Moreno-Lafont Martha Cecilia, López-Santiago Rubén, Paredes-Cervantes Vladimir, Estrada-Aguilera Ariel, Santos-Argumedo Leopoldo
Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, Mexico.
Arch Med Res. 2003 May-Jun;34(3):184-93. doi: 10.1016/S0188-4409(03)00020-1.
T-cell proliferation is a standard method to evaluate cellular immune responses against intracellular infectious agents. The present study was undertaken to look for expression of an early activation marker (CD69) and proliferation using a nonradioactive method to evaluate cellular immune response against a salt-extractable antigen from Brucella melitensis 16M (RCM-BM) in patients suffering from brucellosis.
Expression of CD69 on membrane of CD4+ and CD8+ T-cells was determined by flow cytometry. Lymphoproliferation was determined by tritiated thymidine and 5-bromo-2'-deoxyuridine (BrdU) incorporation using liquid scintillation counter or flow cytometry, respectively, to evaluate DNA synthesis.
Thirty healthy donors and 24 patients suffering from brucellosis were included in this study. In all cases, incubation with mitogen induced expression of CD69 and proliferation of both CD4+ and CD8+ T-cells. In contrast, only brucellosis patients responded with expression of CD69 and proliferation against RCM-BM antigen from Brucella melitensis (p < 0.001).
Methods used in this study were useful to evaluate immune response against specific antigen or polyclonal stimulation. CD4+ and CD8+ T cells from patients became equally activated and proliferated in response to RCM-BM antigen. Our data suggest that both T-cell subpopulations play an important role in immune response against Brucella.
T细胞增殖是评估针对细胞内感染因子的细胞免疫反应的标准方法。本研究旨在寻找一种早期激活标志物(CD69)的表达情况,并采用非放射性方法评估布鲁氏菌病患者针对羊种布鲁氏菌16M盐提取物抗原(RCM - BM)的细胞免疫反应中的增殖情况。
通过流式细胞术测定CD4⁺和CD8⁺T细胞膜上CD69的表达。分别使用液体闪烁计数器或流式细胞术通过氚化胸腺嘧啶核苷和5-溴-2'-脱氧尿苷(BrdU)掺入来测定淋巴细胞增殖,以评估DNA合成。
本研究纳入了30名健康供者和24名布鲁氏菌病患者。在所有情况下,与有丝分裂原孵育均诱导了CD69的表达以及CD4⁺和CD8⁺T细胞的增殖。相比之下,只有布鲁氏菌病患者对羊种布鲁氏菌的RCM - BM抗原出现CD69表达和增殖反应(p < 0.001)。
本研究中使用的方法有助于评估针对特定抗原或多克隆刺激的免疫反应。患者的CD4⁺和CD8⁺T细胞在对RCM - BM抗原的反应中同样被激活并增殖。我们的数据表明,这两个T细胞亚群在针对布鲁氏菌的免疫反应中均发挥重要作用。