Preclinical evaluation of innate immunity to baculovirus gene therapy vectors in whole human blood

TY - JOUR

T1 - Preclinical evaluation of innate immunity to baculovirus gene therapy vectors in whole human blood

AU - Georgopoulos, Lindsay J

AU - Elgue, Graciela

AU - Sanchez, Javier

AU - Dussupt, Vincent

AU - Magotti, Paola

AU - Lambris, John D

AU - Tötterman, Thomas H

AU - Maitland, Norman J

AU - Nilsson, Bo

PY - 2009/9

Y1 - 2009/9

N2 - Interactions of gene therapy vectors with human blood components upon intravenous administration have a significant effect on vector efficacy and patient safety. Here we describe methods to evaluate these interactions and their effects in whole human blood, using baculovirus vectors as a model. Opsonisation of baculovirus particles by binding of IgM and Ob was demonstrated, which is likely to be the cause of the significant blood cell-associated virus that was detected. Preventing formation of the complement C5b-9 (membrane attack) complex maintained infectivity of baculovirus particles as shown by studying the effects of two specific complement inhibitors, Compstatin and a C5a receptor antagonist. Formation of macroscopic blood clots after 4 h was prevented by both complement inhibitors. Pro- and anti-inflammatory cytokines Il-1 beta, IL-6, IL-8 and TNF-alpha were produced at variable levels between volunteers and complement inhibitors showed patient-specific effects on cytokine levels. Whilst both complement inhibitors could play a role in protecting patients from aggressive inflammatory reactions, only Compstatin maintained virus infectivity. We conclude that this ex vivo model, used here for the first time with infectious agents, is a valuable tool in evaluating human innate immune responses to gene therapy vectors or to predict the response of individual patients as part of a clinical trial or treatment. The use of complement inhibitors for therapeutic viruses should be considered on a patient-specific basis. (C) 2009 Elsevier Ltd. All rights reserved.

AB - Interactions of gene therapy vectors with human blood components upon intravenous administration have a significant effect on vector efficacy and patient safety. Here we describe methods to evaluate these interactions and their effects in whole human blood, using baculovirus vectors as a model. Opsonisation of baculovirus particles by binding of IgM and Ob was demonstrated, which is likely to be the cause of the significant blood cell-associated virus that was detected. Preventing formation of the complement C5b-9 (membrane attack) complex maintained infectivity of baculovirus particles as shown by studying the effects of two specific complement inhibitors, Compstatin and a C5a receptor antagonist. Formation of macroscopic blood clots after 4 h was prevented by both complement inhibitors. Pro- and anti-inflammatory cytokines Il-1 beta, IL-6, IL-8 and TNF-alpha were produced at variable levels between volunteers and complement inhibitors showed patient-specific effects on cytokine levels. Whilst both complement inhibitors could play a role in protecting patients from aggressive inflammatory reactions, only Compstatin maintained virus infectivity. We conclude that this ex vivo model, used here for the first time with infectious agents, is a valuable tool in evaluating human innate immune responses to gene therapy vectors or to predict the response of individual patients as part of a clinical trial or treatment. The use of complement inhibitors for therapeutic viruses should be considered on a patient-specific basis. (C) 2009 Elsevier Ltd. All rights reserved.

KW - Baculoviridae

KW - Complement C3b

KW - Complement C5a

KW - Complement Membrane Attack Complex

KW - Cytokines

KW - Gene Therapy

KW - Genetic Vectors

KW - Humans

KW - Immunity, Innate

KW - Immunoassay

KW - Immunoglobulin M

KW - Peptides, Cyclic

UR - http://www.scopus.com/inward/record.url?scp=68749103477&partnerID=8YFLogxK

U2 - 10.1016/j.molimm.2009.07.008

DO - 10.1016/j.molimm.2009.07.008

M3 - Article

C2 - 19665799

VL - 46

SP - 2911

EP - 2917

JO - Molecular immunology

JF - Molecular immunology

SN - 0161-5890

IS - 15

ER -