Molecular cloning of NCCRP-1 gene from orange-spotted grouper (Epinephelus coioides) and characterization of NCCRP-1(+) cells post Cryptocaryon irritans infection.

Abstract

Nonspecific cytotoxic cells (NCCs) are an important cytotoxic cell population in the innate teleost immune system. The receptor designated "NCC receptor protein 1" (NCCRP-1) has been reported to be involved in the recognition and activation of NCCs. In this study, the full-length cDNA of Epinephelus coioides NCCRP-1 (ecnccrp-1) was cloned. The open reading frame (ORF) of ecnccrp-1 is 699 bp, encoding a 232 amino acid protein that includes proline-rich motifs at the N-terminus and is related to the F-box associated family. Although a bioinformatics analysis showed that EcNCCRP-1 had no signal peptide or transmembrane helices, a polyclonal antibody directed against recombinant EcNCCRP-1 efficiently labeled a membrane protein in the head kidney, detected with Western blot analysis, which indicated that the protein localized to the cell surface. RT-PCR showed that the constitutive expression of ecnccrp-1 was higher in the lymphoid organs, such as the trunk kidney, spleen, head kidney, and thymus, and lower in brain, heart, fat, liver, muscle, and skin. After infection with Cryptocaryon irritans, the transcription of ecnccrp-1 was analyzed at the infected sites (skin and gills) and in the systemic immune organs (head kidney and spleen). At the infected sites, especially the skin, ecnccrp-1 expression was upregulated at 6h post infection, reaching peak expression on day 3 post the primary infection. However, the expression patterns differed in the systemic immune organs. In the spleen, ecnccrp-1 was gradually increased in the early infection period and decreased sharply on day 3 post the primary infection, whereas in the head kidney, the transcription of ecnccrp-1 was depressed during almost the whole course of infection. An immunohistochemical analysis showed that EcNCCRP-1(+) cells accumulated at the sites of infection with C. irritans. These results suggested that NCCs were involved in the process of C. irritans infection in E. coioides.

• Authors: Xia-Zi Huang, Yan-Wei Li, Yong-Zhan Mai, Xiao-Chun Luo, Xue-Ming Dan, An-Xing Li
• Journal: Developmental and comparative immunology (Dev Comp Immunol) Vol. 46 Issue 2 Pg. 267-78 (Oct 2014) ISSN: 1879-0089 [Electronic] United States
• PMID: 24844613 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Published by Elsevier Ltd.

Chemical References

• Fish Proteins
• Receptors, Antigen
• nonspecific cytotoxic cell receptor protein 1

Topics

• Amino Acid Sequence
• Animals
• Base Sequence
• Cells, Cultured
• Ciliophora (physiology)
• Ciliophora Infections (immunology, metabolism, veterinary)
• Cloning, Molecular
• Conserved Sequence
• Fish Diseases (immunology, metabolism, parasitology)
• Fish Proteins (biosynthesis, genetics)
• Gene Expression (immunology)
• Host-Parasite Interactions
• Molecular Sequence Data
• Organ Specificity
• Perciformes (genetics, immunology, parasitology)
• Phylogeny
• Receptors, Antigen (biosynthesis, genetics)