Enhanced antilymphoma efficacy of CD19-redirected influenza MP1-specific CTLs by cotransfer of T cells modified to present influenza MP1

Blood. 2005 Feb 15;105(4):1622-31. doi: 10.1182/blood-2004-03-1208. Epub 2004 Oct 26.

Abstract

To enhance the in vivo antitumor activity of adoptively transferred, CD19-specific chimeric antigen receptor (CAR)-redirected cytotoxic T lymphocytes (CTLs), we studied the effect of restimulating CAR(+) CTLs through their endogenous virus-specific T-cell antigen receptor (TcR) by the cotransfer of engineered T-cell antigen-presenting cells (T-APCs). Using influenza A matrix protein 1 (MP1) as a model antigen, we show that ex vivo-expanded CD4(+) and CD8(+) T-APCs expressing a hygromycin phosphotransferase-MP1 fusion protein (HyMP1) process and present MP1 to autologous human leukocyte antigen (HLA)-restricted, MP1-specific CD4(+) and CD8(+) CTL precursors. The MP1-specific CTLs are amenable to subsequent genetic modification to express a CD19-specific CAR, designated CD19R, and acquire HLA-unrestricted reactivity toward CD19(+) leukemia and lymphoma tumor targets while maintaining HLA-restricted MP1 specificity. The restimulation of MP1xCD19 dual-specific CTLs in vivo by the adoptive transfer of irradiated HyMP1(+) T-APCs resulted in the enhanced antilymphoma potency of bispecific effector cells, as measured by elimination of the biophotonic signal of established firefly luciferase-expressing Burkitt lymphoma xenografts in nonobese diabetic/severe combined immunodeficiency (NOD/scid) animals compared with control groups restimulated by Hy(+)MP1(neg) T-APCs. Engineered T-APCs are a novel and versatile antigen-delivery system for generating antigen-specific T cells in vitro and enhancing the in vivo effector functioning of CAR-redirected antitumor effector cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adoptive Transfer / methods
  • Animals
  • Antigen Presentation* / genetics
  • Antigen-Presenting Cells / immunology
  • Antigen-Presenting Cells / metabolism
  • Antigen-Presenting Cells / virology
  • Antigens, CD19 / biosynthesis
  • Antigens, CD19 / physiology*
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / virology
  • CD8-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / virology
  • Cell Line
  • Cell Line, Transformed
  • Coculture Techniques
  • Cytotoxicity Tests, Immunologic / methods
  • Epitopes, T-Lymphocyte / genetics
  • Epitopes, T-Lymphocyte / immunology*
  • Gene Transfer Techniques
  • Humans
  • Influenza A virus / genetics
  • Influenza A virus / immunology
  • K562 Cells
  • Lymphocyte Activation / genetics
  • Lymphoma / genetics
  • Lymphoma / immunology*
  • Lymphoma / therapy*
  • Lymphoma / virology
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Phosphotransferases (Alcohol Group Acceptor) / genetics
  • Plasmids
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • T-Lymphocytes / transplantation*
  • T-Lymphocytes / virology
  • T-Lymphocytes, Cytotoxic / immunology*
  • T-Lymphocytes, Cytotoxic / metabolism
  • T-Lymphocytes, Cytotoxic / virology
  • Viral Matrix Proteins / biosynthesis
  • Viral Matrix Proteins / genetics
  • Viral Matrix Proteins / immunology*
  • Viral Matrix Proteins / metabolism

Substances

  • Antigens, CD19
  • Epitopes, T-Lymphocyte
  • M1 protein, Influenza A virus
  • Viral Matrix Proteins
  • Phosphotransferases (Alcohol Group Acceptor)
  • hygromycin-B kinase