Crucial role for T cell-intrinsic IL-18R-MyD88 signaling in cognate immune response to intracellular parasite infection
- Universidade Federal do Rio de Janeiro, Brazil
- The Rockefeller University, United States
- Universidade Federal Fluminense, Brazil
- Universidade de São Paulo, Brazil
- Instituto Nacional de Ciência e Tecnologia de Vacinas, CNPq-MCT, Brazil
- Emory University School of Medicine, United States
Figures
Figure 1 with 1 supplement
Lower expansion of IFN-γ+CD4+ Myd88-/- cells in infected mixed BM chimeras.
(A, B) Survival curves of mice infected ip with 2 × 103 blood trypomastigotes of the Y strain. (A) Myd88−/− (CD45.2+)→WT (B6 x B6.SJL F1, CD45.1+CD45.2+) and WT (B6.SJL, CD45.1+)→WT (B6 x B6.SJL F1, …
Figure 1—figure supplement 1
Mixed bone-marrow chimeras.
(A) Experimental design for mixed BM chimera construction. Irradiated WT (B6 x B6.SJL F1) mice were reconstituted with a 1:1 mix of WT (B6.SJL, CD45.1+) and Myd88−/− (CD45.2+) BM cells. (B) …
Figure 2 with 1 supplement
Lower expansion of IFN-γ+CD4+ Myd88 T cells in infected Rag2−/− mice reconstituted with WT and Myd88−/− splenocytes.
(A) Gate strategy and dot plot analysis of IFN-γ+CD4+ T cells among CD4+CD45.2+ (Myd88−/−) or among CD4+CD45.1+ (B6) plus CD4+CD45.2+CD45.1+(B6xB6.SJL F1) (WT) T cells in Rag2−/− mice, reconstituted …
Figure 2—figure supplement 1
Lower expansion of IFN-γ+CD8+ Myd88-/- cells in infected Rag2−/− mice reconstituted with WT and Myd88−/−splenocytes.
(A) Gate strategy and dot plot analysis of IFN-γ+CD4+ T cells among CD8+CD45.2+ (Myd88−/−) or among CD8+CD45.1+ (B6) plus CD8+CD45.2+CD45.1+(B6xB6.SJL F1) (WT) T cells. (B–C) Mean frequencies and …
Figure 3 with 1 supplement
Gene expression profiles of WT and Myd88−/− CD4+T cells sorted from mixed BM chimeras infected with T. cruzi.
(A) Experimental design: CD4+CD45.1+ cells (WT) and CD4+CD45.2+ cells (Myd88−/−) were sorted (99% of purity) from the pool of splenocytes obtained from infected mixed BM chimeric mice at day 14 pi …
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Figure 3—source data 1
Table 1. Differential expressed genes between WT and Myd88−/− CD4+ T cells sorted from infected BM-mixed chimeras.
- https://doi.org/10.7554/eLife.30883.008
Figure 3—figure supplement 1
Gene networks with the genes in core from GSEA analysis.
Interactions among the genes were assessed and visualized by Ingenuity Pathway Analysis (IPA) database. Darker colors represent higher expression in (A) Myd88−/− (blue) or in (B) WT (red) CD4+ T …
Figure 4 with 1 supplement
IL-18R, but not IL-1R, is upregulated in CD4+T cells upon activation following infection.
(A) Gate strategy and mean percentage of CD44hiCD4+ and T-bet+CD44hiCD4+ cells gated on CD4+ T splenocytes from non-infected controls and infected B6 mice at day 14 pi. (B) Representative dot plot …
Figure 4—figure supplement 1
Kinetics of IL-1β and IL-18 levels in the serum, as well as of the appearance of IFN-γ+CD4+T cells in the spleen of infected mice.
(A, B) Serum levels of (A) IL-1β and (B) IL-18. (C) Mean percentages of IFN-γ+CD4+ T cells gated on splenic CD4+ T lymphocytes. Symbols represent the mean value of 4 individually analyzed B6 mice, …
Figure 5
Effects of the absence of T cell-intrinsic MyD88, IL-1R or IL-18R signaling on IFN-γ production by spleen CD4+ T cells of infected mixed BM chimeras.
