Kazak Esra, Oliveira Sergio Costa, Goral Güher, Akalin Halis, Yilmaz Emel, Heper Yasemin, Oral Haluk Barbaros
Department of Medical Microbiology, Immunology Unit, Uludag University Faculty of Medicine, Bursa, Turkey, e-mail:
Iran J Immunol. 2010 Sep;7(3):132-41.
Because of high morbidity of the brucellosis in humans and the potential use of the microorganism as an agent of biologic warfare, protection of effective vaccines and specific diagnostic reagents become necessary to eradicate brucellosis.
In this study we aimed to investigate the cytokine responses and changes in peripheral blood lymphocyte subgroups of acute brucellosis patients in response to L7/L12 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) recombinant proteins derived from Brucella abortus.
levels of IFN-γ, IL-4 and IL-10 secreted from PBMCs of 25 acute brucellosis patients and 15 healthy controls, stimulated with Phytohemagglutinin (PHA), L7/L12 or GAPDH were measured by ELISA. Furthermore alterations in lymphocyte subgroups in response to these Brucella antigens were determined by flow cytometry.
Extracellular IFN-γ levels were found to be elevated after stimulation with L7/L12 in patients with acute brucellosis, whereas no significant changes were found in IL-4 and IL-10 levels. Similar data was also obtained with GAPDH, but the stimulation of IFN-γ production was not observed in all patients and was not as strong as that observed for L7/L12. Moreover, when the distribution of lymphocytes subgroups (CD3+, CD3+ CD4+, CD3+ CD8+, CD4+ CD25+, CD3+ CD69+ and CD3+ CD152+) was evaluated, it was found that the stimulation with L7/L12 and GAPDH only led to an increase in the percentage of CD3+ CD69+ lymphocytes.
These data indicate that Brucella abortus L7/L12 or GAPDH induce a Th1 type immune response in acute brucellosis patients. Additionally, these recombinant proteins, especially L7/L12, may be used in new vaccine preparations and diagnostic tests.
由于人类布鲁氏菌病发病率高，且该微生物有被用作生物战剂的潜在可能，因此研发有效的疫苗和特异性诊断试剂对于根除布鲁氏菌病很有必要。
本研究旨在调查急性布鲁氏菌病患者对流产布鲁氏菌来源的L7/L12和甘油醛-3-磷酸脱氢酶（GAPDH）重组蛋白的细胞因子反应及外周血淋巴细胞亚群的变化。
采用酶联免疫吸附测定法（ELISA）检测25例急性布鲁氏菌病患者和15名健康对照外周血单个核细胞（PBMC）在植物血凝素（PHA）、L7/L12或GAPDH刺激下分泌的γ干扰素（IFN-γ）、白细胞介素-4（IL-4）和白细胞介素-10（IL-10）水平。此外，通过流式细胞术确定这些布鲁氏菌抗原刺激后淋巴细胞亚群的变化。
急性布鲁氏菌病患者经L7/L12刺激后，细胞外IFN-γ水平升高，而IL-4和IL-10水平无显著变化。GAPDH刺激也得到类似结果，但并非所有患者均观察到IFN-γ产生的刺激，且不如L7/L12刺激强烈。此外，评估淋巴细胞亚群（CD3+、CD3+ CD4+、CD3+ CD8+、CD4+ CD25+、CD3+ CD69+和CD3+ CD152+）的分布时发现，L7/L12和GAPDH刺激仅导致CD3+ CD69+淋巴细胞百分比增加。
这些数据表明，流产布鲁氏菌L7/L12或GAPDH在急性布鲁氏菌病患者中诱导Th1型免疫反应。此外，这些重组蛋白，尤其是L7/L12，可用于新型疫苗制备和诊断检测。
