Generation of peptide-specific CD8+ T cells by phytohemagglutinin-stimulated antigen-mRNA-transduced CD4+ T cells

Hiroaki Naota, Yoshihiro Miyahara, Satoshi Okumura, Kiyotaka Kuzushima, Yoshiki Akatsuka, Atsunori Hiasa, Shigehisa Kitano, Toshitada Takahashi, Atsushi Yuta, Yuichi Majima, Hiroshi Shiku

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)54-66
Number of pages13
JournalJournal of Immunological Methods
Volume314
Issue number1-2
DOIs
Publication statusPublished - 31-07-2006
Externally publishedYes

Fingerprint

Phytohemagglutinins
T-Lymphocytes
Antigens
Messenger RNA
Peptides
Testicular Neoplasms
Antigen-Presenting Cells
Clone Cells
CD8 Antigens
Enzyme-Linked Immunospot Assay
HLA-A Antigens
HLA-B Antigens
Immunotherapy
Epitopes
Lymphocytes
Proteins

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Naota, Hiroaki ; Miyahara, Yoshihiro ; Okumura, Satoshi ; Kuzushima, Kiyotaka ; Akatsuka, Yoshiki ; Hiasa, Atsunori ; Kitano, Shigehisa ; Takahashi, Toshitada ; Yuta, Atsushi ; Majima, Yuichi ; Shiku, Hiroshi. / Generation of peptide-specific CD8+ T cells by phytohemagglutinin-stimulated antigen-mRNA-transduced CD4+ T cells. In: Journal of Immunological Methods. 2006 ; Vol. 314, No. 1-2. pp. 54-66.
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Naota, H, Miyahara, Y, Okumura, S, Kuzushima, K, Akatsuka, Y, Hiasa, A, Kitano, S, Takahashi, T, Yuta, A, Majima, Y & Shiku, H 2006, 'Generation of peptide-specific CD8+ T cells by phytohemagglutinin-stimulated antigen-mRNA-transduced CD4+ T cells', Journal of Immunological Methods, vol. 314, no. 1-2, pp. 54-66. https://doi.org/10.1016/j.jim.2006.05.009

Generation of peptide-specific CD8+ T cells by phytohemagglutinin-stimulated antigen-mRNA-transduced CD4+ T cells. / Naota, Hiroaki; Miyahara, Yoshihiro; Okumura, Satoshi; Kuzushima, Kiyotaka; Akatsuka, Yoshiki; Hiasa, Atsunori; Kitano, Shigehisa; Takahashi, Toshitada; Yuta, Atsushi; Majima, Yuichi; Shiku, Hiroshi.

In: Journal of Immunological Methods, Vol. 314, No. 1-2, 31.07.2006, p. 54-66.

Research output: Contribution to journalArticle

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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

PY - 2006/7/31

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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.

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