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Rapid, Large-Scale Generation of Highly Pure Cytomegalovirus-Specific Cytotoxic T Cells for Adoptive Immunotherapy
- Source :
- Journal of Hematotherapy & Stem Cell Research. 12:93-105
- Publication Year :
- 2003
- Publisher :
- Mary Ann Liebert Inc, 2003.
-
Abstract
- Adoptive transfer of donor-derived cytomegalovirus (CMV)-specific cytotoxic T cell (CTL) clones can restore immunity in allogeneic stem cell transplant recipients, providing protection against CMV disease. Current methods for selecting and expanding CMV-specific T cell clones are technically difficult, making adoptive T cell therapy impractical for routine clinical use. In this study, we describe a method for ex vivo generation and expansion of high-purity CMV-specific CTL using peptide-pulsed dendritic cells as antigen-presenting cells. Generation of CMV-specific CTL in numbers sufficient for clinical use in the time span of 4 weeks was accomplished in 6 of 8 CMV-seropositive donors. Examination of pp65 specificity by HLA/peptide tetramer staining demonstrated that a purity of greater than 95% peptide-specific cells could be obtained after two weekly stimulations and retained after further expansion for 3-4 weeks. Median expansion of total cell number was greater than 500-fold and expansion of peptide-specific CTL by tetramer staining was greater than 1.7 x 10(5)-fold. Four weeks after initiating CTL culture, we were able to generate greater than 10(9) total cells that specifically lysed target cells loaded with CMV peptide and cells infected with CMV. This simple and rapid method for generating high-purity CMV-specific CTL for adoptive immunotherapy is currently being examined for routine clinical use for allogeneic stem cell transplantation.
- Subjects :
- Adoptive cell transfer
Time Factors
Immunology
Adoptive immunotherapy
Cell Culture Techniques
Congenital cytomegalovirus infection
Antigen-Presenting Cells
Cytomegalovirus
Genes, MHC Class I
Enzyme-Linked Immunosorbent Assay
chemical and pharmacologic phenomena
Biology
Immunotherapy, Adoptive
Monocytes
Immunophenotyping
Viral Matrix Proteins
Immunity
medicine
Humans
Cytotoxic T cell
Models, Genetic
food and beverages
virus diseases
Dendrites
Dendritic Cells
Hematology
Flow Cytometry
Phosphoproteins
medicine.disease
Virology
CTL
Stem cell
Peptides
Stem Cell Transplantation
T-Lymphocytes, Cytotoxic
Subjects
Details
- ISSN :
- 15258165
- Volume :
- 12
- Database :
- OpenAIRE
- Journal :
- Journal of Hematotherapy & Stem Cell Research
- Accession number :
- edsair.doi.dedup.....33381c8e99dfdafa34317a857272ca44