禽网状内皮组织增生病病毒感染对SPF雏鸡血液和局部淋巴组织CD4+/CD8+细胞及相关细胞因子表达的影响

doi:10.11843/j.issn.0366-6964.2020.028

摘要/Abstract

摘要： 旨在探讨禽网状内皮组织增生病病毒（reticuloendotheliosis virus，REV）对SPF雏鸡血液和局部淋巴组织中T淋巴细胞数量以及相关细胞因子表达的影响。将96只1日龄SPF雏鸡随机分为REV感染组和对照组，应用流式细胞术、酸性α-醋酸萘酯酶染色（acid α-naphthyl acetate esterase，ANAE）、荧光定量PCR等方法对上述指标进行检测。试验数据表明，与对照组相比，感染组雏鸡血液CD4⁺T淋巴细胞数量在第7~35天显著降低、CD8⁺T淋巴细胞数量在第7~28天显著升高，CD4⁺/CD8⁺比值在第7~28天显著降低（均P<0.05或P<0.01）；局部淋巴组织哈德尔腺（Hader’s gland，HG）、派伊尔结（Peyer’s patch，PP）和盲肠扁桃体（caecal tonsil，CT）中ANAE⁺T淋巴细胞数量均显著降低（均P<0.05或P<0.01）；GH、PP和CT中细胞因子IL-2、IFN-γ和TNF-α 转录量都有不同程度升高。本研究表明，REV感染引起雏鸡血液中CD4⁺ T淋巴细胞数量降低、CD4⁺/CD8⁺T淋巴细胞比例失衡、局部淋巴组织中T淋巴细胞数量相对减少及细胞因子TNF-α转录持续升高与REV造成感染雏鸡细胞免疫功能显著降低密切相关。

关键词: 禽网状内皮组织增生病病毒, 局部淋巴组织, 细胞免疫

Abstract: The study aimed to investigate the effects of avian reticuloendotheliosis virus (REV) on the number of T lymphocytes in blood and the expression of related cytokines in local lymphoid tissues of SPF chicks. Ninety-six 1-day-old SPF chicks were randomly divided into REV-infected group and control group. The above indexes were detected by Flow Cytometry, acid α-naphthyl acetate esterase (ANAE) and Quantitative Real-time PCR. The results showed that compared with the control group, the number of CD4⁺ T lymphocytes in the infected group was significantly decreased at day 7-35, and the number of CD8⁺ T lymphocytes was significantly increased at day 7-28, the CD4⁺/CD8⁺ ratio was significantly decreased at day 7-28 (all P<0.05 or P<0.01); the number of ANAE⁺ T lymphocytes in the local lymphoid tissues of Hader's gland (HG), Peyer's patch (PP) and caecal tonsil (CT) were significantly decreased (all P<0.05 or P<0.01); the transcription levels of cytokines IL-2, IFN-γ and TNF-α in HG, PP and CT were increased in various degrees. These data indicated that REV infection caused a decrease in the number of CD4⁺ T lymphocytes in the peripheral blood of chickens, an imbalance of the ratio of CD4⁺/CD8⁺ T lymphocytes, a relative decrease in the number of T lymphocytes in local lymphoid tissues, and a continuous increase in the expression of cytokines TNF-α are closely related to the significantly reduced cellular immune function of infected chickens caused by REV.

Key words: reticuloendotheliosis virus, local lymphoid tissue, cellular immunity

中图分类号:

S858.315.3

刘丹华, 郑世民, 刘晓静, 吕晓萍, 高雪丽, 刘超男. 禽网状内皮组织增生病病毒感染对SPF雏鸡血液和局部淋巴组织CD4⁺/CD8⁺细胞及相关细胞因子表达的影响[J]. 畜牧兽医学报, 2020, 51(6): 1447-1454.

LIU Danhua, ZHENG Shimin, LIU Xiaojing, Lü Xiaoping, GAO Xueli, LIU Chaonan. Effects of Avian Reticuloendotheliosis Virus Infection on the CD4⁺/CD8⁺ Ratio and the Expression of Related Cytokines in SPF Chicks[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1447-1454.

