黄牛呼吸道CD3~+淋巴细胞分布的研究     

李乙江  蚌德新  郭兆廷  余绍荣  丁宏  闫信权  张书光  杨科  郭兆卫  罗祥波  李佳赟  杨倩 《中国畜牧兽医》2018,(5)

CD3~+是T细胞群的重要表面标志,呼吸道黏膜下分布的CD3~+淋巴细胞作为抗感染黏膜免疫的基础,在保护机体抵抗呼吸道感染中发挥重要作用。为了解黄牛呼吸道CD3~+淋巴细胞和淋巴组织的分布,本研究运用HE染色法和免疫组织学方法对5头7岁健康婆罗门黄牛的鼻黏膜、气管、肺内支气管及肺脏组织的CD3~+淋巴细胞和淋巴组织的分布进行了研究。结果显示,黄牛的鼻黏膜、气管、肺内支气管和肺脏组织中均分布有CD3~+淋巴细胞,且主要分布于黏膜上皮间及其下方固有层中及腺体周围;在鼻黏膜和肺脏组织中分布有CD3~+淋巴细胞形成的弥散淋巴组织。牛呼吸道中CD3~+淋巴细胞数量在鼻黏膜和肺脏组织中分布最多,气管黏膜、肺内支气管黏膜次之。本试验结果明确了CD3~+淋巴细胞在黄牛呼吸道中的分布谱,表明黄牛呼吸道具备引发局部黏膜免疫的基础条件,为牛呼吸道黏膜免疫及呼吸道疾病防治研究提供了数据支撑。  相似文献   

猪呼吸道淋巴组织发育的亚显微结构变化     

杨倩  庾庆华  李玉磊 《畜牧兽医学报》2006,37(4):374-378

应用组织学和电镜技术研究猪呼吸道发育过程中淋巴组织的变化。结果表明：扁桃体和咽部是呼吸道进入机体的第一个淋巴组织集中的部位，弥散淋巴组织在出生时就存在，淋巴小结不明显；20日龄时扁桃体中淋巴组织增生，淋巴小结清晰可见；120日龄淋巴小结数量增加，紧靠鳞状上皮密集排列，淋巴小结发育很好，并出现生发中心。扁桃体复层鳞状上皮中含有大量的上皮内淋巴细胞。气管叉是呼吸道进入机体的第二个淋巴组织集中的部位，出生时气管叉外膜中淋巴组织直接与气管支气管淋巴结相连，淋巴组织明显可见。20日龄时气管叉外膜中淋巴组织已分开，形成气管叉外膜密集的淋巴组织和气管支气管淋巴结两个部分。120日龄时气管叉处淋巴组织特别发达，黏膜上皮中上皮内淋巴细胞数量也显著增加。肺内气管和细支气管固有膜中均有较多的淋巴细胞，其中浆细胞数量增加，上皮中仍存在少量的上皮内淋巴细胞。本试验结果提示猪呼吸道是黏膜免疫较理想的诱导位点和效应位点，新生仔猪通过鼻腔免疫可提高呼吸道局部黏膜免疫力。  相似文献   

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   总被引：1，自引：1，他引：0  

刘丹华  郑世民  刘晓静  吕晓萍  高雪丽  刘超男 《畜牧兽医学报》2020,51(6):1447-1454

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。  相似文献   

板蓝根多糖对缺乳仔鼠免疫器官发育及T淋巴细胞亚群的影响     

张婷婷  陶孟杰  黄鹏  张学武  肖传斌 《中国畜牧兽医》2013,40(8):97-101

试验应用流式细胞术检测缺乳仔鼠CD3⁺、CD4⁺和CD8⁺ T淋巴细胞含量来研究添加不同剂量的板蓝根多糖(RIP)对缺乳仔鼠免疫器官及T淋巴细胞亚群的影响。结果表明:①RIP可增加缺乳仔鼠胸腺指数和脾脏指数。对胸腺指数和脾脏指数效果最好的RIP剂量随日龄增大而减小。②RIP可以增加缺乳仔鼠外周血CD3⁺、 CD4⁺ 、CD8⁺ T淋巴细胞数量。7~28日龄时初乳组(A组)、缺乳+中RIP组(C组)和缺乳+高RIP组(D组)3组CD3⁺ T淋巴细胞含量与缺乳组(B组)相比差异显著(P<0.05)。7~21日龄时初乳组(A组)、缺乳+中RIP组(C组)和缺乳+高RIP组(D组)3组CD4⁺ T淋巴细胞含量与缺乳组(B组)相比差异显著(P<0.05)。各处理组CD3⁺、 CD4⁺、CD8⁺ T淋巴细胞含量及CD4⁺/CD8⁺比值随着日龄的增加有所下降。适宜剂量的RIP可促进缺乳仔鼠免疫器官发育,提高T淋巴细胞亚群的水平。  相似文献   

