Comparison study for identifying promoter allelic polymorphism in interleukin 10 and tumor necrosis factor alpha genes

Cytokines such as tumor necrosis factor (TNF)-alpha and Interleukin (IL)-10 play significant roles in autoimmunity and transplantation tolerance. Alle lic polymorphisms that occur in the regulatory regions of these cytokine ge nes are closely associated with acute and chronic transplant rejection. The presence of a G-to-A polymorphism at position -308 in the promoter region of the TNF-alpha gene can increase transcription six- to sevenfold. Likewis e, the G-A polymorphism at position -1082 of the IL-10 promoter results in lower levels of IL-10 protein. Accordingly, a genotype that dictates the pr oduction of high levels of TNF-alpha with low IL-10 capabilities is most li kely to generate an inflammatory environment that is less receptive to the transplant. The potential for determining a patient's haplotype before tran splantation may be an effective way of monitoring the post-transplant statu s of such patients. A variety of methodologies that address the detection o f mutations have been used both in research and clinical diagnostic tests. This study analyzes the genetic variations in cytokines using two methodolo gies: the traditional allele-specific oligonucleotide (ASO) polymerase chai n reaction (PCR) and the newer and more flexible Invader technology. The se nsitivity and specificity of the Invader assay for simultaneous investigati on of multiple targets makes it a useful tool in such analyses.