论文部分内容阅读
旨在探明猪细小病毒样颗粒(PPV-VLPs)被猪脾树突状细胞(DC)捕获后,被提呈的方式。首先通过磁性筛选的方法从非免疫猪和免疫猪的脾分离CD172a+CD11R+DC及CD4-CD8+T细胞。DC分别与伯氨喹、放线菌酮、氯喹、乳胞素、布雷菲德菌素A及亮抑肽酶、胃酶抑素等作用1h后,再与细小病毒样颗粒PPV-VLPs-E290在37℃作用4h,应用CD8+T细胞的细胞毒性分析检测DC对PPV-VLPs-E290的提呈情况。乳酸脱氢酶(LDH)释放试验检测结果显示,伯氨喹及亮抑肽酶对DC提呈PPV-VLPs-E290的效率无明显影响,胃酶抑素部分抑制,而氯喹、放线菌酮、布雷菲德菌素A及乳胞素则可使DC提呈PPV-VLPs-E290的效率明显下降。结果表明,DC通过交叉提呈的方式提呈PPV-VLPs-E290。晚期内体的酸性环境以及蛋白酶的水解均参与了PPV-VLPs-E290的提呈过程。
The purpose of this study was to determine whether PPV-VLPs could be expressed after capture by porcine splenic dendritic cells (DCs). CD172a + CD11R + DC and CD4-CD8 + T cells were first isolated from the spleens of non-immunized and immunized pigs by magnetic screening. DC with primaquine, cycloheximide, chloroquine, lactacystin, brefeldin A and aprotinin, pepstatin and other effects 1h, then with parvovirus-like particles PPV-VLPs-E290 At 37 ℃ for 4h, CD8 + T cell cytotoxicity assay was used to detect the presence of PPV-VLPs-E290. Lactate dehydrogenase (LDH) release test results showed that primaquine and aprotinin on PPD-PPV-VLPs-E290 did not have a significant effect on the efficiency of pepstatin partially inhibited, and chloroquine, cycloheximide , Brefeldin A and lactacystin significantly reduced the efficiency of DC presentation of PPV-VLPs-E290. The results showed that DC presented PPV-VLPs-E290 by cross-presentation. Late endosome acidic environment and proteolytic hydrolysis are involved in PPV-VLPs-E290 presentation process.