Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2017 Jul 17;12(7):e0180768.
doi: 10.1371/journal.pone.0180768. eCollection 2017.

Anti-Neu5Gc and anti-non-Neu5Gc antibodies in healthy humans

Bingsi Gao  1   2   3 , Cassandra Long  1 , Whayoung Lee  1 , Zhongqiang Zhang  1   4 , Xiaotian Gao  5 , Doug Landsittel  6 , Mohamed Ezzelarab  1 , David Ayares  7 , Yuliang Huang  3 , David K C Cooper  1 , Yi Wang  2 , Hidetaka Hara  1
Affiliations

Anti-Neu5Gc and anti-non-Neu5Gc antibodies in healthy humans

Bingsi Gao et al. PLoS One. .

Abstract

Our group previously investigated the levels of anti-Gal and anti-nonGal IgM and IgG in a cohort of 75 healthy humans of various backgrounds, and found some significant differences related to factors such as age, gender, ABO blood group, diet, vaccination history, and geographic location during childhood. We have now expanded our cohort (n = 84) to investigate the levels of anti-Neu5Gc and anti-nonGal/nonNeu5Gc antibodies in healthy humans. Anti-nonGal and anti-nonGal/nonNeu5Gc human IgM and IgG binding to pRBCs and pAECs from GTKO/CD46 and GTKO/CD46/Neu5GcKO pigs were measured by flow cytometry. Anti-Gal and anti-Neu5Gc IgM and IgG levels were measured by ELISA. In summary, (i) the great majority (almost 100%) of humans had anti-Neu5Gc IgM and IgG antibodies that bound to pAECs and approximately 50% had anti-Neu5Gc antibodies that bound to pRBCs, (ii) there was significantly less human antibody binding to pig cells that did not express either Gal or Neu5Gc compared with those that did not express Gal alone, (iii) the levels of both IgM and IgG binding to GTKO/CD46/Neu5GcKO pRBCs and pAECs were low, (iv) the level of anti-Neu5Gc IgG was higher in men than women, (v) the level did not change with age or diet, and there was some variability associated with (vi) previous vaccination history and (vii) the geographic region in which the individual spent his or her childhood. Our study confirms that human antibody binding to RBCs and AECs from GTKO/CD46/Neu5GcKO pigs is greatly reduced compared to binding to GTKO/CD46 cells. However, all humans appear to have a low level of antibody that binds to pAECs that is not directed to either Gal or Neu5Gc. Our findings require consideration in planning clinical trials of xenotransplantation.

