Calcitonin gene-related peptide as an endogenous vasodilator: immunoblockade studies in vivo with an anti-calcitonin gene-related peptide monoclonal antibody and its Fab' fragment.
1. Calcitonin gene-related peptide (CGRP) is localized in perivascular sensory neurons and is a potent vasodilator. We investigated the utility of immunoblockade as an in vivo technique for probing the role of CGRP as an endogenous vasodilator. 2. The effects of an anti-CGRP monoclonal antibody (MAb; coded C4.19) and its Fab' fragment on CGRP-induced changes in blood pressure and skin blood flow were studied in pentobarbitone-anaesthetized rats. Antidromic skin vasodilatation in the rat hind paw was measured by laser Doppler fluxmetry. 3. The dose-response relationship for the hypotensive effect of intravenous rat alpha CGRP (r alpha CGRP) was similarly shifted rightward by MAb C4.19 IgG (1 mg/rat; intravenously) and Fab' fragment (2 mg/rat; intravenously). The C-terminal fragment of human alpha CGRP (h alpha CGRP8-37) also blocked the hypotensive effect of r alpha CGRP. 4. MAb C4.19 Fab' fragment (2 mg/rat; intravenously) and h alpha CGRP8-37 (100 nmol/kg; intravenously), but not MAb C4.19 IgG (up to 3 mg/rat; intravenously) or normal mouse Fab' fragment (2 mg/rat; intravenously), blocked the increased skin blood flow response to antidromic stimulation of the saphenous nerve. 5. The mean percentage changes in skin blood flow parameters due to MAb C4.19 Fab' fragment were significantly different from those due to normal mouse Fab' fragment (unpaired t-test; P < 0.05) but not from those due to h alpha CGRP8-37. 6. The results demonstrate the pharmacokinetic advantage of Fab' fragment over IgG for immunoblockade studies in vivo and support the role of CGRP in mediating skin vasodilatation.
Clinical Pharmacology Unit, University of Cambridge Clinical School, Addenbrooke's Hospital, U.K.
K K
KK
Tan
M J
MJ
Brown
R J
RJ
Hargreaves
S L
SL
Shepheard
D A
DA
Cook
R G
RG
Hill
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000494:pharmacology*; D000911:Antibodies, Monoclonal; D001794:Blood Pressure / Q000187:drug effects* / Q000502:physiology; D015740:Calcitonin Gene-Related Peptide / Q000037:antagonists & inhibitors / Q000502:physiology*; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology*; D007074:Immunoglobulin G / Q000494:pharmacology*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012039:Regional Blood Flow; D012867:Skin / Q000098:blood supply; D014664:Vasodilation / Q000187:drug effects / Q000502:physiology*
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D015740:Calcitonin Gene-Related Peptide