Ding, Z.-c. | Search bioRxiv

bioRxiv2026

IL13 critically contributes to the antitumor activity of polyfunctional CD4+ T cells driven by constitutively active Stat5

The presence of polyfunctional T cells capable of concurrently producing multiple cytokines correlates with favorable clinical outcomes in patients with cancer. We previously reported that overexpression of constitutively active Stat5a (CASTAT5) confers polyfunctionality on tumor-specific CD4 T cells, enabling durable antitumor immunity following adoptive cell therapy. However, the mechanisms underlying the enhanced antitumor effects of CASTAT5-modified T cells remain unclear. Here, we identify interleukin-13 (IL13) as a critical effector cytokine contributing to the multifaceted antitumor activity of CASTAT5-conditioned polyfunctional CD4 T cells (C5-CD4). C5-CD4 T cells exhibit a prominent T helper 2 (Th2) cytokine profile, accompanied by extensive epigenomic remodeling at the Th2 cytokine loci, as revealed by Hi-C and histone-modification CUT&Tag assays. Neutralization of IL13 diminished the antitumor efficacy of C5-CD4 T cells. Furthermore, targeted Il13 disruption substantially impaired tumor control and was associated with reduced donor T cell expansion and persistence, diminished systemic cytokine and chemokine responses, and attenuated activation of endogenous tumor-reactive CD8 T cells. Nonetheless, IL13 overexpression alone in donor CD4 T cells failed to fully recapitulate the therapeutic activity of C5-CD4 T cells, indicating that IL13 is a critical contributor but is insufficient to account for the full antitumor activity conferred by CASTAT5. Single-cell RNA sequencing associated donor-derived IL13 with increased representation of tumor-infiltrating myeloid cells and IL13-signaling in monocytes and macrophages exhibiting tissue-repair-associated transcriptional programs. Moreover, Il13 disruption diminished the therapeutic efficacy of CASTAT5-expressing CD19-targeted CAR-T cells in a mouse B-cell lymphoma model. Reanalysis of a human CD19CAR-T infusion-product dataset further revealed an association between IL13 expression and type 2 cytokine signatures linked to prolonged clinical response, suggesting a potential contribution of IL13 to the efficacy of human CD19CAR-T therapy. Together, these findings uncover an unexpected antitumor role for IL13 and establish it as a critical node linking polyfunctional donor T cells with endogenous immune cells to promote sustained tumor immunity.

immunology↗

bioRxiv2024

Single-cell transcriptome profiling of the myeloid cells repopulating after chemotherapy identifies a neutrophil-like monocyte subset with pro-tumor activities

Patients with cancer often receive chemotherapy to control tumor progression and reduce disease symptoms. Cytotoxic chemotherapeutic agents not only kill rapidly growing cancer cells but also reduce normal cells including myeloid cells, the main innate immune population involved in fighting infections and repairing tissue damages. Rapid loss of myeloid cells caused by chemotherapy triggers myelopoiesis, a process in which the hematopoietic stem and progenitor cells in the bone marrow regenerate myeloid cells, including monocytes, neutrophils, dendritic cells and macrophages, to reconstitute the myeloid cell compartment. We previously reported that chemotherapy with an alkylating agent cyclophosphamide (CTX) in mice leads to repopulation of myeloid cells that acquire immunosuppressive activities within the monocyte subset. However, detailed information on the cellular composition and molecular identity of these chemotherapy- induced immunosuppressive monocytes is lacking. Here, we investigated how the various myeloid cell subsets in the bone marrow of mice respond to CTX chemotherapy through single-cell RNA sequencing analysis (scRNAseq). We found that myeloid progenitor cells and monocytes were reduced 2 days after chemotherapy but rebounded and surpassed their pretreatment levels by day 7. Further scRNAseq analysis of pre-enriched monocytes revealed that the monocyte population was heterogenous, and that chemotherapy tilted myelopoiesis towards the production of neutrophil-like monocytes (NeuMo). We identified Cxcr4 and Cx3cr1 as suitable markers for isolation of chemotherapy-induced NeuMo and demonstrated that these cells were suppressive to T cells. Together with the evidence that CTX-induced monocytes can promote breast cancer metastasis in mice, our data reveal the heterogeneity of the monocytes reemerging after chemotherapy and identify the NeuMo subset as a potential therapeutic target for enhancing the efficacy of chemotherapy in cancer.

immunology↗