RRC ID 45153
著者 Takeda A, Tsubaki T, Sagae N, Onda Y, Inada Y, Mochizuki T, Okumura K, Kikuyama S, Kobayashi T, Iwamuro S.
タイトル Bacterial toxin-inducible gene expression of cathelicidin-B1 in the chicken bursal lymphoma-derived cell line DT40: functional characterization of cathelicidin-B1.
ジャーナル Peptides
Abstract Chicken cathelicidin-B1 (chCATH-B1) is a major host defense peptide of the chicken bursa of Fabricius (BF). To investigate the mechanisms of chCATH-B1 gene expression in the BF, we focused on the DT40 cell line derived from chicken bursal lymphoma as a model for analysis. A cDNA encoding chCATH-B1 precursor was cloned from DT40 cells. The nucleotide sequence of the cDNA was identical with that of the BF chCATH-B1. A broth dilution analysis showed that the synthetic chCATH-B1 exhibited a significant defensive activity against both Escherichia coli and Staphylococcus aureus. A scanning microscopic analysis demonstrated that chCATH-B1 inhibited bacterial growth through membrane destruction with formation of blebs and spheroplasts. Limulus amoebocyte lysate assay and electromobility shift assay results revealed that chCATH-B1 bound to lipopolysaccharide (LPS) and lipoteichoic acid (LTA), which are the surface substances of the E. coli and S. aureus cell, respectively. A chemotactic assay results revealed that chCATH-B1 showed mouse-derived P-815 mastocytoma migrating activity dose-dependently but with a higher concentration, resulting in a loss of the activity. A semi-quantitative real-time RT-PCR analysis revealed that LPS stimulated chCATH-B1 gene expression in a dose-dependent manner and that the LPS-inducible chCATH-B1 gene expression was inhibited by the administration of dexamethasone. The chCATH-B1 mRNA levels in DT40 cells were also increased by the administration of bacterial LTA. The results indicate that bacterial toxins induce chCATH-B1 gene expression in the chicken BF and the peptide expressed in the organ would act against pathogenic microorganisms not only directly but also indirectly by attracting mast cells.
巻・号 59
ページ 94-102
公開日 2014-9-1
DOI 10.1016/j.peptides.2014.06.012
PII S0196-9781(14)00185-5
PMID 24984089
MeSH Animals Antimicrobial Cationic Peptides / genetics* Antimicrobial Cationic Peptides / metabolism Antimicrobial Cationic Peptides / pharmacology* Bacterial Toxins / chemistry Bacterial Toxins / pharmacology* Cell Line Chickens Cloning, Molecular Dose-Response Relationship, Drug Escherichia coli / drug effects Gene Expression Regulation / drug effects* Gene Expression Regulation / genetics Lymphoma / genetics* Lymphoma / metabolism Microbial Sensitivity Tests RNA, Messenger / drug effects RNA, Messenger / genetics Staphylococcus aureus / drug effects Structure-Activity Relationship
IF 2.843
引用数 1
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY ENDOCRINOLOGY & METABOLISM PHARMACOLOGY & PHARMACY
リソース情報
ヒト・動物細胞 P-815(RCB1167) COS-7(RCB0539)