Among the assays that have been designed to assess T-cell responses, intracellular cytokine staining (ICS) combined with flow cytometry is well-suited in the frame of clinical trials. This assay can be used starting from isolated peripheral blood mononuclear cells (PBMC) or from whole blood
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Human peripheral basophils extended phenotype shows a high expression of CD244 immuno-regulatory receptor J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-22 Anne-Emmanuelle Berger; Coralie Durrieu; Charles Dzviga; Jean-Luc Perrot; Claude Lambert
Introduction Basophils play a major physio-pathological role in hypersensitivity related diseases. Basophils express high affinity Immunoglobulin (Ig) E receptors (FcεRI), IgG and complement regulatory. Basophils also have immunoregulatory activity through interaction with T cells. The aim of this study was to look for the expression of markers reflecting the activation status of peripheral Basophil
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Relationship between circulating tumor-associated autoantibodies and clinical outcomes in advanced-stage NSCLC patients receiving PD-1/−L1 directed immune checkpoint inhibition. J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-09 Imad Tarhoni; Connor J. Wakefield; Revathi Kollipara; Mary Jo Fidler; Marta Batus; Philip Bonomi; Jeffrey A. Borgia
Background Durable tumor regressions are observed in a subset of advanced-stage non-small cell lung cancer (NSCLC) patients receiving PD-1/−L1 targeted immune checkpoint inhibitors (or ‘immunotherapy’) alone or in combination with chemotherapy. However, the majority of advanced NSCLC patients receiving these agents do not experience long-term disease control. Existing methods to identify patients most
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Systematic approach in macrophage polarization experiments: Maintaining integrity and reproducibility using flow cytometry and sample preparation J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-19 Jennifer R. McCall; Kathryn Sausman
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Development of recombinant NS1-NS3 antigen based indirect ELISA for detection of bluetongue antibodies in sheep J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-04 Nihar Nalini Mohanty; Divakar Hemadri; Archana Munivenkatarayappa; Namrata Shetty; Vinutha Subramanyam; Sanchay Kumar Biswas; Mohammed Mudassar Chanda; Sathish B. Shivachandra
Bluetongue is an insect borne (Culicoides) viral disease of small ruminants. The virus blankets the globe with a wide serotypic variation, numbered from 1 to 28. In India 21 different serotypes have been reported to be circulating across the various agro-climatic zones of the country. Non-structural proteins (NSPs) of bluetongue virus have always remained ideal target for differentiation of infected
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Monocyte monolayer assay in pre-transfusion testing: A magic key in transfusing patients with recurrent bad cross-match due to alloimmunization J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-16 Hebat Allah N. El-sayed; Maha R. Abdollah; Shereen N. Raafat; Dina Ragab
Background The monocyte monolayer assay (MMA) is an in-vitro assay that can predict the outcome of blood transfusion of antigen positive units when serologically compatible blood is not available. Materials and methods Fifty-four patients testing positive by the antibody screening test using gel agglutination were further examined by the alloantibody identification panel to determine alloantibody specificity
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Development and characterization of an indirect ELISA to detect SARS-CoV-2 spike protein-specific antibodies J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-04 Verena Krähling; Sandro Halwe; Cornelius Rohde; Dirk Becker; Susanne Berghöfer; Christine Dahlke; Markus Eickmann; Meryem S. Ercanoglu; Lutz Gieselmann; Astrid Herwig; Alexandra Kupke; Helena Müller; Petra Neubauer-Rädel; Florian Klein; Christian Keller; Stephan Becker
The current Severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) pandemic is a public health emergency of international concern. Sensitive and precise diagnostic tools are urgently needed. In this study, we developed a SARS-CoV-2 spike (S1) protein enzyme-linked immunosorbent assay (ELISA) to detect SARS-CoV-2-specific antibodies. The SARS-CoV-2 S1 ELISA was found to be specific [97
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Impact of complement and difference of cell-based assay and ELISA in determination of neutralization capacity against mumps and measles virus J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-04 Marija Brgles; Tihana Kurtović; Maja Lang Balija; Ana Hećimović; Tatjana Mušlin; Beata Halassy
