Cancino-Díaz M E, Pérez-Salazar J E, Domínguez-López L, Escobar-Gutiérrez A, Granados-Arreola J, Jiménez-Zamudio L, Burgos-Vargas R, García-Latorre E
Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Hospital General de México SSA, DF, México.
J Rheumatol. 1998 Sep;25(9):1756-64.
To study the antibody response of HLA-B27+ patients with ankylosing spondylitis (AS) and their first degree relatives to the 60 kDa protein of Klebsiella pneumoniae and to characterize this protein.
Sera from 84 individuals were analyzed by ELISA to determine the titer of antibodies against the 60 kDa protein of K. pneumoniae. Subjects were divided into 3 categories: Group 1: 44 HLA-B27+ AS related individuals (35 patients, 9 healthy controls); Group 2: 28 healthy B27- AS related individuals; and Group 3: 12 healthy B27- non-AS related subjects. The 60 kDa protein of K. pneumoniae was induced at 45 degrees C and purified by electroelution from sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was characterized as a GroEL-like heat shock protein (HSP). The recognition of GroEL-like protein was confirmed by immunoblot of 2 dimension electrophoresis. The response to GroEL-like protein from other bacteria and the response to lipopolysaccharide (LPS) was also analyzed by immunoblot.
HLA-B27+ individuals (Group 1), independent of their disease status, showed a significant higher response to the 60 kDa protein of K. pneumoniae than HLA-B27- subjects from Groups 2 and 3 (p < 0.0001). This protein was characterized as a HSP of the GroEL family and designated HSP60Kp. The GroEL of other enterobacteria as well as that of Mycobacterium leprae were recognized by HLA-B27+ individuals by immunoblot, whereas HLA-B27- individuals did not. LPS was not recognized by HLA-B27 positive or negative subjects.
These findings suggest a relationship between HLA-B27 and the response to a GroEL-like protein that could have implications in AS.
研究强直性脊柱炎(AS)患者及其一级亲属中HLA - B27阳性者对肺炎克雷伯菌60 kDa蛋白的抗体反应,并对该蛋白进行特性分析。
采用酶联免疫吸附测定(ELISA)分析84名个体血清中抗肺炎克雷伯菌60 kDa蛋白的抗体效价。受试者分为3组:第1组:44名HLA - B27阳性且与AS相关的个体(35例患者,9名健康对照);第2组:28名B27阴性且与AS相关的健康个体;第3组:12名B27阴性且与AS无关的健康受试者。肺炎克雷伯菌60 kDa蛋白在45℃诱导表达,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS - PAGE)后电洗脱纯化。其被鉴定为一种类GroEL热休克蛋白(HSP)。通过二维电泳免疫印迹法确认对类GroEL蛋白的识别。还通过免疫印迹法分析了对其他细菌类GroEL蛋白的反应以及对脂多糖(LPS)的反应。
第1组HLA - B27阳性个体,无论其疾病状态如何,对肺炎克雷伯菌60 kDa蛋白的反应显著高于第2组和第3组的HLA - B27阴性受试者(p < 0.0001)。该蛋白被鉴定为GroEL家族的热休克蛋白,命名为HSP60Kp。通过免疫印迹法,HLA - B27阳性个体可识别其他肠道杆菌以及麻风分枝杆菌的GroEL,而HLA - B27阴性个体则不能。HLA - B27阳性或阴性受试者均不能识别LPS。
这些发现提示HLA - B27与对类GroEL蛋白的反应之间存在关联,这可能与AS发病机制有关。