EOMES and IL-10 regulate anti-tumor activity of PD-1⁺CD4⁺T-cells in B-cell Non-Hodgkin lymphoma

Abstract

41The transcription factor Eomesodermin (EOMES) promotes IL-10 production of CD4 + T-cells, which has 42 been linked to immunosuppressive and cytotoxic activities. We detected EOMES-expressing CD4 + T-43 cells in lymph node samples of patients with chronic lymphocytic leukemia (CLL) or diffuse large B-cell 44 lymphoma. This was in line with an observed expansion of EOMES-positive CD4 + T-cells in leukemic Eµ-45 TCL1 mice, a well-established model of CLL, and upon adoptive transfer of TCL1 leukemia in mice. 46Transcriptome and flow cytometry analyses revealed that EOMES does not only drive the transcription 47 of IL-10, but rather controls a unique differentiation program in CD4 + T-cells. Moreover, EOMES was 48 necessary for the accumulation of a specific CD4 + T-cell subset that expresses IFNγ and IL-10, as well 49 as inhibitory receptors, like PD-1 and LAG3. T-cell transfer studies in leukopenic Rag2 -/mice showed 50 that EOMES-deficient CD4 + T-cells were inferior in controlling TCL1 leukemia development compared 51 to wildtype T-cells, even though expansion of Eomes -/-CD4 + T-cells was observed. We further showed 52 that control of TCL1 leukemia was driven by IL-10 receptor-mediated signals, as Il10rb-deficient CD4 + 53 T-cells showed impaired anti-leukemia activity. Altogether, our data suggest that IL-10 producing PD-54

EOMES is essential for antitumor activity of CD8+ T cells in chronic lymphocytic leukemia

“…Of note, increased levels of EOMES-expressing T cells in both effector and memory subsets of TCL1 AT mice, in contrast to non-leukemic control mice were detected (Fig. 2D, E), which confirms previously published data [20,28]. We subsequently assessed EOMES expression levels together with the exhaustion markers PD-1, LAG3, TGIT, and CD244.…”

“…The importance of EOMES in adaptive immune control of tumors is further exemplified by a subset of CD4 + T cells that expresses EOMES and shows cytotoxic activity [43]. In line with this, we have recently identified an EOMES + CD4 + T-cell subset, which is increased in CLL and coexpresses PD-1 and LAG3 [28]. Similarly, as observed for CD8 + T cells, EOMES deficiency in CD4 + T cells resulted in faster CLL progression in TCL1 AT mice.…”

“…To verify that these observed effects were intrinsic to CD8 + T cells and not induced by other Eomes-deficient cells in Eomes −/− BM chimera, such as previously described for CD4 + T cells [28], we transferred Eomes −/− or WT CD8 + T cells in Rag2 −/− mice and injected these mice 1 day later with Eµ-TCL1 leukemic cells. Similar to the results in the BM chimeric mice, leukemia development was significantly enhanced in mice that received Eomes-deficient CD8 + T cells compared to mice with WT T cells, with higher absolute numbers of CLL cells present in the blood, as well as an increased spleen weight and tumor content in this organ (Fig.…”

“…To analyze the role of EOMES in T-cell-mediated control of CLL, we generated Eomes −/− BM chimeric mice, as previously described [20,28]. Interestingly, mice with an Eomesdeficient hematopoietic system showed a higher tumor burden compared to Eomes WT chimeric mice at the experimental endpoint, 4 weeks after TCL1 AT.…”

“…Analyzed mice were males and females of 47-49 weeks of age. The TCL1 AT mouse model has been previously described [28]. Briefly, 2-3 months old C57BL/ 6 female mice were i.v.…”