An anti-human recombinant tumor necrosis factor alpha (TNF?) monoclonal antibody recognizes an epitope in feline TNF?

Abstract

-It is likely that the murine response to human recombinant TNFα (hrTNFα) may generate antibodies (Ab) to epitopes present in TNFα from other species. Here, we demonstrate that F5 antihrTNFα monoclonal antibody (mAb) recognizes feline TNFα while E8 anti-hrTNFα mAb failed to do so. In order to demonstrate that E8 and F5 mAb recognize different epitopes in the hrTNFα molecule, a constant concentration of E8 and variable concentrations of F5 were incubated with solid phase bound hrTNFα. Binding of E8 and F5 to hrTNFα was determined with anti-µ and γ chain specific Ab. F5 bound equally to hrTNFα in the presence or absence of E8 and the same amount of E8 bound to hrTNFα, in spite of the presence of F5. When using the E8 and F5 mAb for capturing the TNFα from the equine, canine, feline and bovine species, in supernatants of an ex vivo lipopolysaccharide (LPS)-stimulated whole blood cell culture, we only detected the feline TNFα by F5 mAb (p = 0.001). By a cytotoxic assay on L929 fibroblasts, we indeed demonstrated the feline TNFα production after the LPS stimulus. In an inhibition assay, the human and feline cytokines competed for F5, although the inhibition of native human TNFα binding to F5 was significant but only about 20% (p = 0.001). In conclusion, most likely the F5 anti-hrTNFα mAb recognizes an epitope in feline TNFα. Its immunomodulatory potential in the feline model remains to be studied. monoclonal antibodies / tumor necrosis factor alpha / cross-reactivity

Three-color flow cytometry detection of virus-specific CD4+ and CD8+ T cells in the cat

“…MAb 6401.1111 specifically stained feline TNFa-producing murine cells and also S. aureus Enterotoxin B-stimulated feline T cells, thus providing formal evidence for cross-reactivity. Cross-reactivity between a mAb against human TNFa with the feline homologue has also recently been reported by Aguirre et al (2003). By using allophycocyanin-conjugated mAb 6401.1111 in combination with PE-and FITC-conjugated mAbs against feline CD4 and CD8, respectively, antiviral T cells could be specifically identified in the peripheral blood and/or the spleens of FIPV-infected and of FCV-vaccinated cats.…”

Therapeutic effect of an anti-human-TNF-alpha antibody and itraconazole on feline infectious peritonitis

“…Generally, mAbs against human cytokines show no cross-reactive neutralizing activity against animal cytokines. However, Aguirre et al reported that human and feline TNFalpha have similar neutralizing epitopes [26]. Furthermore, the deduced amino acid sequence of human TNF-alpha is 89.7% identical to that of feline TNF-alpha, and 10 of 12 residues that interact with ADA are present in both [27].…”

Age‐associated changes to pathogen‐associated molecular pattern‐induced inflammatory mediator production in dogs

“…Whole blood culture was performed using a modification of a previously described technique 27–29 . The collected blood was diluted with modified Roswell Park Memorial Institute media c (200 U penicillin/mL, 200 μg streptomycin/mL, and 200 mM l ‐glutamine) in a 1:2 dilution and added to the wells of a 24 well plate.…”