Anti-inflammatory effects of infliximab in mice are independent of tumour necrosis factor α neutralization.
Infliximab (IFX) has been used repeatedly in mouse preclinical models with associated claims that anti-inflammatory effects are due to inhibition of mouse tumour necrosis factor (TNF)-α. However, the mechanism of action in mice remains unclear. In this study, the binding specificity of IFX for mouse TNF-α was investigated ex vivo using enzyme-linked immunosorbent assay (ELISA), flow cytometry and Western blot. Infliximab (IFX) did not bind directly to soluble or membrane-bound mouse TNF-α nor did it have any effect on TNF-α-induced nuclear factor kappa B (NF-κB) stimulation in mouse fibroblasts. The efficacy of IFX treatment was then investigated in vivo using a TNF-α-independent Trichuris muris-induced infection model of chronic colitis. Infection provoked severe transmural colonic inflammation by day 35 post-infection. Colonic pathology, macrophage phenotype and cell death were determined. As predicted from the in-vitro data, in-vivo treatment of T. muris-infected mice with IFX had no effect on clinical outcome, nor did it affect macrophage cell phenotype or number. IFX enhanced apoptosis of colonic immune cells significantly, likely to be driven by a direct effect of the humanized antibody itself. We have demonstrated that although IFX does not bind directly to TNF-α, observed anti-inflammatory effects in other mouse models may be through host cell apoptosis. We suggest that more careful consideration of xenogeneic responses should be made when utilizing IFX in preclinical models.
Faculty of Applied Medical Sciences, King AbdulAziz University, Jeddah, Saudi Arabia.|Faculty of Biology Medicine and Health, University of Manchester, Manchester.
M B
MB
Assas
Manchester Royal Infirmary, Manchester, UK.
S E
SE
Levison
School of Biological Sciences, Faculty of Medicine Biology and Health, University of Manchester, Manchester, UK.
M
M
Little
School of Biological Sciences, Faculty of Medicine Biology and Health, University of Manchester, Manchester, UK.
H
H
England
School of Biological Sciences, Faculty of Medicine Biology and Health, University of Manchester, Manchester, UK.
L
L
Battrick
School of Biological Sciences, Faculty of Medicine Biology and Health, University of Manchester, Manchester, UK.
J
J
Bagnall
School of Biological Sciences, Faculty of Medicine Biology and Health, University of Manchester, Manchester, UK.
J T
JT
McLaughlin
School of Biological Sciences, Faculty of Medicine Biology and Health, University of Manchester, Manchester, UK.
P
P
Paszek
School of Biological Sciences, Faculty of Medicine Biology and Health, University of Manchester, Manchester, UK.
K J
KJ
Else
School of Biological Sciences, Faculty of Medicine Biology and Health, University of Manchester, Manchester, UK.
J L
JL
Pennock
D000818:Animals; D019138:Antibodies, Blocking / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects; D002478:Cells, Cultured; D003092:Colitis / Q000188:drug therapy* / Q000469:parasitology; D000939:Epitopes; D005347:Fibroblasts / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D000069285:Infliximab / Q000494:pharmacology / Q000627:therapeutic use*; D008264:Macrophages / Q000187:drug effects* / Q000469:parasitology; D008297:Male; D051379:Mice; D008806:Mice, Inbred AKR; D018345:Mice, Knockout; D011485:Protein Binding; D014257:Trichuriasis / Q000188:drug therapy*; D014258:Trichuris / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D019138:Antibodies, Blocking; D000939:Epitopes; D014409:Tumor Necrosis Factor-alpha; D000069285:Infliximab
antibody; inflammation; macrophage; mucosa
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