免疫细胞在牦牛附睾和输精管的分布规律     

张倩  崔燕  余四九  何俊峰  潘阳阳  王萌 《畜牧兽医学报》2022,53(6):1925-1933

旨在研究免疫细胞在健康雄性牦牛附睾和输精管的分布。采用免疫组织化学和实时荧光定量(qRT-PCR)方法对幼龄(5~6月龄)及成年(3~4岁)牦牛附睾(头、体、尾)和输精管中CD68⁺巨噬细胞、CD3⁺ T淋巴细胞、CD79α⁺ B淋巴细胞、IgA⁺和IgG⁺浆细胞的分布特征及其表面标志分子的表达水平进行研究。结果显示：CD68⁺巨噬细胞、CD3⁺ T淋巴细胞、CD79α⁺ B淋巴细胞、IgA⁺和IgG⁺浆细胞主要分布在附睾管和输精管的上皮和间质;另外,CD68和CD3 mRNA和蛋白水平在各年龄组牦牛附睾头和附睾体显著高于附睾尾和输精管(P < 0.05),而CD79α、IgA和IgG mRNA和蛋白水平在附睾尾和输精管显著高于附睾头和附睾体(P < 0.05);此外,在成年牦牛附睾和输精管CD3、CD79α、IgA、IgG、CD68 mRNA和蛋白水平均显著高于幼龄牦牛(P < 0.05)。综上提示,牦牛附睾头可能主要是细胞免疫发生的位点,而附睾尾和输精管则主要进行体液免疫应答,此外,成年牦牛附睾和输精管的局部免疫可能更完善,以上数据为进一步研究高原牦牛局部生殖免疫和病理提供了形态学资料。  相似文献   

口蹄疫灭活病毒配合枯草芽孢杆菌对提高牛呼吸道先天免疫力的研究     

《中国预防兽医学报》2018,(10)

本研究采用O型口蹄疫灭活病毒配合枯草芽孢杆菌喷鼻气雾免疫健康牛,并设单独使用枯草芽孢杆菌组、灭活病毒、常规疫苗免疫组和空白对照组,评价O型口蹄疫灭活病毒配合免疫增强剂枯草芽孢杆菌通过喷鼻气雾免疫健康牛对其呼吸道先天免疫力的影响。通过免疫组织化学染色显示,免疫后灭活病毒配合免疫增强剂试验组牛鼻黏膜与肺内支气管黏膜中TLR7的表达和CD3~+淋巴细胞呈极显著增多(p0.01);而常规口蹄疫灭活苗对其TLR7的表达和CD3~+淋巴细胞数量的影响不显著(p0.05)。ELISA检测结果显示,与对照组相比,灭活病毒配合免疫增强剂试验组牛呼吸道组织中IL-12水平呈极显著增加(p0.01),IL-4和IL-10水平无明显差异(p0.05);而常规口蹄疫灭活苗对牛呼吸道组织中IL-4、IL-10和IL-12的影响不明显(p0.05)。表明O型口蹄疫灭活病毒配合枯草芽孢杆菌能够提高牛呼吸道先天免疫力。本研究为研发口蹄疫灭活病毒的鼻腔疫苗提供重要的试验数据和参考。  相似文献   

