黑色素瘤细胞裂解物通过巨噬细胞抗原递呈诱导淋巴细胞活化体外实验  

Melanoma Cell Lysate Induced Lymphocyte Activation by Macrophages Antigen Presentation in Vitro

作　　者：孙宇[1] 杨玉霞 车姣子 赵明珍 贾利颖 SUN Yu;YANG Yuxia;CHE Jiaozi;ZHAO Mingzhen;JIA Liying(Affiliated Hospital of Chengde Medical University,Chengde 067000,China;Chengde Central Hospital,Chengde 067000,China)

机构地区：[1]承德医学院附属医院,河北承德067000 [2]承德市中心医院,河北承德067000

出　　处：《皮肤科学通报》2025年第1期49-56,共8页Dermatology Bulletin

基　　金：承德市科技计划项目(202204A155)。

摘　　要：目的探讨巨噬细胞对黑色素瘤细胞裂解物抗原的抗原递呈作用。方法反复冻融法制备小鼠B16F10黑色素瘤细胞裂解物抗原,根据其浓度的不同分为4组:对照组、25μg/mL组、50μg/mL组和100μg/mL组,分别作用于C57BL/6小鼠腹腔巨噬细胞,流式细胞术检测巨噬细胞MHC-Ⅰ分子H-2Kb/H-2Db和MHC-Ⅱ分子I-A/I-E以及协同刺激分子CD80和CD86的表达。制备C57BL/6小鼠免疫淋巴细胞,与上述巨噬细胞共培养,收集未贴壁淋巴细胞,再与B16F10黑色素瘤细胞共培养,CCK8法检测淋巴细胞增殖率和黑色素瘤细胞增殖抑制率。结果几乎所有巨噬细胞均阳性表达H-2Kb/H-2Db,各组间差异无统计学意义(P>0.05);而与对照组相比,100μg/mL组H-2Kb/H-2Db表达强度明显增高,差异有统计学意义(P<0.05)。与对照组相比,25μg/mL组、50μg/mL组和100μg/mL组的I-A/I-E阳性表达率、CD80和CD86阳性表达率以及淋巴细胞对B16F10黑色素瘤细胞的增殖抑制率增高,差异均有统计学意义(P<0.05)。与对照组相比,50μg/mL组和100μg/mL组的淋巴细胞增殖率增高,差异有统计学意义(P<0.05)。结论巨噬细胞对黑色素瘤细胞裂解物抗原具有抗原递呈作用,可诱导淋巴细胞的抗黑色素瘤免疫反应。Objective To investigate the antigen-presenting effect of macrophages on melanoma cell lysate antigen.Methods Mouse B16F10 melanoma cell lysate antigen was prepared by repeated freeze-thaw method and divided into four groups according to its concentration:control,25μg/mL,50μg/mL and 100μg/mL groups,which applied to C57BL/6 mouse peritoneal macrophages respectively.The expression of MHC-I molecules H-2Kb/H-2Db,MHC-II molecules I-A/I-E and costimulatory molecules CD80 and CD86 in macrophages were detected by flow cytometry.C57BL/6 mouse immunized lymphocytes were prepared,co-cultured with the above macrophages,unadherent lymphocytes were collected,and then co-cultured with B16F10 melanoma cells.The proliferation rate of lymphocytes and inhibition rate of melanoma cell proliferation were detected by CCK8 method.Results Almost all macrophages expressed positive H-2Kb/H-2Db,and there was no significant difference among all groups(P>0.05),while the expression intensity of H-2Kb/H-2Db in 100μg/mL group was significantly different from that in control group(P<0.05).The positive expression rate of I-A/I-E,the positive expression rate of CD80 and CD86,and the proliferation inhibition rate of lymphocytes against B16F10 melanoma cells in 25μg/mL,50μg/mL and 100μg/mL groups were significantly different from the control group(P<0.05).The lymphocyte proliferation rate of 50μg/mL group and 100μg/mL group was significantly different from that of the control group(P<0.05).Conclusion Macrophages has antigen-presenting effect on melanoma cell lysate antigen,and are able to induce anti-melanoma immune response of lymphocytes.

关 键 词：黑色素瘤 MHC 协同刺激分子 淋巴细胞活化 抗原递呈 抗肿瘤免疫 

分 类 号：R751[医药卫生—皮肤病学与性病学]