PB1040 - Procoagulant effects of Bothrops atrox and Bothrops lanceolatus venoms: in vitro study

Tuesday, June 25, 2024

13:45 – 14:45 ICT

Room: Exhibition Hall

Co-Author(s)

Sébastien Larréché, MD, PhD

MD, PhD
Hôpital d'instruction des armées Bégin, Université Paris Cité, Inserm UMR-S 1144
Saint-Mandé, Ile-de-France, France

Co-Author(s)

AB

Aurore Bousquet

PharmD
Hôpital d'instruction des armées Bégin
Saint-Mandé, Ile-de-France, France
LC

Lucie Chevillard

PhD
Université Paris Cité, Inserm UMR-S 1144
Paris, Ile-de-France, France
RG

Rabah Gahoual

PhD
Université Paris Cité, CNRS UMR 8258, Inserm
Paris, Ile-de-France, France
Georges Jourdi, PharmD, PhD

Associate Professor in haematology
Inserm Research institute
Paris, Ile-de-France, France
AD

Anne-Laure Dupart

PharmD
Hôpital d'instruction des armées Bégin
Saint-Mandé, Ile-de-France, France
CB

Christilla Bachelot-Loza, PhD

PhD
Université Paris Cité, Inserm UMR-S 1140
Paris, Ile-de-France, France
PG

Pascale Gaussem

PharmD, PhD
Université Paris Cité, Inserm UMR-S1140, Hôpital Européen Georges Pompidou
Paris, Ile-de-France, France
JC

Jean-Philippe Chippaux

MD, PhD
IRD, Université Paris Cité, UMR 261 MERIT
Paris, Ile-de-France, France
BM

Bruno Mégarbane

MD, PhD
Université Paris Cité, Inserm UMR-S 1144, Hôpital Lariboisière
Paris, Ile-de-France, France

Presenting Author(s)

Virginie Siguret

PharmD, PhD
Université Paris Cité, Inserm UMR-S 1140, Hématologie biologique, Hôpital Lariboisière (AP-HP)
Paris, Ile-de-France, France

Background: As observed with other continental Bothrops, B. atrox bite leads to local and systemic bleeding with hypocoagulability state. Conversely, envenomation by B. lanceolatus, an endemic species in Martinique, is unusually associated with systemic bleeding, but rather with thrombotic complications. Pathophysiology of these thrombosis remains unknown since B. lanceolatus venom was previously shown to be devoid of any in vitro procoagulant activity.

Aims: This study aimed (i) to describe the composition of B. atrox and B. lanceolatus venoms; (ii) to study their effect on whole blood coagulation using rotational thromboelastometry (ROTEM); (iii) to evaluate their dependency on co-factors (i.e., calcium and phospholipids); and (iv) to determine the effectiveness of four antivenoms (Bothrofav™, Inoserp™ South America, Antivipmyn™ TRI, and PoliVal-ICP™) on any potential venom procoagulant activity.

Methods: After tryptic digestion, snake venom was analyzed using ultraperformance liquid chromatography–tandem mass spectrometry. Coagulation was triggered in human whole blood (WB) after adding calcium and different concentrations of venom and assessed using ROTEM. Clotting time was measured on an automated coagulation analyzer (STA-R Max™) in human platelet-poor plasma (PPP) spiked with different venom concentrations, in the presence of calcium and/or phospholipids, and in the absence or presence of antivenom.

Results: B. atrox and B. lanceolatus venoms have comparable proteomes and dose-dependent procoagulant activities either in PPP or WB. B. lanceolatus effect was completely dependent on calcium but less dependent on phospholipids compared with B. atrox. The four tested antivenoms were able to neutralize the procoagulant effect of both Bothrops venoms, albeit with marked differences: Bothrofav™ having the most potent neutralizing effect.

Conclusion(s): Our findings were surprising in view of the opposite clinical complications resulting from envenomation by these two species. In vivo studies are needed to understand the exact pathophysiology of systemic bleeding and thrombosis associated with Bothrops bites.