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ICOS and Bcl6-dependent pathways maintain a CD4 T cell population with memory-like properties during tuberculosis.

Moguche AO, Shafiani S, Clemons C, Larson RP, Dinh C, Higdon LE, Cambier CJ, Sissons JR, Gallegos AM, Fink PJ, Urdahl KB - J. Exp. Med. (2015)

Bottom Line: When transferred into uninfected animals, these cells persist, mount a robust recall response, and provide superior protection to Mtb rechallenge when compared to terminally differentiated Th1 cells that reside preferentially in the lung-associated vasculature.Thus, the molecular pathways required to maintain Mtb-specific CD4 T cells during ongoing infection are similar to those that maintain memory CD4 T cells in scenarios of antigen deprivation.These results suggest that vaccination strategies targeting the ICOS and Bcl6 pathways in CD4 T cells may provide new avenues to prevent TB.

View Article: PubMed Central - HTML - PubMed

Affiliation: Seattle Biomedical Research Institute (renamed Center for Infectious Disease Research), Seattle, WA 98109 Department of Immunology, University of Washington School of Medicine, Seattle, WA 98104.

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ESAT-6–specific PD-1+CD4 T cells express memory associated markers and survive in the absence of antigen via ICOSL signaling. Mice were infected as described in Fig. 1. (A) Representative flow cytometry histograms show expression of the indicated markers by naive CD44low (gray), ESAT-6 tetramer-binding PD-1+KLRG1− (blue) or PD-1−KLRG1+ (red) CD4 T cells (day 120 after infection). (B) PD-1+KLRG1− or PD-1−KLRG1+ CD4 T cells sorted from lungs of Mtb-infected mice (day 119 after infection) were adoptively transferred (105 cells/recipient) into uninfected WT or ICOSL−/− mice. The graph shows the number of donor cells recovered from the spleens of WT recipients of donor PD-1+ cells (blue), WT recipients of donor KLRG1+ cells (red), and ICOSL−/− recipients of donor PD-1+ cells (black) at days 1, 8, and 28 after transfer. (C) Flow cytometry plots denote PD-1 and KLRG1 expression by donor-derived (transferred as PD-1+KLRG1− or PD-1−KLRG1+ cells) CD4 T cells in the spleens of WT recipients (28 d after transfer) as described in B. The graph depicts the frequency of cells within each donor-derived population expressing PD-1 (blue) or KLRG1 (red) in individual mice. (D) Flow cytometry plots denote PD-1 and KLRG1 expression by donor-derived PD-1+KLRG1− cells in the spleens of WT or ICOSL−/− recipients (28 d after transfer). The graph below depicts the frequency of PD-1+ cells within the donor-derived CD4 T cell population in individual mice of each group. The mean ± SEM are shown for each group and statistical significance was determined by a two-tailed Student’s t test (**, P < 0.01; ****, P < 0.0001). Data in A are representative of three independent experiments with four to five mice per group. Transfer of PD-1+ and KLRG1+ cells into WT mice was performed twice with three to five mice per group and per time point whereas transfer of PD-1+ cells into ICOSL−/− mice was done once with four to five mice in each of the three time points.
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fig5: ESAT-6–specific PD-1+CD4 T cells express memory associated markers and survive in the absence of antigen via ICOSL signaling. Mice were infected as described in Fig. 1. (A) Representative flow cytometry histograms show expression of the indicated markers by naive CD44low (gray), ESAT-6 tetramer-binding PD-1+KLRG1− (blue) or PD-1−KLRG1+ (red) CD4 T cells (day 120 after infection). (B) PD-1+KLRG1− or PD-1−KLRG1+ CD4 T cells sorted from lungs of Mtb-infected mice (day 119 after infection) were adoptively transferred (105 cells/recipient) into uninfected WT or ICOSL−/− mice. The graph shows the number of donor cells recovered from the spleens of WT recipients of donor PD-1+ cells (blue), WT recipients of donor KLRG1+ cells (red), and ICOSL−/− recipients of donor PD-1+ cells (black) at days 1, 8, and 28 after transfer. (C) Flow cytometry plots denote PD-1 and KLRG1 expression by donor-derived (transferred as PD-1+KLRG1− or PD-1−KLRG1+ cells) CD4 T cells in the spleens of WT recipients (28 d after transfer) as described in B. The graph depicts the frequency of cells within each donor-derived population expressing PD-1 (blue) or KLRG1 (red) in individual mice. (D) Flow cytometry plots denote PD-1 and KLRG1 expression by donor-derived PD-1+KLRG1− cells in the spleens of WT or ICOSL−/− recipients (28 d after transfer). The graph below depicts the frequency of PD-1+ cells within the donor-derived CD4 T cell population in individual mice of each group. The mean ± SEM are shown for each group and statistical significance was determined by a two-tailed Student’s t test (**, P < 0.01; ****, P < 0.0001). Data in A are representative of three independent experiments with four to five mice per group. Transfer of PD-1+ and KLRG1+ cells into WT mice was performed twice with three to five mice per group and per time point whereas transfer of PD-1+ cells into ICOSL−/− mice was done once with four to five mice in each of the three time points.

