First Efficacy and Safety Results with the Antibody Containing Leukocyte Inhibition Module in Cardiac Surgery Patients with Neutrophil Hyperactivity

Abstract

Systemic administration of immune modulating antibodies may play an important role in reducing neutrophil hyperactivity, for example, in patients undergoing cardiac surgery with extracorporeal circulation or in trauma patients. However, this strategy has extremely high costs and is often associated with severe adverse effects. We developed the Leukocyte-Inhibition-Module (LIM), an extracorporeal circulation (ECC) device housing a polyurethane matrix with covalently bound Fas (CD95; APO-1) stimulating antibodies to rapidly prevent neutrophil hyperactivation. A feasibility study with 14 patients undergoing cardiac surgery with the use of immunogenic ECC without (n = 5) and with (n = 9) LIM (venous line) was performed. Our data show that the usually observed ECC associated perioperative increase in neutrophils (control) was prevented by LIM (p = 0.023). Moreover, the increase of the proinflammatory markers tumor necrosis factor (TNF)-alpha and polymorphonuclear elastase was limited by LIM (p = 0.038 and p = 0.002). In both groups, no significant changes in liver enzymes or in clotting were detected after surgery, and up to 12 months follow up, no unusual complications were reported. This study shows for the first time to our knowledge the feasibility, efficacy, and safety of a new cost effective, immune management strategy in patients with aberrant immune activation by exposing the blood stream to immobilized agonistic anti-Fas antibodies.

Mcl-1-Mediated Impairment of the Intrinsic Apoptosis Pathway in Circulating Neutrophils from Critically Ill Patients Can Be Overcome by Fas Stimulation

“…However, experimental systemic application of agonistic antiFas Abs has toxic effects, e.g., in liver and lung (36). Therefore, immobilized agonistic anti-Fas IgM (clone CH-11) on solid biocompatible PU as a carrier has been used in this study (22,23).…”

“…To overcome this neutrophil apoptosis resistance, activation of the extrinsic apoptosis pathway by specific neutrophil Fas cross-linking remains an attractive strategy to transiently decrease neutrophil activity (21). To avoid the systemic application of agonistic Fas effector molecules resulting in nonspecific side effects, an extracorporeal immune therapy with agonistic anti-Fas IgM Abs immobilized on biocompatible carriers has been proposed to selectively target neutrophils within the circulating blood (22,23).…”

Extracorporeal immune therapy with immobilized agonistic anti-Fas antibodies leads to transient reduction of circulating neutrophil numbers and limits tissue damage after hemorrhagic shock/resuscitation in a porcine model

“…The proof of concept of such an approach had been previously shown in patients undergoing cardiac surgery [24,25]. …”

“…It is known that adequate cross-linking of Fas (APO-1, CD95) on the neutrophil surface membrane stimulates proapoptotic signaling pathways [26,27] but probably may also lead to cellular changes independent from apoptosis [28]. In this regard, we could show earlier that neutrophils rapidly become inactive following contact with membrane bound FasL [29] or with immobilized agonistic anti-Fas IgM antibody [24]. Moreover, evidence has been obtained that the transient contact of technetium-labelled neutrophils with immobilized anti-Fas IgM leads to their rapid sequestration in the spleen [22].…”

“…Therefore, an extracorporeal mini circuit was developed for the use in a porcine hemorrhagic shock model. As the functional unit, a down-scaled adaptation of the anti-Fas containing leukocyte inhibition module (LIM) as it was used previously for the integration in heart-lung machines [24] was connected to the circuit. The module allows Fas specific inactivation of circulating neutrophils at a flow of 300 ml/min.…”