参考文献 30

[1]	GAO S, JIANG H, SUN J, et al. Integrated analysis of miRNA and mRNA expression profiles in spleen of specific pathogen-free chicken infected with avian reticuloendotheliosis virus strain SNV[J]. Int J Mol Sci, 2019, 20(5):1041.
[2]	WOŹNIAKOWSKI G, FRANT M, MAMCZUR A. Avian reticuloendotheliosis in chickens-an update on disease occurrence and clinical course[J]. J Vet Res, 2018, 62(3):257-260.
[3]	宏虎, 陈溥言, 蔡宝祥. 禽网状内皮组织增殖病病毒的分离鉴定[J]. 中国畜禽传染病, 1988(2):1-3.HONG H, CHEN P Y, CAI B X. Isolation and identification of avian reticuloendotheliosis virus[J]. Chinese Journal of Preventive Veterinary Medicine, 1988(2):1-3. (in Chinese)
[4]	SUN G R, ZHANG Y P, ZHOU L Y, et al. Co-infection with Marek's disease virus and reticuloendotheliosis virus increases illness severity and reduces Marek's disease vaccine efficacy[J]. Viruses, 2017, 9(6):158.
[5]	FU L S, WANG X Y, ZHAI J, et al. Changes in apoptosis, proliferation and T lymphocyte subtype on thymic cells of SPF chickens infected with reticuloendotheliosis virus[J]. Mol Immunol, 2019, 111:87-94.
[6]	LÓPEZ-YGLESIAS A H, BURGER E, ARAUJO A, et al. T-bet-independent Th1 response induces intestinal immunopathology during Toxoplasma gondii infection[J]. Mucosal Immunol, 2018, 11(3):921-931.
[7]	HWANG E S, SZABO S J, SCHWARTZBERG P L, et al. T helper cell fate specified by kinase-mediated interaction of T-bet with GATA-3[J]. Science, 2005, 307(5708):430-433.
[8]	BI Y L, XU L, QIU L L, et al. Reticuloendotheliosis virus inhibits the immune response acting on lymphocytes from peripheral blood of chicken[J]. Front Physiol, 2018, 9:4.
[9]	MARTENS E C, NEUMANN M, DESAI M S. Interactions of commensal and pathogenic microorganisms with the intestinal mucosal barrier[J]. Nat Rev Microbiol, 2018, 16(8):457-470.
[10]	NISHIMORI A, KONNAI S, OKAGAWA T, et al. In vitro and in vivo antivirus activity of an anti-programmed death-ligand 1(PD-L1) rat-bovine chimeric antibody against bovine leukemia virus infection[J]. PLoS One, 2017, 12(4):e0174916.
[11]	刘文超. REV感染对雏鸡血液主要类型细胞及CyclinD1、p27 mRNA表达的影响[D]. 哈尔滨:东北农业大学, 2016.LIU W C. The influence of main types cells and CyclinD1, p27 mRNA expression in blood of chicks infected with REV[D]. Harbin:Northeast Agricultural University, 2016. (in Chinese)
[12]	RAD F R, AJDARY S, OMRANIPOUR R, et al. Comparative analysis of CD4+ and CD8+ T cells in tumor tissues, lymph nodes and the peripheral blood from patients with breast cancer[J]. Iran Biomed J, 2015, 19(1):35-44.
[13]	HE H Q, GENOVESE K J, KOGUT M H. Modulation of chicken macrophage effector function by TH1/TH2 cytokines[J]. Cytokine, 2011, 53(3):363-369.
[14]	CARPENTER G W, BREEDEN L, CARR D J J. Acute exposure to morphine suppresses cytotoxic T-lymphocyte activity[J]. Int J Immunopharmacol, 1995, 17(12):1001-1006.
[15]	OSTROUMOV D, FEKETE-DRIMUSZ N, SABOROWSKI M, et al. CD4 and CD8 T lymphocyte interplay in controlling tumor growth[J]. Cell Mol Life Sci, 2018, 75(4):689-713.
