Induction of WT1-specific human CD8+ T cells from human HSCs in HLA class I Tg NOD/SCID/IL2rgKO mice.

Abstract

Induction of specific immune response against therapy-resistant tumor cells can potentially improve clinical outcomes in malignancies. To optimize immunotherapy in the clinic, we aimed to create an in vivo model enabling us to analyze human cytotoxic T-lymphocyte (CTL) responses against human malignancies. To this end, we developed NOD/SCID/IL2rgKO (NSG) mice expressing the HLA class I molecules HLA-A*0201 and A*2402. In the bone marrow (BM) and spleen of HLA class I transgenic (Tg) NSG mice transplanted with cord blood hematopoietic stem cells (HSCs), we found human memory CD8(+) T cells and antigen-presenting cells. To evaluate antigen-specific human CTL responses, we immunized HLA class I Tg NSG mice using polyinosinic:polycytidylic acid mixed Wilms tumor 1 (WT1) peptides, with or without WT1 peptide-loaded autologous dendritic cells. After immunization, the frequencies of HLA-restricted WT1-specific CTLs increased significantly in the spleen. Next, we transplanted the WT1-specific T-cell receptor (WT1-TCR) gene-transduced human HSCs into HLA class I Tg NSG newborn mice. WT1 tetramer-positive CD8(+) T cells differentiated from WT1-TCR-transduced HSCs in the recipients' BM, spleen, and thymus. Upon stimulation with WT1 peptide in vitro, these CTLs produced interferon-γ and showed lytic activity against leukemia cells in an antigen-specific, HLA-restricted manner. HLA class I Tg NSG xenografts may serve as a preclinical model to develop effective immunotherapy against human malignancies.

• Authors: Yuho Najima, Mariko Tomizawa-Murasawa, Yoriko Saito, Takashi Watanabe, Rintaro Ono, Toshiki Ochi, Nahoko Suzuki, Hiroshi Fujiwara, Osamu Ohara, Leonard D Shultz, Masaki Yasukawa, Fumihiko Ishikawa
• Journal: Blood (Blood) Vol. 127 Issue 6 Pg. 722-34 (Feb 11 2016) ISSN: 1528-0020 [Electronic] United States
• PMID: 26702062 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2016 by The American Society of Hematology.

Chemical References

• HLA-A*02:01 antigen
• HLA-A*24:02 antigen
• HLA-A2 Antigen
• HLA-A24 Antigen
• Il2rg protein, mouse
• Interleukin Receptor Common gamma Subunit
• WT1 Proteins
• WT1 protein, human

Topics

• Animals
• Animals, Newborn
• CD8-Positive T-Lymphocytes (metabolism, physiology)
• Cell Differentiation (genetics, immunology)
• Cells, Cultured
• Female
• HLA-A2 Antigen (genetics, metabolism)
• HLA-A24 Antigen (genetics, metabolism)
• Hematopoietic Stem Cells (physiology)
• Humans
• Interleukin Receptor Common gamma Subunit (genetics)
• K562 Cells
• Mice
• Mice, Inbred NOD
• Mice, SCID
• Mice, Transgenic
• T-Lymphocytes, Cytotoxic (immunology, metabolism)
• WT1 Proteins (metabolism)