柔嫩艾美耳球虫对藏鸡和隐性白羽鸡致病性差异分析     

李建梅  刘梅  沈欣悦  戴亚斌  俞燕 《中国畜牧兽医》2019,46(10):3058-3064

为进一步明确柔嫩艾美耳球虫对球虫易感性差异鸡种的致病性,本研究以1×10⁵个柔嫩艾美耳球虫孢子化卵囊的剂量分别感染对球虫具有抗性的藏鸡和易感的隐性白羽鸡,接种后观察记录各组鸡的临床表征、血便记分、死亡率、增重、盲肠病变记分、卵囊产量,并于感染前和感染后3、6和8 d每个品种分别随机选择5只鸡采集抗凝血,应用流式细胞仪（FCAS）检测外周血CD4⁺T和CD8⁺T淋巴细胞亚群数量。结果显示,柔嫩艾美耳球虫感染后藏鸡相对增重率高于隐性白羽鸡,死亡率、血便记分和盲肠病变记分均低于隐性白羽鸡,但卵囊产量高于隐性白羽鸡。外周血T淋巴细胞亚群变化方面,感染前,藏鸡CD4⁺T淋巴细胞数及CD4⁺/CD8⁺比值均高于隐性白羽鸡。感染后第3天,藏鸡CD4⁺、CD8⁺ T淋巴细胞数及CD4⁺/CD8⁺比值下降,隐性白羽鸡CD8⁺ T淋巴细胞数略有下降,CD4⁺T淋巴细胞数及CD4⁺/CD8⁺比值上升,但CD4⁺/CD8⁺比值仍显著低于藏鸡（P<0.05）。感染后第6天,2个品种鸡CD4⁺ T淋巴细胞数及CD4⁺/CD8⁺比值均下降,其中藏鸡表现为显著下降（P<0.05）,而隐性白羽鸡仅CD4⁺/CD8⁺比值显著降低（P<0.05）,隐性白羽鸡CD8⁺ T淋巴细胞数显著升高（P<0.05）。感染后第8天,2个品种鸡CD4⁺/CD8⁺比值显著下降（P<0.05）,藏鸡CD8⁺ T淋巴细胞数显著高于隐性白羽鸡（P<0.05）,但CD4⁺/CD8⁺比值显著低于隐性白羽鸡（P<0.05）。结果表明,球虫对藏鸡和隐性白羽鸡的致病性存在差异,这种差异与T淋巴细胞介导的免疫应答反应密切相关。  相似文献   

不同日龄鸡喉黏膜淋巴组织中的T和B淋巴细胞分布检测     

杨树宝  张英楠 《中国动物检疫》2021,38(11):131-135

为了解鸡喉黏膜不同时期的免疫状态,采集不同日龄海兰白鸡的喉组织,利用免疫组织化学方法研究CD3+ T淋巴细胞和Bu-1+ B淋巴细胞的出现、定位分布及数量变化过程。结果显示：4日龄时,CD3+ T淋巴细胞和Bu-1+ B淋巴细胞都首次出现在喉黏膜中,而且都主要分布在喉腔底壁的正中黏膜嵴中;7日龄时喉黏膜中CD3+ T淋巴细胞和Bu-1+ B淋巴细胞急剧增多,并形成淋巴聚集物,CD3+细胞占据淋巴聚集物的中下部区域,而Bu-1+细胞主要位于淋巴聚集物外周;14日龄时Bu-1+细胞虽仍主要分布于CD3+细胞的外周,但分界不如7日龄时明显,有部分Bu-1+细胞穿插分布于CD3+细胞之间;21日龄和35日龄时,喉黏膜固有层中淋巴细胞更为集中;56日龄时,黏膜固有层底端出现主要由Bu-1+ B淋巴细胞构成的生发中心,而CD3+ T淋巴细胞则分布在生发中心周围的滤泡间区域。在数量变化上,随着日龄增长,CD3+ T淋巴细胞和Bu-1+ B淋巴细胞数量均逐渐增加,且14日龄之前CD3+细胞数量多于Bu-1+细胞,而此后各日龄Bu-1+细胞的数量均显著多于CD3+细胞（P＜0.05）。结果表明,鸡喉黏膜中T、B淋巴细胞的分布和数量均呈现日龄相关性变化,并且其变化可以反映出14日龄之前喉黏膜以细胞免疫为主,而之后则倾向于体液免疫。本试验为进一步研究家禽喉黏膜的免疫机制奠定了基础。  相似文献   

口蹄疫灭活病毒配合枯草芽胞杆菌鼻腔免疫增强牛呼吸道黏膜免疫应答     

李乙江  朱明旺  普仕华  郭云然  杨永幸  李世钰  创向辉  杨倩 《畜牧兽医学报》2018,(2)

