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. 2019 Sep 14;20:100–109. doi: 10.1016/j.isci.2019.09.012

Figure 2.

Figure 2

CXCR5+ Primary Effectors Differentiate into Highly Competent Memory Cells

CXCR5+ or CXCR5- OT-1 cells were sorted on day 3.5 after infection and transferred to recipient mice (see Figure S1 for the experimental design). The progeny of CXCR5+ or CXCR5- OT-1 cells was examined at different time points after infection.

(A) The expression of CD127 and KLRG-1 on day 10 post-infection by CXCR5+ and CXCR5- OT-1.

(B) The expression of CXCR5 on day 10 post-infection by CXCR5+ and CXCR5- OT-1.

(C) The expression of CD44 and CD62L by the progeny of CXCR5+ and CXCR5- OT-1 cells, on day 42 post-infection, i.e., at the memory stage.

(D) The expression of PD-1 by the progeny of CXCR5+ and CXCR5- OT-1 cells, on day 42 post-infection, i.e., at the memory stage. PD-1 MFI are also shown from day 7 to day 42 post-infection.

(E) The expression of Bcl-6 by the progeny of CXCR5+ and CXCR5- OT-1 cells, on day 42 post-infection, i.e., at the memory stage. Bcl-6 MFI are also shown at day 42 post-infection.

(F) The frequency in CD8+ T cells of OT-1 memory cells derived from the progeny of CXCR5+ or CXCR5- OT-1 cells.

(G and H) Mice adoptively transferred with CXCR5+ or CXCR5- OT-1 cells were challenged with rLm-OVA on day 42, i.e., at the memory stage. Owing to the possible presence of reacting endogenous memory CD8+ T cells, secondary OT-1 effectors were sorted 4 days after challenge (day 46) and injected into naive wild-type mice infected at the same time with 2 × 105 colony-forming unit rLm-OVA. OT-1 cells and bacterial burden were analyzed 4 days after transfer (day 50, see Figures S1, 2G, and 2H). (G) Day 46, from left to right: the percentage in CD8+ T cells of OT-1 cells, the percentage of OT-1 cells expressing granzyme B, the corrected geometric MFI of granzyme B in OT-1 cells (expressed as [MFIOT-1 – MFInon-OT-1]), and the corrected geometric MFI of IL-21R in OT-1 cells (expressed as [MFIOT-1 – MFInon-OT-1]). (H) The bacterial burden measured in the spleen 4 days after infection (day 50). The data are from five independent experiments. Mann-Whitney test (E–G) and Kruskal-Wallis test (H) were used for statistical analysis. p.i., post-infection. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.