Neutralizing antibodies against mumps and measles virus are considered a correlate of protection against these diseases. Measurement of neutralizing antibodies is mostly performed using plaque reduction neutralization assay or 50% cell culture infective dose (CCID50) neutralization assay, but there are attempts for measuring neutralizing antibodies using enzyme-linked immunosorbent assay (ELISA) which
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Depletion of CD45RA+ T cells: Advantages and disadvantages of different purification methods J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-06 Melanie Bremm; Theresa Krastel; Claudia Cappel; Olga Zimmermann; Lisa-Marie Pfeffermann; Verena Katzki; Halvard Bonig; Richard Schäfer; Eva Rettinger; Michael Merker; Sebastian Bremm; Kirsten Schaefer; Thomas Klingebiel; Jan Soerensen; Peter Bader; Sabine Huenecke
Background Recently, new advances were made regarding the depletion of CD45RA+ naïve T cells from haploidentical grafts as they are suspected to be the most alloreactive. Methods Within this project we investigated CD45RA-depletion from G-CSF mobilized PBSC by two different purification strategies according to GMP, specifically direct depletion of CD45RA+ cells (one-step approach), or CD34-positive
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Graphene oxide-gold nanoparticle-aptamer complexed probe for detecting amyloid beta oligomer by ELISA-based immunoassay J. Immunol. Methods (IF 1.901) Pub Date : 2020-12-14 Jing Zhao; Wenlong Chang; Lu Liu; Xiaoming Xing; Chao Zhang; Huihong Meng; Subash C.B. Gopinath; Thangavel Lakshmipriya; Yeng Chen; Yonggang Liu
Highly sensitive and easy detection method for Alzheimer's disease (AD) with a suitable biomarker is mandatory for preventing the factors resulting from AD. This research reports a modified ELISA with graphene for the detection of AD biomarker amyloid beta (Aβ) oligomer. Gold nanoparticle (AuNP) conjugated aptamer was used as the capture probe and attached on ELISA-graphene oxide surface through the
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Quantitative assessment of NFκB transcription factor activity J. Immunol. Methods (IF 1.901) Pub Date : 2021-01-01 Terrence T.J. Hunter; David Fear; Paul Lavender; Jo Spencer; Mark Peakman; Mohammad A.A. Ibrahim
The Nuclear Factor Kappa B (NFκB) pathway is an important signalling pathway in the immune system. Single gene defects in the NFκB pathway are described in a number of immunodeficiency diseases. These conditions provide a unique opportunity to investigate the mechanisms of NFκB function and how genetic mutations that disrupt this function lead to human disease. Here we describe a robust method for
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Validation of a monoclonal antibody directed against the human sphingosine 1-phosphate receptor type 1 J. Immunol. Methods (IF 1.901) Pub Date : 2020-12-30 Andreas V. Thuy; Jefri Jeya Paul; Cynthia Weigel; Anke C. Ziegler; Orlando Guntinas-Lichius; Markus H. Gräler
The sphingosine 1-phosphate receptor type 1 (S1PR1) has several important functions, including stabilizing endothelial barrier and maintaining lymphocyte circulation. These functions are critically dependent on the regulation of S1PR1 cell surface expression. Currently available antibodies against human S1PR1 are not able to pick up cell surface expression on living cells by flow cytometry due to intracellular
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Detection of engineered T cells in FFPE tissue by multiplex in situ hybridization and immunohistochemistry J. Immunol. Methods (IF 1.901) Pub Date : 2020-12-29 Jocelyn H. Wright; Li-Ya Huang; Stephanie Weaver; L. Diego Archila; Megan S. McAfee; Alexandre V. Hirayama; Aude G. Chapuis; Marie Bleakley; Anthony Rongvaux; Cameron J. Turtle; Savanh Chanthaphavong; Jean S. Campbell; Robert H. Pierce
Identifying engineered T cells in situ is important to understand the location, persistence, and phenotype of these cells in patients after adoptive T cell therapy. While engineered cells are routinely characterized in fresh tissue or blood from patients by flow cytometry, it is difficult to distinguish them from endogenous cells in formalin-fixed, paraffin-embedded (FFPE) tissue biopsies. To overcome
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Multiplex bead binding assays using off-the-shelf components and common flow cytometers J. Immunol. Methods (IF 1.901) Pub Date : 2020-12-25 Takamitsu Hattori; Akiko Koide; Tatyana Panchenko; Larizbeth A. Romero; Kai Wen Teng; Alexis D. Corrado; Shohei Koide
The ability to quantify protein-ligand interactions in an accurate and high-throughput manner is important in diverse areas of biology and medicine. Multiplex bead binding assays (MBBAs) are powerful methods that allows for simultaneous analysis of many protein-ligand interactions. Although there are a number of well-established MBBA platforms, there are few platforms suitable for research and development