口蹄疫病毒(foot-and-mouth disease virus,FMDV)主要通过呼吸道传播,切断呼吸道传播途径能有效的防御口蹄疫。本研究应用O型口蹄疫灭活病毒配合枯草芽胞杆菌通过喷鼻气雾免疫牛来探讨对牛呼吸道黏膜免疫应答及系统免疫应答的影响。结果显示,免疫后牛鼻黏膜与肺内支气管黏膜中的IgA分泌细胞的数量极显著增多(P0.01),而常规口蹄疫灭活苗对IgA分泌细胞数量的影响不大(P0.05);同时,ELISA结果显示鼻、气管、肺和肺内支气管中IL-12、TNF-α水平均极显著增加(P0.01),尤其是在气管和肺内支气管中最为明显,但IL-6无明显差异(P0.05)。从免疫第3天开始牛鼻液、唾液中口蹄疫特异性抗体SIgA以及血清中O型口蹄疫病毒特异性抗体均显著增加(P0.01或P0.05),且维持至3个月。本研究结果表明O型口蹄疫灭活病毒配合枯草芽胞杆菌喷鼻后可刺激牛呼吸道局部体液免疫和细胞免疫,同时还可诱导全身系统免疫应答,本研究为口蹄疫灭活病毒鼻腔免疫提供理论基础和应用前景。  相似文献   

不同日龄鸡肺脏中CD4~+、CD8~+ T淋巴细胞分布规律     

张英楠  徐晶  栾维民  杨树宝 《中国畜牧兽医》2019,(10)

为了解鸡肺脏不同时期的免疫状态,选择不同日龄海兰白鸡的肺脏组织,利用免疫组织化学方法研究CD4~+和CD8~+T淋巴细胞的出现、定位分布及数量变化过程。结果显示,CD8~+细胞最初于18 d胚胎中出现,而CD4~+T淋巴细胞于孵化后1日龄雏鸡中出现。4日龄时,在初级支气管与次级支气管交汇处有淋巴细胞的聚集物,即形成了明显的支气管相关性淋巴组织(BALT)。在定位分布上,各日龄CD4~+T淋巴细胞主要占据BALT的中央区域,而CD8~+T淋巴细胞主要环绕在外周。从56日龄开始,CD8~+T细胞较CD4~+T细胞分布范围更为广泛,在三级支气管气道内壁中、房间隔中、气体交换区和小叶间结缔组织中均有分布,散布在整个肺脏中。在数量变化上,随着日龄增长,CD4~+T淋巴细胞和CD8~+T淋巴细胞数量均逐渐增加,且35日龄之前CD4~+T细胞数量多于CD8~+T细胞,而此后各日龄CD8~+T细胞的数量均显著多于CD4~+细胞。以上结果表明,鸡肺脏中CD4~+和CD8~+ T淋巴细胞的分布和数量均呈日龄相关性变化,且该变化说明鸡35日龄之前肺脏以体液免疫为主,而之后则倾向于细胞免疫。  相似文献   

The Localization Changes of CD4+ and CD8+ T Lymphocytes in Chicken Lung at Different Ages     

ZHANG Yingnan  XU Jing  LUAN Weimin  YANG Shubao 《中国畜牧兽医》2019,46(10):3052-3057

The aim of this study was to investigate the immune state of chicken lung in different periods.With lung tissue of Hy-line White chicken at different ages,the distribution and quantity changes of CD4⁺ and CD8⁺T lymphocytes in lung were studied using immunohistochemistry staining.The results showed that CD8⁺T lymphocytes appeared firstly in embryonic at 18 d,while CD4⁺T lymphocytes appeared at 1-day-old chicken.At 4-day-old,there were aggregates of lymphocytes at the junction of the primary and secondary bronchi,which formed obvious broncho-associated lymphoid tissue (BALT).CD4⁺T lymphocytes of each age mainly occupied the central area of BALT,while CD8⁺T lymphocytes mainly surrounded the periphery.Since 56 days old,CD8⁺T lymphocytes are distributed in the inner wall of third-order bronchial airway,atrial septum,gas exchange area and interlobular connective tissue,and are distributed throughout the lung.In terms of quantity change,with the growth of daily age,the number of CD4⁺T lymphocytes and CD8⁺T lymphocytes gradually increased,and the number of CD4⁺ T lymphocytes was more than that of CD8⁺ T lymphocytes before 35 days of age,while the number of CD8⁺ T lymphocytes was significantly more than that of CD4⁺ T lymphocytes at the same age thereafter.The results showed that the distribution and number of CD4⁺ and CD8⁺T lymphocytes in the lungs of chickens were correlated with the age,and the changes could reflect that the lungs before the age of 35 days were dominated by humoral immunity,while the lungs after that tended to be cellular immunity.  相似文献   

