| description: |
The general aims of our project are to detect possible associations between polymorphisms of the IL-10, IL-19, IL-20 and IL-24 genes and vitiligo and to assess IL-10, IL-19, IL-20 and IL-24 gene expression (mRNA and protein) variations in skin between healthy controls and patients with vitiligo. For the detection of SNPs in IL-10 gene the Amplification Refractory Mutation System-PCR (ARMS-PCR) method will be employed and for genotyping of polymorphisms of IL-19, IL-20 and IL-24 genes the tetra-primer ARMS-PCR method will be applied. Described methods have been successfully used by our group for screening IL-10-related cytokines gene SNPs in psoriasis. To analyze the gene expression levels of IL-10-related cytokines we will use two different methods. We will apply TaqMan-QRT-PCR method with gene specific primers to examine the mRNA expression levels of described cytokines. Second approach is the protein expression analysis of IL-10 and IL-20 in different cell cultures (keratinocytes and melanocytes) by using ELISA method. Previously genotyped patients will be examined to investigate the functional significance of various genotypes/haplotypes of IL-10, IL-19, IL-20 and IL-24 genes in determining the susceptibility and severity of vitiligo. No polymorphisms of the genes of IL-10-related cytokines have been investigated in vitiligo up to the present time. There is no also information about the expression levels of IL-10, IL-19, IL-20 and IL-24 in skin in patients with vitiligo. Genotypic and cytokine expression analyses may enable to understand the functional basis of the involvement of IL-10, IL-19, IL-20 and IL-24 genes in the susceptibility to vitiligo. |