PMID- 31340822 OWN - NLM STAT- MEDLINE DCOM- 20200709 LR - 20210308 IS - 1479-5876 (Electronic) IS - 1479-5876 (Linking) VI - 17 IP - 1 DP - 2019 Jul 24 TI - A strategy to protect off-the-shelf cell therapy products using virus-specific T-cells engineered to eliminate alloreactive T-cells. PG - 240 LID - 10.1186/s12967-019-1988-y [doi] LID - 240 AB - BACKGROUND: The use of "off-the-shelf" cellular therapy products derived from healthy donors addresses many of the challenges associated with customized cell products. However, the potential of allogeneic cell products to produce graft-versus-host disease (GVHD), and their likely rejection by host alloreactive T-cells are major barriers to their clinical safety and efficacy. We have developed a molecule that when expressed in T-cells, can eliminate alloreactive T-cells and hence can be used to protect cell therapy products from allospecific rejection. Further, expression of this molecule in virus-specific T-cells (VSTs) should virtually eliminate the potential for recipients to develop GVHD. METHODS: To generate a molecule that can mediate killing of cognate alloreactive T-cells, we fused beta-2 microglobulin (B2M), a universal component of all human leukocyte antigen (HLA) class I molecules, to the cytolytic endodomain of the T cell receptor zeta chain, to create a chimeric HLA accessory receptor (CHAR). To determine if CHAR-modified human VSTs could eliminate alloreactive T-cells, we co-cultured them with allogeneic peripheral blood mononuclear cells (PBMC), and assessed proliferation of PBMC-derived alloreactive T-cells and the survival of CHAR-modified VSTs by flow cytometry. RESULTS: The CHAR was able to transport HLA molecules to the cell surface of Daudi cells, that lack HLA class I expression due to defective B2M expression, illustrating its ability to complex with human HLA class I molecules. Furthermore, VSTs expressing CHAR were protected from allospecific elimination in co-cultures with allogeneic PBMCs compared to unmodified VSTs, and mediated killing of alloreactive T-cells. Unexpectedly, CHAR-modified VSTs eliminated not only alloreactive HLA class I restricted CD8 T-cells, but also alloreactive CD4 T-cells. This beneficial effect resulted from non-specific elimination of activated T-cells. Of note, we confirmed that CHAR-modified VSTs did not affect pathogen-specific T-cells which are essential for protective immunity. CONCLUSIONS: Human T-cells can be genetically modified to eliminate alloreactive T-cells, providing a unique strategy to protect off-the-shelf cell therapy products. Allogeneic cell therapies have already proved effective in treating viral infections in the stem cell transplant setting, and have potential in other fields such as regenerative medicine. A strategy to prevent allograft rejection would greatly increase their efficacy and commercial viability. FAU - Quach, David H AU - Quach DH AUID- ORCID: 0000-0003-1800-9559 AD - Center for Cell and Gene Therapy, Texas Children's Hospital, Houston Methodist Hospital and Baylor College of Medicine, 1102 Bates Ave, Suite 1770, Houston, TX, 77030, USA. FAU - Becerra-Dominguez, Luis AU - Becerra-Dominguez L AD - Center for Cell and Gene Therapy, Texas Children's Hospital, Houston Methodist Hospital and Baylor College of Medicine, 1102 Bates Ave, Suite 1770, Houston, TX, 77030, USA. FAU - Rouce, Rayne H AU - Rouce RH AD - Center for Cell and Gene Therapy, Texas Children's Hospital, Houston Methodist Hospital and Baylor College of Medicine, 1102 Bates Ave, Suite 1770, Houston, TX, 77030, USA. AD - Department of Pediatrics, Baylor College of Medicine, Houston, TX, 77030, USA. FAU - Rooney, Cliona M AU - Rooney CM AD - Center for Cell and Gene Therapy, Texas Children's Hospital, Houston Methodist Hospital and Baylor College of Medicine, 1102 Bates Ave, Suite 1770, Houston, TX, 77030, USA. cmrooney@txch.org. AD - Department of Pediatrics, Baylor College of Medicine, Houston, TX, 77030, USA. cmrooney@txch.org. AD - Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX, 77030, USA. cmrooney@txch.org. AD - Department of Molecular Virology and Immunology, Baylor College of Medicine, Houston, TX, 77030, USA. cmrooney@txch.org. LA - eng GR - P30 CA125123/CA/NCI NIH HHS/United States GR - P50 CA126752/CA/NCI NIH HHS/United States GR - 5T32DK60445-9/NH/NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20190724 PL - England TA - J Transl Med JT - Journal of translational medicine JID - 101190741 RN - 0 (HLA Antigens) RN - 0 (Histocompatibility Antigens Class I) RN - 0 (Receptors, Antigen, T-Cell) RN - 0 (beta 2-Microglobulin) SB - IM MH - Animals MH - CD4-Positive T-Lymphocytes/cytology MH - CD8-Positive T-Lymphocytes/cytology MH - Cell Proliferation MH - *Cell- and Tissue-Based Therapy MH - Coculture Techniques MH - Graft Rejection/immunology/*prevention & control MH - Graft vs Host Disease/immunology/*prevention & control MH - HLA Antigens/immunology MH - Hematopoietic Stem Cell Transplantation MH - Histocompatibility Antigens Class I/immunology MH - Humans MH - Leukocytes, Mononuclear/cytology MH - Lymphocyte Activation MH - Mice MH - Protein Domains MH - Receptors, Antigen, T-Cell/immunology MH - Regenerative Medicine/*methods MH - T-Lymphocytes/*cytology MH - beta 2-Microglobulin/metabolism PMC - PMC6657103 OTO - NOTNLM OT - Allogeneic T-cells OT - Alloreactive T-cells OT - Cellular therapy OT - Graft-vs-host disease OT - Immunotherapy OT - Off-the-shelf OT - Regenerative medicine OT - Rejection OT - Tolerance OT - Virus-specific T-cell COIS- Baylor College of Medicine (DHQ and CMR are listed as inventors) has filed a patent application using the CHAR for cell therapy. EDAT- 2019/07/26 06:00 MHDA- 2020/07/10 06:00 PMCR- 2019/07/24 CRDT- 2019/07/26 06:00 PHST- 2019/03/14 00:00 [received] PHST- 2019/07/17 00:00 [accepted] PHST- 2019/07/26 06:00 [entrez] PHST- 2019/07/26 06:00 [pubmed] PHST- 2020/07/10 06:00 [medline] PHST- 2019/07/24 00:00 [pmc-release] AID - 10.1186/s12967-019-1988-y [pii] AID - 1988 [pii] AID - 10.1186/s12967-019-1988-y [doi] PST - epublish SO - J Transl Med. 2019 Jul 24;17(1):240. doi: 10.1186/s12967-019-1988-y.