EBAG9 controls CD8+ T cell memory formation responding to tumor challenge in mice
Armin Rehm, … , Gerald Willimsky, Uta E. Höpken
Armin Rehm, … , Gerald Willimsky, Uta E. Höpken
Published April 28, 2022
Citation Information: JCI Insight. 2022;7(11):e155534. https://doi.org/10.1172/jci.insight.155534.
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Research Article Immunology Oncology

EBAG9 controls CD8+ T cell memory formation responding to tumor challenge in mice

Abstract

Insight into processes that determine CD8+ T cell memory formation has been obtained from infection models. These models are biased toward an inflammatory milieu and often use high-avidity CD8+ T cells in adoptive-transfer procedures. It is unclear whether these conditions mimic the differentiation processes of an endogenous repertoire that proceed upon noninflammatory conditions prevailing in premalignant tumor lesions. We examined the role of cytolytic capacity on CD8+ T cell fate decisions when primed by tumor cells or by minor histocompatibility antigen–mismatched leukocytes. CD8+ memory commitment was analyzed in Ebag9-deficient mice that exhibited enhanced tumor cell lysis. This property endowed Ebag9–/– mice with extended control of Tcl-1 oncogene–induced chronic lymphocytic leukemia progression. In Ebag9–/– mice, an expanded memory population was obtained for anti-HY and anti–SV-40 T antigen–specific T cells, despite unchanged effector frequencies in the primary response. By comparing the single-cell transcriptomes of CD8+ T cells responding to tumor cell vaccination, we found differential distribution of subpopulations between Ebag9+/+ and Ebag9–/– T cells. In Ebag9–/– cells, these larger clusters contained genes encoding transcription factors regulating memory cell differentiation and anti-apoptotic gene functions. Our findings link EBAG9-controlled cytolytic activity and the commitment to the CD8+ memory lineage.

Authors

Armin Rehm, Anthea Wirges, Dana Hoser, Cornelius Fischer, Stefanie Herda, Kerstin Gerlach, Sascha Sauer, Gerald Willimsky, Uta E. Höpken

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Figure 4

CD8+ T cells derived from Ebag9-deficient MataHari mice develop a stronger HY-specific memory response.

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CD8+ T cells derived from Ebag9-deficient MataHari mice develop a strong...
(A) Female mice (CD45.1+) were transferred (i.v.) with 4× 104 to 5 × 104 sorted CD8+TCR Vβ 8.3+ CD62Lhi T lymphocytes derived from Ebag9+/+ × MataHari (+/+) or Ebag9–/– × MataHari (–/–) female donors (CD45.2+). One day later, recipients were immunized i.p. with 5 × 106 male splenocytes. The frequency of CD8+CD45.2+CD44hi T cells as percentages of all CD8+ T cells was determined >50–56 days after immunization, without in vitro restimulation. (B) The percentage of CD44hi T cells among all CD45.2+CD8+ T cells is reported. (C) Total numbers of CD8+CD45.2+CD44hi memory T cells in spleen are depicted. Each dot represents data from 1 congenic recipient mouse (CD45.1). WT mice, n = 6; KO mice, n = 5. Bars indicate mean percentage values; n = 2 independent experiments. A Mann-Whitney test was used for this analysis. (D) As in A, congenic female mice were transferred with CD8+TCR Vβ 8.3+ CD62Lhi T lymphocytes sorted from Ebag9+/+ × MataHari (+/+) or Ebag9–/– × MataHari (–/–) female donors (CD45.2+). Animals were immunized as before, and splenic MataHari cells were detected by the expression of CD8+CD45.2+ at day 7. Graph presents the frequencies of CD8+CD45.2+ cells among all splenocytes; the bars indicate mean values. (E) Total numbers of this population; n = 3 independent experiments. WT mice, n = 9; KO mice, n = 11. (D and E) Student’s t test was used. (F) Top: A representative dot plot shows the gating strategy for detection of MataHari T cells. In some cases, recipient mice were congenic for CD90.1+. Differentiation of CD45.2+CD8+ or CD90.2+CD8+ population was further analyzed by anti-CD44, anti-KLRG1, and anti-CD127 staining. Overlay histograms show shifts relative to CD8+ naive T cells. Bottom: Bar charts report quantitation of marker expression. Bars show mean values; n = 3 independent experiments with 7–10 WT and 6–12 KO animals. Student’s t test was applied.