Ann Lab Med.  2015 Nov;35(6):611-617. 10.3343/alm.2015.35.6.611.

Increase in Anti-Gal IgM Level is Associated With Early Graft Failure in Intraportal Porcine Islet Xenotransplantation

• Kang HJ
• Lee H
• Park EM
• Kim JM
• Shin JS
• Kim JS
• Park CG
• Kim SJ

• KMID: 2363264
• DOI: http://doi.org/10.3343/alm.2015.35.6.611

Abstract

BACKGROUND
Anti-Gal is a major antibody induced in non-human primates (NHPs) after xenotransplantation. To understand the mechanism of graft rejection, we investigated the association between anti-Gal responses and graft failure in NHP recipients of porcine islet transplantation (PITx).
METHODS
Intraportal PITx was performed in 35 diabetic NHPs, and graft function was monitored. Early graft failure (EGF) was defined as loss of graft function within a month after PITx. Seven, 19, nine NHPs received immunosuppression (IS) without CD40 pathway blockade (Group I), with anti-CD154 (Group II), and with anti-CD40 (Group III), respectively. The anti-Gal levels on day 0 and day 7 of PITx were measured by ELISA.
RESULTS
The frequency of EGF was significantly lower in Group II (26.3%) than in Group I (100%, P=0.0012) and Group III (77.8%, P=0.0166). While levels of anti-Gal IgG in Group I and anti-Gal IgM in Group III increased on day 7 compared with day 0 (P=0.0156 and 0.0273), there was no increase in either on day 7 in Group II. The ratio of anti-Gal IgM or IgG level on day 7 to that on day 0 (Ratio7/0) was significantly higher in recipients with EGF than without EGF (P=0.0009 and 0.0027). ROC curve analysis of anti-Gal IgM Ratio7/0 revealed an area under the curve of 0.789 (P=0.0003).
CONCLUSIONS
IS with anti-CD154 suppressed anti-Gal responses and prevented EGF in PITx. Anti-Gal IgM Ratio7/0, being associated with EGF, is a predictive marker for EGF.

Keyword

Pig; Non-human primate; Islets; Xenotransplantation; Antibody; Gal; Early graft failure

MeSH Terms

Animals
Antibodies/blood/immunology
Antigens, CD40/immunology
Area Under Curve
CD40 Ligand/immunology
Disaccharides/*immunology
Epidermal Growth Factor/blood
Graft Rejection/*immunology
Immunoglobulin G/blood
Immunoglobulin M/*blood
Immunosuppressive Agents/therapeutic use
*Islets of Langerhans Transplantation
Macaca mulatta
ROC Curve
Swine
Transplantation, Heterologous
Antibodies
Antigens, CD40
CD40 Ligand
Disaccharides
Epidermal Growth Factor
Immunoglobulin G
Immunoglobulin M
Immunosuppressive Agents

Figure

• 

  Fig. 1 Histogram of graft survival in 35 non-human primate recipients after porcine islet transplantation.

• 

  Fig. 2 Receiver operating characteristic (ROC) curves of anti-Gal IgM Ratio7/0 and IgG Ratio7/0 for the prediction of early graft failure (loss of graft function within a month) in porcine islet transplantation (PITx) analyzed from the data of all recipients (n=35, A) and from the data of the recipients receiving CD40 pathway blockade (n=28, B). Using an in-house ELISA, the levels of anti-Gal IgG and IgM were quantitatively measured in the plasma samples obtained from the rhesus monkey recipients prior to PITx (day 0) and on day 7 (±2) of PITx. The values of anti-Gal IgM Ratio7/0 and IgG Ratio7/0 for each recipient were calculated from the equation: (antibody level on day 7)/(antibody level on day 0). The area under the ROC curve (AUC) with P value, sensitivity, and specificity at a given optimal criterion are summarized.Abbreviation: CI, confidence interval.

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