Mentions: Although PD-1+ Mtb-specific CD4 T cells have Tfh-like features, further analysis of cell surface and intracellular molecules revealed a mixed profile, with expression of both effector and memory markers (Fig. 5 A). CD62L expression reflected an effector phenotype and was low in both PD-1+KLRG1− and PD-1−KLRG1+ tetramer-binding populations. Although CCR7 and CD127 expression was slightly higher among PD-1+KLRG1− compared with PD-1−KLRG1+ cells, this expression did not approach the levels usually observed on central memory cells that develop when antigen is cleared. In contrast, as we had previously observed for ICOS and CD69 (Fig. 4 A), CXCR3, Ly6C, and CD43 showed markedly distinct expression patterns on these two populations (Fig. 5 A). Most ESAT-6–specific PD-1+KLRG1− cells were CXCR3+, Ly6C−, and CD43+, whereas most PD-1−KLRG1+ cells exhibited the opposite phenotype. In addition, PD-1+KLRG1− cells, like memory cells (Marshall et al., 2011), were intermediate for T-bet expression, whereas PD-1−KLRG1+ cells, like Th1 effector cells, exhibited high T-bet expression (Fig. 3 D). Our results reveal that ESAT-6–specific PD-1+ CD4 T cells, while expressing some markers of effector T cells, also displayed features of memory T cells. In contrast, their KLRG1+ counterparts exhibit characteristics of terminally differentiated effector Th1 cells.


ICOS and Bcl6-dependent pathways maintain a CD4 T cell population with memory-like properties during tuberculosis.

Moguche AO, Shafiani S, Clemons C, Larson RP, Dinh C, Higdon LE, Cambier CJ, Sissons JR, Gallegos AM, Fink PJ, Urdahl KB - J. Exp. Med. (2015)

