原发性肝癌经皮微波凝固治疗前后局部免疫活性细胞功能检测

目的：了解原发性肝癌经皮微波凝固治疗前、后局部免疫活性细胞功能变化。方法：C地38例病理诊断原发性肝癌，并接受超声导引下经皮微波凝固治疗的患者。分别治疗前及治疗后17d，超声导引下经皮用18G组织切割针于病灶及其周边肝组织取活检标本，取出的组织标本石蜡包埋，采用特异性单克隆抗体免疫组织化学染色，检测CD3^ 、CD56^ 、CD68^ 细胞及T淋巴细胞表面Fas配体；并在光镜下观察，用病理图像分析仪测量治疗前后、后阳性细胞直径、阳性细胞占单位面积百分比、T淋巴细胞Fas-L阳性表达率及治疗前后巨噬细胞内次级溶酶体变化。结果：治疗前肿瘤内仅有少量免疫细胞浸润，多数浸润的CD3^ 和CD56^ 细胞最大径小于10μm，CD68^ 细胞最大径小于18μm。治疗后病灶内浸润的CD3^ 、CD56^ 和CD68^ 细胞数量较治疗前明显增加，细胞体积明显增大（同治疗前相比CD3^ 细胞和CD56^ 细胞t和P值分别为3．48，－4．76和0．025，0．000，巨噬细胞t和P值分别为－2．46和0．028）。最大径大于10μm的CD3^ 和CD56^ 细胞分别由治疗前的10．4％和20．1％增至24．9％和30．2％，最大径大于18μm的CD68^ 细胞由10．2％增至33．4％。T淋巴细胞Fas-L阳性率由治疗前的7．2％增高至20．1％（P＜0．01，巨噬细胞内次级溶酶体和T淋巴细胞内细胞器明显增多。结论：原发性肝癌经皮微波凝固治疗提高局部浸润免疫细胞的功能。

Functional assessment of infiltrating immunocytes in patients with primary hepatocellular carcinoma after percutaneous microwave coagulation therapy

OBJECTIVE: To assess the function local immune cells in patients with primary hepatocellular carcinoma (HCC) after percutaneous microwave coagulation therapy (PMCT). METHODS: Thirty-eight patients with histologically proved primary HCC underwent ultrasound guided PMCT. Specimens were taken from the lesion site before and 17 days after PMCT respectively using 18-guage core needle through US-guide biopsy, embedded in paraffin, and stained by immunohistochemistry. The panel of monoclonal antibodies of CD3, CD56, CD68 and Fas-L were used to detect the CD3+, CD56+, CD68+ cells and T lymphocyte Fas-ligand. The positive cells were detected under light microscopy. Their diameter and area and the Fas-L expression rate of T lymphocytes and changes of secondary lysosomes in macrophages were measured by computer. RESULTS: Before PMCT, only a few infiltrating immunocytes were seen in the tumor specimens; the diameter of most CD3+ and CD56+ cells was less than 10 microns and the diameter of CD68+ cells was less than 18 microns. The amount and volume of CD3+, CD56+, and CD68+ cells significantly increased after PMCT (t = 3.48, P = 0.025 for CD3+ cells, t = -4.76, P = 0.000 for CD56+ cells, and t = -2.46, P = 0.028 for CD68+ cells). The percentage of CD3+ and CD56+ cells with the largest diameter > 10 microns increased from 10.4% and 20.1% respectively before PMCT to 24.9% and 30.2% respectively after PMCT. The percentage of CD68+ cells with the largest diameter > 18 microns increased from 10.2% before PMCT to 33.4% after PMCT. The Fas-l expression rate of T lymphocytes increased from 7.2% to 20.1% (t = -19.12, P = 0.000). The secondary lysosomes and cellular debris within microphages and the cellular organs in T lymphocytes significantly increased. CONCLUSION: The function of intratumaral infiltrating immunocytes is significantly enhanced after PMCT.