论文标题:卡介苗与结核菌对大鼠外周血T 淋巴细胞亚群的影响
Effect of Bacillus Calmette-Guerin(BCG) Vaccine or(/and) Tubercle Germs on Rat T Lymphocyte Subsets of Peripheral Blood
论文作者 李光明
论文导师 雷建平,论文学位 硕士,论文专业 呼吸内科
论文单位 江西医学院,点击次数 966,论文页数 48页File Size405k
2004-05-01论文网 http://www.lw23.com/lunwen_102397/ 卡介苗;分支杆菌;结核;肺;外周血;大鼠;T淋巴细胞亚群;流式细胞仪
Mycobacterium Tuberculosis; BCG vaccine; Pulmonary; Peripheral blood; Rat; T lymphocyte subgroups; FCM
目的： 观察卡介苗或结核菌对大鼠外周血T淋巴细胞亚群的影响并与肺组织T淋巴细胞亚群作比较，并探讨卡介苗的免疫保护作用及结核菌对免疫的损害。对象与方法： 将40只SD大鼠随机分为正常对照组、卡介苗组、结核菌组及卡介苗结核菌组（每组10只）。卡介苗组及卡介苗结核菌组腹部皮内接种活卡介苗（BCG）0.01mg， 三个月后结核菌组及卡介苗结核菌组经尾静脉注入标准人型结核菌株H37Rv 0.01mg；同时，另外两组大鼠注入相同体积的生理盐水作对照。用流式细胞仪检测双色直标单克隆抗体标记的外周血T淋巴细胞亚群及肺组织T淋巴细胞亚群，并对肺部病灶进行病理及抗酸染色检查。结果 外周血T淋巴细胞亚群组间比较：前三个月，接种卡介苗组与非接种卡介苗组间T淋巴细胞亚群无显著性差异(P>0.05)。结核菌接种后一个月，卡介苗组CD8值低于正常组，结核菌组与卡介苗结核菌组CD8值、CD3值低于正常组(P<0.05)。结核菌接种后两个月，结核菌组CD4值与CD4/CD8比值低于正常组，卡介苗组CD8值低于其余三组、DP（CD3+CD4+CD8+）值低于正常组、CD4/CD8比值高于结核菌组而低于卡介苗结核菌组(P<0.05)。结核菌接种后三个月，结核菌组与卡介苗组CD8值低于正常组(P<0.05)。外周血与肺组织中T淋巴细胞亚群比较：卡介苗组、结核菌组及卡介苗结核菌血CD8值低于肺组织，正常组与结核菌组血CD4值高于肺组织，卡介苗结核菌组血CD3值低于肺组织，卡介苗组及卡介苗结核菌组血CD4/CD8比值低于肺组织，卡介苗组血DP值低于肺组织(P<0.05)。四组大鼠外周血T淋巴细胞亚群变化：正常组CD8值在三个月后开始升高，六个月时达最高水平，之后下降；CD4/CD8比值变化与CD8值相反(P<0.05)。卡介苗组CD3值、CD8值、DN（CD3+CD4-CD8-）值在接种卡介苗后下降，三个月时降到最低水平，六个月时大致恢复正常，之后又下降；CD4值及CD4/CD8比值在五个月后下降(P<0.05)。结核菌组CD3值与CD8值变化较一致，接种结核菌后一个月略下降，二个月时大致恢复正常，但CD8值升高明显，之后又下降；接种结核菌后二个月CD4值降到最低水平，之后逐渐回升(P<0.05)。卡介苗结核菌组在接种结核菌后CD3值、CD4值、CD8值、DN值变化幅度大致介于卡介苗组与结核菌组之间。结论： 卡介苗与结核菌对大鼠T淋巴细胞亚群的影响不同。卡介苗主要影响大鼠外周血CD3+CD4-CD8+细胞与CD3+CD4-CD8-细胞，结核菌对外周血CD3+CD4+CD8-细胞和CD3+CD4-CD8+细胞都有影响。卡介苗组大鼠肺组织中CD3+CD4-CD8+细胞和CD3+CD4+CD8+细胞明显高于血，结核苗组大鼠肺组织中CD3+CD4-CD8+细胞值高于血。推测CD3+CD4-CD8-细胞可能来源于CD4细胞，并向CD3+CD4-CD8+细胞或CD3+CD4+CD8+细胞转变。CD3+CD4-CD8+细胞与CD3+CD4+CD8+细胞可能在卡介苗的免疫保护中起主要作用，CD4细胞作用不大。
Objective To observe dynamically the effect of Bacillus Calmette-Guerin (BCG) vaccine or(/and) Mycobacterium tuberculosis on rat T lymphocyte subsets of peripheral blood and compare with T lymphocyte subsets of lung tissues, and evaluate the immunological function of BCG vaccine and the immunological impairment of Mycobacterium tuberculosis.Methods 40 SD rats were randomized into four groups as normal control group, BCG vaccine inoculated group (BCG group), Mycobacterium tuberculosis group (tuberculosis group) and BCG vaccine inoculated and Mycobacterium tuberculosis injected group (BCG-tuberculosis group). The BCG group and the BCG-tuberculosis group were injected with the live BCG vaccine 0.01mg in the abdominal derma at the beginning, after three months the normal control group and the BCG-tuberculosis group were injected in the tail vein with standard person type Mycobacterium tuberculosis H37Rv 0.01mg. For comparison, another two groups were injected simultaneously with the same volume normal saline. T lymphocyte of peripheral blood and lung tissues was measured by flow cytometry that labeled with bicolor direct-scale monoclonal antibody, and pulmonary lesions were examined