Dual HLA class I and class II restricted recognition of alloreactive T lymphocytes mediated by a single T cell receptor complex

Abstract

The alloreactive human T cell clone MBM15 was found to exhibit dual specificity recognizing both an antigen in the context of the HLA class I A2 molecule and an antigen in the context of the HLA class II DR1. We demonstrated that the dual reactivity that was mediated via a single clonal T cell population depended on specific peptide binding. For complete recognition of the HLA-A2-restricted specificity the interaction of CD8 with HLA class I is essential. Interestingly, interaction of the CD8 molecule with HLA class I contributed to the HLA-DR1-restricted specificity. T cell clone MBM15 expressed two in-frame T cell receptor (TCR) V␣ transcripts (V␣1 and V␣2) and one TCR V␤ transcript (V␤13). To elucidate whether two TCR complexes were responsible for the dual recognition or one complex, cytotoxic T cells were transduced with retroviral vectors encoding the different TCR chains. Only T cells transduced with the TCR V␣1V␤13 combination specifically recognized both the HLA-A2 ؉ and HLA-DR1 ؉ target cells, whereas the V␣2V␤13 combination did not result in a TCR on the cell surface. T he function of the major histocompatibility complex (MHC) molecules is to bind and display peptides to T lymphocytes. Allogeneic MHC molecules can induce strong T cell responses, which is reflected by the mixed lymphocyte reaction in vitro and the high incidence of graft rejection and graft versus host disease after transplantation of organs or hematopoietic cells over MHC barriers. T cell recognition of allogeneic MHC is often peptide specific, resembling self-MHC-restricted T cell recognition of foreign antigens (1-3). However, alloreactive T cells are heterogeneous in their degree of peptide specificity (4), and a minor population might be peptide independent or recognize motifs shared by many peptides (5). Several alloreactive T cell clones have been described to be crossreactive, recognizing two unrelated peptides in the context of two different allogeneic MHC class I molecules (6, 7). In addition, alloreactive cytotoxic T lymphocytes recognizing an endogenously processed peptide binding to allogeneic MHC molecules and recognizing a different peptide in the context of self-MHC class I have been described (8). In one instance, crossreactive cytotoxic T lymphocytes showing dual recognition for both HLA class I and class II molecules also have been reported (9, 10). Those authors postulated that based on the shared structural motif between the HLA-B27 and the DR2 B5*0101 chain, the reactivity pattern reflected presentation of identical or structurally related peptides by HLA-B27 and HLA-DR2. In several of these previously mentioned studies, cold-target inhibition experiments were performed to confirm that one clonal T cell population was mediating the crossreactivity. However, whether the crossreactivity of these alloreactive T cell clones was mediated via one or two T cell receptor (TCR) ␣␤ complexes was not investigated. This hypothesis may be possible because 20% of peripheral human T cells and 10% of mouse T cells express two dif...

Differential activation of the death receptor pathway in human target cells induced by cytotoxic T lymphocytes showing different kinetics of killing

“…Generation of retroviral supernatant and retroviral transduction of EBV-LCL were performed as previously described. 33 Transduced cells were purified by FACS ® sorting based on marker gene expression using a FACSVantage (Becton Dickinson, Mountain View, CA, USA).…”

HLA class II restricted T-cell receptor gene transfer generates CD4+ T cells with helper activity as well as cytotoxic capacity

“…[21][22][23] Previously, we demonstrated the retroviral transfer of a dual-specific TCR recognizing both an antigenic peptide in the context of HLA class I and an antigenic peptide in the context of HLA class II. 21 Furthermore, an MHC class II restricted chicken OVA 323À339 peptide-specific TCR was functionally transferred to murine CD4 + T helper cells. 22,23 Effective immune responses have been shown to depend on both CD4 + and CD8 + T lymphocytes.…”

“…[15][16][17][18][19][20] However, the specificity of HLA class II restricted T lymphocytes can also be transferred to other T cells. [21][22][23] Previously, we demonstrated the retroviral transfer of a dual-specific TCR recognizing both an antigenic peptide in the context of HLA class I and an antigenic peptide in the context of HLA class II. 21 Furthermore, an MHC class II restricted chicken OVA 323À339 peptide-specific TCR was functionally transferred to murine CD4 + T helper cells.…”

Minor histocompatibility antigen-specific T cells with multiple distinct specificities can be isolated by direct cloning of IFNγ-secreting T cells from patients with relapsed leukemia responding to donor lymphocyte infusion

“…The anti-HLA-A2-specific CTL clone MBM15 served as a positive control. 28 Effector T cells were irradiated (10 Gy) to abrogate T-cell proliferation without affecting cytolytic activity. As target cells enriched CD34 þ CML progenitor cells or unfractionated CML mononuclear cells obtained from the CML patients prior to transplantation, or CD34 þ HPC purified from the donors were used.…”