米糠多糖对免疫抑制鸡外周血CD4~+和CD8~+ T淋巴细胞亚群的调节作用

为探讨米糠多糖对免疫抑制鸡免疫功能的调节作用,采用流式细胞技术检测了环磷酰胺和传染性法氏囊病毒诱导免疫抑制鸡在服用米糠多糖(剂量为150 mg/kg)和不服用米糠多糖情况下,外周血中CD4+和CD8+T淋巴细胞亚群数量的动态变化。结果显示,米糠多糖能够抑制环磷酰胺和传染性法氏囊病毒处理鸡外周血中CD4+和CD8+T淋巴细胞数量的降低;提高健康雏鸡外周血中CD4+T淋巴细胞比率,对CD8+T淋巴细胞作用不明显。     

PD-L1+ HeLa细胞对人外周血活化T细胞的免疫调节作用

目的 探讨人宫颈癌细胞表达PD-L1对人外周血T细胞的免疫调节作用.方法 采用PD-L1质粒转染人宫颈癌HeLa细胞株,将PD-L1+ HeLa细胞与PHA刺激活化的人外周血T淋巴细胞混合培养,MTT法检测T细胞的增殖水平,流式细胞术分析T细胞凋亡率和CD8+/CD4+T细胞比例;ELISA检测PD-L1+ HeLa细胞与PHA刺激活化的人外周血T淋巴细胞混合培养上清液IFN-γ的含量.结果 MTT检测显示与PD-L1+ HeLa混合培养组的T细胞增殖光密度值为(0.31±0.02),明显低于与PD-L1 - HeLa混合培养组的T细胞和对照组T细胞,分别为[(0.62±0.02)和(0.59±0.03),P＜0.01];T细胞凋亡率明显高于与PD-L1 - HeLa混合培养组和对照组T细胞,分别为(32.7%、17.9%和18.3%);CD8 +/CD4+T细胞比值稍低于PD-L1 - HeLa混合培养组和对照组T细胞(分别为0.86、0.91和0.89);与PD-L1+ HeLa混合的T细胞培养上清液IFN-γ的含量低于与PD-L1 - HeLa混合培养组和对照组T细胞,分别为[(801.1±20.5) pg/ml、(1 006.2±30.2 )pg/ml和(1 070.2±113.3) pg/ml,P＜0.01].结论 PD-L1明显抑制人外周血活化T细胞增殖促进其凋亡并下调其免疫功能.     

捻转血矛线虫谷胱甘肽过氧化物酶体外对山羊外周血单核细胞功能的影响

本文探讨了捻转血矛线虫的谷胱甘肽过氧化物酶(HC29)体外对山羊外周血单核细胞(PBMCs)功能的影响。颈静脉采取无捻转血矛线虫感染的山羊血液,分离单核细胞后加入终浓度为5μg/mL的HC29重组蛋白进行体外培养,通过免疫荧光抗体技术观察重组蛋白与单核细胞的结合情况;HC29刺激山羊外周血单核细胞,用qPCR技术测定各试验组单核细胞中IL-2、IL-4、IL-10、IL-17、IFN-γ和TGF-βmRNA的转录水平;重组蛋白刺激单核细胞,采用CCK-8法测定细胞的增值情况;将重组蛋白与巨噬细胞共培养,测定巨噬细胞吞噬和分泌NO的情况。结果表明,重组HC29能够与外周血单核细胞结合并刺激细胞因子IL-10和IFN-γ的表达水平显著提高(P<0.01),能够引起单核细胞增值(P<0.01),巨噬细胞吞噬作用增强(P<0.01)和NO分泌增加(P<0.01)。结果表明HC29是捻转血矛线虫的一个重要抗原,主要诱导Th1类免疫反应。     