pathomorphologically and pasted on the sheet to do acid-fast stain. Results T lymphocyte subsets is no significant difference between the BCG vaccine inoculated group and the non-BCG vaccine inoculated group( P>0.05). The first month after the injection of Mycobacterium tuberculosis, the level of CD8 in the BCG group is lower than the normal control group, the level of CD8 and CD3 in the tuberculosis group and the BCG-tuberculosis group are lower than the normal control group (P<0.05). The second month after injection , the level of CD4 and CD4/ CD8 ratio in the tuberculosis group are lower than th e normal control group, the level of CD8 in the BCG group is lower than another three groups, but DP（CD3+CD4+CD8+ ）higher than the normal control group, CD4/CD8 ratio higher than the tuberculosis group and lower than the BCG-tuberculosis group(P<0.05). The third month after injection, the level of CD8 in the normal control group is higher than the tuberculosis group and the BCG group (P<0.05). the level of CD8 of peripheral blood in the BCG group ，the tuberculosis group and the BCG-tuberculosis group is lower than lung tissues. The level of CD4 of peripheral blood in the normal control group and the tuberculosis group is higher than lung tissues, the level of CD3 in the BCG-tuberculosis group lower than lung tissues. The CD4/CD8 ratio of peripheral blood in the BCG group and the BCG-tuberculosis group is lower than lung tissues. the level of DP（CD3+CD4+CD8+ ）of peripheral blood in the BCG group is lower than lung tissues.The dynamic variety of peripheral blood T lymphocyte in the four groups: the level of CD8 in the normal control group begins to step up after three months, ascends the tallest level at the sixth month, and then decrease again. The variety of CD4/CD8 ratio is on the contrary (P<0.05). The level of CD3, CD8 and DN( CD3+ CD4- CD8-) in the BCG group declines the lowest level at the third month, recovers roughly the normal level at the sixth month, and then descends again; The level of CD4 and the ratio of CD4 to CD8 descends after five months.( P<0.05) . The level of CD3 and CD8 in the tuberculosis group falls at one month after the injection of Mycobacterium tuberculosis, the level of CD3 resumes normal but CD8 attends the tallest level at the second month, and then decreases again; at the second month after the injection of Mycobacterium tuberculosis, the level of CD4 declines the lowest and then sets up gradually, but is still lower than the normal level( P<0.05). The level of CD3, CD4, CD8 and DN after the injection of Mycobacterium tuberculosis ranges between the tuberculosis group and the BCG group on the whole.Conclusions BCG vaccine protection immunity multiplies the CD4+ T cell, CD8+ T cell and DP T cell of peripheral blood and pulmonary, and Mycobacterium tuberculosis restrains CD4+ T c
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