Work

24 HIV-specific, ex vivo expanded T cell therapy (HXTC) for HIV clearance: feasibility, safety and efficacy

Year: 2017

Type: article

Abstract: in the presence of IL-2 for 3 days.After 3 weeks incubation in IL-2 containing medium in the presence of feeder cells, GFP+ cells were cell sorted and further expanded.Expanded GFP+ cells were analysed by fluorescence microscopy and flow cytometry.Results: From 10,000 to 30,000 CD25+CD134+CD39neg recall Ag-specific CD4+ T cells were isolated by cell sorting.Approximately 3-10% of purified Ag-specific cells were transduced with GFP, by fluorescence microscopy, compared with 50% transduction of positive control SEB-stimulated PBMC.Autologous monocyte-derived dendritic cells (Mo-DC) were used as feeder cells during expansion, and 4-6 weeks was required to obtain ≥2 million GFP+ Ag-specific CD4+ T cells.Using optimized sensitive flow cytometric detection, GFP+ CD4+ T cells were CCR5-negative, and had a Granzyme B+ cytotoxic phenotype.Conclusions: Using the OX40 assay, Ag-specific CD4+ T cells can be isolated for genetic modification of CCR5 expression via stable transduction with lentiviral shRNA.Cell therapy with in vitro expanded Ag-specific T cells has proved beneficial in the clinical setting of opportunistic infections of immune cells, such as EBV and CMV, during lymphopenia following hemopoietic stem cell transplant.Our results suggest that CCR5-negative Ag-specific cytotoxic CD4+ T cells may also be produced in vitro for possible augmentation of immunity, including to HIV-1 itself, in HIV+ subjects.

Source: Journal of Virus Eradication

DOI: 10.1016/s2055-6640(20)30729-9