PCR amplification of interleukin 2 gene in alopecia areata

Alopecia areata or AA is a disease that targets hair follicles in the skin, causing disfiguring hair loss that is histologically marked by accumulation of infiltrating immune cells aroung the affected hair follicles. IL-2 gene has a role in regulating the survival and proliferation of T regulatory cells (Tregs). Tregs play an important role in preventing autoantigen-mediated immune response. Any perturbation in differentiation and proliferation of Tregs might impair their regulatory role which leads to an uncontrolled immune response which destroys follicular cells of hair bulbs. There is limited study on exon 2 of interleukin 2 gene of AA in Malaysia, thus leading to this study which was to design a set of primer to amplify exon 2. The study utilised the availability of pooled DNA sample from healthy volunteers at the facility hence, no ethic approval needed. The primer was designed using NCBI Primer BLAST. Next, the DNA was amplified and best annealing temperature was optimised or determined by gradient PCR. The amplicon was subjected for agarose gel electrophoresis. Then, the amplicon was sent for purification and DNA sequencing. The DNA sequencing result obtained was analysed for presence ofSNP. The DNA concentration was 35.5µg/mL and its purity at A260/280 was 1.83. The best annealing temperature is 57.0 °C. The SNP was absent in the exon 2 of the pooled DNA sample. The exon 2 of Interleukin-2 (IL-2) gene was successfully amplified using newly designed primer set.