Antibodies in passive immunization studies: characteristics and consequences.
Antibodies to neuropeptides or hormones are frequently used in passive immunization studies to unravel their physiological role in signal transfer. In such in vivo experiments antibodies are considered to bind and thereby to biologically inactivate the endogenous substance during its journey from its site of secretion to its site of action (signalling time). However, little is known about the mechanism of action and characteristics of antibodies that determine such biological activity. Since the signalling time in neuronal and hormonal communication is short, the kinetics of antibody binding is an important feature. Here, we present a theoretical framework to describe antibody binding kinetics which can contribute to the design of passive immunization protocols. The specific effects of variation in antibody concentration, dissociation constant, and on-rate constant on these binding kinetics are demonstrated. Simple methods are described to determine these parameters, which may guide the selection of antibodies for passive immunization studies. When time is limited, the on-rate constant and the local antibody concentration are the most important determinants. Several points are illustrated for CRF signal transfer in the rat. CRF signalling time in the hypothalamo-pituitary complex, as established from dye transport experiments, was 3-7 sec. Based on parameters measured for a rat monoclonal antibody to CRF (PFU 83), we computed that half-maximal and full blockades of ether-induced ACTH secretion were associated with approximately 85% and more than 99% binding of CRF, respectively. From the theoretical framework presented in this study we conclude that, in general, the kinetics of antigen binding are sufficiently fast for antibodies to interfere with hormonal and probably nonsynaptic neuronal signal transfer. However, interference with fast signalling processes (less than 10 msec), which may occur in the brain, is unlikely.
Van Oers
J W
JW
Department of Pharmacology, Medical Faculty, Free University, Amsterdam, The Netherlands.
Tilders
F J
FJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000936:Antigen-Antibody Complex; D003346:Corticotropin-Releasing Hormone / Q000097:blood* / Q000276:immunology; D005260:Female; D007030:Hypothalamo-Hypophyseal System / Q000098:blood supply / Q000502:physiology*; D007116:Immunization, Passive; D007700:Kinetics; D008433:Mathematics; D008954:Models, Biological; D051381:Rats; D011919:Rats, Inbred Strains
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D003346:Corticotropin-Releasing Hormone