Adoptive immunotherapy with liver allograft–derived lymphocytes induces anti-HCV activity after liver transplantation in humans and humanized mice
Masahiro Ohira, … , Kazuaki Chayama, Hideki Ohdan
Masahiro Ohira, … , Kazuaki Chayama, Hideki Ohdan
Published October 1, 2009
Citation Information: J Clin Invest. 2009;119(11):3226-3235. https://doi.org/10.1172/JCI38374.
View: Text | PDF
Research Article

Adoptive immunotherapy with liver allograft–derived lymphocytes induces anti-HCV activity after liver transplantation in humans and humanized mice

Abstract

After liver transplantation in HCV-infected patients, the virus load inevitably exceeds pre-transplantation levels. This phenomenon reflects suppression of the host-effector immune responses that control HCV replication by the immunosuppressive drugs used to prevent rejection of the transplanted liver. Here, we describe an adoptive immunotherapy approach, using lymphocytes extracted from liver allograft perfusate (termed herein liver allograft–derived lymphocytes), which includes an abundance of NK/NKT cells that mounted an anti-HCV response in HCV-infected liver transplantation recipients, despite the immunosuppressive environment. This therapy involved intravenously injecting patients 3 days after liver transplantation with liver allograft–derived lymphocytes treated with IL-2 and the CD3-specific mAb OKT3. During the first month after liver transplantation, the HCV RNA titers in the sera of recipients who received immunotherapy were markedly lower than those in the sera of recipients who did not receive immunotherapy. We further explored these observations in human hepatocyte–chimeric mice, in which mouse hepatocytes were replaced by human hepatocytes. These mice unfailingly developed HCV infections after inoculation with HCV-infected human serum. However, injection of human liver–derived lymphocytes treated with IL-2/OKT3 completely prevented HCV infection. Furthermore, an in vitro study using genomic HCV replicon–containing hepatic cells revealed that IFN-γ–secreting cells played a pivotal role in such anti-HCV responses. Thus, our study presents what we believe to be a novel paradigm for the inhibition of HCV replication in HCV-infected liver transplantation recipients.

Authors

Masahiro Ohira, Kohei Ishiyama, Yuka Tanaka, Marlen Doskali, Yuka Igarashi, Hirotaka Tashiro, Nobuhiko Hiraga, Michio Imamura, Naoya Sakamoto, Toshimasa Asahara, Kazuaki Chayama, Hideki Ohdan

×

Figure 5

Anti-HCV activity of IL-2/OKT3–treated liver lymphocytes was dependent on their IFN-γ secretion ability.

Options: View larger image (or click on image) Download as PowerPoint
Anti-HCV activity of IL-2/OKT3–treated liver lymphocytes was dependent o...
(A) IFN-γ was the major cytokine released from the cultured cells. The bar graphs indicate the concentrations of various cytokines (IFN-γ, TNF-α, IL-2, IL-4, IL-5, and IL-10) detected in the coculture supernatant by CBA. Data are presented as mean ± SEM (n = 3). (B) The effects of IL-2 and OKT3 (100 JRU/ml and 1 μg/ml, respectively) on IFN-γ production by stimulated CD3–CD56+ NK, CD3+CD56+ NKT, and CD3+CD56– T cells were evaluated by a combination of cell surface and cytoplasmic mAb staining and subsequent flow cytometric analysis. Histograms represent the log fluorescence intensities obtained upon staining for IFN-γ after gating of each fraction. Dotted lines represent negative control staining with isotype-matched mAbs. Horizontal lines indicate the gated portion of lymphocytes. GMean, geometric mean fluorescent intensity. (C) Blocking of IFN-γ with mAb (100 μg/ml) elucidated the marked role played by IFN-γ in producing the anti-HCV effect. The bar graphs indicate the luciferase activities of the cells in each group. Data are presented as mean ± SEM of a representative triplicate sample.