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In vitro lymphocyte response from vaccinated dogs to canine distemper virus vaccine.

机译：接种犬对犬瘟热病毒疫苗的体外淋巴细胞反应。

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摘要

This thesis is an investigation of the response of lymphocytes from vaccinated dogs to in vitro exposure to canine distemper virus (CDV) vaccine compared to mitogens. The objectives of this study were to develop an in vitro method of measuring lymphocyte responses to CDV antigen in vaccinated dogs, and to determine which subset(s) of lymphocytes respond to CDV antigens in comparison to mitogens.; Blood samples were collected from 15 healthy dogs, previously vaccinated for CDV. Peripheral blood mononuclear cells were separated by density gradient centrifugation and lymphocytes were cultured for 72 hours with CDV modified-live virus vaccine (Onderstepoort strain) 1:10 concentration; concanavalin-A (Con A) 7.5 mug/ml; pokeweed mitogen (PWM) 7.5 mug/ml; or without antigen or mitogen (unstimulated control). Following the culture period, lymphocyte proliferation was measured by trypan blue exclusion and tritiated thymidine incorporation. Flow cytometry was used to quantitate lymphocyte subsets following incubation with monoclonal antibodies to canine CD4, CD8, and CD21-like molecules. Finally, CDV antibody titers of the dogs were determined by virus neutralization.; Significant lymphocyte proliferation (P = 0.001) was found following incubation with CDV antigen, Con A, or PWM compared to unstimulated controls. When lymphocyte subsets were examined by flow cytometry, no significant difference in the proportion of cells labeled with CD4 (P = 0.6772), CD8 (P = 0.829), or CD-21-like (P = 0.112) antibodies were found between CDV antigen exposed lymphocytes and unstimulated controls. There were statistically significant differences in subgroups of lymphocytes stimulated with Con A or PWM (P 0.05). No correlation was found (R 0.70) between the CDV titers of the dogs compared to the lymphocyte proliferation stimulated by CDV vaccine.; These findings suggest CDV vaccine is able to stimulate canine lymphocytes in vitro to proliferate, but does not produce a detectable change in the proportion of lymphocyte subsets as measured by flow cytometry with the monoclonal antibodies used in this study. It is possible that with more specific monoclonal antibodies to detect memory T and B cells or a more sensitive assay, a measurable difference in the proportion of cells responding to CDV antigen stimulation could be identified.
机译：这篇论文是对有疫苗的狗的淋巴细胞对体外暴露于犬瘟热病毒（CDV）疫苗与有丝分裂原的反应的研究。这项研究的目的是开发一种体外方法，测量接种疫苗的狗对CDV抗原的淋巴细胞反应，并确定与有丝分裂原相比，哪些亚群的淋巴细胞对CDV抗原有反应。从15只健康的狗中收集血液样本，这些狗先前已经接种了CDV疫苗。通过密度梯度离心分离外周血单个核细胞，并用浓度为1:10的CDV改良活病毒疫苗（Onderstepoort株）培养淋巴细胞72小时。刀豆球蛋白A（Con A）7.5杯/毫升;商陆有丝分裂原（PWM）7.5杯/毫升;或没有抗原或促分裂原（无刺激的对照）。培养期后，通过锥虫蓝排除法和tri化胸苷掺入法测量淋巴细胞的增殖。与犬CD4，CD8和CD21样分子的单克隆抗体孵育后，使用流式细胞仪定量淋巴细胞亚群。最后，通过病毒中和确定狗的CDV抗体滴度。与未刺激的对照相比，与CDV抗原，Con A或PWM孵育后，发现淋巴细胞显着增殖（P = 0.001）。当通过流式细胞术检查淋巴细胞亚群时，在CDV抗原之间发现CD4（P = 0.6772），CD8（P = 0.829）或CD-21样（P = 0.112）抗体的细胞比例没有显着差异。暴露的淋巴细胞和未刺激的对照。 Con A或PWM刺激的淋巴细胞亚组在统计学上有显着差异（P <0.05）。与CDV疫苗刺激的淋巴细胞增殖相比，在犬的CDV滴度之间没有发现相关性（R <0.70）。这些发现表明CDV疫苗能够在体外刺激犬淋巴细胞增殖，但如本研究中使用的单克隆抗体通过流式细胞术所测量的那样，不会产生可检测的淋巴细胞亚群比例的变化。使用更特异性的单克隆抗体来检测记忆T和B细胞或进行更灵敏的测定，可以确定对CDV抗原刺激有反应的细胞比例存在可测量的差异。

著录项

  • 作者

    Barry, Maureen A.;

  • 作者单位

    University of Guelph (Canada).;

  • 授予单位 University of Guelph (Canada).;
  • 学科 Biology Veterinary Science.
  • 学位 D.V.Sc.
  • 年度 2004
  • 页码 86 p.
  • 总页数 86
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 动物学;
  • 关键词

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