影响猪树突状细胞对猪细小病毒样颗粒提呈方式的因素

doi:10.11843/j.issn.0366-6964.2017.03.014

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (3): 508-514.doi: 10.11843/j.issn.0366-6964.2017.03.014

影响猪树突状细胞对猪细小病毒样颗粒提呈方式的因素

罗尚星¹，申小强¹，顾文源^1,3，范京惠^1*，邸晶美¹，刘宝京¹，代飞¹，孔园园¹，常嫣嫣¹，左玉柱^1,2*

（1.河北农业大学动物医学院，保定071001；2.河北农业大学动物科技学院，保定071001；3.河北省动物疫病预防控制中心，石家庄 050035）

收稿日期:2016-10-09 出版日期:2017-03-23 发布日期:2017-03-22
通讯作者: 左玉柱，副教授，主要从事动物传染病学的教学、科研与社会服务工作，E-mail：zuoyuzhu@163.com；范京惠，博士，副教授，主要从事兽医微生物与免疫学的教学与科研工作，E-mail：jinghui76@163.com
作者简介:罗尚星(1993-)，女，河北张家口人，硕士生，主要从事动物传染病学研究，E-mail：2270989476@qq.com
基金资助:
国家自然科学基金（31101847）；河北农业大学百名青年学术带头人培养计划（0318011）

Studies on Presentation of Recombinant Parvovirus-like Particles by Porcine Splenic Dendritic Cells

LUO Shang-xing¹, SHEN Xiao-qiang¹, GU Wen-yuan^{1, 3}, FAN Jing-hui^1* , DI Jing-mei¹, LIU Bao-jing¹, DAI Fei¹, KONG Yuan-yuan¹, CHANG Yan-yan¹, ZUO Yu-zhu^1,2*

(1. College of Veterinary Medicine, Agricultural University of Hebei, Baoding 071001, China; 2. College of Animal Science and Technology, Agricultural University of Hebei, Baoding 071001, China; 3.Animal Disease Control and Prevention Center of Hebei, Shijiazhuang 050035, China)

Received:2016-10-09 Online:2017-03-23 Published:2017-03-22

摘要/Abstract

摘要：

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1 h后，再与细小病毒样颗粒PPV-VLPs-E290在37 ℃作用4 h，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。

关键词: 病毒样颗粒, 树突状细胞, 提呈方式

Abstract:

In order to decipher the mechanism of PPV-VLP presentation by dendritic cells (DC_s), CD172a⁺ CD11R⁺ DC and CD4^- CD8⁺ T cell were isolated from spleen of unvaccinated and CSFV vaccinated pigs, respectively, using magnetic beads. Purified splenic DCs were preincubated for 1 h with primaquin, cycloheximide, chloroquine, lactacystin, brefeldin A, leupeptin, or pepstatin. Then PPV-VLPs-E290 was added, and the DCs were incubated for 4 h in the continued presence of these different drugs. The presentation assay of PPV-VLPs-E290 was monitored by cytotoxic activity of CD8⁺ T cells, which was measured using the Lactate dehydrogenase (LDH) release assay. The results showed that incubation of DCs with PPV-VLPs-E290 in the presence of primaquin and leupetin did not block PPV-VLPs-E290 presentation. However, the present efficiency of DCs pretreated with chloroquin, cycloheximide, brefeldin A and lactacystin was severely reduced. These results showed that DCs can cross-present PPV-VLPs-E290. Both acidification of late endosomes and protease hydrolysis are necessary for PPV-VLPs-E290 processing.

Key words: parvovirus-like particles, dendritic cells, presentation pathways

中图分类号:

S852.659.2

罗尚星，申小强，顾文源，范京惠，邸晶美，刘宝京，代飞，孔园园，常嫣嫣，左玉柱. 影响猪树突状细胞对猪细小病毒样颗粒提呈方式的因素[J]. 畜牧兽医学报, 2017, 48(3): 508-514.

LUO Shang-xing, SHEN Xiao-qiang, GU Wen-yuan, FAN Jing-hui, DI Jing-mei, LIU Bao-jing, DAI Fei, KONG Yuan-yuan, CHANG Yan-yan, ZUO Yu-zhu. Studies on Presentation of Recombinant Parvovirus-like Particles by Porcine Splenic Dendritic Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(3): 508-514.

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影响猪树突状细胞对猪细小病毒样颗粒提呈方式的因素

Studies on Presentation of Recombinant Parvovirus-like Particles by Porcine Splenic Dendritic Cells

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