石菖蒲对LPS致流产小鼠的保胎作用及子宫免疫细胞的影响

为研究CD4+/CD8+T淋巴细胞和F4/80+巨噬细胞在流产发生机制中的意义,探讨中药石菖蒲的安胎作用机理,本试验选用细菌脂多糖(LPS)给小鼠尾静脉注射(0.1μg/只)制造流产模型,免疫组织化学方法检测小鼠子宫中CD4+,CD8+T淋巴细胞和F4/80+巨噬细胞的数量和分布情况.结果显示,应用LPS诱导流产后小鼠子宫中CD4+T淋巴细胞数量增多,CD8+T淋巴细胞数量无明显变化,CD4+/CD8+升高(P<0.01);F4/80+巨噬细胞数量也显著增多,与对照组比较差异极显著(P<0.01).预先口服保胎中药石菖蒲,能明显抑制LPS的作用,使小鼠流产率和胚胎吸收率降低,CD4+/CD8+比值降低,F4/80+巨噬细胞数量也降低.结果表明,LPS诱导小鼠流产与小鼠子宫中CD4+/CD8+比值和巨噬细胞数量有关,石菖蒲能调节小鼠子宫CD4+/CD8+T淋巴细胞和巨噬细胞数量,起到安胎作用.     

奶牛外周血树突状细胞体外诱导培养与鉴定

通过粒-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素-4(IL-4)体外诱导外周血单核细胞为树突状细胞,为利用树突状细胞免疫疗法治疗奶牛乳房炎奠定基础和提供细胞模型。利用淋巴细胞分离液分离获得奶牛外周血单核细胞,在6孔板内培养2 h后,弃掉含有大量的T细胞和B细胞上清液,贴壁的基本上是单核细胞,磷酸盐缓冲液清洗5次,加入含有GM-CSF和IL-4的2 m L培养基进行3 d诱导。之后,从培养基顶部小心吸弃1.4 m L的培养基,然后再补加含有GM-CSF和IL-4的1.8 m L培养基继续诱导3 d。每天通过显微镜观察细胞形态。第7天经流式检测细胞表面抗原CD11c、CD14、主要组织相容性复合体Ⅱ(MHCⅡ)、CD40、CD80、CD86的表达。结果表明:1)第2天,一些细胞表面可以生长出刺突并伴随着伪足的生长。第3天,细胞表面的刺突和伪足越来越多。第4、5天,一些带有刺突和伪足的细胞开始聚集和融合。第6天,单核细胞基本被诱导为树突状细胞,细胞表面含有大量清晰可见的刺突和伪足。2)经流式检测,CD14、CD11c、MHCⅡ阳性表达细胞分别占诱导细胞的6.8%、65.0%、75.9%,CD80和CD86阳性表达细胞分别占诱导细胞的2.0%和1.2%。综上所述,采用奶牛外周血单核细胞经体外诱导能够获得一定纯度的奶牛树突状细胞。     

传染性支气管炎病毒结构蛋白免疫鸡对CD4+CD8+T淋巴细胞亚群的影响

为了比较传染性支气管炎病毒3种主要结构蛋白免疫鸡后对CD4+、CD8+T淋巴细胞亚群的不同影响.本试验以pVAX1载体为携带工具,制备分别含有IBV主要结构蛋白基因的质粒免疫健康雏鸡,采用流式细胞仪(FACS)对免疫鸡外周血中CD4+、CD8+T淋巴细胞数进行检测.结果显示:各试验组间,携带N蛋白基因的质粒在免疫后3周内CD4+T淋巴细胞数和CD8+T淋巴细胞数均高于其他组,且差异极显著(P＜0.01).由此可见,IBV的N蛋白可明显诱导CD8+T淋巴细胞的CTL免疫作用和CD4+T淋巴细胞的辅助性免疫作用,其细胞免疫原性高于S1蛋白和M蛋白.     

外周血中CD4+、CD8+T、CD4+CD25+Treg在绵羊接种布鲁氏菌M5-90弱毒疫苗后的动态变化

为研究绵羊接种布鲁氏菌弱毒M5-90株后外周血中CD4+、CD8+T、CD4+CD25+Treg细胞的动态变化规律,本研究选择11只健康绵羊,每10 d免疫一次,共免疫3次,分别在免疫前、免疫后10d、20 d、30 d利用流式细胞术检测外周血中CD4+、CD8+T、CD4+CD25+Treg淋巴细胞亚群.在免疫后的第20 d,CD4+T、CD8+T细胞百分含量达到最高水平(P＜0.05)后均缓慢下降;在第10d,CD4+CD25+Treg细胞缓慢升高,至20 d、30 d均显著升高(p＜0.05);在布鲁氏菌M5-90疫苗免疫应答过程中CD4+CD25+Treg细胞参与了机体的免疫反应调控,对CD4+T、CD8+T淋巴细胞的比例进行调节,并且维持CD4+/CD8+比值稳定,起到平衡Th1/Th2细胞间反应的作用.     

