CD28 costimulation and immunoaffinity-based selection efficiently generate primary gene-modified T cells for adoptive...

Area of Science:

• Immunology
• Cell Therapy
• Gene Therapy

Background:

• Allogeneic hematopoietic stem cell transplantation (HSCT) can lead to graft-versus-host disease (GVHD).
• Inducible suicide genes in donor T cells offer a strategy to control antitumor responses and GVHD post-HSCT.
• Existing methods face challenges including extensive ex vivo culture, drug toxicity, and transgene immunogenicity.

Purpose of the Study:

• To develop an efficient transduction and selection strategy for generating large numbers of gene-modified polyclonal T cells for adoptive immunotherapy.
• To overcome limitations of current methods for engineering T cells with suicide genes.

Main Methods:

• Developed a retroviral vector encoding human low-affinity nerve growth factor receptor (LNGFR) for selection and a Fas-based suicide construct (LV'VFas).
• Utilized CD28 ligation with T-cell receptor signaling for efficient T-cell transduction and growth.
• Employed LNGFR-specific microbeads for immunomagnetic selection of gene-modified T cells.

Main Results:

• Achieved efficient transduction and substantial T-cell growth, generating gene-modified T cells with retained clonal diversity and functionality.
• Demonstrated successful selection of large numbers of gene-modified T cells using LNGFR-targeted microbeads.
• The strategy requires only brief ex vivo culture and avoids drug selection.

Conclusions:

• A novel strategy for generating functional, gene-modified polyclonal T cells for adoptive immunotherapy has been identified.
• This method overcomes key obstacles associated with previous suicide gene approaches in T cells.
• The developed approach facilitates the production of therapeutic cell numbers for clinical applications.