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Detergent wash improves vaccinated lymph node handling ex vivo J. Immunol. Methods (IF 1.901) Pub Date : 2020-12-14 Alexander G. Ball; Maura C. Belanger; Rebecca R. Pompano
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Cryopreservation of peripheral blood mononuclear cells for use in proliferation assays: First step towards potency assays J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-10 Morten Juhl; Jan Pravsgaard Christensen; Anders Elm Pedersen; Jens Kastrup; Annette Ekblond
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Development and optimization of a Zika virus antibody-dependent cell-mediated cytotoxicity (ADCC) assay J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-16 Xuemin Chen; Larry J. Anderson; Christina A. Rostad; Lingmei Ding; Lilin Lai; Mark Mulligan; Nadine Rouphael; Muktha S. Natrajan; Courtney McCracken; Evan J. Anderson
Zika virus (ZIKV) has become a global public health issue due to its teratogenicity and ability to cause Guillain-Barré syndrome in adults. Although anti-ZIKV envelope protein neutralizing antibodies correlate with protection, the non-neutralizing function of ZIKV antibodies including antibody-dependent cell-mediated cytotoxicity (ADCC) is incompletely understood. To study the role of ADCC antibodies
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Generation of a cost-effective cell line for support of high-throughput isolation of primary human B cells and monoclonal neutralizing antibodies J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-15 Rachael E. Whaley; Sarah Ameny; Tanvi Arkatkar; Aaron Seese; Abigail Wall; Iram Khan; Joseph J. Carter; Erin M. Scherer; David J. Rawlings; Denise A. Galloway; M. Juliana McElrath; Kristen W. Cohen; Andrew T. McGuire
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The antinuclear antibody dense fine speckled pattern and possible clinical associations: An indication of a proinflammatory microenvironment J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-27 Mia C. Lundgren; Smarika Sapkota; Daniel J. Peterson; John T. Crosson
Background Indirect immunofluorescence (IIF) is the most prevalent screening antinuclear antibody test for systemic autoimmune rheumatic disease (SARD). Certain IIF patterns have known antibody and disease associations, but the dense fine speckled (ANA-DFS) pattern has no confirmed clinical associations. Our objective was to determine the prevalence of SARD among a group of ANA-DFS positive individuals
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Development of an in-house capture ELISA: An attempt to detect CagA antigen in sera of Helicobacter pylori infected patients J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-29 Barik A. Salih; Cebrail Karakus; Duygu Yazici; Zeynep Ulupinar; Fahri Akbas; Fatima Yucel; Esin Akcael; Yusuf Akcan
The CagA protein one of the key virulence factors of Helicobacter pylori plays an important role in the pathogenesis of peptic ulcer diseases. Unfortunately the cagA gene status can only be determined by PCR while serology is an alternative approach to detect antigens or antibodies. Our aim is to detect the CagA antigen in sera of infected subjects by the development of an in-house capture ELISA test
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Development of a lateral flow immunochromatography test for the rapid detection of bovine tuberculosis J. Immunol. Methods (IF 1.901) Pub Date : 2020-12-13 Natalia Alonso; Natanael Griffa; Roberto D. Moyano; Maria L. Mon; María A. Colombatti Olivieri; Soledad Barandiaran; Marcela Martínez Vivot; Gonzalo Fiorini; Ana M. Canal; María P. Santangelo; Mahavir Singh; María I. Romano
Detection of specific antibodies would be a useful test strategy for bovine tuberculosis (bTB) as a complement to the single skin test. We developed a lateral flow immunochromatography (LFIC) test for rapid bTB detection based on the use of a conjugate of gold nanoparticles with a recombinant G protein. After evaluating 3 Mycobacterium bovis (MB) antigens: ESAT-6, CFP-10 and MPB83 for the control line
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Monitoring Ravulizumab effect on complement assays J. Immunol. Methods (IF 1.901) Pub Date : 2020-12-13 Maria A.V. Willrich; Paula M. Ladwig; Mark A. Martinez; Meera R. Sridharan; Ronald S. Go; David L. Murray
Ravulizumab is a new C5 inhibitor therapeutic monoclonal antibody with a longer half-life than eculizumab. Monitoring complete complement blockade by eculizumab has allowed personalized therapy in specific settings. Similar action is expected with ravulizumab. Ravulizumab has 4 different amino acids from eculizumab, which allow greater affinity for the FcRn immunoglobulin receptor and change the affinity
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Development of competitive inhibition ELISA as an effective potency test to analyze human rabies vaccines and assessment of the antigenic epitope of rabies glycoprotein J. Immunol. Methods (IF 1.901) Pub Date : 2020-12-09 Dipen Soni; Itishree Sahoo; Asha D. Mallya; Praveen Kamthe; Ashish Sahai; Sunil Kumar Goel; Prasad S. Kulkarni; Rajeev M. Dhere
