Clofazimine, but Not Isoniazid or Rifampicin, Augments Platelet Activation in vitro

Ronald Anderson; Annette J Theron; Jan G Nel; Chrisna Durandt; Moloko C Cholo; Charles Feldman; Gregory R Tintinger

doi:10.3389/fphar.2018.01335

Clofazimine, but Not Isoniazid or Rifampicin, Augments Platelet Activation in vitro

Front Pharmacol. 2018 Nov 20:9:1335. doi: 10.3389/fphar.2018.01335. eCollection 2018.

Authors

Ronald Anderson^{1

2}, Annette J Theron¹, Jan G Nel^{3

4}, Chrisna Durandt^{1

2}, Moloko C Cholo¹, Charles Feldman⁵, Gregory R Tintinger⁶

Affiliations

¹ Department of Immunology, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa.
² Institute for Cellular and Molecular Medicine, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa.
³ Department of Haematology, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa.
⁴ Tshwane Academic Division, National Health Laboratory Service, Pretoria, South Africa.
⁵ Department of Internal Medicine, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.
⁶ Department of Internal Medicine, Faculty of Health Sciences, Steve Biko Academic Hospital, University of Pretoria, Pretoria, South Africa.

Abstract

Although the inclusion of the cationic amphiphilic, anti-mycobacterial agent, clofazimine, in the chemotherapeutic regimens of patients with multidrug-resistant tuberculosis (TB) has contributed to improved outcomes, concerns remain about the cardiotoxic potential of this agent. Accordingly, the current study was undertaken with the primary objective of investigating the effects of clofazimine, on the reactivity of human platelets in vitro, a seemingly unexplored, mechanism of cardiotoxicity. Platelet-rich plasma (PRP) prepared from the blood of healthy, adult humans was treated with clofazimine (0.625-10 mg/L), or the primary anti-TB agents, isoniazid and rifampicin (at final concentrations of 5 and 10 mg/L), followed by addition of either adenosine 5'-diphosphate (ADP) or thrombin and measurement of platelet activation according to the magnitude of expression of CD62P (P-selectin), as well as the CD62P-mediated formation of heterotypic neutrophil:platelet (NP) aggregates, using flow cytometry. Clofazimine, but neither isoniazid nor rifampicin, caused dose-related potentiation of both ADP- and thrombin-activated expression of CD62P by platelets, achieving statistical significance at threshold concentrations of 0.625 and 2.5 mg/L, respectively, as well as significant formation of N:P aggregates. These stimulatory effects of clofazimine on platelet activation were partly attenuated by pre-treatment of PRP with the membrane-stabilizing agent, α-tocopherol, possibly consistent with a membrane-disruptive mechanism. In conclusion, clofazimine, at concentrations within the therapeutic range, augments platelet activation in vitro, probably by a mechanism linked to membrane destabilization. If operative in vivo, these pro-thrombotic activities of clofazimine may predispose for development of microvascular occlusion, exacerbating an already existing high risk for development of TB-associated cardiovascular disease.

Keywords: P-selectin; adenosine 5′-diphosphate; clofazimine; isoniazid; neutrophils; platelets; rifampicin; thrombin.