黄芪多糖对PCV2阳性猪外周血免疫细胞及相关细胞因子的影响

检测外周血T淋巴细胞、嗜中性粒细胞、CD4+、CD8+细胞百分数及IL-2、IL-4和IFN-γ含量的变化,研究黄芪多糖(APS)对PCV2阳性猪免疫功能的调节作用。结果显示,APS能显著增加外周血T淋巴细胞、NBT阳性细胞、CD4+T细胞百分数及IL-2、IL-4、IFN-γ含量,显著降低CD8+T细胞百分数,推测APS通过上调CD4+细胞百分数、CD4+/CD8+及Th1反应恢复正常免疫机能。     

左旋咪唑对鸡离体外周血淋巴细胞的作用

本文报道了左旋咪唑对鸡离体外周血淋巴细胞（PBL）增殖及伴刀豆球蛋白A（ConA)诱导细胞（CIC）活性的作用；左旋咪唑以剂量依赖性方式促进鸡PBL的增殖；与ConA对促进鸡PBL增殖呈现协同作用；对脂多糖（LPS）激活鸡PBL具有明显促增殖作用；可逆转磷酸组胺和肾上腺素对ConA和LPS激活鸡PBL增殖的抑制作用；低浓度左旋咪唑不能逆转氢化可的松对ConA激活鸡PBL增殖的抑制作用，但高浓度（100-400μg/ml）可明显逆转氢化可的松的抑制作用；左旋咪唑可明显逆转氢化可的松的对LPS激活鸡PBL增殖的抑制作用；可促进CD4^ 细胞增殖，抑制CD8^ 细胞增殖，使CD4^ /CD8^ 细胞比值升高，CIC活性增强。     

AA肉鸡免疫禽流感油乳剂灭活疫苗细胞免疫应答的变化

为了解肉鸡生长期内免疫前后的细胞免疫反应,本实验采用CD3/CD4/CD8三色流式细胞术、MTT法和Griess法依次检测了AA肉鸡免疫禽流感灭活疫苗和非免疫状态下外周血、胸腺和脾脏中T淋巴细胞及其亚群的相对含量、外周血中T淋巴细胞对PHA-P的增殖反应能力和诱导巨噬细胞NO分泌量的变化。结果表明,非免疫状态下,AA肉鸡外周血CD4+、CD8+、CD3+T淋巴细胞相对百分含量在7日龄时均为最低,脾脏中CD4+T淋巴细胞相对百分含量在整个试验期一直呈下降趋势,免疫后外周血、脾脏和胸腺中CD4+、CD8+、CD3+T淋巴细胞相对百分含量均明显高于非免疫组,且脾脏CD4+T淋巴细胞呈阶段性上升状态;各时期淋巴细胞刺激指数(SI)值和巨噬细胞NO分泌量检测结果表明,免疫组均高于同期对照组(p<0.05)。研究结果还表明:肉鸡在7日龄时细胞免疫水平最低,灭活疫苗可以激发肉鸡产生较强的细胞免疫应答。     

Heartworm in dogs in Canada in 1984

In late December 1984, 1853 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1984 and 35% of them responded. Veterinarians reported that 97,794 dogs were blood-tested to check for microfilariae and 1417 dogs (1.45% of those tested) were found with heartworm. Another 34 dogs were amicrofilaremic, but were diagnosed as having heartworm disease, to give the total number diagnosed in 1984 as 1451 (1.48%). Heartworm was reported from all provinces except Prince Edward Island and Newfoundland but most (1310) of the cases were in Ontario. In Quebec, 126 cases were reported mostly from west of Montreal.

Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 27% of the cases were observed with clinical signs of heartworm disease and 72% had a history of not having left Canada. Southwestern Ontario continued to be the primary focus of the infection.

     

Heartworm in dogs in Canada in 1983

In late December 1983, 2 800 veterinarians across Canada were sent a questionnaire in order to assess the status of heartworm disease in Canada in 1983 and 26% of them responded. Veterinarians reported that 59 504 dogs were blood-tested to check for microfilariae and 771 dogs (1.30% of those tested) were found with Dirofilaria immitis. Heartworm disease was diagnosed in all provinces except New Brunswick and Newfoundland but most (733) of the cases were in Ontario.