(A) Representative dot plots showing gating strategy for CD4+ T cells and for CD45.1+ (B6.SJL WT) or CD45.2+ (B6 WT or Myd88−/− or Il1r1−/− or Il18r1−/−) splenocytes from mixed BM chimeras. (B) …
Figure 6 with 5 supplements
Effects of the absence of T cell-intrinsic MyD88, IL-1R or IL-18R signaling on the proliferation and phenotype of CD4+ and CD8+spleen T cells from infected mixed BM chimeras.
(A, C, G, E and I) Mean frequencies and (B, D, F, H and J) absolute cell numbers of: (A and B) BrdU+CD4+ and (C and D) CCR5+CD4+ cells gated on WT CD45.1+ or KO CD45.2+ CD4+ T cells, as in Figure 5A.…
Figure 6—figure supplement 1
Representative dot plots and gate strategy of BrdU+CD4+T cell analysis from chimeric mice.
Representative dot plots showing anti-BrdU staining in CD4+CD45.1+(B6.SJL, WT) or CD4+CD45.2+ (B6 WT or Myd88−/− or Il1r1−/−or Il18r1−/−) splenocytes from control or infected mixed BM chimeras, …
Figure 6—figure supplement 2
Representative dot plots and gate strategy of CCR5+CD4+ T cell analysis from chimeric mice.
Representative dot plots showing CCR5 expression on CD4+CD45.1+ (B6.SJL, WT) or CD4+CD45.2+ (B6 WT or Myd88−/− or Il1r1−/−or Il18r1−/−) splenocytes, from control or infected mixed BM chimeras, gated …
Figure 6—figure supplement 3
Representative dot plots and gate strategy of IFN-γ+CD8+ T cell analysis from chimeric mice.
(A) Gate strategy for CD45.1+ (B6.SJL WT) and CD45.2+ (B6 WT or Myd88−/− or Il1r1−/−or Il18r1−/−) expression on CD8+ T cells. (B) Representative dot plots of IFN-γ expression gated on CD8+CD45.1+ …
Figure 6—figure supplement 4
Representative dot plots and gate strategy of GzB+CD8+ T cell analysis from chimeric mice.
Representative dot plots showing anti-GzB staining in CD8+CD45.1+(B6.SJL, WT) or CD8+CD45.2+(B6 WT or Myd88−/− or Il1r1−/− or Il18r1−/−) splenocytes from control or infected mixed BM chimeras, gated …
Figure 6—figure supplement 5
Representative dot plots and gate strategy of CD44hiCD4+ T cell analysis from chimeric mice.
Representative dot plots showing anti-CD44 staining on CD4+CD45.1+(B6.SJL, WT) or CD4+CD45.2+(B6 WT or Myd88−/− or Il1r1−/− or Il18r1−/−) splenocytes from control or infected mixed BM chimeras, …
Figure 7
Il18r1−/− mice are highly susceptible to infection with T. cruzi and display lower frequency and absolute numbers of Th1 cells.
(A) Parasitemia curve, (B) survival and (C) parasite load in the myocardium at day 14 pi of Il18r1−/−, Myd88−/− and WT B6 mice infected with 2 × 103 blood trypomastigotes of the Y strain. Survival …
Figure 8 with 1 supplement
The adoptive transfer of purified WT CD4+ T cells protects Il18r1−/− and Myd88−/− mice against infection with T. cruzi.
(A and C) Survival; (B and D) mean parasitemia levels at day 9 pi of Il18r1−/− (A and B) or Myd88−/− mice (C and D), which received or not 1.3 × 106 CD4+ T cells purified from infected WT B6 mice. …
Figure 8—figure supplement 1
Adoptive transfer of WT, but not of Ifng−/−CD4+ T cells diminished parasitemia in infected Myd88−/− mice.
(A) Sorting of CD4+ T cells: Representative frequency of CD4+CD3+ T cells after enrichment by negative selection with magnetic beads (pre-sorting, left) followed by FACS sorting (post-sorting, …
Author response image 1
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.30883.021