The potency of all modern tissue culture human rabies vaccines is measured based on the National Institute of Health (NIH) potency test that is laborious, time-consuming, involves large test variations and requires sacrifice of large number of animals. To circumvent these limitations, several researchers and WHO expert working groups have discussed development of alternative in vitro methods to replace
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Immunogenicity evaluating of the SLNs-alginate conjugate against Pseudomonas aeruginosa J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-28 Hossein Afshari; Masoud Maleki; Mozhdeh Hakimian; Roghaye Ahmadlou Tanha; Mojtaba Salouti
P. aeruginosa is of particular importance due to its numerous pathogens and the spread of its multidrug-resistant strains around the world. Hence there is a need to develop an effective vaccine to prevent the diseases with P. aeruginosa. The aim of present study was to evaluate the immunogenicity of alginate (Alg) antigen in conjugation with SLN as a candidate for nanovaccine against P. aeruginosa
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Development of magnetic particle-based chemiluminescence immunoassay for measurement of human procalcitonin in serum J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-12 Minjing Liao; Jiao Zheng; Ye Xu; Yilan Qiu; Chuan Xia; Zhihong Zhong; Lihui Liu; Hongrong Liu; Rushi Liu; Songyue Liang
Background Serum procalcitonin (PCT) has been recognized as a primary biomarker in bacterial infections, and monitoring its concentration could help to evaluate the prognosis of sepsis and guide the antibiotic administration. We aimed to establish a fast and accurate immunoassay for PCT quantitation. Methods Our newly developed monoclonal antibodies (mAbs) against human PCT were preliminarily evaluated
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A streamlined proliferation assay using mixed lymphocytes for evaluation of human mesenchymal stem cell immunomodulation activity J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-16 Maryanne C. Herzig; Christopher P. Delavan; Katherine J. Jensen; Carolina Cantu; Robbie K. Montgomery; Barbara A. Christy; Andrew P. Cap; James A. Bynum
Background Mesenchymal stromal cells (MSCs) have been proposed for treatment of acute respiratory distress syndrome (ARDS), graft versus host disease (GVHD), wound healing and trauma. A consensus is building that immunomodulation by MSCs is important for therapeutic potential. MSCs suppress peripheral blood mononuclear cell (PBMC) proliferation in vitro, potentially reflecting an ability to suppress
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Development and verification of an enzyme-linked immunosorbent assay for the quantification of toxoid A and toxoid B from Clostridioides difficile J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-19 S. Anwar; D. Bryan; P. Rigsby; T. Dougall; S. Rijpkema
Clostridioides difficile (C. difficile) is the most common cause of nosocomial antibiotic associated diarrhoea. The incidence of C. difficile infection (CDI) has been rising worldwide over the last 20 years with consequent rises in morbidity, mortality and healthcare costs, although the incidence has fallen in the UK over the last few years. Confirmation of diagnosis and early intervention are critical
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A method for expansion and retroviral transduction of mouse regulatory T cells J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-20 Dan Wu; May Q. Wong; Jens Vent-Schmidt; Dominic A. Boardman; Theodore S. Steiner; Megan K. Levings
Adoptive cell therapy with genetically modified regulatory T cells (Tregs) is under clinical investigation for the treatment of transplant rejection and various autoimmune conditions. A limitation of modelling this approach in mice is the lack of optimized protocols for expanding and transducing mouse Tregs. Here we describe a protocol for purifying, expanding and retrovirally transducing mouse Tregs
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Comparative analyses of SARS-CoV-2 binding (IgG, IgM, IgA) and neutralizing antibodies from human serum samples J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-28 Livia Mazzini; Donata Martinuzzi; Inesa Hyseni; Linda Benincasa; Eleonora Molesti; Elisa Casa; Giulia Lapini; Pietro Piu; Claudia Maria Trombetta; Serena Marchi; Ilaria Razzano; Alessandro Manenti; Emanuele Montomoli
A newly identified coronavirus, named SARS-CoV-2, emerged in December 2019 in Hubei Province, China, and quickly spread throughout the world; so far, it has caused more than 49.7 million cases of disease and 1,2 million deaths. The diagnosis of SARS-CoV-2 infection is currently based on the detection of viral RNA in nasopharyngeal swabs by means of molecular-based assays, such as real-time RT-PCR.
