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Γδ T Cells Affect IL-4 Production and B-Cell Tolerance PNAS PLUS

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Γδ T Cells Affect IL-4 Production and B-Cell Tolerance PNAS PLUS γδ T cells affect IL-4 production and B-cell tolerance PNAS PLUS Yafei Huanga, Ryan A. Heiserb, Thiago O. Detanicoa, Andrew Getahunb, Greg A. Kirchenbaumb, Tamara L. Caspera, M. Kemal Aydintuga, Simon R. Cardingc, Koichi Ikutad, Hua Huanga, John C. Cambierb,a, Lawrence J. Wysockia,b, Rebecca L. O’Briena,b, and Willi K. Borna,b,1 aDepartment of Biomedical Research, National Jewish Health, Denver, CO 80206; bDepartment of Immunology & Microbiology, University of Colorado Health Sciences Center, Aurora, CO 80045; cThe Institute of Food Research and Norwich Medical School, University of East Anglia, Norwich Research Park, Norwich NR4 7JT, United Kingdom; and dLaboratory of Biological Protection, Department of Biological Responses, Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan Edited* by Philippa Marrack, Howard Hughes Medical Institute, National Jewish Health, Denver, CO, and approved November 7, 2014 (received for review August 6, 2014) γδ T cells can influence specific antibody responses. Here, we re- In the current study, we further tested this idea by examining port that mice deficient in individual γδ T-cell subsets have altered antibody levels and B cells in nonimmunized mice genetically levels of serum antibodies, including all major subclasses, some- deficient either in individual γδ T-cell subsets or in all γδ T cells. times regardless of the presence of αβ T cells. One strain with The focus on antibodies derives from our earlier observation that a partial γδ deficiency that increases IgE antibodies also displayed mutant mice selectively deficient in Vγ4 and Vγ6 (B6.TCR- − − − − increases in IL-4–producing T cells (both residual γδ T cells and αβ Vγ4 / /6 / ) produce substantially more IgE antibody than WT − − T cells) and in systemic IL-4 levels. Its B cells expressed IL-4–regu- controls or mice deficient in all γδ T cells (B6.TCR-δ / ) (10). lated inhibitory receptors (CD5, CD22, and CD32) at diminished Here, we report that deficiency in individual γδ T-cell subsets levels, whereas IL-4–inducible IL-4 receptor α and MHCII were in- (16, 17) can change antibody production and B-cell activation in creased. They also showed signs of activation and spontaneously nonimmunized mice to a degree that jeopardizes self-tolerance. formed germinal centers. These mice displayed IgE-dependent fea- However, the effect cannot simply be ascribed to an altered γδ tures found in hyper-IgE syndrome and developed antichromatin, T-cell balance. Instead, it correlates with functional changes that antinuclear, and anticytoplasmic autoantibodies. In contrast, mice γδ γδ occur within the remaining T cells themselves, when they are deficient in all T cells had nearly unchanged Ig levels and did not no longer restrained by normal γδ cross-talk. Our data show that develop autoantibodies. Removing IL-4 abrogated the increases in this cross-talk controls the amount of IL-4 produced by a subset IgE, antichromatin antibodies, and autoantibodies in the partially of γδ T cells and other T cells, resulting in downstream effects on γδ-deficient mice. Our data suggest that γδ T cells, controlled by antibodies, B cells, and self-tolerance. their own cross-talk, affect IL-4 production, B-cell activation, and B-cell tolerance. Results Genetic γδ T-Cell Deficiencies Change Systemic Antibody Levels in gammadelta T cell | interleukin-4 | autoimmunity | immunoglobulin | Nonimmunized Mice. Having found that IgE antibody responses tolerance in ovalbumin/alum-immunized and nonimmunized mice are sen- γδ γδ sitive to the functional balance within the T-cell compartment he role of T cells within the vertebrate immune system is (10), we wondered if this balance also affects other preimmune Tnot yet fully understood, but it has become clear that they antibodies. We therefore examined a panel of background- exert a strong influence on the immune responses. These cells matched mouse strains with genetic deficiencies in TCR-γ or represent a system of specialized subsets with different de- TCR-δ genes for systemic antibody levels (Fig. 1). The panel velopmental kinetics, tissue distributions, and functional roles − − includes C57BL/6 mice (WT control), B6.TCR-δ / mice (lack- (1). Moreover, at least some of the subsets appear to balance γδ γ −/− γ pos γδ each other’s influence on the immune system (2). Like αβ T cells ing all T cells) (18), B6.TCR-V 1 mice (lacking V 1 and B cells, γδ T cells express antigen receptors encoded by rearranging genes (3, 4), which enable adaptive responses to Significance antigenic challenge. Following such stimulation in the course of diseases, γδ T-cell populations can undergo large changes in size This study changes our understanding of the relationship