Address reprint requests to Dr Shuji Miyagawa, Division of Organ Transplantation, Department of Molecular Therapeutics, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan
(E-mail: [email protected])

Read the full text

About

PDF

Tools

Share a link

Email
Wechat
Bluesky

Abstract

Abstract: Background: The pig pancreas is considered to be the most suitable source of islets for xenotransplantation in patients with type I diabetes. The objective of this study was to assess the antigenicity of neonatal porcine islet-like cell clusters (NPCC), including the Galα1–3Galβ1–4GlcNAc-R (α-Gal) and Hanganutziu–Deicher (H–D) antigens, and the pathway involved in human complement activation. The efficiency of expression of human decay-accelerating factor (DAF: CD55) on NPCC by adenoviral transduction was also examined, and the functional capacity of DAF was also estimated.

Methods: The deposition of human natural antibodies, immunoglobulin (Ig)G and IgM, and the expression of α-Gal and H–D antigens on NPCC were investigated by FACS analysis. The downregulation in the antigenicity to human natural antibodies, including the α-Gal and H–D antigens on NPCC by treatment with tunicamycin, PDMP and neuraminidase were also examined. In addition, complement-mediated islet lysis was examined using factor D-deficient and C1-deficient sera. An adenovirus encoding DAF under the control of the cytomegalovirus promoter, Ad.pCMV-DAF, was then constructed, and used for transducing NPCC. The amelioration of complement-dependent cytotoxicity of the NPCC by the transduced DAF was assessed as an in vitro hyperacute rejection model of a pig to human xenograft.

Results: The NPCC clearly expressed the α-Gal epitope, and the human natural antibodies, IgG and IgM, and the anti-H–D antibody also reacted with the NPCC. Treatment of NPCC with tunicamycin led to a drastic reduction in the extent of deposition of IgG, indicating the importance of N-linked sugars on the islets, presumably related to α-Gal expression on N-linked sugars. Neuraminidase treatment indicated the presence of, not only the H–D antigen, but also other sialic acid antigens which reacted with the human natural antibody, especially IgG. The complement deposition of factor B on NPCC was clear, and the alternative pathway-mediated NPCC killing accounted for approximately 30% of that by the total complement pathway. On the other hand, approximately 90% of the NPCC could be transduced to express DAF by the adenovector, Ad.pCMV-DAF. The expressed DAF showed an approximately 50–62% suppression in complement-dependent NPCC lysis.

Conclusion: The origin of the antigenicity of NPCC is mainly N-linked sugars including α-Gal and sialic acid antigens, and NPCC expressed the transduced molecule in high efficiency by the adenovector.

