Li Qin, Du Guan-huan, Tang Guo-yao
Department of Oral Medicine, Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratary of Stomatology, Shanghai 200011, China.
Shanghai Kou Qiang Yi Xue. 2011 Feb;20(1):16-20.
To establish a correlated cocultured model of keratinocytes and T lymphocytes in vitro to simulate the immune response circumstance of oral lichen planus(OLP) lesions.
Keratinocytes prepared with enzyme digestion from normal oral mucosa and T lymphocytes prepared with negative isolation procedure by magnetic beads were co-cultured. Proliferation of T lymphocytes in cocultured cells was detected using MTT assay and the expression of IFN-γ was detected by ELISA. All the results were analyzed with independent samples t test using SAS6.12 software package.
The proliferation level of T lymphocytes in the cocultured group of T lymphocytes and keratinocytes which expressed PD-L1 and PD-L2 was higher than that in the group of T lymphocytes (P < 0.05). The expression of IFN-γ in the cocultured group was significantly higher than that in the group of T lymphocytes (P < 0.01).
The cocultured model of T lymphocytes and keratinocytes which express PD-L1 and PD-L2 can simulate the immune response circumstance of OLP lesions to a certain extent. Supported by National Natural Science Foundation of China (30872888),Research Fund of Science and Technology Commission of Shanghai Municipality(08DZ2271100) and Shanghai Leading Academic Discipline Project (S30206).
建立角质形成细胞与T淋巴细胞体外共培养模型,以模拟口腔扁平苔藓(OLP)病损的免疫反应环境。
采用酶消化法从正常口腔黏膜制备角质形成细胞,采用磁珠阴性分选法制备T淋巴细胞,并进行共培养。采用MTT法检测共培养细胞中T淋巴细胞的增殖情况,采用ELISA法检测干扰素-γ(IFN-γ)的表达。所有结果采用SAS6.12软件包进行独立样本t检验分析。
表达程序性死亡配体-1(PD-L1)和程序性死亡配体-2(PD-L2)的角质形成细胞与T淋巴细胞共培养组中T淋巴细胞的增殖水平高于T淋巴细胞组(P<0.05)。共培养组中IFN-γ的表达明显高于T淋巴细胞组(P<0.01)。
表达PD-L1和PD-L2的角质形成细胞与T淋巴细胞共培养模型可在一定程度上模拟OLP病损的免疫反应环境。 本研究受国家自然科学基金(30872888)、上海市科委科研基金(08DZ2271100)和上海市重点学科项目(S30206)资助。