be- and subset composition (5). The γδ T-cell populations also tween T cells and B cells. Although it is known that T cells change during ontogeny and due to interindividual genetic dif- provide help for specific B-cell responses, it is unclear if and to ferences (6, 7). Conceivably, such changes might alter γδ T-cell what extent T cells also influence preimmune B-cell functions. balance and, with it, γδ T-cell influence on other immune cells. We show here that γδ T cells modulate systemic antibody levels In mice and humans, it was found that functional attributes of in nonimmunized mice, including all major subclasses and es- γδ T cells segregate with expressed γδ T-cell receptors (TCRs) pecially IgE antibodies. One mouse strain deficient in certain γδ (8, 9), although functional differentiation has also been observed T cells developed various autoantibodies, whereas mice de- within or across TCR-defined subsets and correlated with other ficient in all γδ T cells had relatively normal antibodies. Based markers, such as CD27 and CD8 (10, 11). The murine TCR-γ on these and other findings, we conclude that γδ T cells, locus contains seven Vγ genes, six of which are functional and influenced by their own cross-talk, affect IL-4 production, B-cell expressed on the cell surface (3, 12). In the normal mouse activation, and B-cell tolerance. INFLAMMATION spleen, the largest γδ T-cell population expresses Vγ1, followed IMMUNOLOGY AND by Vγ4pos cells and smaller populations expressing Vγ2 and Vγ7 Author contributions: Y.H. and W.K.B. designed research; Y.H., R.A.H., T.O.D., A.G., and pos pos T.L.C. performed research; R.A.H., T.O.D., A.G., G.A.K., S.R.C., K.I., H.H., J.C.C., L.J.W., and (13). Vγ5 and Vγ6 cells are not present in substantial R.L.O. contributed new reagents/analytic tools; Y.H., M.K.A., L.J.W., R.L.O., and W.K.B. numbers. In earlier studies relying on cell transfer and targeted analyzed data; and W.K.B. wrote the paper. inactivation with antibodies, we and others (8, 10, 14, 15) found The authors declare no conflict of interest. pos pos that splenic Vγ1 and Vγ4 cells exert opposite influences on *This Direct Submission article had a prearranged editor. host responses to infection, allergic sensitization, and malig- 1To whom correspondence should be addressed. Email: [email protected]. γδ nancy. The data suggested that these two T-cell subsets bal- This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. ance each other in their influence on the immune responses (2). 1073/pnas.1415107111/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1415107111 PNAS | Published online December 22, 2014 | E39–E48 Downloaded by guest on October 1, 2021 Fig. 1. Influence of γδ T cells on systemic antibody levels. Results of antibody ELISAs in sera from 8- to 12-wk-old female mice, including C57BL/6 (B6), B6.TCR-δ−/− (δ−/−), B6.TCR-Vγ1−/− (Vγ1−/−), and B6.TCR-Vγ4−/−/Vγ6−/− (Vγ4−/−/6−/−) mice, are shown. Total serum Ig (A) and ELISA of Ig subclasses (B–H)asin- dicated (n = 10–23 mice per group). *P < 0.05; **P < 0.01; ***P < 0.001; NS, not significant. − − − − T cells) (17), and B6.TCR-Vγ4 / /6 / mice (lacking Vγ4pos and a αβ T cell-deficient background (Fig. 2). Similar changes in the Vγ6pos γδ T cells) (16, 19). In the absence of all γδ T cells, total antibodies would indicate independence from αβ T cells, whereas absolute serum Ig levels were somewhat decreased (approxi- absent or different changes would indicate a requirement for mately twofold relative to WT mice). However, when only Vγ1pos them. Deficiency in Vγ4pos and Vγ6pos cells on an αβ-deficient − − − − − − − − cells were missing, total Ig levels were decreased more drastically background (B6.TCR-β / /Vγ4 / /6 / vs. B6.TCR-β / )stillmuch (approximately sixfold). In marked contrast, the absence of increased total Ig, IgM, IgG3, IgG2b, and IgG2c, as well as IgA Vγ4pos plus Vγ6pos cells was associated with an increase in total to a lesser degree (Fig. 2 A–C, E, F, and H), as had been found in Ig levels (approximately fourfold relative to WT mice) (Fig. 1A). the comparison of αβ T cell-sufficient mice (Fig. 1), suggesting that Similar patterns were seen within Ig subclasses, although the the γδ effect on these antibodies is largely αβ T cell-independent. extent of the changes varied. Thus, IgM, IgG3, IgG2c, and IgA However, it no longer increased IgG1 and IgE (Fig. 2 D and G), levels were not substantially affected by the absence of all γδ T revealing that the γδ effects on these antibody subclasses are αβ cells (Fig. 1 B, C, F,andH), although at least one of the partial γδ T cell-dependent. Despite much lower levels in the αβ T cell- deficiencies changed all of these antibodies. The largest serum an- deficient mice, antichromatin antibodies were still significantly tibody changes in the partially γδ T cell-deficient mice were seen increased by this partial γδ deficiency (Fig.
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