Local immunity of the respiratory mucosal system in chickens and turkeys   总被引：1，自引：0，他引：1  

Smiałek M  Tykałowski B  Stenzel T  Koncicki A 《Polish journal of veterinary sciences》2011,14(2):291-297

This review article presents fundamental mechanisms of the local mucosal immunity in selected regions of the respiratory tract in healthy birds and in some pathological conditions. The respiratory system, whose mucosa come into direct contact with microorganisms contaminating inhaled air, has some associated structures, such as Harderian gland (HG), conjunctive-associated lymphoid tissue (CALT) and paranasal glands (PG), whose participation in local mechanisms of the mucosal immunity has been corroborated by numerous scientific studies. The nasal mucosa, with structured clusters of lymphoid tissue (NALT - nasal-associated lymphoid tissue) is the first to come into contact with microorganisms which contaminate inhaled air. Lymphoid nodules, made up of B cells with frequently developed germinal centres (GC), surrounded by a coat of CD4+ cells, are the major NALT structures in chickens, whereas CD8+ cells are situated in the epithelium and in the lamina propria of the nasal cavity mucosa. Studies into respiratory system infections (e.g. Mycoplasma gallisepticum) have shown the reactivity of the tracheal mucosa to infection, despite a lack of essential lymphoid tissue. Bronchus-associated lymphoid tissue (BALT) takes part in bronchial immune processes and its structure, topography and ability to perform defensive function in birds is largely age-dependent. Mature BALT is covered by a delicate layer of epithelial cells, called follicle-associated epithelium (FAE). Germinal centres (GC), surrounded by CD4+ cells are developed in most mature BALT nodules, while CD8+ lymphocytes are dispersed among lymphoid nodules and in the epithelium, and they are rarely present in GC. Macrophages make up the first line of defence mechanisms through which the host rapidly responds to microorganisms and their products in the respiratory mucosal system. Another very important element are polymorphonuclear cells, with heterophils being the most important of them. Phagocytic cells obtained from lung lavages in birds are referred to as FARM (free avian respiratory macrophages). Their number in chickens and turkeys is estimated to be 20 times lower than that in mice and rats, which indicates a deficit in the first-line of defence in the birds' respiratory system. There are numerous B cells and antibody secreting cells (ASC) present throughout the respiratory system in birds. Their role comes down to perform antigen-specific protection by producing antibodies (IgM, IgY or IgA class) as a result of contact with pathogenic factors.  相似文献   

玉屏风多糖对小鼠派氏结形态结构及其T细胞亚群的影响     

陈刚  周兆海  梁浩钊  杨鸿  卢玉葵  邓桦 《中国畜牧兽医》2019,46(1):313-319

通过研究玉屏风多糖（YPF-P）对小鼠派氏结（Peyer's patches,PPs）形态结构及其T细胞亚群的影响,探讨其对小鼠肠黏膜的免疫调节作用。选取96只SPF小鼠随机分为6组,分别为空白对照组（0.2 mL生理盐水）、YPF-P阳性对照组（YPF-P 200 mg/kg）、环磷酰胺（cyclophosvnamide,Cy）免疫抑制组（80 mg/kg Cy）及YPF-P低、中、高剂量组（100、200、400 mg/kg YPF-P）,饲喂1周,取小肠PPs,常规切片HE染色后应用图像分析技术检测PPs形态结构的变化;体外培养小鼠PPs淋巴细胞,采用流式细胞术研究PPs中T淋巴细胞亚群的变化。结果表明,YPF-P对小鼠小肠PPs的生长发育具有促进作用,Cy可极显著降低小鼠小肠PPs面积、小肠纵切面面积及PPs面积和小肠纵切面面积的比值（P < 0.01）,低、中、高剂量YPF-P可在一定程度上缓解Cy对小鼠小肠的损伤作用。同时,YPF-P可显著或极显著提高PPs中CD3⁺、CD4⁺ T淋巴细胞及CD4⁺/CD8⁺（P < 0.05; P < 0.01）。结果显示,YPF-P能提高Cy诱导的免疫抑制小鼠的肠黏膜免疫功能,并能促进PPs中相关T淋巴细胞增殖,对小鼠肠黏膜功能具有增强作用。  相似文献   

