Cervical cancer (CC) is a leading cause of cancer-related deaths among women, primarily driven by persistent high-risk human papillomavirus (HPV) infection. This study explores HPV prevalence, genotypes, and their associations with peripheral blood immune factors and vaginal microenvironment balance in a CC screening population. A total of 2467 women undergoing CC screening from January to June 2024 were included. Among these, 227 tested positive for HPV (infection rate: 9.20 %). Using random sampling and propensity score matching, 100 HPV-positive and 100 HPV-negative individuals were selected. The HPV-positive group was further divided into HPV16/18 and other HPV subtypes. Samples from cervical cells, secretions, and peripheral blood were analyzed for HPV genotyping, vaginal microenvironment parameters (pH, cleanliness, Trichomonas, hydrogen peroxide-producing bacteria), Th1/Th2 cytokines (IL-6, IL-10, TNF-α), and T cell subsets (CD4 HPV-positive patients exhibited higher vaginal pH, abnormal vaginal cleanliness, and elevated hydrogen peroxide-producing bacteria compared to HPV-negative patients (P < 0.05). HPV16/18-positive individuals had greater vaginal alterations than other HPV subtypes. CD4 HPV, particularly HPV16/18, is associated with immune imbalance and vaginal microenvironment disruptions, highlighting the need for tailored strategies to improve CC prevention and diagnosis.