Genetic and Immunological Impact of the HRES-1/Rab4 Locus in SLE

Systemic lupus erythematosus (SLE) is a chronic inflammatory disease characterized by circulating antinuclear autoantibodies and dysfunction of T and B lymphocytes. Both genetic and environmental factors are believed to influence development of the disease. We detected and cloned the HRES-1 human endogenous retrovirus, mapped it to chromosome 1 at q42, newly identified six haplotypes in the long terminal repeat (LTR), and revealed an association of polymorphic HindIII653C-containing alleles with SLE. Thus, HRES-1 or a gene in linkage disequilibrium with this locus may influence autoimmunity in SLE. A newly discovered 2,986-base antisense transcript encodes exon 1 of a 24 kD protein, HRES-1/Rab4, that regulates surface expression of CD4, and to a lesser extent, expression of the transferrin receptor (TFR) through endosome recycling. Overexpression of HRES-1/Rab4 reduces surface expression of CD4 by inhibition of endocytic recycling and targets CD4 for lysosomal degradation, while dominant-negative HRES-1/Rab4S27N has the opposite effect both in Jurkat cells and peripheral blood T cells. HRES-1/Rab4 and CD4 protein levels inversely correlate both in healthy and lupus PBL. CD4 protein levels are reduced, while HRES-1/Rab4 expression is increased in lupus T cells having at least one HindIII653C in the HRES-1 LTR. CD4 plays essential roles in formation of the immunological synapse (IS) during normal T-cell activation by a cognate MHC class II peptide complex. The key intracellular transducer of T-cell activation, Lck, is brought to the IS via binding to CD4. TCR? chain binds to the TFR. Abnormal T cell responses in SLE have been associated with reduced TCR? chain and Lck levels in the lipid rafts of the IS. Although the regulatory roles of Rab GTP-ases in endosome trafficking are well recognized, their involvement in T-cell activation is largely unknown. Under Specific Aim 1, we will test the hypothesis that HRES1/Rab4 regulates the composition of lipid rafts, the assembly of the T cell synapse, and the functional outcomes of T-cell activation in peripheral blood T cells and Jurkat cells when stimulated with CD3 or superantigen. Under Specific Aim 2 we will determine the role of increased HRES-1/Rab4 expression in the altered lipid raft composition of lupus T cells. Under Specific Aim 3, we will test the hypothesis that the HindIIIG653C allele, alone or in combination with other genetic factors of lupus-associated haplotypes, enhances the expression of HRES-1/Rab4.