The Role of Human Endogenous Retrovirus (HERV) K Encoded Deoxyuridine Triphosphate Nucleotidohydrolase in Psoriasis
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Publisher:The Ohio State University
Series/Report no.:The Ohio State University. Department of Microbiology Honors Theses; 2011
Psoriasis is a chronic inflammatory disease that affects 2-3% of the worldwide population. It is characterized by an epidermal hyperproliferation and an inflammatory cell infiltrate into the epidermis and dermis that results in the development of psoriatic lesions. Presently, the drugs used for treatment are generally immunosuppressive agents that either suppress the activation of immune response cells or inhibit cytokine/chemokine molecules that promote the inflammatory microenvironments characteristic of psoriasis pathophysiology. The PSORS1 locus, located in the major histocompatibility complex on chromosome 6p21, has been identified as the strongest genetic determinant of psoriasis, accounting for 35 to 50% of the heritability of the disease. It has been reported that the target region of the PSORS1 harbors a fragment of a human endogenous retrovirus K (HERV-K), which encodes for a deoxyuridine triphosphate nucleotidohydrolase (dUTPase). Our hypothesis is that the HERV-K dUTPase protein triggers a Th1, Th17 cytokine response by activating NF-κB through toll-like receptor 2 (TLR2). The goal of my study was to identify the region of the HERV-K dUTPase responsible for activating TLR2. To accomplish this I constructed three specific deletion constructs (∆ 60-171; ∆90-171; and ∆110-171) of the consensus HERV-K dUTPase sequence via PCR amplification and cloned these DNA fragments into the pTRC his vector for expression and subsequent purification of the recombinant proteins. The recombinant proteins were purified using cobalt affinity chromatography. The purified recombinant proteins will be used to determine structural characteristics of the recombinant protein and to determine whether these mutants proteins activate a NF-κB recorder gene in human embryonic kidney (HEK-293) cells expressing TLR2. We believe that successful completion of our studies will enable our research group to pursue development of a vaccine to generate antibodies that specifically target our identified region and thus provide a novel alternative treatment option for psoriasis patients.