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Bulk RNA Sequencing of MCMV-Specific M45 Tetramer-Positive CD8⁺ T Cells from Wild-Type and B cell-deficient JHT mice

ABSTRACT: To assess the impact of B cell deficiency on MCMV-specific CD8⁺ T cells, equal numbers of M45 tetramer-positive CD8⁺ T cells were sorted from wild-type and B cell–deficient JHT mice. The isolated antigen-specific CD8⁺ T cells were subjected to bulk RNA sequencing to compare transcriptional profiles between the two groups.

ORGANISM(S): Mus musculus

PROVIDER: GSE315617 | GEO | 2026/01/18

REPOSITORIES: GEO

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Project description:Reconstitution of cytomegalovirus (CMV)-specific immunity following transplant remains a primary clinical objective to prevent CMV disease, and adoptive immunotherapy of CMV-specific T cells can be an effective therapeutic approach. Due to the persistence of CMV, most CMV-specific CD8+ T cells become terminally differentiated effector cells (TEFF). However, a minor subset retains a memory phenotype (TM). Interestingly, recent studies suggest that CMV-specific CD8+ T cells with different phenotypes may have different abilities to reconstitute sustained immunity following transfer. The immunology of human CMV (HCMV) infections is reflected in the mouse model of MCMV infection. We found that HCMV- and MCMV-specific T cells displayed shared genetic programs, validating the MCMV model for studies of CMV-specific T cells in vivo. After transfer, the proliferative capacity of MCMV-specific TM cells was vastly superior to TEFF cells. Strikingly, TM cells expanded and established sustained and diverse T cell populations even after multiple challenges. Although both TEFF and TM cells could protect Rag-/- mice, only TM cells could consistently survive after transfer into immune replete, latently infected recipients and respond if recipient immunity was lost. These data show that CMV-specific TM cells retain memory function during persistent infection and can re-establish CMV immunity when necessary. C57BL/6 mice were infected intraperitoneally (i.p.) with MCMV strain MW97.01 between 6-16 weeks of age. Splenocytes were isolated from chronically-infected mice and co-stained with three PE-conjugated tetramers loaded with the antigenic peptides from M38, m139 and IE3, all of which promote memory inflation. Cells were then stained with fluorescently conjugated antibodies and sorted on a MoFlo cell sorter. Naïve CD8+ cells were identified as CD44lo. MCMV-specific T cells were identified as CD8+, CD44hi and tetramer binding and then further segregated into memory and effector cells subsets by their expression of KLRG1 and CD127.

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Bulk RNA Sequencing of MCMV-Specific M45 Tetramer-Positive CD8⁺ T Cells from Wild-Type and B cell-deficient JHT mice

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