Antigen-specific in vitro expansion of functional redirected NY-ESO-1-specific human CD8+ T-cells in a cell-free system.

Abstract

BACKGROUND:

Tumors can be targeted by the adoptive transfer of chimeric antigen receptor (CAR) redirected T-cells. Antigen-specific expansion protocols are needed to generate large quantities of redirected T-cells. We aimed to establish a protocol to expand functional active NY-ESO-1-specific redirected human CD8(+) T-cells.

MATERIALS AND METHODS:

The anti-idiotypic Fab antibody A4 with specificity for HLA-A 0201/NY-ESO-1157-165 was tested by competition assays using a HLA-A 0201/NY-ESO-1157-165 tetramer. HLA-A 0201/NY-ESO-1157-165 redirected T-cells were generated, expanded and tested for CAR expression, cytokine release, in vitro cytolysis and protection against xenografted HLA-A 0201/NY-ESO-1157-165-positive multiple myeloma cells.

RESULTS:

A4 demonstrated antigen-specific binding to HLA-A 0201/NY-ESO-1157-165 redirected T-cells. Expansion with A4 resulted in 98% of HLA-A 0201/NY-ESO-1157-165 redirected T-cells. A4 induced strong proliferation, resulting in a 300-fold increase of redirected T-cells. After expansion protocols, redirected T-cells secreted Interleukin-2, (IL-2), interferon gamma (IFNγ) and tumor necrosis factor alpha (TNFα) and lysed target cells in vitro and were protective in vivo.

CONCLUSION:

A4 expanded HLA-A 0201/NY-ESO-1157-165 redirected T-cells with preservation of antigen-specific function.

• Authors: Gopinadh Jakka, Petra C Schuberth, Markus Thiel, Gerhard Held, Frank Stenner, Maries Van Den Broek, Christoph Renner, Axel Mischo, Ulf Petrausch
• Journal: Anticancer research (Anticancer Res) Vol. 33 Issue 10 Pg. 4189-201 (Oct 2013) ISSN: 1791-7530 [Electronic] Greece
• PMID: 24122982 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Neoplasm
• CTAG1B protein, human
• Immunoglobulin Fab Fragments
• Membrane Proteins

Topics

• Animals
• Antibody Affinity
• Antigens, Neoplasm (immunology)
• CD8-Positive T-Lymphocytes (immunology, transplantation)
• Cell Line, Tumor
• Cell Proliferation
• Cell-Free System
• Coculture Techniques
• Cytotoxicity, Immunologic
• HEK293 Cells
• Humans
• Immunoglobulin Fab Fragments (immunology)
• Immunotherapy, Adoptive
• Lymphocyte Activation
• Membrane Proteins (immunology)
• Mice
• Mice, Inbred NOD
• Mice, SCID
• Multiple Myeloma (immunology, pathology, therapy)
• Neoplasm Transplantation
• T-Lymphocytes, Cytotoxic (immunology)
• Tumor Burden