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An in vitro refolding method to produce oligomers of anti-CHIKV, E2-IgM Fc fusion subunit vaccine candidates expressed in E. coli J. Immunol. Methods (IF 1.901) Pub Date : 2020-09-21 Sandeep Kumar; Vikas Kumar Singh; Manohar Vasam; Poonam Shewale Patil; Rajeev K. Dhaked; Abdul S. Ansari; Nirmal K. Lohiya; Deepti Parashar; Suman Tapryal
Recombinant envelope protein-1 (E1) and E2 of Chikungunya virus (CHIKV) has been shown to elicit neutralizing antibodies and a balanced Th1/Th2 response in mice however with limited protection. Recently reported CHIK virus-like particles showed augmented immunity and protection in adult mice in comparison to E1 and E2, however exacerbated the disease in aged subjects. In order to improve the overall
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Objective evaluation of immediate reading skin prick test applying image planimetric and reaction thermometry analyses. J. Immunol. Methods (IF 1.901) Pub Date : 2020-09-19 Ana Laura Mendes Almeida,Edson Luiz Pontes Perger,Ramon Hernany Martins Gomes,Guilherme Dos Santos Sousa,Lucas Hecker Vasques,José Eduardo Petit Rodokas,Jaime Olbrich Neto,Rafael Plana Simões
The skin prick test is used to diagnose patients' sensitization to antigens through a mediated IgE response. It is a practical and quick exam, but its diagnosis depends on instruments for measuring the allergic response and observer's interpretation. The conventional method for inferring about the allergic reaction is performed from the dimensions of the wheals, which are measured using a ruler or
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Production and characterization of a neutralizing antibody against botulinum neurotoxin A J. Immunol. Methods (IF 1.901) Pub Date : 2020-09-30 Xianghua Xiong; Sunhui Lv; Chuxi Fu; Lei Li; Zhijie Sun; Xiaodong Han; Weicai Zhang
As a category A toxic, the botulinum toxin(BoNT) is responsible for human botulism with an estimated lethal dose of 1 ng/kg which greatly increases the potential risk of use as bioweapons. Therefore, the development of anti-BoNT antibodies is urgent. In this paper, the HC domain of BoNT/A was purified and immunized with Balb/c mice. Monoclonal antibodies were screened against BoNT/A from 55 stable
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A novel enzyme immunoassay for the measurement of plasma (1 → 3)-β-D-glucan levels J. Immunol. Methods (IF 1.901) Pub Date : 2020-09-29 Ei-ichiro Sunamura; Manami Iwasaki; Shota Shiina; Shin-ichiro Kitahara; Takuya Yotani; Mitsuhisa Manabe; Osamu Miyazaki
The presence of (1 → 3)-β-D-glucan in human plasma is a marker for fungal infections. Currently, the Limulus amebocyte lysate (LAL)-based assay is widely used for the quantification of plasma (1 → 3)-β-D-glucan. However, it has limitations in clinical use, such as an unstable supply of natural resources, complicated manufacturing process, and low-throughput of the reagents. Alternative assays exploiting
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Generation and characterization of genetically stable heterohybridomas producing foot-and-mouth disease virus-specific porcine monoclonal antibodies J. Immunol. Methods (IF 1.901) Pub Date : 2020-09-28 Michael C. Puckette; Erica Martel; Jacob Rutherford; José Barrera; William Hurtle; Melia Pisano; Lauren Martignette; Mariceny Zurita; John G. Neilan; Chungwon J. Chung
This report covers the methodology for generation of stable heterohybridoma clones producing Foot-and-mouth disease virus (FMDV) reactive porcine monoclonal antibodies (mAbs). Swine received five inoculations of an inactivated O1 Manisa FMDV vaccine prior to the harvest of splenocytes. Due to the lack of a species-specific hybridoma fusion partner, the Sp2/0 murine myeloma cell line was utilized for
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A high-throughput multiplex assay to characterize flavivirus-specific immunoglobulins J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-03 Mélanie Merbah; Suzanne Wollen-Roberts; Zhanna Shubin; Yifan Li; Hongjun Bai; Vincent Dussupt; Letzibeth Mendez-Rivera; Bonnie Slike; Shelly J. Krebs; Kayvon Modjarrad; Nelson L. Michael; Morgane Rolland
Genus Flavivirus, which includes 53 virus species, is the leading cause of arthropod-borne diseases in humans. Diagnosis of these viral diseases is complicated by their overlapping epidemiology and clinical manifestations, and the fact that cross-reactive antibody responses are frequently elicited by individuals in response to infection. We developed a bead-based immunoassay to concomitantly profile
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Intracellular fluoride influences TASK mediated currents in human T cells J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-05 Alexander M. Herrmann; Manuela Cerina; Stefan Bittner; Sven G. Meuth; Thomas Budde
The expression of Kv1.3 and KCa channels in human T cells is essential for maintaining cell activation, proliferation and migration during an inflammatory response. Recently, an additional residual current, sensitive to anandamide and A293, compounds specifically inhibiting currents mediated by TASK channels, was observed after complete pharmacological blockade of Kv1.3 and KCa channels. This finding
