Induction of Aspergillus fumigatus Specific T Cells using Dendritic Cells Pulsed with Asp f16 Recombinant Protein in vitro.
- Author:
Hee Yeun WON
1
;
Eun Kyung KIM
;
Hyun Jung SOHN
;
Wan Shik SHIN
;
Woo Sung MIN
;
Chun Choo KIM
;
Tai Gyu KIM
Author Information
1. Department of Microbiology College of Medicine, The Catholic University of Korea, Seoul 137-701, Korea. kimtg@catholic.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
Invasive aspergillosis;
Aspergillus fumigatus;
Asp f16;
Dendritic cell;
ELISPOT assay
- MeSH:
Antifungal Agents;
Aspergillosis;
Aspergillus fumigatus*;
Aspergillus*;
Clone Cells;
Dendritic Cells*;
Enzyme-Linked Immunospot Assay;
Immunocompromised Host;
Immunotherapy;
Lymphocytes;
T-Lymphocytes*;
T-Lymphocytes, Helper-Inducer;
Viperidae*
- From:Journal of Bacteriology and Virology
2005;35(4):307-314
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Invasive aspergillosis is the most common Aspergillus fumigatus infection in immunocompromised patients. Although the treatment of the invasive aspergillosis has been mostly relied on antifungal agents, there still exists the need for more effective therapy. To develop cellular immunotherapy specific for Aspergillus fumigatus, we generated specific T cells using dendritic cells (DCs) pulsed with an Aspergillus fumigatus derived recombinant protein in vitro and examined their functions. The f16. p2+3 region containing the conserved region of Asp f16 gene was cloned from Aspergillus fumigatus and the recombinant protein was produced in E. coli. IFN-gamma secretion from the T cells stimulated with recombinant f16. p2+3 (rf16. p2+3) was measured by enzyme linked immunospot (ELISPOT) assay and the cytolytic activity of the stimulated T cells by 51Cr release assay. The number of IFN-gamma secreting cells were significantly increased in the peripheral blood mononuclear cells (PBMCs) stimulated with the rf16. p2+3 pulsed DCs (31+/-12 spots/10(4) cells), compared to that of PBMCs directly stimulated with rf16. p2+3 (83+/-15 spots/10(6) cells). IFN-gamma ELISPOT assay using purified CD4+ or CD8+ as responder cells showed that CD4+ T cells (43 spots/10(4) cells) mainly produced IFN-gamma compared with CD8+ T cells (7 spots/10(4) cells). Furthermore, helper T cells specific for rf16. p2+3 could be efficiently generated by the stimulation with DCs for two weeks. However, cytotoxic T lymphocyte activity was not induced. Our results suggest that the rf16. p2+3 protein could be used for the generation of helper T cells in vitro.
