Role of CD28 + PD-1 + T-c cells in Immune Response and Prognosis Prediction in Hepatocellular Carcinoma

WUHAN YANG¹Teng Pan²Yaowen Chen¹Hao Guo¹Yaqi Peng³Chao Wang¹Li Peng^1*Shubin Wang^1*

• ¹Fourth Hospital of Hebei Medical University, Shijiazhuang, China
• ²First Hospital of Shijiazhuang, Shijiazhuang, Hebei Province, China
• ³Second Hospital of Hebei Medical University, Shijiazhuang, Hebei Province, China

Background CD28+PD1+ T-cellsCD28 + PD-1 + Tc cells (CD8 + T cells) constitute a dysfunctional subset of T -cell subset; however, their the underlying mechanisms underlying their dysfunction and their significance in hepatocellular carcinoma (HCC) remain unclear. We aimed to elucidate the prognostic significance and molecular characteristics of CD28+PD1+ T-cellCD28 + PD-1 + Tc cell infiltration in HCC.We established a single-cell HCC transcriptional map, focusing on cell-cell communication and trajectory analysis of CD28+PD1+ T-cellsCD28 + PD-1 + Tc cells. We assessed the correlation between CD28+PD1+ T-cellCD28 + PD-1 + Tc-cell enrichment and prognosis and investigated potential molecular mechanisms using enrichment analyses. Flow cytometry was used to compare CD28+PD1+ T-cellCD28 + PD-1 + Tc-cell infiltration between HCC and adjacent normal tissues and evaluate cytotoxic factors and immune checkpoint expression were evaluated.Overall, 25,644 T-cells were identified from single-cell RNA sequencingscRNA data of from 10 HCC samples and corresponding normal samples. Overall T-cell infiltration decreased was lower in HCC tissues, with a significantly higher increase in CD28+PD1+ T-cellCD28 + PD-1 + Tc-cell infiltration. Bulk RNA sequencing data integration revealed a correlation between higher CD28+PD1+ T-cellCD28 + PD-1 + Tc-cell infiltration and significantly worse prognosis. Flow cytometry confirmed higher CD28+PD1+ T-cellCD28 + PD-1 + Tc-cell enrichment in HCC tissues.