Asp f16 재조합 단백질 감작 수지상 세포를 이용한 시험관 내 Aspergillus fumigatus 특이 T 세포의 유도

• Published in: Journal of bacteriology and virology Vol. 35; no. 4; pp. 307 - 314
• Main Authors: 원희연, Hee Yeun Won, 김은경, Eun Kyung Kim, 손현정, Hyun Jung Sohn, 신완식, Wan Shik Shin, 민우성, Woo Sung Min, 김춘추, Chun Choo Kim, 김태규, Tai Gyu Kim
• Format: Journal Article
• Language: Korean
• Published: 대한바이러스학회 2005; 대한미생물학회
• Subjects: Asp f16; Aspergillus fumigatus; Dendritic cell; ELISPOT assay; Invasive aspergillosis; Dendritic cell; ELISPOT assay; Invasive aspergillosis; Aspergillus fumigatus; Asp f16
• ISSN: 1598-2467; 2093-0429

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 $^{51}Cr$ 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 $(31{\pm}12\;spots/10^4 cells)$, compared to that of PBMCs directly stimulated with rf16. p2+3 $(83{\pm}15\; spots/10^6 \;cells)$. $IFN-{\gamma}$ 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 gene ration of helper T cells in vitro. 본 연구는 침습성 국균증 (invasive aspergillosis)을 일으키는 Aspergillus fumigatus가 주요하게 발현하는 Asp f16의 고정된 염기서열 부분 (rf16. p2+3)을 E. coli에서 재조합 단백질 항원으로 생산하고, 수지상 세포에 감작시켜 시험관 내에서 항원 특이 Th1 세포를 성공적으로 유도할 수 있었으며, ELISPOT 측정법으로 세포성면역반응을 증명하였다. 이러한 결과는 침습성 국균증의 면역학적 기전을 연구하고 면역치료법 개발에 응용될 수 있을 것으로 사료된다.