Transduction and differentiation of dendritic cells from human hematopoietic progenitor cells

M. E. Reeves, S. A. Rosenberg, P. Hwu

Research output: Contribution to journalArticle

Abstract

Dendritic cells (DC) are highly potent antigen presenting cells, and may be able to stimulate effective anti-tumor immunity. Therefore, the ability to constitutively express tumor antigen genes in DC might be a powerful method to uncover new tumor antigens in vitro and to actively immunize in vivo. However, the growth of large numbers of DC has been difficult, and such cells have not been receptive to gene transfer. Since DC can differentiate from hematopoietic progenitor cells (HPC), we are retrovirally transducing human HPC with tumor antigen genes, followed by in vitro differentiation into DC. In initial experiments, human CD34+ HPC were transduced with marker genes (β-galactosidase or murine B7-1), and differentiated in vitro into DC using GM-CSF, TNFa, and stem cell factor. The resulting cell population contained 20-30% DC by morphological and phenotypic analyses, and was functionally far superior to PBMC in allogeneic MLR. By FACS analysis, >5% of cells expressed both the transduced marker gene and the DC phenotype (HB7-2+. class IP, CDla+, hB7-l+. The DC population expanded 20 fold during differentiation. Using this method, the melanoma tumor antigen gene MART-1 was introduced into DC. The MART-1 transduced DC stimulated high levels of cytokine release by MART-1 specific tumor infiltrating lymphocytes (2816 pg/mt IFNy vs 193 pg/ml for controls transduced with an irrelevant gene). This indicates that they were able to express, process, and present a MART-1 epitope on MHC class I molecules. This expression and presentation was stable, persisting beyond two weeks without selection. We are currently trying to raise specific anti-MART-1 CTL by in vitro stimulation of autologous PBL with the MART-1 transduced DC. In addition, we are evaluating the ability of transduced DC to present endogenously expressed antigens on MHC class II molecules.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - 1996
Externally publishedYes

Fingerprint

dendritic cells
Hematopoietic Stem Cells
Dendritic Cells
Genes
Neoplasm Antigens
neoplasms
antigens
hematopoietic stem cells
Tumors
genes
Galactosidases
stem cell factor
Melanoma-Specific Antigens
Gene transfer
galactosidases
Tumor-Infiltrating Lymphocytes
Molecules
genetic markers
Stem Cell Factor
antigen-presenting cells

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Transduction and differentiation of dendritic cells from human hematopoietic progenitor cells. / Reeves, M. E.; Rosenberg, S. A.; Hwu, P.

In: FASEB Journal, Vol. 10, No. 6, 1996.

Research output: Contribution to journalArticle

Reeves, M. E. ; Rosenberg, S. A. ; Hwu, P. / Transduction and differentiation of dendritic cells from human hematopoietic progenitor cells. In: FASEB Journal. 1996 ; Vol. 10, No. 6.
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