ESAT-6–specific PD-1+CD4 T cells express memory associated markers and survive in the absence of antigen via ICOSL signaling. Mice were infected as described in Fig. 1. (A) Representative flow cytometry histograms show expression of the indicated markers by naive CD44low (gray), ESAT-6 tetramer-binding PD-1+KLRG1− (blue) or PD-1−KLRG1+ (red) CD4 T cells (day 120 after infection). (B) PD-1+KLRG1− or PD-1−KLRG1+ CD4 T cells sorted from lungs of Mtb-infected mice (day 119 after infection) were adoptively transferred (105 cells/recipient) into uninfected WT or ICOSL−/− mice. The graph shows the number of donor cells recovered from the spleens of WT recipients of donor PD-1+ cells (blue), WT recipients of donor KLRG1+ cells (red), and ICOSL−/− recipients of donor PD-1+ cells (black) at days 1, 8, and 28 after transfer. (C) Flow cytometry plots denote PD-1 and KLRG1 expression by donor-derived (transferred as PD-1+KLRG1− or PD-1−KLRG1+ cells) CD4 T cells in the spleens of WT recipients (28 d after transfer) as described in B. The graph depicts the frequency of cells within each donor-derived population expressing PD-1 (blue) or KLRG1 (red) in individual mice. (D) Flow cytometry plots denote PD-1 and KLRG1 expression by donor-derived PD-1+KLRG1− cells in the spleens of WT or ICOSL−/− recipients (28 d after transfer). The graph below depicts the frequency of PD-1+ cells within the donor-derived CD4 T cell population in individual mice of each group. The mean ± SEM are shown for each group and statistical significance was determined by a two-tailed Student’s t test (**, P < 0.01; ****, P < 0.0001). Data in A are representative of three independent experiments with four to five mice per group. Transfer of PD-1+ and KLRG1+ cells into WT mice was performed twice with three to five mice per group and per time point whereas transfer of PD-1+ cells into ICOSL−/− mice was done once with four to five mice in each of the three time points.
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fig5: ESAT-6–specific PD-1+CD4 T cells express memory associated markers and survive in the absence of antigen via ICOSL signaling. Mice were infected as described in Fig. 1. (A) Representative flow cytometry histograms show expression of the indicated markers by naive CD44low (gray), ESAT-6 tetramer-binding PD-1+KLRG1− (blue) or PD-1−KLRG1+ (red) CD4 T cells (day 120 after infection). (B) PD-1+KLRG1− or PD-1−KLRG1+ CD4 T cells sorted from lungs of Mtb-infected mice (day 119 after infection) were adoptively transferred (105 cells/recipient) into uninfected WT or ICOSL−/− mice. The graph shows the number of donor cells recovered from the spleens of WT recipients of donor PD-1+ cells (blue), WT recipients of donor KLRG1+ cells (red), and ICOSL−/− recipients of donor PD-1+ cells (black) at days 1, 8, and 28 after transfer. (C) Flow cytometry plots denote PD-1 and KLRG1 expression by donor-derived (transferred as PD-1+KLRG1− or PD-1−KLRG1+ cells) CD4 T cells in the spleens of WT recipients (28 d after transfer) as described in B. The graph depicts the frequency of cells within each donor-derived population expressing PD-1 (blue) or KLRG1 (red) in individual mice. (D) Flow cytometry plots denote PD-1 and KLRG1 expression by donor-derived PD-1+KLRG1− cells in the spleens of WT or ICOSL−/− recipients (28 d after transfer). The graph below depicts the frequency of PD-1+ cells within the donor-derived CD4 T cell population in individual mice of each group. The mean ± SEM are shown for each group and statistical significance was determined by a two-tailed Student’s t test (**, P < 0.01; ****, P < 0.0001). Data in A are representative of three independent experiments with four to five mice per group. Transfer of PD-1+ and KLRG1+ cells into WT mice was performed twice with three to five mice per group and per time point whereas transfer of PD-1+ cells into ICOSL−/− mice was done once with four to five mice in each of the three time points.
Mentions: Although PD-1+ Mtb-specific CD4 T cells have Tfh-like features, further analysis of cell surface and intracellular molecules revealed a mixed profile, with expression of both effector and memory markers (Fig. 5 A). CD62L expression reflected an effector phenotype and was low in both PD-1+KLRG1− and PD-1−KLRG1+ tetramer-binding populations. Although CCR7 and CD127 expression was slightly higher among PD-1+KLRG1− compared with PD-1−KLRG1+ cells, this expression did not approach the levels usually observed on central memory cells that develop when antigen is cleared. In contrast, as we had previously observed for ICOS and CD69 (Fig. 4 A), CXCR3, Ly6C, and CD43 showed markedly distinct expression patterns on these two populations (Fig. 5 A). Most ESAT-6–specific PD-1+KLRG1− cells were CXCR3+, Ly6C−, and CD43+, whereas most PD-1−KLRG1+ cells exhibited the opposite phenotype. In addition, PD-1+KLRG1− cells, like memory cells (Marshall et al., 2011), were intermediate for T-bet expression, whereas PD-1−KLRG1+ cells, like Th1 effector cells, exhibited high T-bet expression (Fig. 3 D). Our results reveal that ESAT-6–specific PD-1+ CD4 T cells, while expressing some markers of effector T cells, also displayed features of memory T cells. In contrast, their KLRG1+ counterparts exhibit characteristics of terminally differentiated effector Th1 cells.

Bottom Line: When transferred into uninfected animals, these cells persist, mount a robust recall response, and provide superior protection to Mtb rechallenge when compared to terminally differentiated Th1 cells that reside preferentially in the lung-associated vasculature.Thus, the molecular pathways required to maintain Mtb-specific CD4 T cells during ongoing infection are similar to those that maintain memory CD4 T cells in scenarios of antigen deprivation.These results suggest that vaccination strategies targeting the ICOS and Bcl6 pathways in CD4 T cells may provide new avenues to prevent TB.

View Article: PubMed Central - HTML - PubMed

Affiliation: Seattle Biomedical Research Institute (renamed Center for Infectious Disease Research), Seattle, WA 98109 Department of Immunology, University of Washington School of Medicine, Seattle, WA 98104.

Show MeSH
Related in: MedlinePlus