Enhanced expression of a cloned and sequenced Ciona intestinalis TNFa-like (CiTNFa) gene during the LPS-induced inflammatory response.

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Abstract

A tumor necrosis factor-alpha (TNFα)-like genefrom Ciona intestinalis (CiTNFα-like) body wall challengedprepared in the presence of detergents. Both soluble andhemocyte-bound CiTNFα-like protein therefore appearedto be modulated by the LPS challengewith bacterial lipopolysaccharide (LPS) was clonedand sequenced 4 h after LPS inoculation. An open readingframe of 936 bp encoding a propeptide of 312 amino acids(35.4 kDa) displaying a transmembrane domain frompositions 7 to 29, a TACE cleavage site, and a maturepeptide domain of 185 amino acids (20.9 kDa), wasdetermined with a predicted isoelectric point of 9.4. Thephylogenetic tree based on deduced amino acid sequencesof invertebrate TNF-like protein and vertebrate TNFssupported the divergence between the ascidian and vertebrateTNF families, whereas D. melanogaster Eiger A andB TNF-like sequences were distinctly separated from thechordate TNFs. Thus, the ascidian TNFα-like cytokine wasupregulated by in vivo LPS challenge supporting its proinflammatoryrole. In the pharynx, increased expressionlevels were found following analysis by real-time polymerasechain reaction, whereas in situ hybridization assayshowed positive hemocytes both in the tissue and incirculating hemocytes. Finally, Western blot with monoclonalantibodies disclosed human TNFα epitopes in a 15-kDaprotein component of the hemolymph serum and in a 43-kDa protein contained in the hemocyte lysate supernatant
Lingua originaleEnglish
pagine (da-a)305-317
Numero di pagine12
RivistaCell and Tissue Research
Volume334
Stato di pubblicazionePublished - 2008

All Science Journal Classification (ASJC) codes

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  • ???subjectarea.asjc.2700.2722???
  • ???subjectarea.asjc.1300.1307???

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