CAJ | 학술논문

目的：观察恶性黑色素瘤B16细胞经重组腺病毒感染后，表达在B16细胞膜上的SEA和CD80体外能否诱导免疫学反应。方法：B16细胞分别经空载体腺病毒Ad（空）和重组腺病毒Ad-MMRE-mTERT-B7、Ad-MMRE-mTERT-SEA、Ad-MMRE -mTERT-BIS感染后，和小鼠脾淋巴细胞共培养，然后，采用Brdu 酶联免疫法（ ELISA）检测淋巴细胞增殖；流式细胞术检测T淋巴细胞亚群增殖；ELISA法检测细胞因子IL-2、TNF-α和IFN-γ的产生。结果：与感染空载体腺病毒Ad（空）和未感染B16细胞相比，经重组腺病毒感染的B16细胞体外能够显著诱导脾淋巴细胞的增殖，其中CD3+CD4+和CD3+CD8+T淋巴细胞增殖显著；细胞因子TNF-α、IFN-γ和IL-2的产生显著升高；CTLs对肿瘤细胞的杀伤活性显著增强；表达双基因的B16细胞体外诱导免疫应答的能力高于单基因过表达B16细胞。结论：B16细胞经重组腺病毒感染后，表达在细胞膜上的SEA和CD80具有免疫学活性，本研究为今后进一步采用所构建重组腺病毒进行恶性黑色素瘤的免疫基因治疗提供了实验依据。
목적：관찰악성흑색소류B16세포경중조선병독감염후，표체재B16세포막상적SEA화CD80체외능부유도면역학반응。방법：B16세포분별경공재체선병독Ad（공）화중조선병독Ad-MMRE-mTERT-B7、Ad-MMRE-mTERT-SEA、Ad-MMRE -mTERT-BIS감염후，화소서비림파세포공배양，연후，채용Brdu 매련면역법（ ELISA）검측림파세포증식；류식세포술검측T림파세포아군증식；ELISA법검측세포인자IL-2、TNF-α화IFN-γ적산생。결과：여감염공재체선병독Ad（공）화미감염B16세포상비，경중조선병독감염적B16세포체외능구현저유도비림파세포적증식，기중CD3+CD4+화CD3+CD8+T림파세포증식현저；세포인자TNF-α、IFN-γ화IL-2적산생현저승고；CTLs대종류세포적살상활성현저증강；표체쌍기인적B16세포체외유도면역응답적능력고우단기인과표체B16세포。결론：B16세포경중조선병독감염후，표체재세포막상적SEA화CD80구유면역학활성，본연구위금후진일보채용소구건중조선병독진행악성흑색소류적면역기인치료제공료실험의거。
Objective:To observe whether Staphylococcus enterotoxin A( SEA)and CD80 expressing on the surface of B16 cells could induce immune response after infection with recombinant adenoviruses. Methods:Mouse spleen lymphocytes were cocultured with B16 cells after infection with empty adenoviruse vector Ad( empty)or the recombi-nant adenoviruses of Ad-MMRE-mTERT-B7,Ad-MMRE-mTERT-SEA or Ad-MMRE-mTERT-BIS. Then,lymphocyte proliferation was assayed by BrdU incorporation using a commercial kit,flow cytometric analysis was used to detect proliferation of T lymphocyte subgroups. The production of IL-2,TNF-α and IFN-γ was tested by enzyme-linked immunosorbent assay. Results:Compared with B16 infected with empty adenovirus or without infec-tion,B16 infected with recombinant adenoviruses could significantly induce proliferation of spleen lymphocytes and T lymphocyte subgroups,and increase IL-2,TNF-αand IFN-γproduction. The antitumor response was significantly stronger in B16 cells after dual-gene expression than that in B16 cells after single-gene expression. Conclusion:The results indicate that SEA and CD80 expressed on the membrane of B16 cells after infection with recombinant ade-novirus has immune activities. This study provides some experimental evidence for further immune genetherapy against malignant melanoma with the recombinant adenovirus.