Comparative transcriptome analysis of normal and CD44-deleted mouse brain under chronic infection with Toxoplasma gondii
Introduction
Toxoplasmosis is an important zoonosis caused by T. gondii, an obligate intracellular apicomplexan parasite with a wide host range in almost all the warm-blood animals including humans (Tenter et al., 2000; Montoya and Liesenfeld, 2004; Liu et al., 2015). Over two billion people in the world were infected with T. gondii (Robert-Gangneux et al., 2012; Galvan-Ramirez Mde et al., 2012) . Generally, T. gondii infection is asymptomatic for humans with normal immunity, but is severe or even fatal for immune-compromised individuals or for the fetus (Luft and Remington, 1992; Dubey, 2004; Almeida et al., 2015; Arantes et al., 2015; Machala et al., 2015). Clinical infection is common and asymptomatic during clinical toxoplasmosis, usually with the longtime existing of cysts in the brain of patients (Hakimi et al., 2017) . However, cysts in the brain can be re-activated in practice and cause serious damage to the host when the immunity decreases (Krishnamurthy and Konstantinou, 2017).
CD44 is a highly expressed cell adhesion molecule with hyaluronan (HA) as a principal ligand, a glycosaminoglycan that is a major component of the extracellular matrix (ECM) (Toole, 1997; Ponta et al., 2003; Su et al., 2017; Rios de la Rosa et al., 2017). Previous studies indicated that infection with T. gondii contributes to the increasing expression of CD44 of T, B, NK cells, and macrophages in mice (Blass et al., 2001). The role of CD44 in the inflammatory process is conducive to the transfer of leukocytes into local inflammatory site (Lesley et al., 1993; Pure and Cuff, 2001; Ehlers et al., 2018; Dong et al., 2018). Besides, CD44 can also regulate cytotoxicity, strengthen production of pro-inflammatory cytokines and chemokines, and provide costimulation for T cells (Matsumoto et al., 1998; Lee-Sayer et al., 2018). Furthermore, CD44 has an important role in the regulation of IFN-γ production which contributes to immunopathology during T. gondii infection (Blass et al., 2001; Saraav et al., 2019, Goldszmid et al., 2007). However, knowledge about the molecular mechanism of CD44 during the chronic infection of T. gondii is still unclear, clarifying this mechanism should provide a foundation for understanding the host's immune defense against T. gondii infection.
In the present study, the post infection total gene expression patterns between normal mouse brain and CD44-deleted mouse brain were investigated using RNA-seq strategies to understand on a transcriptional level the possible function of CD44 in the brain during chronic infection with T. gondii. Our findings indicated genes related with immune responses, such as immunoglobulins, MHC and chemokines ligands, were significantly up-regulated in mouse after T. gondii infection. The transcriptome data based on RNA-seq demonstrated that the immune responses were significantly stimulated in normal C57 mouse brain after chronic infection with T. gondii, but not for CD44-deleted mouse brain. The present study should provide basis for understanding the possible function of CD44 in the mouse brain after chronic infection with T. gondii, as well as contribute to the development of novel targets for the prevention and control of toxoplasmosis.
Section snippets
Mice
Ten-week-old male wild-type C57BL/6 female mice and CD44-deleted C57Bl/6 female mice in the present were obtained from Hubei Provincial Center for Diseases Control and Prevention (Wuhan, China). All the mice were kept under SPF condition of the experimental animal center of Hubei Province. The study was performed under the instructions and approval of Laboratory Animals Research Centre of Hubei province in P. R. China and the ethics committee of Huazhong Agricultural University (Permit number:
Illumina sequencing and read mapping
For each group, more than 3.3 × 107 raw sequences were generated (Table S2). By elimination of low-quality reads, poly-N reads, 3′ adaptor sequences, and the short reads, 32–48 million good sequences with average length of 150 bp were obtained from each group. For each group, more than 96% sequences were high-quality, indicating that the cDNA library quality was suitable for subsequent analysis.
Then, the clean reads were compared with the Mouse (M. musculus) and T. gondii genome, respectively.
Discussion
T. gondii infection results in the appearance of a population of CD44. Furthermore, administration of anti-CD44 to infected mice prevents the development of a CD4+ T cell-dependent, infection-induced inflammatory response in small intestine characterized by the over-expression of IFN-γ(Blass et al., 2001; Saraav et al., 2019). Thus, there is an urgent need to explore the function of CD44 in mouse during Toxoplasma infection. In this research, the gene expression profile between normal and
Conclusion
In summary, the present study is helpful to understand the response of the host to chronic T. gondii infection, and the results of the study serves as a model to investigate the important role of CD44 in assisting the host against chronic infection of T. gondii. CD44 was not a necessary cytokine for the host against T. gondii infection, but compared normal and CD44-deleted mice after T. gondii infection, four types of genes containing immunoglobulins, MHC II antigens, chemokines ligands and
Author contributions
RF conceived and designed the study. SYL and BH drafted the manuscript. SYL, BH, CHY, JY, LXW, XD, XKD performed the experiments and JLZ analyzed the data. All the authors read and approved the final manuscript.
Ethics approval and consent to participate
The study was performed under the instructions and approval of Laboratory Animals Research Centre of Hubei province in P. R. China and the ethics committee of Huazhong Agricultural University (Permit number: 4200695757).
Consent for publication
Not applicable.
Availability of data and materials
The datasets supporting the conclusions detailed in this article are available in the SRP repository (SRP155079, https://www.ncbi.nlm.nih.gov/sra/SRP155079).
Funding
This work was supported by the National Natural Science Foundation of China (R.F., No. 31572510), the Natural Science Foundation of Hubei Province (Grant No. 2017CFA020) and Da Bei Nong Group Promoted Project for Young Scholar of HZAU (Grant No. 2017DBN001).
Declaration of Competing Interest
The authors declare that they have no competing interests. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
Not applicable.
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The first two authors contributed equally to this paper