Effect of C . albicans on IL - 6 Gene Expression of Human Peripheral Blood Lymphocytes.
- Author:
Hee Sun KIM
;
Young Sun LEE
;
Sung Kwang KIM
;
Haw Sun CHA
- Publication Type:Original Article
- Keywords:
C. albicans;
IL-6;
Human peripheral blood lymphocytes
- MeSH:
Blotting, Northern;
Dactinomycin;
Enzyme-Linked Immunosorbent Assay;
Gene Expression*;
Hot Temperature;
Humans*;
Interleukin-6;
Lymphocytes*;
Polymerase Chain Reaction;
Reverse Transcription;
RNA, Messenger;
Yeasts;
Zymosan
- From:Korean Journal of Immunology
1997;19(2):263-276
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
The gene expressions of Interleukin-6 (IL-6) from human peripheral blood lymphocytes (HPBL) stimulated by C. albicans were investigated by using ELISA (Enzyme linked immunosorbent assay), reverse transcription polymerase chain reaction (RT-PCR) and northern blotting. HPBL (1 X 10'/ml) obtained from normal human peripheral blood lymphocytes were cultured with live C. albicans (LCA) or heat killed C. albicans type A 311 (KCA, 3 X 10/ml) for various times (0.5, 1, 4, 8, 18, 24, 48 and 72 hours). On the purpose of this experiment, we also used lipopolysacchalide (LPS, 10 ug/ml), zymosan (1, 10, 100 ug/ml) as a polysacchaide component of the wall of yeast cells or TNFa (50, 100 ng/ml) as a IL-6 inducers. For observation of the level of IL-6 gene expression, actinomycin D (AD, 5 pg/ml) or cyclohexamide (CHX, 25 ug/ml) was added to HPBL stimulated with LCA for 0.5, 2, 4 hours and the HPBL were assessed for IL-6 mRNA. The highest value for IL-6 activity by LCA were observed at 48 hours reaction, but in the case of KCA, highest value of IL-6 activity was observed at 72 hours reaction and the value was also higher (500 pg/ml) than that of LCA (188 pg/ml). 1L-6 mRNA induced by LCA were detected up to 48 hours but in the case of KCA, the band for IL-6 mRNA were far stronger and appeared until lately than that of LCA. Therefore, the results of IL-6 gene expression agreed with that of ELISA.
