肿瘤抗原诱导的T淋巴细胞膀胱灌注预防膀胱癌复发

目的探讨一种预防膀胱癌术后复发的新方法.方法由手术切除的膀胱癌组织制备肿瘤可溶性抗原,以卡介苗为佐剂,诱导人外周血单个核细胞成为细胞毒性T细胞(CTL).用培养8～10d的CTL及其培养上清为37例膀胱癌切除术后患者进行膀胱腔内灌注治疗.定期膀胱镜复查,SAP免疫组化法检测外周血T淋巴细胞亚群分布的变化,ELISA法测定血清及尿液中IL-2等细胞因子含量,观察疗效.结果患者外周血CD3+、CD4+T细胞亚群的比例增加显著(P＜0.01);CD4+/CD8+T细胞比例明显提高(1.60±0.20,P＜0.01)血清及尿液中白细胞介素2、肿瘤坏死因子α、干扰素γ的水平均较治疗前明显提高(P＜0.01);组织学观察显示膀胱粘膜呈轻度非特异性炎症反应.随访6～26个月(中位时间为16个月),复发率2.7%,未见有严重不良反应.结论CTL及其培养上清膀胱腔内灌注治疗能够提高膀胱癌术后患者膀胱局部和全身的免疫水平,有效降低膀胱癌的术后复发率.

Intravesical instillation of cytotoxic T-lymphocytes induced by soluble tumor antigen in the prophylaxis of recurrence of bladder cancer

OBJECTIVE: To explore a new method for preventing the recurrence of bladder cancer. METHODS: With the adjuvant of BCG, cytotoxic T-lymphocytes (CTLs) were induced from human peripheral blood mononuclear cells (PBMC) by bladder tumor soluble antigen which was extracted through salt-infiltrating biochemical procedures from autologous bladder tumor. Intravesical instillation of CTLs together with their supernatants after culture for 8 to 10 days was performed in 37 patients with superficial bladder cancer after surgery. The effects of the treatment were evaluated by endocytoscopy, pathological examinations and ELISA assays for the levels of cytokines IL-2, TNF-alpha and IFN-gamma in the patients' serum and urine. The distribution of T-lymphocyte subpopulations was also monitored by SAP immuno-histochemical techniques. RESULTS: The proportion of CD3+ and CD4+ T-lymphocyte subpopulations increased significantly while CD8+ T-lymphocyte subpopulation was stable. The ratio of CD4+ to CD8+ increased to 1.60 +/- 0.29. The levels of cytokines IL-2, TNF-alpha and IFN-gamma in serum and urine increased significantly, especially the level of TNF-alpha in urine. Pathological studies revealed that non-specific inflammation is the major change in the bladder mucosa after therapy. Follow-up for 6-26 months showed that 1 patient havd moderate heterogeneity in bladder mucosa. The recurrence rate was 2.7%. Side effects were not found except transient lower fever or hematuria. CONCLUSION: Intravesical insitillation of CTLs could effectively prevent the recurrence of superficial bladder cancer after the improvement of immunological reactivity in the whole body.