Lipid-mediated gene transfer of viral IL-10 prolongs vascularized cardiac allograft survival by inhibiting donor-specific cellular and humoral immune responses.

TitleLipid-mediated gene transfer of viral IL-10 prolongs vascularized cardiac allograft survival by inhibiting donor-specific cellular and humoral immune responses.
Publication TypeJournal Article
Year of Publication1998
AuthorsDeBruyne LA, Li K, Chan SY, Qin L, Bishop DK, Bromberg JS
JournalGene Ther
Date Published1998 Aug
KeywordsAnimals, Antibody Formation, Female, Gene Expression, Gene Transfer Techniques, Genetic Therapy, Genetic Vectors, Graft Rejection, Heart Transplantation, Immunity, Cellular, Immunosuppression, Interleukin-10, Liposomes, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Reverse Transcriptase Polymerase Chain Reaction, Transgenes, Transplantation, Homologous

The gene encoding the immunosuppressive cytokine viral interleukin-10 (vIL-10) was introduced into BALB/c (H-2d) vascularized cardiac allografts by perfusing the graft vasculature with DNA-liposome complexes, utilizing the experimental cationic lipid gamma AP DLRIE/DOPE and a plasmid encoding vIL-10 under the control of the HCMVie promoter. The DNA to lipid ratio and DNA dose were critical factors in obtaining optimal biologic effects. Gene transfer of vIL-10 with a 3:1 DNA to lipid weight ratio using 375 micrograms DNA significantly prolonged allograft survival in MHC-mismatched C57BL/6 (H-2b) recipients (16.00 days) compared with both unmodified allografts (8.14 days) and vIL-10 antisense controls (8.28 days). Enhanced graft survival was specific to vIL-10 expression since treatment with antisense plasmid or anti-vIL-10 monoclonal antibody (mAb) abrogated the effect. Prolonged survival was associated with a novel histology characterized by a moderate mononuclear infiltrate, edema, and diffuse fibrillar/collagen deposition in the interstitium. Despite these morphologic changes, myocytes remained viable and vessels were patent. Limiting dilution analysis revealed transient infiltration of IL-2 secreting, donor-reactive, helper T lymphocytes (HTL) and cytotoxic T lymphocytes (CTL) in vIL-10 expressing grafts on day 7, that decreased significantly by day 14. Similarly, vIL-10 gene transfer inhibited the accumulation of donor-specific HTL and CTL in the spleen, compared with antisense controls. Prolonged survival was also associated with a marked decrease in IgM and IgG alloantibody production, with little to no IgG isotype switching. These results show that viral IL-10 gene transfer inhibits graft rejection in a clinically relevant model by inhibiting donor-specific cellular and humoral immune responses.

Alternate JournalGene Ther
PubMed ID10326031
Grant ListP60-AR20557 / AR / NIAMS NIH HHS / United States
R01 AI-31946 / AI / NIAID NIH HHS / United States
T32 AI-07413 / AI / NIAID NIH HHS / United States
Related Faculty: 
Lihui Qin, M.D., Ph.D.

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