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目的:评价经皮冷冻消融治疗局限性前列腺癌前后机体抗肿瘤免疫反应。方法:10例局限性前列腺癌患者行经皮冷冻消融治疗。分别于冷冻治疗前1d、治疗后2周采取外周血,分离外周血单个核细胞(PBMCs)。穿刺活检制备前列腺自身肿瘤和非肿瘤组织溶解物。酶联免疫吸附测定(ELISA)法检测血清TNF-α、IFN-γ、IL-4、IL-10水平,以IFN-γ/IL-4计算Th1/Th2比值。酶联免疫斑点(ELISPOT)试验检测不同组织溶解物刺激下分泌IFN-γ的T细胞数。以人前列腺癌LNCaP细胞和肾癌GRC-1细胞为靶细胞,乳酸脱氢酶(LDH)释放试验检测细胞毒T淋巴细胞(CTL)特异性杀伤活性。结果:与术前比较,冷冻治疗后TNF-α、IFN-γ明显升高,IL-4、IL-10轻度下降,Th1/Th2比值明显升高(P<0.01)。肿瘤组织溶解物刺激后IFN-γ+T细胞数明显增多(P<0.01)。非肿瘤组织溶解物刺激后,IFN-γ+T细胞数无明显变化。冷冻治疗后细胞毒CTL对人前列腺癌细胞株LNCaP特异杀伤活性明显增强,而对人肾癌细胞株GRC-1无明显杀伤活性。随访3～6个月,仅1例患者肿瘤复发。结论:经皮冷冻治疗前列腺癌可诱导机体产生肿瘤特异性免疫反应。
OBJECTIVE: To evaluate the anti-tumor immune response of percutaneous frozen ablation in the treatment of localized prostate cancer. Methods: Percutaneous frozen ablation was performed in 10 patients with localized prostate cancer. Peripheral blood was collected at 1 day before cryotherapy and 2 weeks after treatment, and peripheral blood mononuclear cells (PBMCs) were isolated. Puncture biopsy to prepare prostate autologous tumor and non-tumor tissue lysate. Serum levels of TNF-α, IFN-γ, IL-4 and IL-10 were detected by enzyme linked immunosorbent assay (ELISA), and Th1 / Th2 ratio was calculated by IFN- γ / IL-4. ELISPOT assay was used to detect the number of T cells secreting IFN-γ stimulated by different tissue lysates. The cytotoxic T lymphocytes (CTLs) specific killing activity of LNCaP cells in human prostate cancer and GRC-1 cells in renal carcinoma were detected by the lactate dehydrogenase (LDH) release assay. Results: Compared with the preoperative value, the levels of TNF-α and IFN-γ were significantly increased, the levels of IL-4 and IL-10 decreased slightly and the Th1 / Th2 ratio increased significantly (P <0.01). The number of IFN-γ + T cells was significantly increased after tumor tissue lysate stimulation (P <0.01). There was no significant change in the number of IFN-γ + T cells after stimulation with non-tumor lysate. After cytotoxic CTL treatment, the cytotoxicity of CTL on human prostate cancer cell line LNCaP was significantly enhanced, but no obvious cytotoxicity on human renal cell carcinoma cell line GRC-1. Follow-up 3 to 6 months, only 1 patient with tumor recurrence. Conclusion: Percutaneous freezing treatment of prostate cancer can induce tumor-specific immune response.