Anti-idiotypic antibody facilitates scFv chimeric immune receptor gene transduction and clonal expansion of human lymphocytes for tumor therapy Journal Article
| Authors: | Cheung, N. K. V.; Guo, H. F. ; Modak, S.; Cheung, I. Y. |
| Article Title: | Anti-idiotypic antibody facilitates scFv chimeric immune receptor gene transduction and clonal expansion of human lymphocytes for tumor therapy |
| Abstract: | Chimeric immune receptors (CIR) transduced into lymphocytes link target recognition by single chain antibody Fv (scFv) to activation through CD28/TCRζ signaling. As surrogate antigens, anti-idiotypic antibodies may facilitate gene-transduction and clonal expansion of human lymphocytes for in vivo tumor therapy. The murine monoclonal antibody (MAb) 8H9 reacts with a novel antigen widely expressed on solid tumors. A CIR consisting of human CD8-leader sequence, 8H9-scFv, CD28 (transmembrane and cytoplasmic domains), and TCR-ζ chain was constructed, ligated into the pMSCVneo vector, and used to transfect the packaging line GP+envAM12 bearing an amphotropic envelope. Rat anti-idiotypic MAb 2E9 (IgG2a) was used to clone retroviral producer line as well as to expand gene-modified primary human lymphocytes. Sequential enrichments using either affinity chromatography or cell sorting using anti-idiotypic MAb 2E9 significantly improved the percentage of producer clones positive for surface 8H9-scFv and the efficiency of their supernatant in transducing the indicator cell line K562. By 3 weeks of in vitro culture, >95% of transduced primary human lymphocytes were CIR-positive. Upon periodic stimulation with 2E9, these lymphocytes underwent >10 6-fold expansion by 6 months in culture. They mediated antigen-specific non-MHC restricted cytokine release and tumor cytotoxicity, and inhibited human xenograft engraftment in SCID mice. Anti-idiotypic antibody may provide a useful tool for optimizing gene transduction of CIR fusion constructs into primary human lymphocytes and their continual expansion in vitro. |
| Keywords: | signal transduction; controlled study; protein expression; unclassified drug; human cell; nonhuman; cd8 antigen; lymphocyte proliferation; mouse; animals; mice; cancer immunotherapy; animal experiment; animal model; mice, scid; tumor cells, cultured; transfection; time factors; genetic transduction; animalia; genetic vectors; monoclonal antibody; cloning, molecular; cancer inhibition; t lymphocyte receptor; lymphocyte activation; antibodies, monoclonal; immunoglobulin variable region; genetic transfection; recombinant fusion proteins; antigen specificity; receptors, antigen, t-cell; signal peptide; lymphocyte clone; t-lymphocytes, cytotoxic; retrovirus vector; murinae; transplantation, heterologous; rats; neoplasm transplantation; cytokine release; clone cells; adoptive immunotherapy; immunotherapy, adoptive; single chain fragment variable antibody; cd28 antigen; cell selection; retroviridae; chimeric protein; cell mediated cytotoxicity; cell surface; affinity chromatography; supernatant; idiotypic antibody; antibodies, anti-idiotypic; monoclonal antibody 8h9; unidentified retrovirus; humans; human; priority journal; article; monoclonal antibody 2e9 |
| Journal Title: | Hybridoma and Hybridomics |
| Volume: | 22 |
| Issue: | 4 |
| ISSN: | 1536-8599 |
| Publisher: | Mary Ann Liebert, Inc |
| Date Published: | 2003-08-01 |
| Start Page: | 209 |
| End Page: | 218 |
| Language: | English |
| PUBMED: | 14511566 |
| PROVIDER: | scopus |
| DOI: | 10.1089/153685903322328938 |
| DOI/URL: | |
| Notes: | Export Date: 12 September 2014 -- Source: Scopus |
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