Human antibody reactivity against xenogeneic N-glycolylneuraminic acid and galactose-α-1,3-galactose antigen

Sunghoon Hurh, Bohae Kang, Inho Choi, Bumrae Cho, Eun Mi Lee, Hwajung Kim, Young June Kim, Yun Shin Chung, Jong Cheol Jeong, Jong-Ik Hwang, Jae Young Kim, Byeong Chun Lee, Charles D. Surh, Jaeseok Yang, Curie Ahn

Research output: Contribution to journalArticle

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Abstract

Background: Despite the development of α1,3-galactosyl transferase-knockout (GTKO) pigs, acute humoral xenograft rejection caused by antibodies against non-Gal antigens, along with complement activation, are hurdles that need to be overcome. Among non-Gal antigens, N-glycolylneuraminic acid (Neu5Gc) is considered to play an important role in xenograft rejection in human. Methods: We generated human embryonic kidney 293 (HEK293) cells that expressed xenogeneic Neu5Gc (HEK293-pCMAH) or α1,3Gal (HEK293-pGT) antigen and investigated the degree of human antibody binding and complement-dependent cytotoxicity (CDC) against these antigens using 100 individual human sera. Results: Both IgM and IgG bound to α1,3Gal, while only IgG bound to Neu5Gc. Of the ABO blood groups, the degree of IgG binding to α1,3Gal was highest for blood group A. The degree of CDC against HEK293-pCMAH cells was significantly lower than that against HEK293-pGT cells. However, CDC against HEK293-pCMAH cells was significantly higher than that against control HEK293 cells. In addition, the severity of CDC against HEK293-pCMAH cells positively correlated with that against GTKO pig aortic endothelial cells (PAECs), suggesting that Neu5Gc is the main antigen in GTKO PAECs. Similar to antibody-binding activity, only IgG binding correlated with CDC against HEK293-pCMAH cells. The most common subclass of IgGs against Neu5Gc was IgG1, which typically induces strong complement activation. Conclusions: We showed that IgG-mediated CDC was detected in Neu5Gc-overexpressed HEK293 cells incubated with human sera; however, this antibody reactivity to Neu5Gc was highly variable among individuals. Our results suggest that additional modifications to the CMAH gene should be considered for widespread use of pig organs for human transplants.

Original languageEnglish
Pages (from-to)279-292
Number of pages14
JournalXenotransplantation
Volume23
Issue number4
DOIs
Publication statusPublished - 2016 Jul 1

Fingerprint

Galactose
Antigens
Antibodies
Kidney
Immunoglobulin G
Swine
Transferases
Complement Activation
N-glycolylneuraminic acid
Blood Group Antigens
Heterografts
Endothelial Cells
Serum
Immunoglobulin M

Keywords

  • antibody
  • antibody isotype
  • complement activation
  • Neu5Gc
  • xenoantigen

ASJC Scopus subject areas

  • Immunology
  • Transplantation

Cite this

Human antibody reactivity against xenogeneic N-glycolylneuraminic acid and galactose-α-1,3-galactose antigen. / Hurh, Sunghoon; Kang, Bohae; Choi, Inho; Cho, Bumrae; Lee, Eun Mi; Kim, Hwajung; Kim, Young June; Chung, Yun Shin; Jeong, Jong Cheol; Hwang, Jong-Ik; Kim, Jae Young; Lee, Byeong Chun; Surh, Charles D.; Yang, Jaeseok; Ahn, Curie.

In: Xenotransplantation, Vol. 23, No. 4, 01.07.2016, p. 279-292.

Research output: Contribution to journalArticle

Hurh, S, Kang, B, Choi, I, Cho, B, Lee, EM, Kim, H, Kim, YJ, Chung, YS, Jeong, JC, Hwang, J-I, Kim, JY, Lee, BC, Surh, CD, Yang, J & Ahn, C 2016, 'Human antibody reactivity against xenogeneic N-glycolylneuraminic acid and galactose-α-1,3-galactose antigen', Xenotransplantation, vol. 23, no. 4, pp. 279-292. https://doi.org/10.1111/xen.12239
Hurh, Sunghoon ; Kang, Bohae ; Choi, Inho ; Cho, Bumrae ; Lee, Eun Mi ; Kim, Hwajung ; Kim, Young June ; Chung, Yun Shin ; Jeong, Jong Cheol ; Hwang, Jong-Ik ; Kim, Jae Young ; Lee, Byeong Chun ; Surh, Charles D. ; Yang, Jaeseok ; Ahn, Curie. / Human antibody reactivity against xenogeneic N-glycolylneuraminic acid and galactose-α-1,3-galactose antigen. In: Xenotransplantation. 2016 ; Vol. 23, No. 4. pp. 279-292.
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AU - Hurh, Sunghoon

AU - Kang, Bohae

AU - Choi, Inho

AU - Cho, Bumrae

AU - Lee, Eun Mi

AU - Kim, Hwajung

AU - Kim, Young June

AU - Chung, Yun Shin

AU - Jeong, Jong Cheol

AU - Hwang, Jong-Ik

AU - Kim, Jae Young

AU - Lee, Byeong Chun

AU - Surh, Charles D.

AU - Yang, Jaeseok

AU - Ahn, Curie

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N2 - Background: Despite the development of α1,3-galactosyl transferase-knockout (GTKO) pigs, acute humoral xenograft rejection caused by antibodies against non-Gal antigens, along with complement activation, are hurdles that need to be overcome. Among non-Gal antigens, N-glycolylneuraminic acid (Neu5Gc) is considered to play an important role in xenograft rejection in human. Methods: We generated human embryonic kidney 293 (HEK293) cells that expressed xenogeneic Neu5Gc (HEK293-pCMAH) or α1,3Gal (HEK293-pGT) antigen and investigated the degree of human antibody binding and complement-dependent cytotoxicity (CDC) against these antigens using 100 individual human sera. Results: Both IgM and IgG bound to α1,3Gal, while only IgG bound to Neu5Gc. Of the ABO blood groups, the degree of IgG binding to α1,3Gal was highest for blood group A. The degree of CDC against HEK293-pCMAH cells was significantly lower than that against HEK293-pGT cells. However, CDC against HEK293-pCMAH cells was significantly higher than that against control HEK293 cells. In addition, the severity of CDC against HEK293-pCMAH cells positively correlated with that against GTKO pig aortic endothelial cells (PAECs), suggesting that Neu5Gc is the main antigen in GTKO PAECs. Similar to antibody-binding activity, only IgG binding correlated with CDC against HEK293-pCMAH cells. The most common subclass of IgGs against Neu5Gc was IgG1, which typically induces strong complement activation. Conclusions: We showed that IgG-mediated CDC was detected in Neu5Gc-overexpressed HEK293 cells incubated with human sera; however, this antibody reactivity to Neu5Gc was highly variable among individuals. Our results suggest that additional modifications to the CMAH gene should be considered for widespread use of pig organs for human transplants.

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