Monkeypox (Mpox) was declared a public health emergency of international concern by WHO, and there is growing evidence of an association between mpox and HIV infection. However, the underlying mechanism is largely unknown. In this study, using integrated bioinformatics analysis, the underlying mechanisms and important genes involved in the crosstalk between HIV and Mpox were investigated. CHMP2A was shown to be significantly upregulated after Mpox infection and in HIV-infected CD8+ T cells utilizing DEG and weighted gene co-expression network (WGCNA) analysis. Following that, the HIV scRNA-seq dataset was examined further to establish the expression level and location of the CHMP2A gene. CHMP2A was shown to be significantly expressed in CD8+ exhaustion T cells, and its expression increased in CD8+ exhaustion T cells from HIV patients. Interestingly, pseudotime analysis revealed that CHMP2A was linked to the differentiation of CD8+ effector memory T cells into CD8+ exhaustion T cells. In addition, TAF7 and IRF3 were identified as possible upstream transcription factors that regulate CHMP2A by pySCENIC analysis. Finally, applying virtual screening of 4381 approved drugs and molecular dynamics simulations analysis, ZINC000027990463 has the potential to be a drug that targets CHMP2A. Overall, our results reveal key biomarkers that are potentially involved in the crosstalk between Mpox and HIV, which provides new insights into the mechanisms of disease co-infection. Further investigation is needed to determine the function of CHMP2A in CD8+ T cells following infection.