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Virus-specific regulatory T cells ameliorate encephalitis by repressing effector T cell functions from priming to effector stages.

Zhao J, Zhao J, Perlman S - PLoS Pathog. (2014)

Bottom Line: In addition, M133 Tregs diminished microglia activation and decreased the number and function of Tconv in the infected brain.Thus, virus-specific Tregs inhibited pathogenic CD4 T cell responses during priming and effector stages, particularly those recognizing cognate antigen, and decreased mortality and morbidity without affecting virus clearance.These cells are more suppressive than bulk Tregs and provide a targeted approach to ameliorating immunopathological disease in infectious settings.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, University of Iowa, Iowa City, Iowa, United States of America.

ABSTRACT
Several studies have demonstrated the presence of pathogen-specific Foxp3+ CD4 regulatory T cells (Treg) in infected animals, but little is known about where and how these cells affect the effector T cell responses and whether they are more suppressive than bulk Treg populations. We recently showed the presence of both epitope M133-specific Tregs (M133 Treg) and conventional CD4 T cells (M133 Tconv) in the brains of mice with coronavirus-induced encephalitis. Here, we provide new insights into the interactions between pathogenic Tconv and Tregs responding to the same epitope. M133 Tregs inhibited the proliferation but not initial activation of M133 Tconv in draining lymph nodes (DLN). Further, M133 Tregs inhibited migration of M133 Tconv from the DLN. In addition, M133 Tregs diminished microglia activation and decreased the number and function of Tconv in the infected brain. Thus, virus-specific Tregs inhibited pathogenic CD4 T cell responses during priming and effector stages, particularly those recognizing cognate antigen, and decreased mortality and morbidity without affecting virus clearance. These cells are more suppressive than bulk Tregs and provide a targeted approach to ameliorating immunopathological disease in infectious settings.

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M133 Tregs preferentially inhibit proliferation of M133-specific Tconvs in an in vitro suppression assay.(A) Experimental design. M133, S358 and S510-specific T cells were obtained from infected mice, labeled as described in Materials and Methods and used as responder cells. M133-specific Tregs were pre-activated in vitro for 24 hours and co-cultured with the labeled responders and irradiated antigen presenting cells (CHB3 cells). Cells were stimulated with the indicated peptides for 66 hours and then analyzed for Violet dilution. (B) Representative histograms showing proliferation of responders in the absence (open) or presence (shaded) of M133 Tregs. (C) Summary of data from 3 rJ2.2 and 3 rJ2.2.MY135Q-infected mice. *P<0.05, **P<0.01, ***P<0.001, M133 or S358 suppression compared to S510 suppression; #P<0.05, ###P<0.001, M133 suppression compared to S358 suppression.
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ppat-1004279-g009: M133 Tregs preferentially inhibit proliferation of M133-specific Tconvs in an in vitro suppression assay.(A) Experimental design. M133, S358 and S510-specific T cells were obtained from infected mice, labeled as described in Materials and Methods and used as responder cells. M133-specific Tregs were pre-activated in vitro for 24 hours and co-cultured with the labeled responders and irradiated antigen presenting cells (CHB3 cells). Cells were stimulated with the indicated peptides for 66 hours and then analyzed for Violet dilution. (B) Representative histograms showing proliferation of responders in the absence (open) or presence (shaded) of M133 Tregs. (C) Summary of data from 3 rJ2.2 and 3 rJ2.2.MY135Q-infected mice. *P<0.05, **P<0.01, ***P<0.001, M133 or S358 suppression compared to S510 suppression; #P<0.05, ###P<0.001, M133 suppression compared to S358 suppression.

Mentions: To further buttress the conclusion that M133 Tregs preferentially suppressed the M133 Tconv response, we performed an in vitro suppression assay (Figure 9). Because S358 and S510 TCR Tg mice were not available, we obtained CD4 and CD8 T cells from infected mice and labeled them with Violet. Since these cells had been activated in vivo, M133 Tregs, isolated from uninfected M133 TCR Tg mice were therefore pre-activated in vitro with anti-CD3 and anti-CD28 antibodies. In all cases, M133 peptide was included in the in vitro suppression assay to activate M133 Tregs. Consequently, in order to distinguish proliferation of M133 and S358 CD4 or S510 CD8 T cells, the latter cells were obtained from mice infected with virus lacking expression of the M133 epitope (rJ2.2.MY135Q). As shown in Figure 9B and C, M133 Tregs preferentially suppressed the proliferation of M133 Tconv. In agreement with the results shown in Figure 8E and our previous results [27], M133 Tregs inhibited the proliferation of S358-specific CD4 T cells more potently than that of S510-specific CD8 T cells.


