Original Article
The Introduction of Human Heme Oxygenase-1 and Soluble Tumor Necrosis Factor-α Receptor Type I With Human IgG1 Fc in Porcine Islets Prolongs Islet Xenograft Survival in Humanized Mice

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Apoptosis during engraftment and inflammation induce poor islet xenograft survival. We aimed to determine whether overexpression of human heme oxygenase-1 (HO-1) or soluble tumor necrosis factor-α receptor type I with human IgG1 Fc (sTNF-αR-Fc) in porcine islets could improve islet xenograft survival. Adult porcine islets were transduced with adenovirus containing human HO-1, sTNF-αR-Fc, sTNF-αR-Fc/HO-1 or green fluorescent protein (control). Humanized mice were generated by injecting human cord blood–derived CD34+ stem cells into NOD-scid-IL-2Rγnull mice. Both HO-1 and sTNF-αR-Fc reduced islet apoptosis under in vitro hypoxia or cytokine stimuli and suppressed RANTES induction without compromising insulin secretion. Introduction of either gene into islets prolonged islet xenograft survival in pig-to-humanized mice transplantation. The sTNF-αR-Fc/HO-1 group showed the best glucose tolerance. Target genes were successfully expressed in islet xenografts. Perigraft infiltration of macrophages and T cells was suppressed with decreased expression of RANTES, tumor necrosis factor-α and IL-6 in treatment groups; however, frequency of pig-specific interferon-γ–producing T cells was not decreased, and humoral response was not significant in any group. Early apoptosis of islet cells was suppressed in the treatment groups. In conclusion, overexpression of HO-1 or sTNF-αR-Fc in porcine islets improved islet xenograft survival by suppressing both apoptosis and inflammation. HO-1 or sTNF-αR-Fc transgenic pigs have potential for islet xenotransplantation.

basic (laboratory) research / science
islet transplantation
xenotransplantation
cytokines / cytokine receptors
innate immunity
rejection
cell death: apoptosis
xenoantigen

Abbreviations

Ad-
adenovirus containing
AUC
area under the curve
CHX
cycloheximide
ELISA
enzyme-linked immunosorbent assay
ELISPOT
Enzyme-Linked ImmunoSpot assay
GFP
green fluorescent protein
HO-1
heme oxygenase-1
HRP
horseradish peroxidase
IFN
interferon-γ
IP-10
interferon-γ–induced protein 10
IPGTT
intraperitoneal glucose tolerance test
LPS
lipopolysaccharide
MOI
multiplicity of infection
NSG
NOD-scid IL-2Rγnull
RT-PCR
reverse transcriptase–polymerase chain reaction
SEM
standard error of the mean;
sTNF-αR
soluble tumor necrosis factor-α receptor type I
sTNF-αR-Fc
soluble tumor necrosis factor-α receptor type I with human IgG1 Fc
TNF-α
tumor necrosis factor-α;
TUNEL
terminal deoxynucleotidyl transferase dUTP nick end labeling

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