Therapeutic Anticoagulation with Argatroban and Heparins Reduces Granulocyte Migration: Possible Impact on ECLS-Therapy?

Andre Bredthauer,Manuel Kopfmueller,Diane Bitzinger,Sophie-Marie Pfaehler,Walter Petermichl,Michael Gruber,Timo Seyfried,Andreas Redel,Karla Lehle

doi:10.1155/2020/9783630

Abstract

Introduction Anticoagulants such as argatroban and heparins (low-molecular-weight and unfractionated) play an immense role in preventing thromboembolic complications in clinical practice. Nevertheless, they can also have a negative effect on the immune system. This study is aimed at investigating the influence of these substances on polymorphonuclear neutrophils (PMNs), whose nonspecific defense mechanisms can promote thrombogenesis. Methods Blood samples from 30 healthy volunteers were investigated, whereby PMNs were isolated by density gradient centrifugation and incubated with 0.8 μg/mL of argatroban, 1.0 U/mL of low-molecular-weight heparin (LMWH), 1.0 U/mL of unfractionated heparin (UFH), or without drug (control). A collagen-cell mixture was prepared and filled into 3D μ-slide chemotaxis chambers (IBIDI® GmbH, Germany). Stimulation was initiated by using a chemokine gradient of n-formyl-methionine-leucyl-phenylalanine (fMLP), and microscopic observation was conducted for 4.5 hours. The cells' track length and track straightness, as well as the number of attracted granulocytes, level of ROS (reactive oxygen species) production, and NET (neutrophil extracellular traps) formation, were analyzed and categorized into migration distances and time periods. Results All three anticoagulants led to significantly reduced PMN track lengths, with UFH having the biggest impact. The number of tracks observed in the UFH group were significantly reduced compared to the control group. Additionally, the UFH group demonstrated a significantly lower track straightness compared to the control. ROS production and NET formation were unaffected. Conclusion Our data provide evidence that anticoagulants have an inhibitory effect on the extent of PMN migration and chemotactic migration efficiency, thus indicating their potential immune-modulatory and prothrombotic effects.

Highlights

Anticoagulants such as argatroban and heparins play an immense role in preventing thromboembolic complications in clinical practice
The lowest value was found in the unfractionated heparin (UFH) group (n = 204 tracks) with a median of 19.3 μm (IQR = 34:7 μm), which differed from the low-molecular-weight heparin (LMWH) and the argatroban groups (p < 0:001) (Figure 1(b))
This study investigated the impact of the clinically used anticoagulants UFH, the LMWH Enoxaparin, and the direct thrombin-inhibitor argatroban on isolated granulocytes in terms of granulocyte migration, time dependency of reactive oxygen species (ROS) production and NET formation, and vitality

Summary

Introduction

Anticoagulants such as argatroban and heparins (low-molecular-weight and unfractionated) play an immense role in preventing thromboembolic complications in clinical practice. They can have a negative effect on the immune system. Blood samples from 30 healthy volunteers were investigated, whereby PMNs were isolated by density gradient centrifugation and incubated with 0.8 μg/mL of argatroban, 1.0 U/mL of low-molecular-weight heparin (LMWH), 1.0 U/mL of unfractionated heparin (UFH), or without drug (control). The number of tracks observed in the UFH group were significantly reduced compared to the control group. Clot formation within the membrane lung or the extracorporeal circuit is the most common technical complication, with an incidence of up to 22% [6] during ECLS therapy resulting in high mortality rates despite technical improvements and more sophisticated anticoagulation regimens [7]. Leukocyte deposits can be found in thrombi formed on gas exchange membranes of membrane lungs [11, 12]

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