The gene expression of Interleukin-6 (IL-6) from human peripheral blood lymphocytes(HPBL) stimulated by C. albicans were investigated by using ELISA(Enzyme linked immunosorbent assay) and reverse transcription polymerase chain reaction(RT-PCR).
HPBL(1×10E6/ml) obtained from normal human peripheral blood lymphocytes were cultured with live C. albicans(LCA) or heat killed C. albicans type A 311(KCA, 3×10E7/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, 10μg/ml) or TNFα as a stimulants. For observation of the level of IL-6 gene expression, actinomycin D(AD, 5μg/ml) or cyclohexamide(CHX, 25μg/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 and LPS, highest value was observed at 72 hours reaction and the value was also higher than of
LCA.
IL-6 induced by LCA were detected up to 48 hours but in the case of KCA, the band for IL-6 were far stronger and appeared until lately than that of LCA. Therefore, the results of IL-6 gene expression agreed with that of ELISA, TNFα induced gene
expression of IL-6 up to 24 hours compared width up to 18 hours of HPBL without any stimuiatants. AD have the inhibition activity for the gene expression of IL-6 but CHX showed a superinduction activity.
These results suggested that KCA were stronger inducer of the IL-6 than LCA and it may be caused by change of the cell wall components, and C. albicans elicits a detectable IL-6 mRNA response and production from the HPBL within the first few
hours
of
stimulation.
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