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Avoiding ambient air in test tubes during incubations of human whole-blood minimizes complement background activation J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-06 Benjamin S. Storm; Dorte Christiansen; Tom Eirik Mollnes; Erik Waage Nielsen
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Distinct and overlapping effects of β2-glycoprotein I conformational variants in ligand interactions and functional assays J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-06 Gábor Szabó; Krisztina Pénzes; Bernadett Torner; Miklós Fagyas; Tünde Tarr; Pál Soltész; Gréta Kis; Miklós Antal; János Kappelmayer
One of the most abundant coagulation proteins is β2-glycoprotein I (β2GPI) that is present in humans at a concentration of around 200 mg/L. Its physiological role is only partially understood, but it adopts several different structural forms the majority of which are the open and closed forms. We isolated native (circular) β2GPI and converted it into an open conformation. The effectiveness of these
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An algorithmic approach to serological work-up of ABO sub-groups which present as ABO discrepancies in resource constraint settings J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-13 Aseem K. Tiwari; Divya Setya; Dinesh Arora; Swati Pabbi Mehta; Geet Aggarwal; Subhasis Mitra
Background ABO subgroups or weaker variants of A or B are group A or B subjects whose erythrocytes give a weak or negative reaction serologically with anti-A or Anti – B antisera respectively. Occurrence of these subgroups may lead to an ABO discrepancy which often puts transfusion services in a quandary. ABO subgroups which present as ABO discrepancies can be missed if reverse grouping is not performed
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An innovative method for characterizing neutralizing antibodies against antibody-derived therapeutics J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-13 Annelies Coddens; Veerle Snoeck; Lieselot Bontinck; Marie-Ange Buyse; Samuel O. Pine
Detection of anti-drug antibodies (ADA) that have a neutralizing capacity is an important aspect of immunogenicity evaluation during development of biotherapeutics, but developing and validating neutralizing antibody (NAb) assays that show direct interference of a biologic function is a challenging and resource-intensive activity. In particular, the need for adequate drug and target tolerance often
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A novel co-culture assay to assess anti-tumor CD8+ T cell cytotoxicity via luminescence and multicolor flow cytometry J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-15 Verónica Olivo Pimentel; Ala Yaromina; Damiënne Marcus; Ludwig J. Dubois; Philippe Lambin
T cell immunotherapies have shown great promise in patients with advanced cancer disease, revolutionizing treatment. T cell cytotoxicity is crucial in its efficacy, therefore developing ex vivo methods testing tumor and T cell interactions is pivotal. Increasing efforts have been made in developing co-culture assays with sophisticated materials and platforms aiming to mimic the tumor microenvironment
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An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS? J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-15 Gerald Pfister; Salman M. Toor; Varun Sasidharan Nair; Eyad Elkord
Flow cytometry and fluorescence-activated cell sorting have become invaluable tools to analyze and isolate specific cell populations in a wide range of biomedical research and clinical applications. In countless approaches worldwide, scientists are using single cell analyses to better understand the significance and variation within different cellular populations, and fluorescence-activated cell sorting
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Guidelines for analysis of low-frequency antigen-specific T cell results: Dye-based proliferation assay vs 3H-thymidine incorporation J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-03 Daniela Di Blasi; Iris Claessen; Annelies W. Turksma; Josine van Beek; Anja ten Brinke
It is generally recognized that dysregulation of the immune system plays a critical role in many diseases, including autoimmune diseases and cancer. T cells play a crucial role in maintaining self-tolerance, while loss of immune tolerance and T cell activation can lead to severe inflammation and tissue damage. T cell responses have a key role in the effectiveness of vaccination strategies and immunomodulating
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Chemically modified mRNA nucleofection of primary human T cells J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-06 Nikolaus Thuille; Tajana Sajinovic; Kerstin Siegmund; Gottfried Baier
Here we show that an approach of in-vitro transcribed mRNA nucleofection expands the range of transfection of primary human T cells. It represents a reproducible and time-efficient technology, and is thus an ideal tool in basic research involving highly controlled in-vitro experiments with a gene of interest aiming at identifying its biological human T cell function.