Perforin expression can define CD8 positive lymphocyte subsets in pigs allowing phenotypic and functional analysis of natural killer, cytotoxic T, natural killer T and MHC un-restricted cytotoxic T-cells     

Denyer MS  Wileman TE  Stirling CM  Zuber B  Takamatsu HH 《Veterinary immunology and immunopathology》2006,110(3-4):279-292

In this study we have used the expression of perforin to characterize subsets of porcine cytotoxic lymphocytes. Perforin positive lymphocytes expressed both CD2 and CD8, most were small dense lymphocytes (SDL) and up to 90% were CD3 negative. However, the numbers of perforin positive T-cells increased with the age of the animal and their populations increased after specific antigen stimulation in vitro. The remaining perforin positive lymphocytes were large and granular and contained more CD3⁺CD5⁺CD6⁺ T-cells (−40%) of which a substantial proportion also co-expressed CD4. Perforin was expressed in subpopulations of both CD8 and CD8β lymphocytes, but was not expressed in γδ T-cells or monocyte/macrophages. The perforin positive CD3⁻ subset was phenotypically homogeneous and defined as CD5⁻CD6⁻CD8β⁻CD16⁺CD11b⁺. This population had NK activity and expressed mRNA for the NK receptor NKG2D, and adaptors DAP10 and DAP12. Perforin positive T-cells (CD3⁺) could be divided into at least three subsets. The first subset was CD4⁻CD5⁺CD6⁺CD11b⁻CD16⁻ most were small dense lymphocytes with cytotoxic T-cell activity but not all expressed CD8β. The second subset was mainly observed in the large granular lymphocytes. Their phenotype was CD4⁺CD5⁺CD6⁺CD8β⁺CD16⁻CD11b⁻ and also showed functional CTL activity. Thus not all of double positive T-cells are memory helper T-cells. The third subset did not express the T-cell co-receptor CD6, but up to half of them expressed another T-cell co-receptor CD5. The majority of this subset expressed CD11b and CD16, thus the third perforin positive T-cell subset was CD3⁺CD4⁻CD5⁺CD6⁻CD8β^±CD11b⁺CD16⁺, and possessed MHC-unrestricted cytotoxicity and LAK activity.  相似文献   

鸡传染性支气管炎病毒S1蛋白抗原表位多肽的预测、鉴定及免疫效果的初步研究     

范文胜  朱丹  张愉  廖健淇  王露  雍璐  韦平  磨美兰 《畜牧兽医学报》2020,51(9):2257-2264

旨在对鸡传染性支气管炎病毒（IBV）H120株S1蛋白的B细胞及T细胞可能的表位进行预测并合成相应的多肽,然后免疫小鼠,并分析其免疫效果,以验证候选表位多肽的免疫原性。利用表位预测软件筛选并化学合成针对IBV H120株S1蛋白的5条表位多肽,然后免疫小鼠,通过间接ELISA、中和试验和流式细胞技术检测各表位多肽诱导的特异性抗体、中和抗体和外周血T细胞亚群。ELISA检测结果显示,5条表位多肽均具有良好的反应原性,免疫小鼠的血清效价高低顺序依次为Pep76-106 > Pep240-257 > Pep511-537 > Pep403-421 > Pep135-172;中和试验结果表明,5条多肽免疫小鼠的血清中和滴度均高于空白对照组,其高低顺序依次为Pep240-257=Pep403-421=Pep511-537>Pep76-106=Pep135-172;流式检测结果表明,5条多肽免疫小鼠在CD3⁺、CD4⁺CD8^-、CD8⁺CD4^- T淋巴细胞水平上均极显著高于空白对照组（P<0.01）,CD3⁺T及CD4⁺CD8^-T淋巴细胞数大小顺序依次为Pep403-421 > Pep240-257 > Pep76-106 > Pep511-537 > Pep135-172,CD8⁺CD4^-T淋巴细胞数大小顺序依次为Pep403-421 > Pep76-106 > Pep511-537 > Pep240-257 > Pep135-172。合成的5条表位多肽中,Pep240-257、Pep76-106和Pep403-421可以诱导体液免疫,Pep403-421可以诱导细胞免疫,其中,Pep403-421可以同时诱导细胞免疫及体液免疫。本研究结果为深入了解S1蛋白的免疫学特性以及研发诊断试剂和有效表位疫苗奠定了基础。  相似文献   

Prediction,Identification and Preliminary Study on Immune Efficacy of S1 Protein Epitope-polypeptides of Avian Infectious Bronchitis Virus     

