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Phenotypic and Functional Analysis of Non-Activated and Allo-Activated Eomesodermin+ CD8+ T Cells in Humans and Nonhuman Primates

A. Perez Gutierrez1, A. W. Thomson2, D. M. Metes2, M. B. Ezzelarab2

1Transplantation Institute, University of Chicago Medicine, Chicago, IL, 2Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, PA

Meeting: 2019 American Transplant Congress

Abstract number: D149

Keywords: Primates, T cells, Tumor necrosis factor (TNF)

Session Information

Date: Tuesday, June 4, 2019

Session Name: Poster Session D: Lymphocyte Biology: Signaling, Co-Stimulation, Regulation

Session Time: 6:00pm-7:00pm

 Presentation Time: 6:00pm-7:00pm

Location: Hall C & D

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*Purpose: The T-box transcription factors Eomesodermin (Eomes) and T-bet play a critical role in differentiation and function of CD8+ memory T cells (Tmem). We have previously shown, that in humans, high Eomes expression by allo-reactive CD8+T cells correlates with enhanced effector function. Our purpose was to evaluate the phenotypic and functional differences of alloreactive Eomeshi CD8+ T cells in human and nonhuman primates.

*Methods: Healthy human and juvenile rhesus monkey T cells were purified followed by co-culture with allogeneic T-cell depleted peripheral blood mononuclear cells (PBMC). CD8+Tmem subsets were identified based on CD45RA/CD28 expression. Eomes expression by nonactivated and allo-activated CD8+ T cells was evaluated. The expression of T-bet, cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), Programmed cell death-1 (PD-1) and effector cytokines (TNFα and IFNγ) were assessed.

*Results: Before allo-stimulation, human and monkey Eomeshi CD8+T cells comprised high percentage of CD45RA-CD28+ central memory T cells, while Eomeslo CD8+T cells comprised higher percentage of CD45RA+CD28+ naïve T cells. Additionally, human but not monkey Eomeshi CD8+T cells exhibited higher percentages of TNFα/IFNγ double positive cells, than Eomeslo CD8+T cells.T-bet expression was higher in human, but lower in monkey Eomeshi, compared to Eomeslo CD8+T cells. After allo-stimulation, the percentages of Tmem subsets was variable in both humans and monkeys CD8+ T cells. While both human and monkey Eomeshi CD8+T-cells exhibited significantly higher TNFα/IFNγ double positive cells, than Eomeshi CD8 T cells, only human but not monkey Eomeshi CD8+T cells express significantly higher T-bet than Eomeslo CD8+ T cells. In proliferating cells, high percentages of TNFα/INFγ double positive cells were observed in human Eomeshi Tbethi and monkey EomeshiTbetlo CD8+T cell subsets.

CTLA4 expression was significantly upregulated by CD8+T cells after allo-stimulation in both human and monkeys, where in monkeys, but not humans, an inverse correlation between Eomes and CTLA4 expression was observed by allo-stimulated CD8+ T cells. While co-expression of PD-1 and CTLA4 was comparable between Eomeshi and Eomeslo CD8+T cells in humans, the co-expression of PD-1 and CTLA4 was significantly lower in Eomeshi compared to Eomeslo monkey CD8+T cells

*Conclusions: In both humans and monkeys, higher Eomes expression correlates with effector T cell phenotype after allo-stimulation. However, concomitant expression of T-bet and the exhaustion/activation markers CTLA4 and PD-1 was different. The role of Eomes in the development and maintenance of allo-reactive Tmem may not be identical in human and nonhuman primates.

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To cite this abstract in AMA style:

Gutierrez APerez, Thomson AW, Metes DM, Ezzelarab MB. Phenotypic and Functional Analysis of Non-Activated and Allo-Activated Eomesodermin+ CD8+ T Cells in Humans and Nonhuman Primates [abstract]. Am J Transplant. 2019; 19 (suppl 3). https://atcmeetingabstracts.com/abstract/phenotypic-and-functional-analysis-of-non-activated-and-allo-activated-eomesodermin-cd8-t-cells-in-humans-and-nonhuman-primates/. Accessed January 18, 2021.

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