TY - JOUR
T1 - Generation of peptide-specific CD8+ T cells by phytohemagglutinin-stimulated antigen-mRNA-transduced CD4+ T cells
AU - Naota, Hiroaki
AU - Miyahara, Yoshihiro
AU - Okumura, Satoshi
AU - Kuzushima, Kiyotaka
AU - Akatsuka, Yoshiki
AU - Hiasa, Atsunori
AU - Kitano, Shigehisa
AU - Takahashi, Toshitada
AU - Yuta, Atsushi
AU - Majima, Yuichi
AU - Shiku, Hiroshi
N1 - Funding Information:
We thank Ms. Junko Suzuki for technical assistance. This work was supported in part by Scientific Research on Priority Areas (nos. 17016031 and 17016089) and Grants-in-Aid for Scientific Research (no. 17591025), from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.
PY - 2006/7/31
Y1 - 2006/7/31
N2 - Functional analysis of antigen-specific CD8+ T cells is important for understanding the immune response in various immunological disorders. To analyze CD8+ T cell responses to a variety of antigens with no readily defined peptides available, we developed a system using CD4+ phytohemagglutinin (PHA) blasts transduced with mRNA for antigen molecules. CD4+ PHA blasts express MHC class I and II, and also CD80 and CD86 and are thus expected to serve as potent antigen presenting cells. EGFP mRNA could be transduced into and the protein expressed by more than 90% of either LCL or CD4+ PHA blasts. Its expression stably persisted for more than 2 weeks after transduction. In experiments with HLA-A*2402 restricted CD8+ CTL clones for either EBNA3A or a cancer-testis antigen, SAGE, mRNA-transduced lymphoid cells were appropriate target cells in ELISPOT assays or 51Cr releasing assays. Finally, using CD4+ PHA blasts transduced with mRNA of a cancer-testis antigen MAGE-A4, we successfully generated specific CTL clones that recognized a novel HLA-B*4002 restricted epitope, MAGE-A4223-231. Messenger RNA-transduced CD4+ PHA blasts are thus useful antigen presenting cells for analysis of CD8+ T cell responses and induction of specific T cells for potential immunotherapy.
AB - Functional analysis of antigen-specific CD8+ T cells is important for understanding the immune response in various immunological disorders. To analyze CD8+ T cell responses to a variety of antigens with no readily defined peptides available, we developed a system using CD4+ phytohemagglutinin (PHA) blasts transduced with mRNA for antigen molecules. CD4+ PHA blasts express MHC class I and II, and also CD80 and CD86 and are thus expected to serve as potent antigen presenting cells. EGFP mRNA could be transduced into and the protein expressed by more than 90% of either LCL or CD4+ PHA blasts. Its expression stably persisted for more than 2 weeks after transduction. In experiments with HLA-A*2402 restricted CD8+ CTL clones for either EBNA3A or a cancer-testis antigen, SAGE, mRNA-transduced lymphoid cells were appropriate target cells in ELISPOT assays or 51Cr releasing assays. Finally, using CD4+ PHA blasts transduced with mRNA of a cancer-testis antigen MAGE-A4, we successfully generated specific CTL clones that recognized a novel HLA-B*4002 restricted epitope, MAGE-A4223-231. Messenger RNA-transduced CD4+ PHA blasts are thus useful antigen presenting cells for analysis of CD8+ T cell responses and induction of specific T cells for potential immunotherapy.
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U2 - 10.1016/j.jim.2006.05.009
DO - 10.1016/j.jim.2006.05.009
M3 - Article
C2 - 16828790
AN - SCOPUS:33746905935
SN - 0022-1759
VL - 314
SP - 54
EP - 66
JO - Journal of immunological methods
JF - Journal of immunological methods
IS - 1-2
ER -