Virus-specific regulatory T cells ameliorate encephalitis by repressing effector T cell functions from priming to effector stages.

Zhao J, Zhao J, Perlman S - PLoS Pathog. (2014)

M133 Tregs preferentially inhibit proliferation of M133-specific Tconvs in an in vitro suppression assay.(A) Experimental design. M133, S358 and S510-specific T cells were obtained from infected mice, labeled as described in Materials and Methods and used as responder cells. M133-specific Tregs were pre-activated in vitro for 24 hours and co-cultured with the labeled responders and irradiated antigen presenting cells (CHB3 cells). Cells were stimulated with the indicated peptides for 66 hours and then analyzed for Violet dilution. (B) Representative histograms showing proliferation of responders in the absence (open) or presence (shaded) of M133 Tregs. (C) Summary of data from 3 rJ2.2 and 3 rJ2.2.MY135Q-infected mice. *P<0.05, **P<0.01, ***P<0.001, M133 or S358 suppression compared to S510 suppression; #P<0.05, ###P<0.001, M133 suppression compared to S358 suppression.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4125232&req=5

ppat-1004279-g009: M133 Tregs preferentially inhibit proliferation of M133-specific Tconvs in an in vitro suppression assay.(A) Experimental design. M133, S358 and S510-specific T cells were obtained from infected mice, labeled as described in Materials and Methods and used as responder cells. M133-specific Tregs were pre-activated in vitro for 24 hours and co-cultured with the labeled responders and irradiated antigen presenting cells (CHB3 cells). Cells were stimulated with the indicated peptides for 66 hours and then analyzed for Violet dilution. (B) Representative histograms showing proliferation of responders in the absence (open) or presence (shaded) of M133 Tregs. (C) Summary of data from 3 rJ2.2 and 3 rJ2.2.MY135Q-infected mice. *P<0.05, **P<0.01, ***P<0.001, M133 or S358 suppression compared to S510 suppression; #P<0.05, ###P<0.001, M133 suppression compared to S358 suppression.
Mentions: To further buttress the conclusion that M133 Tregs preferentially suppressed the M133 Tconv response, we performed an in vitro suppression assay (Figure 9). Because S358 and S510 TCR Tg mice were not available, we obtained CD4 and CD8 T cells from infected mice and labeled them with Violet. Since these cells had been activated in vivo, M133 Tregs, isolated from uninfected M133 TCR Tg mice were therefore pre-activated in vitro with anti-CD3 and anti-CD28 antibodies. In all cases, M133 peptide was included in the in vitro suppression assay to activate M133 Tregs. Consequently, in order to distinguish proliferation of M133 and S358 CD4 or S510 CD8 T cells, the latter cells were obtained from mice infected with virus lacking expression of the M133 epitope (rJ2.2.MY135Q). As shown in Figure 9B and C, M133 Tregs preferentially suppressed the proliferation of M133 Tconv. In agreement with the results shown in Figure 8E and our previous results [27], M133 Tregs inhibited the proliferation of S358-specific CD4 T cells more potently than that of S510-specific CD8 T cells.

Bottom Line: In addition, M133 Tregs diminished microglia activation and decreased the number and function of Tconv in the infected brain.Thus, virus-specific Tregs inhibited pathogenic CD4 T cell responses during priming and effector stages, particularly those recognizing cognate antigen, and decreased mortality and morbidity without affecting virus clearance.These cells are more suppressive than bulk Tregs and provide a targeted approach to ameliorating immunopathological disease in infectious settings.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, University of Iowa, Iowa City, Iowa, United States of America.

ABSTRACT
Several studies have demonstrated the presence of pathogen-specific Foxp3+ CD4 regulatory T cells (Treg) in infected animals, but little is known about where and how these cells affect the effector T cell responses and whether they are more suppressive than bulk Treg populations. We recently showed the presence of both epitope M133-specific Tregs (M133 Treg) and conventional CD4 T cells (M133 Tconv) in the brains of mice with coronavirus-induced encephalitis. Here, we provide new insights into the interactions between pathogenic Tconv and Tregs responding to the same epitope. M133 Tregs inhibited the proliferation but not initial activation of M133 Tconv in draining lymph nodes (DLN). Further, M133 Tregs inhibited migration of M133 Tconv from the DLN. In addition, M133 Tregs diminished microglia activation and decreased the number and function of Tconv in the infected brain. Thus, virus-specific Tregs inhibited pathogenic CD4 T cell responses during priming and effector stages, particularly those recognizing cognate antigen, and decreased mortality and morbidity without affecting virus clearance. These cells are more suppressive than bulk Tregs and provide a targeted approach to ameliorating immunopathological disease in infectious settings.

Show MeSH
Related in: MedlinePlus