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Generation of cytotoxic T lymphocytes specific for the prostate and breast tissue antigen TARP
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Oncology, Radiology and Clinical Immunology, Clinical Immunology.
2004 (English)In: The Prostate, ISSN 0270-4137, Vol. 61, no 2, 161-170 p.Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Expansion of cytotoxic T lymphocytes (CTL) directed against peptide epitopes from antigens that are specifically expressed by normal and neoplastic prostate epithelial cells has during the last years emerged as an interesting therapeutic approach to treat advanced prostate cancer. TCRgamma alternate reading frame protein (TARP) is a protein that in males is specifically expressed by normal prostate epithelial cells and prostate cancer cells. We have evaluated TARP for human leukocyte antigen (HLA)-A*0201-restricted peptides capable of triggering TARP-specific CTL. METHODS: Dendritic cells (DC) were pulsed either with synthetic peptides derived from the natural amino acid sequence of TARP or with cognate peptides having enhanced affinity for HLA-A*0201 due to an N-terminal anchor residue substitution. The peptide-pulsed DC were used to stimulate autologous T cells ex vivo. RESULTS: We were able to generate T cells against TARP(27-35) and TARP(4-13) and their mutated counterparts TARP(V28L)(27-35) and TARP(P5L)(4-13). The use of affinity-enhanced peptides resulted in the generation of T cells recognizing target cells displaying either wild-type or mutated peptide. We further show that TARP-specific T cells can be tetramer-sorted and subsequently expanded to large numbers by general T cell stimulation, with retained specificity and activity. Sorted and expanded T cells, obtained by stimulation with TARP(P5L)(4-13), exert moderate lysis of the TARP-expressing prostate cancer cell line, LNCaP, and breast cancer cell line, MCF-7, indicating that the TARP(4-13) epitope may be endogenously processed and presented by TARP-positive, HLA-A*0201-positive cells. CONCLUSIONS: Our findings suggest that synthetic TARP peptides, such as TARP(P5L)(4-13), may play a role in prostate and breast cancer immunotherapy.

Place, publisher, year, edition, pages
2004. Vol. 61, no 2, 161-170 p.
Keyword [en]
Antigens; Neoplasm/*immunology, Breast Neoplasms/*immunology, Cell Line; Tumor, Dendritic Cells/immunology, Female, Humans, Male, Nuclear Proteins/*immunology, Prostatic Neoplasms/*immunology, T-Lymphocytes; Cytotoxic/*immunology
National Category
Medical and Health Sciences
URN: urn:nbn:se:uu:diva-92711DOI: 10.1002/pros.20091PubMedID: 15305339OAI: oai:DiVA.org:uu-92711DiVA: diva2:165888
Available from: 2005-03-17 Created: 2005-03-17 Last updated: 2010-02-05Bibliographically approved
In thesis
1. Adoptive T Cell Therapy of Viral Infection and Cancer: Ex vivo Expansion of Cytomegalovirus- and Prostate Antigen-specific T Cells
Open this publication in new window or tab >>Adoptive T Cell Therapy of Viral Infection and Cancer: Ex vivo Expansion of Cytomegalovirus- and Prostate Antigen-specific T Cells
2005 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The main focus of my thesis has been to develop protocols for generating antigen-specific cytotoxic T lymphocytes (CTLs) and T helper cells (TH) for adoptive transfer to treat cytomegalovirus (CMV) disease and prostate cancer. CMV viremia is a severe complication in immunocompromised stem cell transplanted patients. Prostate cancer is a leading cause of death for men in Western countries. Although different in nature, CMV-infected cells and prostate cancer cells can both be eliminated through specific activation of the adaptive immune system.

To generate CMV pp65-specific T cells, I utilized dendritic cells (DCs) modified with an HLA-A*0201/pp65495-503 peptide, a recombinant adenovirus coding for pp65, in vitro transcribed pp65 mRNA and a recombinant pp65 protein. Peptide stimulation yielded large numbers of peptide-specific CD8+ T cells with high lytic activity while adenovirus or mRNA stimulation resulted in the expansion of CTLs against multiple pp65 epitopes. The recombinant protein activated primarily CD4+ TH cells. Stimulation with DCs co-modified with pp65 mRNA and pp65 protein simultaneously generated both pp65-specific CTLs and TH cells. Such T cells would cover all pp65 epitopes while avoiding potential virus related biohazards. The mRNA/protein combinatory approach can be used to stimulate T cells ex vivo from virtually all stem cell donors for adoptive T cell transfer.

I have identified two immunogenic HLA-A*0201-restricted peptide epitopes from the prostate tissue antigen TARP. Repeated stimulations with TARP peptide-pulsed DCs yielded up to 20% TARP-directed CD8+ T cells even when starting from undetectable frequencies (<0.01%). The T cells could be sorted to 99% purity and expanded 1000-fold with retained specificity and activity. We also detected TARP-directed CD8+ T cells in the blood of prostate cancer patients. Therefore, TARP seems to have potential as antigen in DC vaccination or adoptive T cell therapy of prostate cancer.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2005. 65 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 15
Immunology, Immunotherapy, Adoptive T cell therapy, Dendritic cell, T cell, Tetramer, Cytomegalovirus, Transplantation, pp65, Prostate cancer, TARP, Prostate antigens, Immunologi
National Category
Immunology in the medical area
urn:nbn:se:uu:diva-4821 (URN)91-554-6167-0 (ISBN)
Public defence
2005-04-15, Rudbecksalen, Rudbecklaboratoriet, Dag Hammarskölds väg 20, Uppsala, 13:15
Available from: 2005-03-17 Created: 2005-03-17Bibliographically approved

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