[16]	付礼胜, 侯宁, 王晓燕, 等. SPF雏鸡感染REV后中枢免疫器官细胞凋亡及相关因子的变化[J]. 畜牧兽医学报, 2017, 48(10):1983-1989.FU L S, HOU N, WANG X Y, et al. Changes of cell apoptosis and relevant factor in central immune organs of SPF chicks infected with reticuloendotheliosis virus[J]. Acta Veterinaria et Zootechnica Sinica, 2017, 48(10):1983-1989. (in Chinese)
[17]	WANG G H, WANG Z Z, ZHUANG P P, et al. Exosomes carring gag/env of ALV-J possess negative effect on immunocytes[J]. Microb Pathog, 2017, 112:142-147.
[18]	SANCHEZ-GUZMAN D, LE GUEN P, VILLERET B, et al. Silver nanoparticle-adjuvanted vaccine protects against lethal influenza infection through inducing BALT and IgA-mediated mucosal immunity[J]. Biomaterials, 2019, 217:119308.
[19]	AHLUWALIA B, MAGNUSSON M K, ÖHMAN L. Mucosal immune system of the gastrointestinal tract:maintaining balance between the good and the bad[J]. Scand J Gastroenterol, 2017, 52(11):1185-1193.
[20]	SHACKLETT B L. Mucosal immunity in HIV/SIV infection:T cells, B cells and beyond[J]. Curr Immunol Rev, 2019, 15(1):63-75.
[21]	WANG T, YIN H Q, LI Y, et al. Vaccination with recombinant adenovirus expressing multi-stage antigens of Toxoplasma gondii by the mucosal route induces higher systemic cellular and local mucosal immune responses than with other vaccination routes[J]. Parasite, 2017, 24:12.
[22]	QIN Y, XU Z C, WANG Y Q, et al. VP2 of Infectious bursal disease virus induces apoptosis via triggering oral cancer overexpressed 1(ORAOV1) protein degradation[J]. Front Microbiol, 2017, 8:1351.
[23]	LAI G H, LIEN Y Y, LIN M K, et al. VP2 of chicken anaemia virus interacts with apoptin for down-regulation of apoptosis through de-phosphorylated threonine 108 on apoptin[J]. Sci Rep, 2017, 7(1):14799.
[24]	WAN X R, YANG R X, LIU G L, et al. Downregulation of cellular prion protein inhibited the proliferation and invasion and induced apoptosis of Marek's disease virus-transformed avian T cells[J]. J Vet Sci, 2016, 17(2):171-178.
[25]	XU H T, YAO Y X, SMITH L P, et al. microRNA-26a-mediated regulation of interleukin-2 expression in transformed avian lymphocyte lines[J]. Cancer Cell Int, 2010, 10:15.
[26]	RAFTERY N, STEVENSON N J. Advances in anti-viral immune defence:revealing the importance of the IFN JAK/STAT pathway[J]. Cell Mol Life Sci, 2017, 74(14):2525-2535.
[27]	ZHENG Y S, CUI Z Z, ZHAO P, et al. Effects of reticuloendotheliosis virus and Marek's disease virus infection and co-infection on IFN-gamma production in SPF chickens[J]. J Vet Med Sci, 2007, 69(2):213-216.
[28]	NI H M, MCGILL M R, CHAO X J, et al. Caspase inhibition prevents tumor necrosis factor-α-induced apoptosis and promotes necrotic cell death in mouse hepatocytes in vivo and in vitro[J]. Am J Pathol, 2016, 186(10):2623-2636.
[29]	CHOI C, HWANG K K, CHAE C. Classical swine fever virus induces tumor necrosis factor-α and lymphocyte apoptosis[J]. Arch Virol, 2004, 149(5):875-889.
[30]	GAO S, WANG Z Z, JIANG H, et al. Transcriptional analysis of host responses related to immunity in chicken spleen tissues infected with reticuloendotheliosis virus strain SNV[J]. Infect Genet Evol, 2019, 74:103932.