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

Abstract

Both cytotoxic T cells and helper T cells are important in immune responses against pathogens and malignant cells.In hematological malignancies which express HLA class II molecules, immunotherapy may be directed to HLA class II restricted antigens. We investigated whether it is possible to engineer HLA class II restricted T cells with both antigenspecific cytolytic activity and the capacity to produce high amounts of cytokines. CD4 + and CD8 + peripheral-bloodderived T cells were retrovirally transduced with the HLA class II restricted minor histocompatibility antigen dead box RNA helicase Y (DBY)-specific TCR. The TCR-transduced CD4 + T cells exerted DBY-specific cytolytic activity, produced Th0, Th1, or Th2 cytokines, and proliferated upon DBY-specific stimulation. TCR-transduced CD8 + T cells exerted cytolytic activity which equaled the level of cytolytic activity of the TCR-transferred CD4 + T cells. Cotransfer of CD4 enhanced the cytolytic activity of the TCR-transduced CD8 + T cells, but introduction of CD4 was not sufficient to generate DBY-specific CD8 + T cells with the capacity to produce high amounts of cytokines. In this study, we demonstrated the feasibility to engineer T cells with antigen-specific cytolytic activity, as well as the ability to produce significant amounts of cytokines, by TCR transfer to CD4 + T cells. Gene Therapy (2005) IntroductionPatients relapsing from leukemia after allogeneic stem cell transplantation can successfully be treated by donor lymphocyte infusions (DLI), due to the graft-versusleukemia (GVL) response of the donor-derived T cells. [1][2][3][4] However, DLI is also associated with graft-versus-hostdisease (GVHD). Several studies have illustrated that depletion of CD8 + T cells from the DLI may preserve the beneficial GVL reactivity without the induction of GVHD, indicating that the CD4 + T lymphocytes may play an essential role in the antileukemic reactivity. [5][6][7][8][9][10] The mechanism by which the CD4 + T cells contribute to the GVL response is, however, not known. CD4 + T cells have been suggested to elicit the antileukemic response via the induction of minor histocompatibility antigen (mHag)-specific CD8 + T cells. 5 In addition, mHag-specific CD4 + T-helper cells have been reported to mature dendritic cells and enhance the expansion of mHag-specific cytotoxic T lymphocytes in vitro. 11 Alternatively, we have demonstrated that CD4 + T cells are capable of exerting direct cytolytic activity against leukemic cells. [12][13][14] Cellular immunotherapeutic approaches based on the adoptive transfer of T lymphocytes may require large cell numbers. Large numbers of T cells with a defined antigen specificity can be acquired by retroviral transfer of the T-cell receptor to peripheral-blood-derived T lymphocytes. Until now, the application of retroviral transfer of TCRs has mainly been limited to the transfer of HLA class I restricted TCRs. [15][16][17][18][19][20] However, the specificity of HLA class II restricted T lymphocytes can also be transferred to other T c...

αβ T-Cell Receptor Engineered γδ T Cells Mediate Effective Antileukemic Reactivity

“…The construction of the retroviral vectors containing the TCRs of the HA2.5 T-cell clone and the DBY-specific JBB4 T-cell clone has been previously described (10,25). The TCR of the HA2.5 T-cell clone was found to consist of AV15S1 and J42 in combination with BV18S1 with J2S7 according to the nomenclature described by Arden et al (26).…”

“…The Moloney murine leukemia virus-based retroviral vector LZRS and packaging cells B-NX-A were used (29). Retroviral supernatant was produced as previously described (10). By reverse transcription-PCR, sequencing and subsequent cloning retroviral vectors were constructed encoding the coreceptor molecules CD4a (DNGF-R), CD8a (eGFP), and CD8h (DNGF-R).…”

The HLA-A*0201-restricted minor histocompatibility antigen HA-1H peptide can also be presented by another HLA-A2 subtype, A*0206

“…22 For TCRAV usage determination, each reaction was carried out with a single sense primer specific for individual alpha families and a primer specific for the constant region of TCRA. 23 The PCR products were size-fractionated on 2% agarose gels. Subsequently, the complementarity-determining region 3 (CDR3) of each positive PCR product was sequenced with corresponding antisense primer as above.…”

Optimization of the HA-1-specific T-cell receptor for gene therapy of hematologic malignancies

“…TCRAV and TCRBV gene usage of the HA-1-specific T-cell clone HA1.M7, determined as previously described, 13 was demonstrated to be AV32 and BV6S4. Wild-type (WT) and modified TCRα and TCRβ chains were cloned separately into the retroviral vector LZRS, either in combination with eGFP (TCRα) or with NGF-R (TCRβ) marker gene.…”