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Bystander Transfer of Functional Human CD40 Ligand from Gene-Modified Fibroblasts to B-Chronic Lymphocytic Leukemia Cells
- Source :
- Human Gene Therapy. 14:545-559
- Publication Year :
- 2003
- Publisher :
- Mary Ann Liebert Inc, 2003.
-
Abstract
- CD40 ligand (CD40L) is a good candidate molecule for the immunotherapy of B cell malignancies including B-chronic lymphocytic leukemia (B-CLL), because it may increase the capacity of the malignant cells to present tumor antigens. However, efforts to manipulate expression of the human CD40L (hCD40L) molecule have foundered on problems associated with lack of consistent gene transfer into the malignant target cells. We now describe a new, highly reproducible method for inducing hCD40L surface expression on malignant B cells, which is dependent on intercellular transfer of the hCD40L protein from donor gene-modified fibroblasts to patient tumor cells. Ten B-CLL samples were cocultured with MRC-5 fibroblasts (a human embryonic lung cell line) previously transduced with an adenoviral vector encoding the hCD40L gene. The malignant cells expressed high levels of surface hCD40L, B7-1, B7-2, and ICAM-1 after coculture. Upregulation of B7-1 and B7-2 was cycloheximide inhibitable and was a consequence of CD40 activation. However, inhibition of protein synthesis had no effect on the ability of B-CLL cells to acquire surface expression of hCD40L. hCD40L surface expression required cell-to-cell contact, but was independent of CD40 engagement. hCD40L transfer was not mediated by membrane fusion. The transferred hCD40L was functionally intact and B-CLL cells expressing this molecule induced increased interferon-gamma production from autologous peripheral blood T lymphocytes. This approach does not use any direct gene transfer to primary leukemia cells and can readily be scaled up for production of clinical B-CLL vaccines.
- Subjects :
- medicine.medical_treatment
Genetic Vectors
CD40 Ligand
Biology
Cancer Vaccines
Adenoviridae
Cell Line
Viral vector
Immunoenzyme Techniques
Antigen
Transduction, Genetic
Antigens, CD
Tumor Cells, Cultured
Genetics
medicine
Humans
Coculture Technique
Molecular Biology
B cell
CD40
Histocompatibility Antigens Class I
Bystander Effect
Immunotherapy
Fibroblasts
medicine.disease
Leukemia, Lymphocytic, Chronic, B-Cell
Embryonic stem cell
Molecular biology
Coculture Techniques
Cell biology
Leukemia
medicine.anatomical_structure
Cell culture
biology.protein
Fibroblast
Molecular Medicine
Genetic Vector
Cancer Vaccine
Human
Subjects
Details
- ISSN :
- 15577422 and 10430342
- Volume :
- 14
- Database :
- OpenAIRE
- Journal :
- Human Gene Therapy
- Accession number :
- edsair.doi.dedup.....cae82fc6685b61e7ee342ec96adf0421