Cg. Benishin et al., PRODUCTION OF POLYCLONAL ANTISERA TO PARATHYROID HYPERTENSIVE FACTOR FROM SPONTANEOUSLY HYPERTENSIVE RATS, Journal of hypertension, 11(3), 1993, pp. 245-251
Objective: Parathyroid hypertensive factor (PHF) is a newly described
hypertensive factor which may cause elevation of blood pressure in app
roximately 40% of North American essential hypertensive patients. PHF
is also found in several animal models of hypertension, including spon
taneously hypertensive rats (SHR), deoxycorticosterone acetate-salt hy
pertensive rats and salt-sensitive Dahl rats. The objective of the pre
sent study was to raise an antibody to PHF, and to use this antibody t
o study the effect of PHF in SHR. Design: Plasma and parathyroid gland
culture media collected from SHR were used in the present study as th
e antigen in the production and analysis of a polyclonal antiserum to
PHF. Methods: PHF is of low molecular weight (approximately 3000 dalto
ns) and is sensitive to inactivation by trypsin. We used PHF partially
purified from parathyroid gland culture media as the antigen to inocu
late mice. The substance was bound to aminophenyl thioether paper disc
s to make it more antigenic. The discs were then emulsified in Freund'
s adjuvant (complete for first inoculation, incomplete for subsequent
inoculations) for intraperitoneal implantation. Production of anti-PHF
antisera was monitored by enzyme-linked immunosorbent assay.Results:
Antisera produced in mice reacted with purified PHF prepared from SHR
plasma as well as with PHF prepared from parathyroid gland culture med
ia. PHF treated with the PHF antiserum produced no characteristic hype
rtensive response in normotensive assay rats. The antisera did not cro
ssreact with two forms of bovine parathyroid hormone, bPTH (1-84), bPT
H (1-34) or with shorter parathyroid hormone fragments, or with any ot
her vasoactive substance tested. Injection of an aliquot of the antise
rum in anesthetized spontaneously hypertensive rats reduced mean arter
ial pressure to a normal range of approximately 110 mmHg. Conclusions:
These results indicate that (1) polyclonal antisera to PHF can be rai
sed in mice and (2) PHF may contribute significantly to the elevated b
lood pressure in SHR.