Manca F, Rossi G, Valle M T, Lantero S, Li Pira G, Fenoglio D, De Bruin J, Costantini M, Damiani G, Balbi B
Department of Immunology, San Martino Hospital-University of Genoa, Italy.
Infect Immun. 1991 Feb;59(2):503-13. doi: 10.1128/iai.59.2.503-513.1991.
To detect possible differences in phenotype and fine specificity for mycobacterial antigens between CD4-positive T cells from peripheral blood (PB) and from inflammatory sites, we identified four patients presenting with a mycobacterial pleural exudate (PE) rich in PPD-specific lymphocytes and with a negative skin test to tuberculin purified protein derivative (PPD) and a negative proliferative response of PB lymphocytes to PPD at the same time. Several weeks after chemotherapy, these patients converted to PPD responsiveness in the periphery, and PPD-specific clones could be generated from PB at this stage. The phenotypic comparison of PE lymphocytes and concomitant PB lymphocytes obtained before treatment showed an increase of CD8 cells and a high frequency of HLA-DR-positive activated T cells in PE. The frequency of tetanus toxoid-specific and Candida albicans-specific proliferating T cells was lower than that of PPD-specific cells in PE but not in PB. PPD-specific clones were derived initially from PE and from PB once the patients had converted to PPD responsiveness. The two sets of clones from each patient were compared for proliferative response to mycobacterial antigen clusters of defined molecular weight ranges. A large number of PE-derived clones (36%) responded to a fraction of 27 to 35 kDa, whereas only one clone from PB responded to the same fraction. The purified antigen P32 (32 kDa), a soluble mycobacterial protein, stimulated PE-derived clones that were responsive to the 37- to 27-kDa fraction but did not stimulate PB-derived clones. The data demonstrate that PE- and PB-derived lymphocytes differ both in phenotype and in fine specificity, suggesting a limited clonal heterogeneity of T cells localizing at the inflammatory site in tuberculous patients without a PPD response in the periphery. Therefore T cells compartmentalized at inflammatory sites provide information that is different from that provided by T cells in the periphery.
为检测外周血（PB）和炎症部位的CD4阳性T细胞在分枝杆菌抗原表型及精细特异性方面可能存在的差异，我们确定了4例患者，他们患有富含PPD特异性淋巴细胞的分枝杆菌性胸腔积液（PE），同时结核菌素纯蛋白衍生物（PPD）皮肤试验阴性，且PB淋巴细胞对PPD的增殖反应阴性。化疗数周后，这些患者外周血转为对PPD有反应，此时可从PB中产生PPD特异性克隆。对治疗前获取的PE淋巴细胞和同期PB淋巴细胞进行表型比较，结果显示PE中CD8细胞增加，且HLA-DR阳性活化T细胞频率较高。在PE中，破伤风类毒素特异性和白色念珠菌特异性增殖T细胞的频率低于PPD特异性细胞，但在PB中并非如此。PPD特异性克隆最初源自PE，患者外周血转为对PPD有反应后也可源自PB。比较了每位患者的两组克隆对分子量范围确定的分枝杆菌抗原簇的增殖反应。大量源自PE的克隆（36%）对27至35 kDa的组分有反应，而PB中只有一个克隆对相同组分有反应。纯化抗原P32（32 kDa），一种可溶性分枝杆菌蛋白，刺激了对37至27 kDa组分有反应的源自PE的克隆，但未刺激源自PB的克隆。数据表明，源自PE和PB的淋巴细胞在表型和精细特异性方面均存在差异，提示在无外周血PPD反应的结核患者中，定位于炎症部位的T细胞克隆异质性有限。因此，分隔在炎症部位的T细胞提供的信息与外周血T细胞提供的信息不同。
