IL-21/IL-21R Promotes the Pro-Inflammatory Effects of Macrophages during C. muridarum Respiratory Infection

Interleukin-21 and its receptors (IL-21/IL-21R) aggravate chlamydial lung infection, while macrophages (M phi) are one of the main cells infected by chlamydia and the main source of inflammatory cytokines. Therefore, it is particularly important to study whether IL-21/IL-21R aggravates chlamydia respiratory infection by regulating M phi. Combined with bioinformatics analysis, we established an IL-21R-deficient (IL-21R (-/-)) mouse model of Chlamydia muridarum (C. muridarum) respiratory tract infection in vivo, studied C. muridarum-stimulated RAW264.7 by the addition of rmIL-21 in vitro, and conducted adoptive transfer experiments to clarify the association between IL-21/IL-21R and M phi. IL-21R (-/-) mice showed lower infiltration of pulmonary total M phi, alveolar macrophages, and interstitial macrophages compared with WT mice following infection. Transcriptomic analysis suggested that M1-related genes are downregulated in IL-21R (-/-) mice and that IL-21R deficiency affects the M phi-mediated inflammatory response during C. muridarum infection. In vivo experiments verified that in IL-21R (-/-) mice, pulmonary M1-type CD80(+), CD86(+), MHC II+, TNF alpha(+), and iNOS(+) M phi decreased, while there were no differences in M2-type CD206+, TGF-alpha+, IL-10+ and ARG1+ M phi. In vitro, administration of rmIL-21 to C. muridarum-stimulated RAW264.7 cells promoted the levels of iNOS-NO and the expression of IL-12p40 and TNF alpha, but had no effect on TGF beta or IL-10. Further, adoptive transfer of M1-like bone marrow-derived macrophages derived from IL-21R (-/-) mice, unlike those from WT mice, effectively protected the recipients against C. muridarum infection and induced relieved pulmonary pathology. These findings help in understanding the mechanism by which IL-21/IL-21R exacerbates chlamydia respiratory infection by promoting the proinflammatory effect of M phi.