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Semi-automated methodology for detection of IgM oligoclonal bands in cerebrospinal fluid and serum samples J. Immunol. Methods (IF 1.901) Pub Date : 2020-10-10 Carmen M. Cabrera; Andrea Gosis
Among the new biomarkers to propose therapeutic decisions in patients suffering from multiple sclerosis (MS) are the IgM oligoclonal bands (OCBs) in cerebrospinal fluid (CSF). At the current time, however, IgM OCBs are detected in laboratories at investigation level and not in the routine practice due to their complexity. For this, we have applied a semi-automated method based on an isoelectrofocusing
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Enrichment of circulating tumor-derived extracellular vesicles from human plasma J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-24 Kathryn E. Yoh; Christopher J. Lowe; Shilpi Mahajan; Rebecca Suttmann; Trung Nguy; Mike Reichelt; Jenny Yang; Rachel Melendez; Yijin Li; Luciana Molinero; Jane Ruppel; Wenfeng Xu; Vicki Plaks
Extracellular vesicles (EVs) are gaining considerable traction within the liquid biopsy arena, as carriers of information from cells in distant sites that may not be accessible for biopsy. Therefore, there is a need to develop methods to enrich for specific EV subtypes, based on their cells of origin. Here we describe the development of an automated method to enrich tumor-derived EVs from plasma using
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Application of a novel drug-tolerant target assay for measuring target engagement when only one epitope remains after therapeutic antibodies bind their targets J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-23 Quincy L. Carter; Robert W. Siegel; Yuewei Qian; Robert J. Konrad
The measurement of proteins with a limited number of available non-overlapping epitopes recognizable by antibodies represents a common challenge for the development of drug-tolerant clinical biomarker assays. For target proteins with two dominant epitopes, only one epitope remains when the other is occupied by the therapeutic antibody. Alternative strategies for overcoming this obstacle have been described
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Using different proteolytic enzymes to digest antibody and its impact on stability of antibody mimetics J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-21 Hanieh Khalili
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Novel immunoassay for diagnosis of ongoing Clostridioides difficile infections using serum and medium enriched for newly synthesized antibodies (MENSA) J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-19 Natalie S. Haddad; Sophia Nozick; Geena Kim; Shant Ohanian; Colleen Kraft; Paulina A. Rebolledo; Yun Wang; Hao Wu; Adam Bressler; Sang Nguyet Thi Le; Merin Kuruvilla; L. Edward Cannon; F. Eun-Hyung Lee; John L. Daiss
Background Clostridioides difficile infections (CDI) have been a challenging and increasingly serious concern in recent years. While early and accurate diagnosis is crucial, available assays have frustrating limitations. Objective Develop a simple, blood-based immunoassay to accurately diagnose patients suffering from active CDI. Materials and methods Uninfected controls (N = 95) and CDI patients (N = 167)
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Purification of murine immunoglobulin E (IgE) by thiophilic interaction chromatography (TIC) J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-14 Natasa Vukovic; Salim Harraou; Sander M.J. van Duijnhoven; Dietmar M. Zaiss; Andrea van Elsas
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Characterization of a rat monoclonal antibody raised against ferroptotic cells J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-12 Sho Kobayashi; Yumi Harada; Takujiro Homma; Chikako Yokoyama; Junichi Fujii
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Production of a full chimeric mouse x pig anti-porcine DEC205 receptor recombinant antibody J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-10 Lorena Bustamante-Córdova; Edgar Alonso Melgoza-González; Héctor Parra-Sánchez; Mónica Reséndiz-Sandoval; Alexel Jesús Burgara-Estrella; Jesús Hernández
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Analytical performance of lateral flow immunoassay for SARS-CoV-2 exposure screening on venous and capillary blood samples J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-07 Margaret A. Black; Guomiao Shen; Xiaojun Feng; Wilfredo F. Garcia Beltran; Yang Feng; Varshini Vasudevaraja; Douglas Allison; Lawrence H. Lin; Tatyana Gindin; Michael Astudillo; Diane Yang; Mandakolathur Murali; A. John Iafrate; George Jour; Paolo Cotzia; Matija Snuderl
