Pre-existing malignancy results in increased prevalence of distinct populations of CD4(+) T cells during sepsis

The presence of pre-existing malignancy in murine hosts results in increased immune dysregulation and risk of mortality following a septic insult. Based on the known systemic immunologic changes that occur in cancer hosts, we hypothesized that the presence of pre-existing malignancy would result in phenotypic and functional changes in CD4(+) T cell responses following sepsis. In order to conduct a non-biased, unsupervised analysis of phenotypic differences between CD4(+) T cell compartments, cohorts of mice were injected with LLC1 tumor cells and tumors were allowed to grow for 3 weeks. These cancer hosts and age-matched non-cancer controls were then subjected to CLP. Splenocytes were harvested at 24h post CLP and flow cytometry and SPADE (Spanning-tree Progression Analysis of Density-normalized Events) were used to analyze populations of CD4(+) cells most different between the two groups. Results indicated that relative to non-cancer controls, cancer mice contained more resting memory CD4(+) T cells, more activated CD4(+) effectors, and fewer naïve CD4(+) T cells during sepsis, suggesting that the CD4(+) T cell compartment in cancer septic hosts is one of increased activation and differentiation. Moreover, cancer septic animals exhibited expansion of two distinct subsets of CD4(+) T cells relative to previously healthy septic controls. Specifically, we identified increases in both a PD-1(hi) population and a distinct 2B4(hi) BTLA(hi) LAG-3(hi) population in cancer septic animals. By combining phenotypic analysis of exhaustion markers with functional analysis of cytokine production, we found that PD-1(+) CD4(+) cells in cancer hosts failed to make any cytokines following CLP, while the 2B4(+) PD-1(lo) cells in cancer mice secreted increased TNF during sepsis. In sum, the immunophenotypic landscape of cancer septic animals is characterized by both increased CD4(+) T cell activation and exhaustion, findings that may underlie the observed increased mortality in mice with pre-existing malignancy following sepsis.