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

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Abstract

A tumor necrosis factor-alpha (TNFα)-like gene from Ciona intestinalis (CiTNFα-like) body wall challenged with bacterial lipopolysaccharide (LPS) was cloned and sequenced 4 h after LPS inoculation. An open reading frame of 936 bp encoding a propeptide of 312 amino acids (35.4 kDa) displaying a transmembrane domain from positions 7 to 29, a TACE cleavage site, and a mature peptide domain of 185 amino acids (20.9 kDa), was determined with a predicted isoelectric point of 9.4. The phylogenetic tree based on deduced amino acid sequences of invertebrate TNF-like protein and vertebrate TNFs supported the divergence between the ascidian and vertebrate TNF families, whereas D. melanogaster Eiger A and B TNF-like sequences were distinctly separated from the chordate TNFs. Thus, the ascidian TNFα-like cytokine was upregulated by in vivo LPS challenge supporting its pro-inflammatory role. In the pharynx, increased expression levels were found following analysis by real-time polymerase chain reaction, whereas in situ hybridization assay showed positive hemocytes both in the tissue and in circulating hemocytes. Finally, Western blot with monoclonal antibodies disclosed human TNFα epitopes in a 15-kDa protein component of the hemolymph serum and in a 43-kDa protein contained in the hemocyte lysate supernatant prepared in the presence of detergents. Both soluble and hemocyte-bound CiTNFα-like protein therefore appeared to be modulated by the LPS challenge.

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Acknowledgements

We thank Mr. G. Miceli for collecting ascidians, and Mr. M. Guarcello for expert maintenance of the aquaria.

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Authors and Affiliations

  1. Dipartimento di Biologia Animale, Laboratory of Marine Immunobiology, Università di Palermo, Via Archirafi 18, 90123, Palermo, Italy

    Nicolò Parrinello, Aiti Vizzini, Vincenzo Arizza, Giuseppina Salerno, Daniela Parrinello, Matteo Cammarata, Francesca Tiziana Giaramita & Mirella Vazzana

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  1. Nicolò Parrinello
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  2. Aiti Vizzini
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  7. Francesca Tiziana Giaramita
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  8. Mirella Vazzana
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Correspondence to Nicolò Parrinello.

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This work was supported by a research grant from the Italian Ministry of University and Scientific Research (PRIN 2006 to N. Parrinello), co-funded by the University of Palermo.

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Parrinello, N., Vizzini, A., Arizza, V. et al. Enhanced expression of a cloned and sequenced Ciona intestinalis TNFα-like (CiTNFα) gene during the LPS-induced inflammatory response. Cell Tissue Res 334, 305–317 (2008). https://doi.org/10.1007/s00441-008-0695-4

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  • DOI: https://doi.org/10.1007/s00441-008-0695-4

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