Objectives We validate the use of a lateral flow immunoassay (LFI) intended for rapid screening and qualitative detection of anti-SARS-CoV-2 IgM and IgG in serum, plasma, and whole blood, and compare results with ELISA. We also seek to establish the value of LFI testing on blood obtained from a capillary blood sample. Methods Samples collected by venous blood draw and finger stick were obtained from
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Blood biomarker algorithms for the diagnosis of mycoplasma pneumoniae respiratory infections J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-07 Per Venge; Staffan Eriksson; Karlis Pauksen
The correct diagnosis of acute infections as to bacteria, mycoplasma or virus is a clinical challenge and has a great impact on the therapeutic decisions. Current diagnostic tests of mycoplasma pneumoniae infections of the respiratory tract such as PCR and serology are either somewhat unreliable or slow and do not entirely meet the clinical needs of accurate and fast diagnosis. The aim of this report
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Positive tissue transglutaminase antibodies with negative endomysial antibodies: Unresolved issues in diagnosing celiac disease J. Immunol. Methods (IF 1.901) Pub Date : 2020-11-07 Maria Infantino; Mario Merone; Mariangela Manfredi; Valentina Grossi; Alessandra Landini; Maria Grazia Alessio; Giulia Previtali; Maria Teresa Trevisan; Brunetta Porcelli; Martina Fabris; Donatella Macchia; Danilo Villalta; Luigi Cinquanta; Federico D'Antoni; Giulio Iannello; Paolo Soda; Nicola Bizzaro
Background The serological screening for celiac disease (CD) is currently based on the detection of anti-transglutaminase (tTG) IgA antibodies, subsequently confirmed by positive endomysial antibodies (EMA). When an anti-tTG IgA positive/EMA IgA negative result occurs, it can be due either to the lower sensitivity of the EMA test or to the lower specificity of the anti-tTG test. This study aimed at
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Methodological comparison of FACS and MACS isolation of enriched microglia and astrocytes from mouse brain. J. Immunol. Methods (IF 1.901) Pub Date : 2020-08-15 Jie Pan,Jun Wan
Microglia and astrocytes, the two innate cells in CNS, are thought to protect and remodel of synapses for proper maintenance and plasticity of neuronal circuits. The two types of cells are the major responders by producing and releasing inflammatory mediators. Isolation of microglia and astrocytes from CNS tissue provides a powerful tool to study basic cell biology and examine the effects of in vivo
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Development and validation of ELISAs for the quantitation of interleukin (IL)-1β, IL-6, IL-8 and IL-10 in ovine plasma. J. Immunol. Methods (IF 1.901) Pub Date : 2020-08-20 Mahé Bouquet,Margaret R Passmore,Louise E See Hoe,John-Paul Tung,Gabriela Simonova,Ai-Ching Boon,John F Fraser
There is growing evidence that inflammation underpins many common diseases. Inflammatory/immunomodulatory/immune mediators, such as cytokines, are key modulators of inflammation and mediate both immune cell recruitment and complex intracellular signalling pathways. Ovine models of disease are increasingly utilized in pre-clinical research, however existing methods for measuring cytokine levels are
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Estimation of ELISA results using a parallel curve analysis. J. Immunol. Methods (IF 1.901) Pub Date : 2020-08-20 Francis Bursa,Ann Yellowlees,Alka Bishop,Angela Beckett,Bassam Hallis,Mary Matheson
We introduce a new method for the analysis of enzyme-linked immunosorbent assay (ELISA) data. The new method can use data near the asymptotes and does not give undue weight to responses on the flatter parts of the dose-response curve. We apply it to simulated data and to two real-world assays and show it is more accurate and more precise than the traditional interpolation method. In particular, the
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Analysis of COVID-19 convalescent plasma for SARS-CoV-2 IgG using two commercial immunoassays. J. Immunol. Methods (IF 1.901) Pub Date : 2020-08-20 Melkon G DomBourian,Kyle Annen,Leah Huey,Gillian Andersen,Patricia A Merkel,Sarah Jung,Samuel R Dominguez,Vijaya Knight
Coronavirus Disease 2019 (COVID-19) convalescent plasma (CCP) was approved by the FDA for use in severe cases of COVID-19 under an emergency Investigational New Drug (IND) protocol. Eligibility criteria for CCP donors includes documentation of evidence of COVID-19 either by viral RNA detection at the time of illness or positive SARS-CoV-2 IgG after recovery if diagnostic testing for COVID-19 was not
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