A C57BL/6N mice model of CP/CPPS established by prostate antigen immunization with DPT and BCG co-administration

Background and objective Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is common in young and middle-aged men, but existing animal models have key limitations. We aimed to establish a C57BL/6N mice model for mechanistic studies of CP/CPPS. Methods We immunized C57BL/6N mice with prostate antigen plus complete Freund’s adjuvant and added DPT, BCG, or DPT+BCG according to group assignment. Von Frey testing, ELISA, flow cytometry, HE staining, and immunohistochemistry were performed to assess pain, cytokines, immune cells, and prostate pathology. Key findings and limitations PAg+CFA+DPT+BCG booster immunization induced the strongest pain-related phenotype, increased IL-1β and TNF-α levels, reduced IL-10, aggravated prostatic inflammation, and increased NLRP3, COX-2, NF-κB, and Caspase-1 expression compared with the NC group. Compared with either monotherapy, the PAg+CFA+DPT+BCG group showed stronger inflammatory and pain-related phenotypes across most readouts. Limitation: The model does not fully replicate human CP/CPPS heterogeneity. Conclusions and clinical implications We established a C57BL/6N mice model with CP/CPPS-like inflammatory and pain-related features and found that DPT+BCG co-administration produced the strongest phenotype. Its compatibility with transgenic mice may facilitate mechanistic studies and preclinical hypothesis testing in CP/CPPS. Patient summary In this study, we established a prostatitis-like model in C57BL/6N mice. Booster immunization with DPT+BCG induced inflammatory and pain-related changes, providing a useful model for future mechanistic studies.