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Retrovirology  2008 

Activation and detection of HTLV-I Tax-specific CTLs by Epitope expressing Single-Chain Trimers of MHC Class I in a rat model

DOI: 10.1186/1742-4690-5-90

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Abstract:

We have established expression vectors which encode SCTs of rat MHC-I (RT1.Al) with Tax180-188 peptide. Human cell lines transfected with the established expression vectors were able to induce IFN-γ and TNF-α production by a Tax180-188-specific CTL line, 4O1/C8. We have further fused the C-terminus of SCTs to EGFP and established cells expressing SCT-EGFP fusion protein on the surface. By co-cultivating the cells with 4O1/C8, we have confirmed that the epitope-specific CTLs acquired SCT-EGFP fusion proteins and that these EGFP-possessed CTLs were detectable by flow cytometric analysis.We have generated a SCT of rat MHC-I linked to Tax epitope peptide, which can be applicable for the induction of Tax-specific CTLs in rat model systems of HTLV-I infection. We have also established a detection system of Tax-specific CTLs by using cells expressing SCTs fused with EGFP. These systems will be useful tools in understanding the role of HTLV-I specific CTLs in HTLV-I pathogenesis.Human T-cell leukemia virus type I (HTLV-I) is etiologically linked to adult T-cell leukemia (ATL) [1,2], a chronic progressive neurological disorder termed HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) [3,4], and various other human diseases [5-8]. ATL is a malignant lymphoproliferative disease affecting a subgroup of middle-aged HTLV-I carriers characterized by the presence of mature T cell phenotype [9]. HTLV-I genome contains a unique 3' region, designated as pX, which encodes the viral transactivator protein, Tax [10]. Because of its broad transactivation capabilities [11], it is speculated that Tax plays a central role in HTLV-I associated immortalization and transformation of T cells, which may lead to the development of ATL.Tax is also known as a major target protein recognized by cytotoxic T lymphocytes (CTL) of HTLV-I carriers [12]. It has been reported that the levels of HTLV-I-specific CTL are quite diverse among HTLV-I carriers and that ATL patients have impaired l

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