Jindal S, Burke E, Bettencourt B, Khandekar S, Young R A, Dwyer D S
PROCEPT, Cambridge, MA 02139.
Immunol Lett. 1994 Feb;39(2):127-35. doi: 10.1016/0165-2478(94)90097-3.
Bacterial hsp60 proteins are major targets of immune responses during infection, and the highly conserved nature of bacterial and mammalian hsp60 has led to speculation that immune reactivity to these stress proteins may be a component of certain autoimmune diseases. We have developed recombinant proteins and monoclonal antibodies to facilitate further study of human hsp60 and its association with disease. The human hsp60 gene was expressed in Escherichia coli and a method for purification of the recombinant protein essentially devoid of E. coli GroEL was developed. Using the purified protein we have generated a number of monoclonal antibodies which are specific for human hsp60 and do not cross-react with its counterparts from E. coli and mycobacteria. A highly sensitive sandwich ELISA was developed to quantitate hsp60 levels and was used to study hsp60 accumulation in cells due to vaccinia virus infection and heat shock. This ELISA will be useful for monitoring hsp60 levels in body fluids or tissues during autoimmune reactions and/or inflammatory responses.
细菌热休克蛋白60（hsp60）是感染期间免疫反应的主要靶点，并且细菌和哺乳动物hsp60的高度保守性质引发了这样的推测：对这些应激蛋白的免疫反应性可能是某些自身免疫性疾病的一个组成部分。我们已经开发了重组蛋白和单克隆抗体，以促进对人hsp60及其与疾病关联的进一步研究。人hsp60基因在大肠杆菌中表达，并开发了一种纯化基本上不含大肠杆菌GroEL的重组蛋白的方法。使用纯化的蛋白，我们产生了许多对人hsp60具有特异性且不与大肠杆菌和分枝杆菌的相应蛋白发生交叉反应的单克隆抗体。开发了一种高度灵敏的夹心酶联免疫吸附测定法（ELISA）来定量hsp60水平，并用于研究由于痘苗病毒感染和热休克导致的细胞中hsp60的积累。这种ELISA将有助于在自身免疫反应和/或炎症反应期间监测体液或组织中的hsp60水平。
