"Bioactivities of a tumour necrosis like factor released by chicken mac" by Silke Rautenschlein, Anuradha Subramanian et al.

Chemical and Biomolecular Engineering Research and Publications

 

Date of this Version

July 1999

Comments

This article was Published in Developmental and Comparative Immunology 23 (1999) 629-640.© 1999 Elsevier Science Ltd. For published vesrion of theis article please visit publishers site

Abstract

To test for tumour necrosis-like factor (TNF) of chickens, supernatants of a lipopolysaccharide (LPS)-stimulated chicken macrophage cell line MQ-NCSU were analysed. A sequence of ion-exchange and gel-permeation chromatography was utilised to isolate TNF-like activity from the culture supernatant. The peak of TNF-like cytotoxic activity corresponded to the fractions with a nlolecular weight of 81 kDa or higher. Polyclonal anti-human TNF-a antiserum cross-reacted by Western blotting with a 17 kDa protein in the TNF-containing fraction ~ ~ n d e denaturing conditions. This result indicated that chicken TNF-like factor in the biologically active form may be a protein multimer of monomers of about 17 kDa. The molecular weight of these monomers is similar to the molecular weight of mammalian TNF-CLC. hicken TNF-like factor stimulated macrophages by inducing morphological changes, enhancing Ia-expression, nitric oxide (NO) production and by synergising with interferon (1FN)-y in the induction of NO release from macrophages. The biological activities were not neutralised by anti-human TNF antiserum. These data suggest that LPS-stimulated chicken macrophages produced a functional homologue to mammalian TNF-CL. This may be structurally quite different from the mammalian TNF molecule. Other factors may have been co-purified with the chicken TNF-like factor having overlapping functions and molecular weight. However, CO-purification of chemokines and interleukin-l, major macrophage derived factors, with the chicken TNF-like factor can be excluded based on the purification strategies.

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