DigiFab interacts with endogenous cardiotonic steroids and reverses preeclampsia-induced Na/K-ATPase inhibition.
Elevated levels of endogenous Na/K-ATPase (NKA) inhibitors, cardiotonic steroids (CTSs) including marinobufagenin (MBG), contribute to pathogenesis of preeclampsia (PE) and represent a target for immunoneutralization by Digibind (Ovine Digoxin Immune Antibody, Glaxo-Smith Kline). Because Digibind is no longer commercially available, we studied whether DigiFab (BTG International Ltd, UK) can substitute Digibind for immunoneutralization of CTS in patients with PE. We compared DigiFab, Digibind, and anti-MBG monoclonal antibody (mAb) with respect to their ability to interact with CTS in PE plasma and to restore NKA activity in erythrocytes from patients with PE. Using immunoassays based on DigiFab, Digibind, and anti-MBG mAb, we studied the elution profile of CTS following high-performance liquid chromatography (HPLC) fractionation of PE plasma. Totally, 7 patients with mild PE (28 ± 2 years; gestational age, 39 ± 0.5 weeks; blood pressure 156 ± 5/94 ± 2 mm Hg) and 6 normotensive pregnant participants (28 ± 1 years; gestational age, 39 ± 0.4 weeks; blood pressure 111 ± 2/73 ± 2 mm Hg) were enrolled. Preeclampsia was associated with a substantial inhibition of erythrocyte NKA (1.47 ± 0.17 vs 2.65 ± 0.16 µmol Pi/mL per h in control group, P < .001). Ex vivo, at 10 µg/mL concentration, which is consistent with the clinical dosing of Digibind administered previously in PE, DigiFab and Digibind as well as anti-MBG mAb (0.5 µg/mL) restored erythrocyte NKA activity. Following HPLC fractionation of pooled PE and control plasma, PE-associated increase in CTS material was detected by Digibind (176 vs 75 pmoles), DigiFab (221 vs 70 pmoles), and anti-MBG mAb (1056 vs 421 pmoles). Therefore, because DigiFab interacts with CTS from PE plasma and reverses PE-induced NKA inhibition, it can substitute Digibind for immunoneutralization of CTS in patients with PE.
Laboratory of Reproduction and Women's Health, Institutes of Neonatology and Heart and Vessels, Almazov Heart, Blood and Endocrinology Center, St. Petersburg, Russia.
Valentina V
VV
Ishkaraeva-Yakovleva
Olga V
OV
Fedorova
Nelly G
NG
Solodovnikova
Elena V
EV
Frolova
Anton M
AM
Bzhelyansky
Igor V
IV
Emelyanov
C David
CD
Adair
Irina E
IE
Zazerskaya
Alexei Y
AY
Bagrov
D000328:Adult; D000911:Antibodies, Monoclonal / Q000627:therapeutic use; D002018:Bufanolides / Q000037:antagonists & inhibitors / Q000097:blood / Q000276:immunology; D002301:Cardiac Glycosides / Q000037:antagonists & inhibitors* / Q000097:blood / Q000276:immunology; D004791:Enzyme Inhibitors / Q000378:metabolism; D004912:Erythrocytes / Q000201:enzymology; D005260:Female; D005865:Gestational Age; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000627:therapeutic use*; D007167:Immunotherapy; D011225:Pre-Eclampsia / Q000201:enzymology / Q000628:therapy*; D011247:Pregnancy; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000097:blood*
D000911:Antibodies, Monoclonal; D002018:Bufanolides; D002301:Cardiac Glycosides; D004791:Enzyme Inhibitors; D007140:Immunoglobulin Fab Fragments; C050199:digoxin antibodies Fab fragments; C093896:marinobufagenin; D000254:Sodium-Potassium-Exchanging ATPase