Down-regulation of Membrane Bound CD27 Expression Mediated by Protein Kinase C-beta and Protein Kinase C-delta in B Cells / 대한류마티스학회지
The Journal of the Korean Rheumatism Association
;
: 33-45, 2006.
Article
in Korean
| WPRIM
| ID: wpr-203398
ABSTRACT
OBJECTIVE:
CD27 is a member of the tumor necrosis factor receptor (TNFR) superfamily and is expressed on T, B, and NK cells. The signaling via CD27 plays pivotal roles in T-T and T-B interaction. CD27 is a useful marker in assessing the number of circulation B cells and B cell subsets because it permits one step identification of the major B cell compartments, CD27- naive and CD27+ memory B cells as well as CD27high plasma cells. We have analyzed the mechanisms underlying the regulation of CD27 expression.METHODS:
Isolation B cells and Raji cells were cultured with PMA. The levels of cell surface CD27 and CD 27 mRNA were analyzed by FACs staining and RT-PCR. Raji cells were cultured with phorbol 12-myristate 13-acetate (PMA), with or without pretreated shedding inhibitor BB3103 and TAPI-1. sCD27 was measured in culture supernatant by ELISA. Cell lysates were analyzed for PKC isotype activation by Western blot. We used PKC inhibitor Ly379196 and rottlen.RESULTS:
Membrane expression of CD27 was down-regulated after PMA stimulation without cytotoxic effect in B cells. Furthermore, PMA treatment could directly reduce CD27 mRNA without intermediate protein synthesis in B cells. In contrast, PMA treatment induced soluble form of CD27 (sCD27), which was shed from the cell surface and was found in PMA treatment B cell culture supernatant. PMA-induced sCD27 proteins were decreased with shedding inhibitor BB3103 and TAPI-1. PMA-induced down regulation of CD27 expressions were quenched with protein kinase C (PKC) inhibitor Staurosporin, PKC-beta inhibitor rottlerin and PKC-delta inhibitor Ly379196.CONCLUSION:
These data suggest that PMA-induced activation of PKC plays a crucial role in down-regulation of the expression of the CD27 and up-regulation of the shedding of the CD27 in human B cells.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasma Cells
/
Protein Kinases
/
Protein Kinase C
/
RNA, Messenger
/
Enzyme-Linked Immunosorbent Assay
/
B-Lymphocytes
/
Killer Cells, Natural
/
Down-Regulation
/
Up-Regulation
/
Blotting, Western
Type of study:
Prognostic study
Limits:
Humans
Language:
Korean
Journal:
The Journal of the Korean Rheumatism Association
Year:
2006
Type:
Article
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