Metyrapone test for hypothalamic-pituitary-adrenocortical function.

Our earlier attempts at immunization with human adrenocorticotropin hormone (ACTH) were unsuccessful and we therefore developed a new strategy including the chemical modification of the hormone by succinic anhydride in order to increase its immunogenicity. This process allowed us to obtain antisera with titers of up to 1/1000 and yielded 39 anti-succinylated ACTH (sACTH)-secreting hybridomas. Subsequently, the epitopes of sACTH were mapped by testing monoclonal antibodies two by two for simultaneous binding to sACTH and for their capacity to recognize its succinylated fragments 1-13, 1-17 and 1-24. The results, obtained with the use of radioactive tracers, were confirmed by and complemented with experiments conducted with biosensor technology. Seven groups of antibodies were defined on the basis of their pattern of reactivity and it was shown that four monoclonal antibodies could bind simultaneously to sACTH. Their dissociation constants (Kd) for sACTH were calculated and ranged from 10(-8) M to 10(-11) M. In order to obtain a fast and sensitive immunoassay for the hormone, we developed a protocol for the chemical modification of ACTH in serum and the most efficient monoclonal antibodies were selected on the basis of the epitope map and of their dissociation constants.

Adrenocorticotropic Hormone↗

PubMed1997

Sindbis virus infection of neonatal mice results in a severe stress response.

Neonatal mice were infected with virus derived from a molecular clone of a laboratory strain of Sindbis virus, TRSB. The resulting acute fatal infection was typified by few if any of the classic hallmarks of encephalitis, very high levels of interferon-alpha/beta (IFNalphabeta), and lesions in the thymus and hematopoietic tissues usually associated with a severe stress response. Infection with an attenuated mutant of TRSB, which harbors a single amino acid change in the E2 surface glycoprotein (TRSBr114), was characterized by encephalitis, reduced mortality, low levels of IFNalphabeta, and no thymic pathology (J. Trgovcich, J. F. Aronson, and R. E. Johnston, 1996, Virology 224, 73-83). Here we report that infection of neonatal mice with TRSB, but not TRSBr114, resulted in induction of high levels of tumor necrosis factor-alpha as well as high and sustained levels of adrenalcorticotropin-releasing hormone and corticosterone. This syndrome of potentially toxic cytokine and stress hormone induction correlates with lethal Sindbis virus infection and constitutes a previously unrecognized aspect of Sindbis virus pathogenesis in mice.

Adrenocorticotropic Hormone↗