References

1 Cooper DK. Clinical xenotransplantion – how close are we? Lancet 2003; 362: 557.
10.1016/S0140-6736(03)14118-9
CAS PubMed Web of Science® Google Scholar
2 Miyagawa S, Hirose H, Shirakura R et al. The mechanism of discordant xenograft rejection. Transplantation 1988; 46: 825.
10.1097/00007890-198812000-00007
CAS PubMed Web of Science® Google Scholar
3 Platt JL. New directions for organ transplantation. Nature 1998; 392(Suppl): 11.
10.1038/32023
CAS PubMed Web of Science® Google Scholar
4 Groth CG, Korsgren O, Tibell A et al. Transplantation of porcine fetal pancreas to diabetic patients. Lancet 1994; 344: 1402.
10.1016/S0140-6736(94)90570-3
CAS PubMed Web of Science® Google Scholar
5 Valdes-Gonzalez RA, Dorantes LM, Garibay GN et al. Xenotransplantation of porcine neonatal islets of Langerhans and Sertoli cells: a 4-year study. Eur J Endocrinol 2005; 153: 419.
10.1530/eje.1.01982
CAS PubMed Web of Science® Google Scholar
6 Murakami H, Nagashima H, Takahagi Y et al. Transgenic pigs expressing human decay-accelerating factor regulated by porcine MCP gene promoter. Mol Reprod Dev 2002; 61: 302.
10.1002/mrd.10043
CAS PubMed Web of Science® Google Scholar
7 Bennet W, Bjorkland A, Sundberg B et al. Expression of complement regulatory proteins on islets of Langerhans: a comparison between human islets and islets isolated from normal and hDAF transgenic pigs. Transplantation 2001; 72: 312.
10.1097/00007890-200107270-00026
CAS PubMed Web of Science® Google Scholar
8 Cooper DKC, Koren E, Oriol R. Oligosaccharides and discordant xenotransplantation. Immunol Rev 1994; 141: 31.
10.1111/j.1600-065X.1994.tb00871.x
CAS PubMed Web of Science® Google Scholar
9 Sandrin MS, Fodor W, Mouhtouris E et al. Enzymatic remodeling of the carbohydrate surface of a xenogenic cell substantially reduces human antibody binding and complement-mediated cytolysis. Nature Med 1995; 1: 1261.
10.1038/nm1295-1261
CAS PubMed Web of Science® Google Scholar
10 Tanemura M, Miyagawa S, Ihara Y et al. Significant downregulation of the major swine xenoantigen by N-acetylglucosaminyltransferase III gene transfection. Biochem Biophys Res Chem 1997; 235: 359.
10.1006/bbrc.1997.6784
CAS PubMed Web of Science® Google Scholar
11 Miyagawa S, Murakami H, Takahagi Y et al. Remodeling of the major pig xenoantigen by N-acetylglucosaminyltransferase III in transgenic pig. J Biol Chem 2001; 276: 39310.
10.1074/jbc.M104359200
CAS PubMed Web of Science® Google Scholar
12 Phelps CJ, Koike C, Vaught TD et al. Production of alpha 1,3-galactosyltransferase-deficient pigs. Science 2003; 299: 411.
10.1126/science.1078942
CAS PubMed Web of Science® Google Scholar
13 Nishimaki T, Kano K, Milgrom F. Hanganutziu–Deicher antigen and antibody in pathologic sera and tissues. J Immunol 1979; 122: 2314.
CAS PubMed Web of Science® Google Scholar
14 Komoda H, Miyagawa S, Kubo T et al. A study of the xenoantigenicity of adult pig islet cells. Xenotransplantation 2004; 11: 237.
10.1111/j.1399-3089.2004.00121.x
CAS Google Scholar
15 Korbutt GS, Elliot JF, Ziliang AO et al. Large scale isolation, growth, and function of porcine neonatal islet cells. J Clin Invest 1996; 97: 2119.
10.1172/JCI118649
CAS PubMed Web of Science® Google Scholar
16 Asaoka H, Nishinaka S, Wakamiya N, Matsuda H, Murata M. Two chicken monoclonal antibodies specific for heterophil Hanganutziu–Deicher antigens. Immunol Lett 1992; 32: 91.
10.1016/0165-2478(92)90205-3
CAS PubMed Web of Science® Google Scholar
17 Fukuta D, Miyagawa S, Kubo T et al. Effect of hybrid complement regulatory proteins on xenogeneic cells. Biochem Biophys Res Commun 2003; 306: 476.
10.1016/S0006-291X(03)00988-4
CAS PubMed Web of Science® Google Scholar
18 Uchibori E, Kitano E, Tsuji T, Kitamura H. Study on CH50 levels in factor D-depleted serum. Rinsho Byori 2002; 50: 815.
CAS PubMed Google Scholar
