以免疫应答为基础的《医学免疫学》综合实验设计及实践

Design and Implementation of a Comprehensive Experiment in Medical Immunology Based on Immune Response

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中文摘要：以往的《医学免疫学》实验课程内容设置相对分散、不同节次之间缺乏联系,不能有效地展现机体内免疫应答这一连续的反应过程.为解决这些问题,南京医科大学基础医学院免疫学系在前期《医学免疫学》实验课的基础上,紧密围绕机体抗感染免疫应答这一主线,制定了综合实验课程方案:(1)小鼠外周血中性粒细胞的吞噬作用、(2)流式细胞术检测小鼠外周血中性粒细胞含量、(3)酶联免疫吸附试验检测小鼠外周血肿瘤坏死因子α(Tumor necrosis factor α,TNF-α)含量.通过建立细菌腹腔免疫小鼠模型,本研究依次串联出这3 个目前免疫学常用且先进的实验方法,同时检测指标契合抗感染免疫应答的时序性和整体性.学生通过显微镜能够直接观察到具有分叶核的中性粒细胞,胞质内可见被吞噬的白色葡萄球菌;白色葡萄球菌腹腔免疫小鼠后外周血中性粒细胞(CD11b+Ly-6G+)占外周血单个核细胞比例显著升高,血清中TNF-α的含量也显著增高.该综合实验的实施有助于学生更系统地理解机体应对细菌感染的早期免疫应答过程:外周血中性粒细胞对细菌的直接吞噬、中性粒细胞的活化增殖及炎性细胞因子(TNF-α)的分泌在机体抗感染过程中的作用;同时通过综合实验改革能够切实提高学生的实验操作和数据分析能力,并有助于培养他们的科研思维能力和团队合作精神.

外文摘要：In the past,the experimental course of Medical Immunology was relatively scattered and there was no connection between different sessions,which could not effectively show the continuous reaction process of immune response in the body.In order to solve these problems,the department of immunology of the School of Basic Medicine,Nanjing Medical University developed a comprehensive experimental course plan on the basis of the previous experimental course of Medical Immunology,closely focusing on the main line of the body's anti-infection immune response:(1)Phagocytosis of neutrophils from the peripheral blood of mice.(2)Detection of neutrophils content in the peripheral blood of mice by flow cytometry.(3)Detection of tumor necrosis factor α(TNF-α)in the peripheral blood of mice by enzyme-linked immunosorbent assay.By establishing a mouse model of bacterial intraperitoneal immunity,the three commonly used and advanced experimental methods in immunology were sequentially linked,and the detection indexes were consistent with the timing and integrity of the anti-infection immune response.The neutrophils with lobulated nuclei and phagocytosis of white staphylococcus were observed by microscope.The proportion of neutrophils(CD11b+Ly-6G+)in peripheral blood mononuclear cells was significantly increased,and the content of TNF-αin serum was also significantly increased in the mice which were immunized with staphylococcus.The implementation of the comprehensive experiment was helpful for students to understand the early immune response process of the body in response to bacterial infection more systematically:the direct phagocytosis of bacteria by neutrophils,the activation and proliferation of neutrophils in peripheral blood,and the role of the secretion of TNF-α in the process of anti-infection in the body.At the same time,comprehensive experiment reform can effectively improve students'ability of experimental operation and data analysis and help to cultivate their scientific research thinking ability and team spirit.

外文关键词：

comprehensive experimentimmune responseneutrophilsphagocytosisflow cytometryenzyme-linked immunosorbent assay

作者：

蔡振明、周泽伟、施冬艳、杨晓帆、张明顺、王慧娟、陈云、卢燕来

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作者单位：

南京医科大学基础医学院免疫学系,江苏南京 211166

关键词：

综合实验免疫应答中性粒细胞吞噬作用流式细胞术实验酶联免疫吸附试验

基金：

南京医科大学教育研究课题南京医科大学教育研究课题南京医科大学教育研究课题南京医科大学研究生教育研究课题

项目编号：

2023LX0102021YJS-ZC032021ZD0022021YJS-ZD002

出版年：

2024

DOI：

10.3969/j.issn.1674-9308.2024.01.004

中国继续医学教育

影响因子：2.564

ISSN：1674-9308

年,卷(期)：2024.16(1)

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