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Glycosylation of Sera Thyroglobulin Antibody in Patients with Thyroid Diseases. Department of Endocrinology, Peking University First Hospital, Beijing 100034, China Ying Gao, Lanlan Zhao, Mingming Liu, Youyuan Huang, Guizhi Lu, Yanming Gao, Xiaohui Guo. Backgrounds.

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slide1
Glycosylation of Sera Thyroglobulin

Antibody in Patients with

Thyroid Diseases

Department of Endocrinology,

Peking University First Hospital,

Beijing 100034, China

Ying Gao,Lanlan Zhao, Mingming Liu, Youyuan Huang, Guizhi Lu, Yanming Gao, Xiaohui Guo

backgrounds
Backgrounds

Spencer CA, et al. J Clin Endocrinol Metab. 2011,96:3615-3627.

Calder EA, et al. Clin Exp Immunol. 1973, 14: 153-8.

  • Thyroglobulin antibody (TgAb) is one of the major autoantibodies in thyroid diseases.
    • In papillary thyroid cancer (PTC)
    • In autoimmune thyroid disease (AITD):

Hashimoto\' thyroiditis (HT) and Graves\' disease (GD)

backgrounds1
Backgrounds

Fab fragments

CH2 domains

Fc fragments

Fuc: fucose

Gal:galactose

Neu5AC: sialic acid

TgAb is predominantly of IgG class

IgG is a glycoprotein with a sugar moiety attached to each of the asparagin 297 residues in the CH2-domains of the two Fc-fragments.

Torrigiani G, et al. Clin Exp Immunol. 1968,3:621-630.

slide4

Backgrounds

Shields RL, et al. J Biol Chem. 2002,277:26733-26740.

Scallon BJ,et al. Mol Immunol. 2007,44:1524-1534.

Kodar K et al. Glycoconj J. 2012,29:57-66.

The absence of core fucose residues in the Fc glycans substantially increases the ADCC activity of IgG.

Increased sialylation of Fc glycans results in decreased ADCC activity.

Alteration of IgG-Fc galactosylation have been described not only in autoimmune diseases, but also in some malignancy diseases.

These glycoforms can differ in their efficacy of effector function activation as it influences binding of IgG molecules to Fc receptors and C1q .

objective
Objective

The aim of our study was to investigate the glycosylation of sera TgAb in patients with different thyroid diseases including HT, GD, and PTC.

materials and methods
Materials and Methods

Eu: euthyroidism; sH: subclinical hypothyroidism; H: hypothyroidism

Sera samples were collected in Peking University First Hospital.

slide7

Materials and Methods

the percentage of fucose positive control

x 100%

the percentage of TgAb IgG positive control

Thyroid function: Chemiluminescence immunoassays

TgAb IgG: antigen specific ELISAs

carbohydrate residues on sera TgAb: Lectin-ELISAs

The relative amount of fucose in each TgAb IgG was calculated as:

Comparisons were carried out by the Mann–Whitney test,ANOVA, Chi-Square test and Kruskal-Wallis H test.

slide8

Results

Table 1. Demographic data, thyroid functional status and TgAb IgG levels of the patients in different groups.

a P < 0.05 vs. GD; b P < 0.05 vs. thyroid cancer group.

slide9

Results

Table2. Demographic data, thyroid functional status and TgAb IgG levels of the patients in HT subgroups.

a P < 0.05 vs. sH.; b P < 0.05 vs. H.

slide10

Results

Fig.1. Comparisons of the relative amount of carbohydrate residues on each sera TgAb from patients with different thyroid diseases.

slide11

Results

Fig.2. Comparisons of the relative amount of carbohydrate residues on each serum TgAb from Hashimoto’s thyroiditis patients with different thyroid functional status.

slide12

Results

Fig.3. The correlation between the relative amount of carbohydrate residues on each TgAb and TgAb IgG in all the patients (n = 109).

slide13

Discussions

The levels of fucosylation and sialylation on TgAb varied in different thyroid diseases.

We speculated that TgAb with lower content of core fucose and terminal sialic acid might have stronger ability to participate in ADCC in HT.

Terminal galactose content of IgG does not affect ADCC but complement dependent cytotoxicity (CDC).

slide14

Discussions

There were no significant differences in the levels of glycosylation on each TgAb among the three HT subgroups.

The levels of glycosylation on TgAb might not representthyrocyte hyperplasia but merely reflect the capacity of inducing thyroid destruction

slide15

Discussions

Vanderpump MP, et al. Clin Endocrinol (Oxf). 1995, 43:55-68.

Parekh R, et al. J Autoimmun. 1989, 2:101-114.

The levels of glycosylation on each TgAb had a negative relationship with TgAb IgG levels

a consequence of elevated Ig synthesis by B cells.

the individuals with thyroid antibodies might be at high risk of developing thyroid failure.

slide16

Conclusions

Glycosylation of sera TgAb varied in the patients with different thyroid diseases.

The levels of glycosylation of TgAb might decrease with increasing TgAb levels.

acknowledgements
Acknowledgements

Supported by

Beijing Natural Science Foundation

Beijing Nova Program

Program for New Century Excellent Talents in University

Sector funds of ministry of health (no. 201002002).

Thank you for your attention!!

fig 1 the biotinylated lectins binding to tg in different oxidation time with sodium periodate
Fig.1. The biotinylated lectins binding to Tg in different oxidation time with sodium periodate.
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Fig.2. TgAb lgG binding to thyroglobulin with a serial dilution (diluted 1:12.5 - 1:800) in different oxidation time with sodium periodate.
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