Fig. 2: Downregulation of TPX2 in CD8 + T cells indicated low antitumor activity and a poor anti-PD-1 response.

A The expression of immune checkpoint markers (PD-1, TIM-3, CTLA-4, and TIGHT) was detected in TPX2^low and TPX2^high (indicated in Fig. 1D) CD8+ T cells by flow cytometry (n = 10). B The proportion of PD-1 + TCF1+ cells among TPX2^low and TPX2^high (indicated in Fig. 1D) CD8+ T cells (n = 10). C Proliferation (Ki67) and apoptosis (A-Caspase-3) were assayed in tumor-infiltrating CD8 + cells in which TPX2 was knocked down (LV-shRNA) or overexpressed (LV-TPX2) (n = 6). D The production of effector cytokines (IFN-γ and TNF-α) was assayed in tumor-infiltrating CD8+ cells in which TPX2 was knocked down (LV-shRNA) or overexpressed (LV-TPX2) (n = 6). *p < 0.05, ***p < 0.001; the two-tailed unpaired Student’s t-test was used to compare two groups. A-Caspase-3 activated Caspase 3, IFN-γ interferon gamma, MKI67 Marker of proliferation Ki67, LAG-3 lymphocyte activation gene 3, LV-NC control lentivirus, LV-shRNA lentivirus used to knock down the human TPX2 gene, LV-TPX2 lentivirus used to overexpress the human TPX2 gene, MFI mean fluorescence intensity, PD-1 programmed death 1, TCF1 T cell factor 1, TIGHT T cell immunoreceptor with Ig and ITIM domains, TIM-3 T cell immunoglobulin and mucin domain containing-3, TNF-α tumor necrosis factor alpha.