Viral Interleukin-10-Engineered Autologous Hematopoietic Stem Cell Therapy: A Novel Gene Therapy Approach to Prevent Graft Rejection

Shashikumar K. Salgar, Dinghua Yang, Phillip Ruiz, Joshua Miller, Andreas G. Tzakis

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

The Epstein-Barr virus-encoded protein BCRF1 (viral interleukin [vIL]-10) is a biologically active homologue of cellular interleukin (IL)-10. In this study, a novel gene therapy approach to prolong allograft survival was designed. Autologous (syngeneic) hematopoietic progenitor/stem cell-enriched (HSC; lineage-ve) population derived from CBA/J mouse bone marrow were transduced with retrovirus encoding vIL-10 gene (vIL-10-HSC), ex vivo; vIL-10-HSC were injected (4-6 X 106 cells intravenously) into lethally (9.5 Gy) or sublethally (4 Gy) irradiated CBA/J mice. Six weeks after vIL-10-HSC administration, vascular heterotopic heart (C57BL/6) transplantation was performed. Ex vivo, the vIL-10-HSC produced 5.4 ± 0.5 ng of vIL-10 protein/2 X 105 cells per 24 hr. In vivo, serum vIL-10 production was 187 ± 205 pg/ml during 3-10 weeks after vIL-10-HSC administration. Cardiac allograft survival was prolonged (p < 0.004) in lethally (71 ± 40 days) and sublethally (114 ± 15 days) irradiated mice that received vIL-10-HSC compared to controls that received unengineered (UE) HSC or vector DNA-engineered HSC (12-16 days). However, secondary skin graft (C57BL/6) survival was not prolonged in cardiac allograft-tolerant animals. In the vIL-10-HSC-administered group, graft histopathology demonstrated mild arteritis/venulitis (grade 0.7) and rejection (grade 1.0). Intragraft expression of costimulatory molecules (B7.1, B7.2), cytokines (IL-2, IL-4, mIL-10, interferon [IFN]-γ), and inducible nitric oxide synthase (iNOS) molecules was markedly lower in vIL-10-HSC-treated tolerant grafts that survived more than 100 days compared to vector DNA-HSC- or UE-HSC-treated controls. Furthermore, T lymphocytes derived from vIL-10-HSC-treated tolerant recipients demonstrated hyporeactivity to donor antigens in mixed lymphocyte cultures. Administration of autologous vIL-10-engineered HSC prior to organ transplantation prolonged cardiac allograft survival significantly.

Original languageEnglish (US)
Pages (from-to)131-144
Number of pages14
JournalHuman gene therapy
Volume15
Issue number2
DOIs
StatePublished - Feb 1 2004

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ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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