Exploiting cytokine secretion to rapidly produce multivirus-specific T cells for adoptive immunotherapy.
Viral infections remain a major cause of morbidity and mortality after hematopoietic stem cell transplantation (HSCT), and conventional small-molecule therapeutics often have modest benefit, high cost, and adverse effects. Adoptive transfer of donor-derived virus-specific T cells has proved feasible and safe after HSCT and to reconstitute immunity against cytomegalovirus, Epstein-Barr virus, and adenovirus. Current protocols to generate these cytotoxic T cell lines are lengthy, taking up to 12 weeks. As viral infections often occur <30 days after HSCT, speedy production of virus-specific cytotoxic T cells lacking alloreactivity is highly desirable. We now describe a modified rapid selection method for production and characterization of CD4 and CD8 T cells specific for cytomegalovirus, Epstein-Barr virus, and adenovirus in a single infusate. We use Ad5f35-pp65/latent membrane protein 2 vectors in a single procedure over a 48-hour time period and manufacture a product suited for clinical use. By simultaneously expanding a portion of the selected product, we can characterize phenotype and function of the infused product and link them with subsequent in vivo outcome.
Fujita
Yuriko
Y
Center for Cell and Gene Therapy, Baylor College of Medicine, the Methodist Hospital, Texas Children's Hospital, Houston, TX 77030, USA.
Leen
Ann M
AM
Sun
Jiali
J
Nakazawa
Yozo
Y
Yvon
Eric
E
Heslop
Helen E
HE
Brenner
Malcolm K
MK
Rooney
Cliona M
CM
D000260:Adenoviruses, Human / Q000276:immunology; D049109:Cell Proliferation; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000276:immunology; D004266:DNA Virus Infections / Q000276:immunology / Q000517:prevention & control*; D005822:Genetic Vectors; D018380:Hematopoietic Stem Cell Transplantation; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive; D007371:Interferon-gamma / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology; D010750:Phosphoproteins / Q000276:immunology; D037182:T-Cell Antigen Receptor Specificity / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000276:immunology
C037662:EBV-associated membrane antigen, Epstein-Barr virus; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D007371:Interferon-gamma