19 Nagaki K, Tsuboi M, Matsumoto M, Kitamura H. A simple one-step hemolytic assay of the first component of human complement (C1) with polyethylene glycol treated human serum: details of the assay procedures and its application to the C1 activation study. J Clin Lab Immunol 1986; 21: 141.
CAS PubMed Web of Science® Google Scholar
20 Platts-Mills TA, Ishizaka K. Activation of the alternate pathway of human complements by rabbit cells. J Immunol 1974; 113: 348.
CAS PubMed Web of Science® Google Scholar
21 Miyagawa S, Shirakura R, Iwata K et al. Effects of transfected complement regulatory proteins, MCP, DAF, and MCP/DAF hybrid, on complement-mediated swine endotherial cell lysis. Transplantation 1994; 58: 834.
10.1097/00007890-199410000-00015
CAS PubMed Web of Science® Google Scholar
22 Bennet W, Sundberg B, Lundgren T et al. Damage to porcine islets of Langerhans after exposure to human blood in vitro, or after intraportal transplantation to cynomologus monkeys: protective effects of sCR1 and heparin. Transplantation 2000; 69: 711.
10.1111/j.1399-3089.2004.00121.x
CAS PubMed Web of Science® Google Scholar
23 Rayat GR, Rajotte RV, Hering BJ, Binette TM, Korbutt GS. In vitro and in vivo expression of Gal alpha-(1,3)Gal on porcine islet cells is age dependent. J Endocrinol 2003; 177: 127.
CAS Google Scholar
24 Irie A, Koyama S, Kozutsumi Y, Kawasaki T, Suzuki A. The molecular basis for the absence of N-glycolylneuraminic acid in humans. J Biol Chem 1998; 273: 15866.
10.1074/jbc.273.25.15866
CAS PubMed Web of Science® Google Scholar
25 Morozumi K, Kobayashi T, Usami T et al. Significance of histochemical expression of Hanganutziu–Deicher antigens in pig, baboon and human tissues. Transplant Proc 1999; 31: 942.
10.1016/S0041-1345(98)01847-8
CAS PubMed Web of Science® Google Scholar
26 Murase A, Miyagawa S, Koma M et al. An attempt to downregulate the Hanganutziu–Deicher antigen by overexpression of glycosyltransferases. Transplant Proc 2000; 32: 2507.
10.1016/S0041-1345(00)01770-X
CAS PubMed Web of Science® Google Scholar
27 Kobayashi T, Yokoyama I, Suzuki A et al. Lack of antibody production against Hanganutziu–Deicher (H–D) antigens with N-glycolylneuraminic acid in patients with porcine exposure history. Xenotransplantation 2000; 7: 177.
10.1034/j.1399-3089.2000.00056.x
CAS PubMed Web of Science® Google Scholar
28 Zhu A, Hurst R. Anti-N-glycolylneuraminic acid antibodies identified in healthy human serum. Xenotransplantation 2002; 9: 376.
10.1034/j.1399-3089.2002.02138.x
CAS PubMed Web of Science® Google Scholar
29 Miwa Y, Kobayashi T, Nagasaka T et al. Are N-glycolylneuraminic acid (Hanganutziu–Deicher) antigens important in pig-to-human xenotransplantation? Xenotransplantation 2004; 11: 247.
10.1111/j.1399-3089.2004.00126.x
CAS PubMed Web of Science® Google Scholar
30 Fujita T. Evolution of the lectin-complement pathway and its role in innate immunity. Nat Rev Immunol 2002; 2: 346.
10.1111/j.0141-9838.2004.00665.x
CAS PubMed Web of Science® Google Scholar
31 Matsushita M, Fujita T. Activation of the classical complement pathway by mannose-binding protein in association with a novel C1s-like serine protease. J Exp Med 1992; 176: 1497.
10.1084/jem.176.6.1497
CAS PubMed Web of Science® Google Scholar
32 Schmidt P, Goto M, Le Mauff B, Anegon I, Korsgren O. Adenovirus-mediated expression of human CD55 or CD59 protects adult porcine islets from complement-mediated cell lysis by human serum. Transplantation 2003; 75: 697.
10.1097/01.TP.0000053249.39753.D6
CAS Google Scholar

Citing Literature

Volume13, Issue5

September 2006

Pages 455-464

A study of the xenoantigenicity of neonatal porcine islet-like cell clusters (NPCC) and the efficiency of adenovirus-mediated DAF (CD55) expression

Abstract

References

Citing Literature

References

Information

About Wiley Online Library

Help & Support

Opportunities

Connect with Wiley

A study of the xenoantigenicity of neonatal porcine islet-like cell clusters (NPCC) and the efficiency of adenovirus-mediated DAF (CD55) expression

Abstract

References

Citing Literature

References

Related

Information