FAN Wensheng  ZHU Dan  ZHANG Yu  LIAO Jianqi  WANG Lu  YONG Lu  WEI Ping  MO Meilan 《畜牧兽医学报》1956,51(9):2257-2264

For the sake of verifying the immunogenicity of candidate epitope-polypeptide, the B and T cell epitopes of S1 protein of avian infectious bronchitis virus (IBV) H120 strain were predicted and the corresponding epitope-polypeptides were synthesized, and then were used to immunize mice, the immune effect was analyzed. Five epitope-polypeptides against S1 protein of IBV H120 strain were selected by epitope prediction software and acquired by chemical synthesis, then were immunized to mice. The specific antibodies, neutralizing antibodies and T lymphocyte subsets induced by each epitope-polypeptides were analyzed by indirect ELISA, neutralization test and flow cytometry. The ELISA results showed that the five epitope-polypeptides had good reactivity. The antibody titers of antisera induced by the five epitope-polypeptides sorted from high to low as follows: Pep76-106, Pep240-257, Pep511-537, Pep403-421, Pep135-172. The neutralization test results showed that the neutralization titers of antisera induced by the five epitope-polypeptides groups in mice were higher than that of the blank control group, and the order of neutralization titers was Pep240-257 = Pep403-421 = Pep511-537 > Pep76-106 = Pep135-172. The flow cytometry results showed that the percentages of CD3⁺, CD4⁺CD8^- and CD8⁺CD4^- T lymphocytes in all the five epitope-polypeptides groups were significantly higher (P<0.01) than those in the blank control group. The number of the CD3⁺ and CD4⁺CD8^- T lymphocytes sorted from large to small as follows: Pep403-421, Pep240-257, Pep76-106, Pep511-537 and Pep135-172. The number of the CD8⁺CD4^- T lymphocytes sorted from large to small as follows: Pep403-421, Pep76-106, Pep511-537, Pep240-257 and Pep135-172. In conclusion, Pep240-257, Pep76-106 and Pep403-421 could induce humoral immunity among the five epitope-polypeptides, while Pep403-421 could induce cellular immunity. Thus, peptide of Pep403-421 could induce cellular immunity and humoral immunity. This study laid a foundation for further understanding the immunological characteristics of the S1 protein and the development of diagnostic reagents and effective epitope vaccines.  相似文献   

新型滴鼻灭活鹦鹉热衣原体疫苗的免疫效力评价     

陈思宇  隋卓君  王占新  李强  陈艺  王飞  陈爽  刘心怡  严专强  何诚 《中国畜牧兽医》2022,49(5):1951-1959

【目的】 探究由霍乱弧菌菌影(Vibrio cholerae ghosts,VCG)和纳米壳聚糖凝胶(Gel)组合制作的一种新型灭活衣原体疫苗复合佐剂是否可增强鹦鹉热衣原体原体(elementary body,EB)灭活抗原诱导动物机体产生的免疫应答。【方法】 将60只7日龄SPF鸡随机分为4个组,分别为:EB＋VCG＋Gel组、EB＋VCG 组、Gel组和EB组。通过滴鼻途径免疫鸡群,免疫2次,每次间隔14 d,免疫后检测鸡血清中IgG抗体水平、淋巴细胞增殖指数、CD4^＋/CD8^＋T 细胞比例、细胞因子(白介素-4(IL-4)、IL-10、IL-12和干扰素-γ(IFN-γ))含量及攻毒后鸡喉头排菌量和肺脏的病理损伤程度。【结果】 与Gel和EB组相比,EB＋VCG＋Gel和EB＋VCG组IgG 抗体水平、淋巴细胞增殖指数、IFN-γ含量和CD4^＋/CD8^＋T细胞比值显著或极显著升高(P<0.05;P<0.01)。此外,与Gel和EB组相比,攻毒后第12天,EB＋VCG＋Gel组喉头排菌量极显著降低(P<0.01),攻毒后第7天,肺脏损伤评分极显著降低(P<0.01)。【结论】 壳聚糖凝胶、VCG、灭活衣原体 EB 组合构成的新型滴鼻衣原体疫苗经鼻腔接种动物后可刺激动物机体产生良好的体液免疫和细胞免疫应答,阻断鹦鹉热衣原体经呼吸道途径传播,防止鹦鹉热衣原体从动物向人群传播。  相似文献