Nonylphenol (NP) is the final biodegradation product of nonylphenol polyethoxylates, which are widely used as surfactants in domestic and industrial products. NP has been reported to have estrogenic activity and shown to have potential reproductive toxicity. However, its influence on immune system function remains unclear. In this study, to determine the immunological effects of NP, the effects of NP on apoptosis and Fas/FasL gene expression in rat thymocyte in vitro were investigated. Thymocytes were treated with NP 0.1, 1, and 10 ppm, respectively. Viable cell numbers were determined by MTT assay. Apoptotic cells were identified by DNA fragment analysis. A semi-quantitative reverse transcriptase-polymerase chain reaction method was used to analyze Fas and FasL mRNA levels. Fas and FasL protein expression was evaluated by flow cytometry. The results showed that NP decreased the cellularity; induced apoptotic death and enhanced the expression of Fas and FasL mRNA as well as proteins in thymocytes. These findings suggest that NP may induce apoptosis by altering the expression of Fas and FasL in thymocytes so as to affect the immune system function.