PubMed Disclaimer

Conflict of interest statement

Competing Interests: David Ayares is an employee of Revivicor, Inc, and receives a salary from Revivicor, Inc. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1
Fig 1. Expression of Gal, Neu5Gc, and hCD46 on wild-type, GTKO/CD46, GTKO/CD46/Neu5GcKO pig and human RBCs and AECs by flow cytometry.
RBCs and AECs from GTKO/CD46 pigs expressed Neu5Gc, but not Gal. AECs from these pigs expressed hCD46, but RBCs did not. Neither Gal nor Neu5Gc was expressed on GTKO/CD46/Neu5GcKO pig RBCs or AECs. Human AECs expressed hCD46, but not Gal or Neu5Gc.
Fig 2
Fig 2. Definitions of anti-pig IgM and IgG antibodies.
Anti-pig antibodies (both IgM and IgG) such as anti-Gal, anti-Neu5Gc, anti-nonGal, and anti-nonGal/nonNeu5Gc were defined according to binding to various pig cells.
Fig 3
Fig 3
[A] Human serum IgM/IgG binding to GTKO/CD46 and GTKO/CD46/Neu5GcKO pRBCs and pAECs (flow cytometry). There was no significant difference in IgM or IgG binding to GTKO/CD46 pRBCs or pAECs (P>0.05). IgM and IgG binding to GTKO/CD46/Neu5GcKO pRBCs were both significantly less than to the equivalent pAECs (P<0.01) although the levels of binding to pAECs were lower. [B] Difference between IgM/IgG binding to GTKO/CD46 and GTKO/CD46/Neu5GcKO pRBCs. There was a significant decrease in human IgM and IgG binding to GTKO/CD46/Neu5GcKO pRBCs when compared with binding to GTKO/CD46 pRBCs (P<0.01). IgM binding decreased in 75% of sera (63/84), though a very small number (3.6%, 3/84) were found to have very slightly increased (statistically insignificant) binding; the relative GM decreased from >8 to <1. IgG binding decreased in 65% of sera (55/84), and increased minimally in 2.4% (2/84); the relative GM was reduced from >5 to <1. [C] Difference between IgM/IgG binding to GTKO/CD46 and GTKO/CD46/Neu5GcKO pAECs. There was significantly less IgM and IgG binding to cells from GTKO/CD46/Neu5GcKO pAECs than to GTKO/CD46 pAECs (p<0.01). IgM binding was reduced in 100% of sera, and IgG in 93% of sera (78/84). Relative GM fell from 4 to 2 (IgM) and from 2 to 1.5 (IgG). No sera showed an increase in IgG binding to GTKO/CD46/Neu5GcKO cells, but 7.2% (6/84) showed no change in binding.
Fig 4
Fig 4
[A] Human IgM/IgG binding to Gal and Neu5Gc (ELISA). The level of anti-Gal IgM/IgG antibodies were higher than those of anti-Neu5Gc antibodies (P<0.0001). The level of anti-Gal and Neu5Gc IgG was significantly higher than that of anti-Gal and anti-Neu5Gc IgM (P<0.0001). [B] Correlation between human serum anti-Gal and anti-Neu5Gc IgM and IgG antibody levels. There is no correlation between anti-Gal level and anti-Neu5Gc level both in IgM and IgG (P>0.05). [C] Correlation of human serum anti-Neu5Gc IgM and IgG antibody with age (ELISA). There was no correlation between either anti-Neu5Gc IgM or IgG levels with age (P>0.05). [D] Difference in human anti-Neu5Gc antibody between gender (ELISA). There was no significant difference in anti-Neu5Gc IgM levels between female and male (P>0.05), but the level of anti-Neu5Gc IgG in males was significantly higher than in females (P<0.05).
Fig 5
Fig 5
[A] Correlation between anti-nonGal and anti-nonGal/nonNeu5Gc IgM and IgG antibody levels. When pRBCs were the target cells, there was no correlation between anti-nonGal IgM and anti-nonGal/nonNeu5Gc IgM (P>0.05). There was significant positive correlation between anti-nonGal IgG and anti-nonGal/nonNeu5Gc IgG (P<0.05). When pAECs were the target cells, there was highly significant positive correlation between anti-nonGal and anti-nonGal/nonNeu5Gc IgM/IgG (P<0.001). [B] Correlation between anti-Neu5Gc, anti-nonGal, and anti-nonGal/nonNeu5Gc IgM. When pRBCs were the target cells, there was a significant negative correlation between anti-Neu5Gc IgM and anti-nonGal/nonNeu5Gc IgM (P<0.05), but no correlation between anti-Neu5Gc IgM and anti-nonGal IgM. When pAECs were the target cells, there was no correlation between anti-nonGal IgM and anti-nonGal/nonNeu5Gc IgM (P>0.05). [C] Correlation between anti-Neu5Gc, anti-nonGal, and anti-nonGal/nonNeu5Gc IgG. When pRBCs were the target cells, there was a highly significant positive correlation between anti-Neu5Gc IgG and anti-nonGal IgG (P<0.001). When pAECs were the target cells, there was a significant positive correlation between anti-Neu5Gc IgG and anti-nonGal IgG (P<0.05). There is no correlation between anti-Neu5Gc IgG and anti-nonGal/nonNeu5Gc IgG both to pRBCs and pAECs (P>0.05).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Galili U, Shohet SB, Kobrin E, Stults CL, Macher BA. Man, apes, and Old World monkeys differ from other mammals in the expression of alpha-galactosyl epitopes on nucleated cells. J Biol Chem. 1988;263:17755–62. . - PubMed
    1. Good AH, Cooper DK, Malcolm AJ, Ippolito RM, Koren E, Neethling FA, et al. Identification of carbohydrate structures that bind human antiporcine antibodies: implications for discordant xenografting in humans. Transplant Proc. 1992;24:559–62. . - PubMed
    1. Oriol R, Ye Y, Koren E, Cooper DK. Carbohydrate antigens of pig tissues reacting with human natural antibodies as potential targets for hyperacute vascular rejection in pig-to-man organ xenotransplantation. Transplantation. 1993;56:1433–42. . - PubMed
    1. Kobayashi T, Cooper DK. Anti-Gal, alpha-Gal epitopes, and xenotransplantation. Subcell Biochem. 1999;32:229–57. . - PubMed
    1. Lexer G, Cooper DK, Rose AG, Wicomb WN, Rees J, Keraan M, et al. Hyperacute rejection in a discordant (pig to baboon) cardiac xenograft model. J Heart Transplant. 1986;5:411–8. . - PubMed

MeSH terms

LinkOut - more resources