Studis On The Effects Of Gliotoxin On The Invasion Of Aspergllius Fumigates To Host Cells And The Changes Of Cellular Phospholipase D And Actin Cytoskeleton

Objective To study the effect of gliotoxin from Aspergillus fumigates on invasion efficiency , phospholipase D (PLD) activation and the organization of actin cytoskeleton in the invasion of A.fumigutas conidia into host cells.Methods The human lung epithelial cells (A549) and mouse macrophage cells(J774) were incubated with three kinds of A.fumigutas (wildtype/B-5233,gliP deleted mutant/gliPΔand which complements defect in gliPR) and gliotoxin in 1, 10, 20, 50, 100, 200, 500, 800ng/ml, the invasion efficiency of A.fumigutas into cells and the activation of PLD in the cells were detected; A549 cells and J774 cells were pretreated with gliotoxin, the PLD expression was identified with western blot; the rearrangement of cellular actin cytoskeleton were observed under the laser confocal scanning microscope with the immuno-fluorescent staining; A549 and J774 cells cytoactivity was measured by microplate-MTT colorimetric assay.Results The invasion efficiency of gliPΔconidia into A549 cells were much lower than B-5233(P<0.05), there was on major difference between B-5233and gliPR (P>0.05). Gliotoxin (100ng/ml) could significantly enhance the internalization of B-5233 conidia into A549 cells(P<0.05),and gliotoxin (200ng/ml) made the internalization achieve to the maximum. However, the influence of gliotoxin on the invasion of conidia to J774 was different from A549. Gliotoxin(20ng/ml) was significantly decrased the internalization of A.fumigutas conidia into J774 cells (P<0.05). The activation and expression of PLD in both A549 cells and J774 cells were increased (p<0.05). Gliotoxin could influence the organization of A549 cellular actin cytoskeleton but didn't affect its cytoactivity. On the contrary ,we found gliotoxin didn't influence the cellular actin cytoskeleton but inhibited the cytoactivity of J774.Conclusion Gliotoxin increased the invasion of A.fumigutas conidia into A549 cells but not decreased the invasion into J774; gliotoxin could enhance the activation and expression of phospholipase D (PLD) of A549 and J774, and resulted in the rearrangement of A549 cellular actin cytoskeleton. Gliotoxin(≥100ng/ml)have toxic effect on J774 cells .