Malassezia spp. are believed to be the causative agents of pityriasis versicolor and are strongly implicated in seborrhoeic dermatitis. The yeast also forms part of the normal human cutaneous microflora. We have previously shown that when Malassezia yeast cells are incubated with human peripheral blood mononuclear cells (PBMCs), they are capable of reducing the levels of pro-inflammatory cytokines produced. In order to test the hypothesis that this immunoevasive phenomenon may be related to the unusually high level of lipid in the Malassezia yeast cell wall, we have compared the immunomodulatory capacity of normal and lipid-depleted yeast cells. Stationary phase yeast cells of Malassezia sympodialis, M. globosa and M. restricta were treated with chloroform/methanol to extract the surface lipids. The lipid-depleted and non-depleted yeast cells were then co-cultured with human PBMCs from three different human donors at a ratio of 20 yeasts per leukocyte for 24 h. The levels of interleukin (IL)-1beta, IL-6 and tumour necrosis factor (TNF)-alpha were then determined by enzyme-linked immunosorbent assay (ELISA). The results demonstrated that extraction of lipid reversed the yeast cell capacity to reduce the levels of pro-inflammatory cytokines. The levels of IL-1beta, IL-6 and TNF-alpha produced in response to lipid-extracted Malassezia of all three species were either no different from or significantly greater (P < 0.05; ANOVA) than the constitutive control levels. These results suggest that the lipid microfibrillar layer of Malassezia may prevent the yeast cells from inducing inflammation and provide an explanation for the normal commensal status of the organism on human skin. The hypothesis that the lipid layer is absent or altered in seborrhoeic dermatitis may provide an explanation for the inflammatory nature of this dermatosis.

Malassezia spp., the causative agents of pityriasis versicolor, are members of the normal human cutaneous microflora. Utilizing a combination of both enzyme‐linked immunosorbent assay (ELISA) and bioassay, we have investigated the ability of both formalin‐preserved and viable Malassezia (serovars A, B and C) to modulate pro‐inflammatory cytokine (IL‐6, IL‐1β and TNF‐α) release by human peripheral blood mononuclear cells (PBMNC) in vitro, over a 48‐h co‐incubation period. The results demonstrated that formalin‐preserved Malassezia (serovars A, B and C) at midexponential phase were generally able to induce a pro‐inflammatory cytokine response at a yeast cell to PBMNC ratio of 1:1. In addition, the results consistently demonstrated that at a yeast cell to PBMNC ratio of 20:1, formalin‐preserved Malassezia, irrespective of serovar, growth phase or PBMNC donor, were capable of significantly (P<0·05) decreasing the release of both immunochemical IL‐6 and IL‐1β plus bioactive IL‐1β and TNF‐α below that of unstimulated culture medium control values. This was apparent following 24‐ and 48‐h co‐incubation times, where maximal cytokine production was detected after 24 h. Similar results were obtained for the effect of viable Malassezia on pro‐inflammatory cytokine release by PBMNC. Our results suggest that a possible inhibitory component, present perhaps within the cell wall of Malassezia, was responsible for this depressive effect on pro‐inflammatory cytokine production.

Malassezia spp., the causative agents of pityriasis versicolor, are members of the normal human cutaneous microflora. Utilizing a combination of both enzyme-linked immunosorbent assay (ELISA) and bioassay, we have investigated the ability of both formalin-preserved and viable Malassezia (serovars A, B and C) to modulate pro-inflammatory cytokine (IL-6, IL-1beta and TNF-alpha) release by human peripheral blood mononuclear cells (PBMNC) in vitro, over a 48-h co-incubation period. The results demonstrated that formalin-preserved Malassezia (serovars A, B and C) at mid-exponential phase were generally able to induce a pro-inflammatory cytokine response at a yeast cell to PBMNC ratio of 1:1. In addition, the results consistently demonstrated that at a yeast cell to PBMNC ratio of 20:1, formalin-preserved Malassezia, irrespective of serovar, growth phase or PBMNC donor, were capable of significantly (P<0.05) decreasing the release of both immunochemical IL-6 and IL-1beta plus bioactive IL-1beta and TNF-alpha below that of unstimulated culture medium control values. This was apparent following 24- and 48-h co-incubation times, where maximal cytokine production was detected after 24 h. Similar results were obtained for the effect of viable Malassezia on pro-inflammatory cytokine release by PBMNC. Our results suggest that a possible inhibitory component, present perhaps within the cell wall of Malassezia, was responsible for this depressive effect on pro-inflammatory cytokine production.

Malassezia spp., the causative agents of pityriasis versicolor, are members of the normal human cutaneous microflora. Utilizing a combination of both enzyme-linked immunosorbent assay (ELISA) and bioassay, we have investigated the ability of both formalin-preserved and viable Malassezia (serovars A, B and C) to modulate pro-inflammatory cytokine (IL-6, IL-1beta and TNF-alpha) release by human peripheral blood mononuclear cells (PBMNC) in vitro, over a 48-h co-incubation period. The results demonstrated that formalin-preserved Malassezia (serovars A, B and C) at mid-exponential phase were generally able to induce a pro-inflammatory cytokine response at a yeast cell to PBMNC ratio of 1:1. In addition, the results consistently demonstrated that at a yeast cell to PBMNC ratio of 20:1, formalin-preserved Malassezia, irrespective of serovar, growth phase or PBMNC donor, were capable of significantly (P<0.05) decreasing the release of both immunochemical IL-6 and IL-1beta plus bioactive IL-1beta and TNF-alpha below that of unstimulated culture medium control values. This was apparent following 24- and 48-h co-incubation times, where maximal cytokine production was detected after 24 h. Similar results were obtained for the effect of viable Malassezia on pro-inflammatory cytokine release by PBMNC. Our results suggest that a possible inhibitory component, present perhaps within the cell wall of Malassezia, was responsible for this depressive effect on pro-inflammatory cytokine production.

The present study explores the UVB protective role of Asperyellone (AY), a secondary metabolite of Aspergillus niger strain AN01. The in vitro UVB protective efficacy of AY was studied using the Human Epidermal keratinocytes cells (HaCaT) cell line. The results suggest the appreciable scavenging of UVBinduced reactive oxygen species in the AY-pretreated cells compared with UVB control. Experimental results on the antioxidant enzymes (Catalase, SOD, LPO, and GPx) profile, histochemical, and molecular analyses support the UVB protective effect of AY in HaCaT cells. Further, the in vivo UVB protective efficacy of AY was studied using animal models and compared with that of commercially available UVB protective agents. Physical, biochemical, and molecular analyses of skin samples emphasized the UVB protective role of AY. Thus, the important beneficial effects of AY have been explored in the present study. K E Y W O R D S apoptotic proteins, Aspergillus niger, asperyellone (AY), epidermal thickening, human epidermal keratinocytes cells (HaCaT) cells, UVB protection J Cell Biochem. 2019;120:10715-10725.wileyonlinelibrary.com/journal/jcb