Heartworm disease was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 31% of the cases were observed with clinical signs of heartworm disease and 64% had a history of not having left Canada. Southwestern Ontario continues to be the focus of the infection and most of the dogs there had not left the province previously.

     

Heartworm in dogs in Canada in 1991

In late November 1991, 1883 clinics in Canada were sent a questionnaire to assess the status of Dirofilaria immitis in dogs in 1991 and there was a 60.0% response. There were 344,031 dogs tested for heart-worm (HW), 627 were found infected and the prevalence of HW infection was 0.18%. There were 417 dogs with HW in Ontario, 116 in Manitoba, 38 in Quebec, 53 in British Columbia, three in Alberta, and one in Nova Scotia. In British Columbia, all of the infected dogs but one were from the Okanagan valley which, as from 1991, is a new focus of infection in Canada. Most dogs with HW had not been on preventive medication in 1990, and the prevalence among dogs tested and unprotected was 0.59%. That prevalence was considerably higher in endemic areas. Companion dogs, over three years of age and maintained primarily outdoors in rural areas, were most frequently infected. One cat was diagnosed with D. immitis and 33 dogs had Dipetalonema reconditium.     

Heartworm in dogs in Canada in 1985

In late December 1985, 1485 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1985 and 44% of them responded. Veterinarians reported that 137,300 dogs were blood-tested to check for microfilariae and 1210 dogs were found with heartworm. Another 36 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number diagnosed in 1985 as 1247 (0.91%).

Heartworm was reported from all provinces except Prince Edward Island, Newfoundland and Saskatchewan but most (1126) of the cases were in Ontario. Southwestern Ontario continued to be the primary focus of the infection in Canada. From Quebec, 91 cases were reported mostly from and around Montreal. From Manitoba, 19 cases were reported from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 28% of the cases were observed with clinical signs of heartworm disease and 78% had a history of not having left Canada.

     

Heartworm in dogs in Canada in 1988

In late November 1988, 1581 small and mixed animal clinics and institutional veterinarians across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1988, and 46% of them responded. Veterinarians reported that 181,577 dogs were blood-tested for heartworm disease and 367 dogs were found with D. immitis microfilariae. Another 60 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number of cases diagnosed in 1988 as 441 (0.24%).     

Heartworm in dogs in Canada in 1989

In late November 1989, 1732 clinics and institutional veterinarians were sent a questionnaire to assess the status of Dirofilaria immitis, and 51.7% responded. Of 247,716 dogs tested, 394 had D. immitis microfilariae and 51 were amicrofilaremic for a total of 445 cases and heartworm prevalence of 0.17%. Most (408) of these dogs had no preventive medication and the prevalence among dogs tested and unprotected was 1.01%. That prevalence was considerably higher in endemic areas. Thirty-seven dogs with heartworm had preventive medication. Heartworm was most frequent in companion dogs over three years of age maintained outdoors in rural areas. About 75% of the cases had never left Canada, 26% had clinical signs and 125 were not treated.

Heartworm was reported from British Columbia, Manitoba, Ontario, Quebec, Nova Scotia and Newfoundland, but 383 cases were in Ontario. South-western Ontario was the primary focus of infection. There were 33 cases in Quebec and 24 in Manitoba, mainly found in and around Metropolitan Montreal and Winnipeg respectively.

     

Heartworm in dogs in Canada in 1987

In late November 1987, 1246 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1987, and 50% of them responded. Veterinarians reported that 165,428 dogs were blood tested for heartworm disease and 511 dogs were found with D. immitis microfilariae. Another 78 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number of cases diagnosed in 1987 as 589 (0.35%).     

Heartworm in dogs in Canada in 1986

In late December 1986, 1224 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1986; 46% of them responded. Veterinarians reported that 150,989 dogs were blood-tested for microfilariae and 869 dogs were found with heartworm. Another 65 dogs were amicrofilaremic but diagnosed with heartworm disease and one was found with heartworm at necropsy to give the total number diagnosed in 1986 as 935 (0.62%).

Heartworm was reported from Manitoba, New Brunswick, Ontario and Quebec, but most (810) of the cases were from Ontario. South-western Ontario continued to be the primary focus of the infection in Canada. There were 103 cases reported from Quebec, mostly from and around Montreal, and 21 cases from Manitoba, from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 33% of the cases were observed with clinical signs of heartworm disease and 81% had a history of not having left Canada.