肿瘤引流淋巴结中免疫活性细胞分布的原位分析

In situ analysis of distribution of immunocompetent cells in tumor’s local draining lymph nodes

目的:观察人乳腺癌和胃癌局部引流淋巴结(LDLN)从无转移、微转移到晚期转移过程中,免疫组织学变化及免疫活性细胞(ICC)的分布特征。方法:采用传统的病理学方法,对22例乳腺癌LDLN(71个)和7例进展期胃癌LDLN(28个)进行组织形态学分类,并以抗穿孔素、抗颗粒酶B、抗CD8、抗CD56、抗CD68、抗S-100、抗CD134及抗CD25单克隆抗体(mAb)进行催化信号放大(Catalyzedsignalamplification,CSA)免疫组化染色,检测肿瘤LNDN中ICC的分布。结果:肿瘤LDLN中以副皮质区增生和窦组织细胞增生为主,细胞毒性T淋巴细胞(CTL)及树突状细胞(DC)的数量,从无转移、微转移到晚期转移过程中有逐渐减少的趋势。在无和微转移的淋巴结内,穿孔素+、颗粒酶B+及S100+DC的数量高于晚期转移淋巴结(P<0.05);而S100+DC不仅数量减少,而且其形态也有变化,呈多角形、星形,并有胞质突起,与周围淋巴细胞接触呈活化状态的DC变为椭圆形,少有胞质突起或呈短突起的静止状态的DC。CD134+细胞及CD25+细胞的数量在晚期转移淋巴结中明显高于无和微转移淋巴结(P<0.01)。ICC在无和微转移的前哨和非前哨淋巴结中的分布无统计学意义(P>0.05)。结论:肿瘤LDLN中ICC分布的变化,提示随着肿瘤的进展,其免疫微环境向抑制机体抗肿瘤免疫的方向偏移。

AIM： To observe the distribution of immunocompetent cells （ICCs） in tumor＇s local draining lymph nodes （LDLN） at different stages of disease including nonmetastasis, micro-metastasis and late metastasis. METHODS： 71 LDLN from 22 breast cancer, 28 LDLN from 7 gastric carcinoma were analysed by using catalyzed signal amplification （CSA） immunohistochemical staining. Monoclonal antibodies （mAbs） to perforin, granzyme B, CD8, CD56, CD68, S-100, CD134 and CD25 were used to detect the amount and functional change of ICCs. RESULTS： Paracortical hyperplasia and sinus histocytosis was maily observed in tumor＇s LDLN. As it was reported, the density of S100 ^＋ dendritic cells （DCs） was decreased as the draining lymph nodes became tumor-containing from the status of tumor free （P〈0.05）, and the morphology of DCs turned to be inactivated. As the lymph nodes were invaded by tumors, the densities of CD134^＋ lymphocytes and CD25 ^＋ cells were significantly increased （ P 〈 0.01 ）. Furthermore, there was a significant trend of decreasing in the number of activated cytotoxic T lymphocytes （ granzyme B ^＋ cells） in LDLN as tumor progressed from non/micro-metastasis, early metastasis to advanced stage metastasis （ P 〈0.01 ）. There was no obvious difference in distribution of ICC between non/micro-metastasis sentinel lymph nodes and non-sentinel lymph nodes. CONCLUSION： The dynamic change of the ICCs in LDLN implied that immune response of tumor bearing host tends to be inhibited with the progress of tumors.