HUMAN IMMUNE-RESPONSE TO MYCOBACTERIUM-TUBERCULOSIS ANTIGENS
HUMAN IMMUNE-RESPONSE TO MYCOBACTERIUM-TUBERCULOSIS ANTIGENS
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DOI：
10.1128/iai.59.2.665-670.1991
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发表时间：
1991-02-01
影响因子：
3.1
通讯作者：
ELLNER, JJ
中科院分区：
文献类型：
--
作者：
HAVLIR, DV;WALLIS, RS;ELLNER, JJ
对人类结核分枝杆菌的免疫优势抗原或保护性抗原知之甚少。细胞介导的免疫对保护是必要的，健康的结核菌素阳性个体对外源性再感染具有相对的抵抗力。我们比较了健康结核菌素阳性个体与结核病患者和结核菌素阴性个体的细胞介导免疫反应的目标。利用硝基纤维素结合结核分枝杆菌培养滤液的t细胞Western blotting（免疫印迹）技术，在健康的结核菌素反应器中鉴定出30、37、44、57、64、71和88 kDa处的t细胞成母活性峰值。其中三个部分（30,64和71 kDa）与先前表征的蛋白一致：抗原6/ α抗原，HSP60和HSP70。观察纯化结核分枝杆菌抗原6/ α抗原和卡介苗HSP60的成母反应。当用纯化抗原6/ α抗原培养时，健康结核菌素反应器的淋巴细胞比健康结核菌素阴性对照或结核病患者表现出更多的[H-3]胸腺嘧啶掺入（分别为8,113 +/- 1,939-DELTA-cpm与645 +/- 425-DELTA-cpm和1,019 +/- 710-DELTA-cpm, P < 0.01）。健康反应者对HSP60也有应答，但应答程度低于抗原6/ α抗原（4276 +/- 1095 - δ -cpm; P < 0.05）。部分纯化的HSP70结合到硝化纤维素纸上，在6名结核患者中的2名中引起了显著的淋巴细胞成母反应，但在8个健康的结核菌素反应器中没有。5个结核素阴性对照的淋巴细胞对14或19 kDa的重组抗原或HSP70均无应答。抗体反应性通常与成胚反应呈负相关：结核血清对结核分枝杆菌培养滤液具有高滴度抗体，在35至180 kDa范围内。这是在健康的结核菌素反应器和结核病患者中首次系统地评估人体对一组天然抗原和重组抗原的反应。通过t细胞Western blot分析，鉴定出了7种能刺激淋巴细胞成母反应的抗原。其中两个可能代表先前表征的蛋白质；其他可能含有免疫优势蛋白，需要进一步鉴定。
Little is known about the immunodominant or protective antigens of Mycobacterium tuberculosis in humans. Cell-mediated immunity is necessary for protection, and healthy tuberculin-positive individuals are relatively resistant to exogenous reinfection. We compared the targets of the cell-mediated immune response in healthy tuberculin-positive individuals to those of tuberculosis patients and tuberculin-negative persons. By using T-cells Western blotting (immunoblotting) of nitrocellulose-bound M. tuberculosis culture filtrate, peaks of T-cell blastogenic activity were identified in the healthy tuberculin reactors at 30, 37, 44, 57, 64, 71 and 88 kDa. Three of these fractions (30, 64, and 71 kDa) coincided with previously characterized proteins: antigen 6/alpha antigen, HSP60, and HSP70, respectively. The blastogenic responses to purified M. tuberculosis antigen 6/alpha antigen and BCG HSP60 were assessed. When cultured with purified antigen 6/alpha antigen, lymphocytes of healthy tuberculin reactors demonstrated greater [H-3]thymidine incorporation than either healthy tuberculin-negative controls or tuberculous patients (8, 113 +/- 1,939-DELTA-cpm versus 645 +/- 425-DELTA-cpm and 1,019 +/- 710-DELTA-cpm, respectively; P < 0.01). Healthy reactors also responded to HSP60, although to a lesser degree than antigen 6/alpha antigen (4,276 +/- 1,095-DELTA-cpm; P < 0.05). Partially purified HSP70 bound to nitrocellulose paper elicited a significant lymphocyte blastogenic response in two of six of the tuberculous patients but in none of the eight healthy tuberculin reactors. Lymphocytes of none of five tuberculin-negative controls responded to recombinant antigens at 14 or 19 kDa or to HSP70. Antibody reactivity generally was inversely correlated with blastogenic responses: tuberculous sera had high titer antibody to M. tuberculosis culture filtrate in a range from 35 to 180 kDa. This is the first systematic evaluation of the human response to a panel of native and recombinant antigens in healthy tuberculin reactors and tuberculous patients. Antigens which stimulated prominent lymphocyte blastogenic responses were identified in seven fractions on T-cell Western blot analysis. Two of these may represent previously characterized proteins; the others may contain immunodominant proteins that will require further characterization.