Fig 2: TMEM, TINT, and TEFF cells reside in distinct areas of the spleen(A–C) TG6-GFP chimeric mice infected with T. gondii and their spleens were analyzed by confocal microscopy after 6 weeks. Sections were stained with antibodies against TCF1 (A), Ki67 (B), and KLRG1 (C) to identify the location of TMEM, T, and TINT/TEFF TG6-GFP cells, respectively. B220 staining was used to identify B cell zones (B) and T cell zones (T) to map the white pulp (WP) and red pulp (RP) regions. In some images, bridging channels (BCs) are marked with dotted lines. Dashed rectangles indicate the area in the magnified regions on the right. Examples of cells staining positively with the indicated marker are highlighted with yellow arrows and negative examples with white arrows.(D) Data summarizing the location of TG6-GFP cells that stained positively for each protein.(E) Data showing the proportion of each GRA6-specific population labeled after i.v. anti-CD8 antibody injection.(F) Summary data showing the location of TMEM, TINT, and TEFF cells in the spleen after adoptive transfer.Images in (A)–(C) and data in (D) are representative of images taken from three or more separate mice. Data in (E) are taken from two separate infections and are representative of data from three separate experiments. Data in (F) are summarized from multiple recipient mice across three infections (TMEM cells, n = 3; TINT cells, n = 3; TEFF cells, n = 4).

Fig 3: CXCR3-deficient CD8+ T cells retain a memory phenotype in the RP(A–C) CXCR3 KO TG6-GFP neonatal chimeras were infected with T. gondii and analyzed by confocal microscopy after 6 weeks. Sections were stained with antibodies against TCF1 (A), Ki67 (B), and KLRG1 (C). B220 staining was used to identify B cell zones (B) to map the T cell zone (T) and RP regions. Yellow arrows highlight examples of cells staining positively for the specified marker; white arrows highlight examples of negative staining.(D) Summary data showing the proportion of GFP cells found in the WP or RP that are identified as positive or negative for expression of the indicated marker. Bar graphs display the proportion of WT or CXCR3 KO GFP cells expressing the protein of interest (TCF1, Ki67, or KLRG1) within the WP, BC, or RP.Data for WT cells are combined from at least three mice across two different experiments, and data for KO cells are combined from four mice taken across two experiments.

Fig 4: CXCR3-deficient CD8+ T cells develop an inflated TMEM cell pool(A) Experimental design.(B) Representative flow cytometry plots of gated GRA6 tetramer+ CD8+ splenocytes at the indicated time points.(C) Summary data showing the proportion of the transferred cells belonging to the WT or CXCR3 KO population.(D) Individual data points for the time point 6 weeks after infection in (C).(E and F) The differentiation profile of WT and CXCR3 KO TG6 cells 3 weeks (E) or 6 weeks (F) after infection. TMEM, TINT, and TEFF cell subsets were identified using Ki67 and KLRG1 staining. Bar graphs display the proportion of the transferred population belonging to each subset. Adjacent plots display the ratios of TMEM:TINT and TINT:TEFF cells in the WT and KO populations. Lines connect populations from the same mouse.(G) Absolute numbers of cells belonging to each population.(H) Experimental design.(I and J) Representative flow plots and summary data for CFSE dilution and KLRG1 expression within the population of transferred WT and KO TMEM cells 3 weeks (I) and 6 weeks (J) after infection.Data in (A)–(G) are combined from three independent experiments. Data in (H)–(J) are combined from two independent experiments.

Fig 5: CD8 T cells present a less activated phenotype following ZIKVBR infection.a–c Representative flow cytometry plots of CD127 and KLRG1 expression by CD8aloCD11ahi CD8 T cells from ZIKVCDN and ZIKVBR-infected mice (a), and frequency (b) and number (c) of CD127loKLRG1hi CD8aloCD11ahi CD8 T cells (circles; TEC) and CD127hiKLRG1lo CD8aloCD11ahi CD8 T cells (squares; MPC) in the spleen 7 dpi with ZIKVCDN or ZIKVBR. In b TEC p = 0.0004 and MPC p = 0.0002. In c TEC p = 0.000049 and MPC p = 0.0103. d–f Representative histogram (d), frequency (e) and number (f) of CD62L+ CD8aloCD11ahi CD8 T cells in the spleen 7 dpi with ZIKVCDN or ZIKVBR. Line on histogram indicates gating strategy used to identify CD62L+ cells. In e p = 0.0003 and in f p = 0.01. g–i Representative histogram (g), frequency (h), and number (i) of granzyme B+ CD8aloCD11ahi CD8 T cells in the spleen 7 dpi with ZIKVCDN or ZIKVBR. Shaded histogram indicates isotype control. Line on histogram indicates gating strategy used to identify granzyme B+ cells. In h p = 0.0026 and in i, p = 0.0005. All data are representative of two independent experiments with n = 3 mice per group and are shown as mean ± SEM. All data were analyzed by a two-tailed, unpaired Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. Red symbols = ZIKVCDN-infected mice, blue symbols = ZIKVBR-infected mice. Source data are provided as a Source Data file.