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Stimulation by means of dendritic cells followed by Epstein-Barr virus-transformed B cells as antigen-presenting cells is more efficient than dendritic cells alone in inducing Aspergillus f16-specific cytotoxic T cell responses. Clin Exp Immunol 2008 Feb;151(2):284-96

Date

11/17/2007

Pubmed ID

18005260

Pubmed Central ID

PMC2276953

DOI

10.1111/j.1365-2249.2007.03544.x

Scopus ID

2-s2.0-37849043608   17 Citations

Abstract

Adoptive immunotherapy with in vitro expanded antigen-specific cytotoxic T lymphocytes (CTLs) may be an effective approach to prevent, or even treat, Aspergillus (Asp) infections. Such lines can be generated using monocyte-derived dendritic cells (DC) as antigen-presenting cells (APC) but requires a relatively high volume of starting blood. Here we describe a method that generates Asp-specific CTL responses more efficiently using a protocol of antigen presented on DC followed by Epstein-Barr virus (EBV)-transformed B lymphoblastoid cell lines (BLCL) as APC. Peripheral blood mononuclear cells were stimulated weekly (2-5x) with a complete pool of pentadecapeptides (PPC) spanning the coding region of Asp f16 pulsed onto autologous mature DC. Cultures were split and stimulated subsequently with either PPC-DC or autologous PPC-pulsed BLCL (PPC-BLCL). Lines from the DC/BLCL arm demonstrated Asp f16-specific cytotoxicity earlier and to a higher degree than lines generated with PPC-DC alone. The DC/BLCL-primed lines showed a higher frequency of Asp f16-specific interferon (IFN)-gamma producing cells but an identical effector cell phenotype and peptide specificity compared to PPC-DC-only-primed lines. Tumour necrosis factor (TNF)-alpha, but not IL-10, appeared to play a role in the effectiveness of BLCL as APC. These results demonstrate that BLCL serve as highly effective APC for the stimulation of Asp f16-specific T cell responses and that a culture approach using initial priming with PPC-DC followed by PPC-BLCL may be a more effective method to generate Asp f16-specific T cell lines and requires less starting blood than priming with PPC-DC alone.

Author List

Zhu F, Ramadan G, Davies B, Margolis DA, Keever-Taylor CA

Author

David A. Margolis MD Interim Chair, Professor in the Pediatrics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Allergens
Antigen Presentation
Antigen-Presenting Cells
Antigens, Fungal
Antigens, Plant
Aspergillus
B-Lymphocytes
Cell Line
Cell Transformation, Viral
Cells, Cultured
Cytotoxicity, Immunologic
Dendritic Cells
Epitopes, T-Lymphocyte
Fungal Proteins
Herpesvirus 4, Human
Humans
Hyphae
Immunophenotyping
Interferon-gamma
Interleukin-10
T-Lymphocytes, Cytotoxic
Tumor Necrosis Factor-alpha