| RRC ID |
45153
|
| Author |
Takeda A, Tsubaki T, Sagae N, Onda Y, Inada Y, Mochizuki T, Okumura K, Kikuyama S, Kobayashi T, Iwamuro S.
|
| Title |
Bacterial toxin-inducible gene expression of cathelicidin-B1 in the chicken bursal lymphoma-derived cell line DT40: functional characterization of cathelicidin-B1.
|
| Journal |
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.
|
| Volume |
59
|
| Pages |
94-102
|
| Published |
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
|
| Times Cited |
1
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
ENDOCRINOLOGY & METABOLISM
PHARMACOLOGY & PHARMACY
|
| Resource |
| Human and Animal Cells |
P-815(RCB1167)
COS-7(RCB0539) |
