Ex vivo expansion of human CD8+ T cells using autologous CD4+ T cell help

• Published in: PloS one Vol. 7; no. 1; p. e30229
• Main Authors: Butler, Marcus O, Imataki, Osamu, Yamashita, Yoshihiro, Tanaka, Makito, Ansén, Sascha, Berezovskaya, Alla, Metzler, Genita, Milstein, Matthew I, Mooney, Mary M, Murray, Andrew P, Mano, Hiroyuki, Nadler, Lee M, Hirano, Naoto
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 2012; Public Library of Science (PLoS)
• Subjects: Adenomatous polyposis coli; Animal models; Antigen-presenting cells; Antigens; Biology; Cancer; CD3 antigen; CD3 Complex - immunology; CD3 Complex - metabolism; CD4 antigen; CD4-Positive T-Lymphocytes - immunology; CD4-Positive T-Lymphocytes - metabolism; CD8 antigen; CD8-Positive T-Lymphocytes - drug effects; CD8-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - metabolism; Cell Proliferation; Cells, Cultured; Coculture Techniques; Cytokines; Cytokines - immunology; Cytokines - metabolism; Expansion; Flow Cytometry; Foxp3 protein; Genetic engineering; Genomics; Granulocytes; Growth factors; Histocompatibility antigen HLA; Humans; Immunoregulation; In vitro methods and tests; Influenza; Interferon; Interferon-gamma - immunology; Interferon-gamma - metabolism; Interleukin 2; Interleukin 21; Interleukin-2 - immunology; Interleukin-2 - metabolism; Interleukin-2 - pharmacology; Interleukins - immunology; Interleukins - metabolism; Interleukins - pharmacology; K562 Cells; Leukemia; Lymphocytes; Lymphocytes T; Lymphocytes, Tumor-Infiltrating - drug effects; Lymphocytes, Tumor-Infiltrating - immunology; Lymphocytes, Tumor-Infiltrating - metabolism; Medical prognosis; Medical research; Medical schools; Medicine; Melanoma; Metastasis; Monoclonal antibodies; Muromonab-CD3 - immunology; Muromonab-CD3 - metabolism; Muromonab-CD3 - pharmacology; Oncology; Receptors, Interleukin-21 - immunology; Receptors, Interleukin-21 - metabolism; Regulation; Rheumatoid arthritis; T cell receptors; T-Lymphocytes - immunology; T-Lymphocytes - metabolism; Tumors; Up-Regulation - drug effects; Womens health; Canada; Toronto Ontario Canada; United States > US; Japan; Massachusetts
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0030229

Using in vivo mouse models, the mechanisms of CD4+ T cell help have been intensively investigated. However, a mechanistic analysis of human CD4+ T cell help is largely lacking. Our goal was to elucidate the mechanisms of human CD4+ T cell help of CD8+ T cell proliferation using a novel in vitro model. We developed a genetically engineered novel human cell-based artificial APC, aAPC/mOKT3, which expresses a membranous form of the anti-CD3 monoclonal antibody OKT3 as well as other immune accessory molecules. Without requiring the addition of allogeneic feeder cells, aAPC/mOKT3 enabled the expansion of both peripheral and tumor-infiltrating T cells, regardless of HLA-restriction. Stimulation with aAPC/mOKT3 did not expand Foxp3+ regulatory T cells, and expanded tumor infiltrating lymphocytes predominantly secreted Th1-type cytokines, interferon-γ and IL-2. In this aAPC-based system, the presence of autologous CD4+ T cells was associated with significantly improved CD8+ T cell expansion in vitro. The CD4+ T cell derived cytokines IL-2 and IL-21 were necessary but not sufficient for this effect. However, CD4+ T cell help of CD8+ T cell proliferation was partially recapitulated by both adding IL-2/IL-21 and by upregulation of IL-21 receptor on CD8+ T cells. We have developed an in vitro model that advances our understanding of the immunobiology of human CD4+ T cell help of CD8+ T cells. Our data suggests that human CD4+ T cell help can be leveraged to expand CD8+ T cells in vitro.