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Streptococcal IdeS: therapeutic potential for Guillain-Barré syndrome.

Takahashi R, Yuki N - Sci Rep (2015)

Bottom Line: We tested whether IdeS cleaved the anti-ganglioside IgG antibodies and inhibited deposition of activated complement component on ELISA plates.IdeS efficiently cleaved IgG and blocked complement activation mediated by anti-GM1, anti-GD1a and anti-GQ1b IgG antibodies.IdeS has therapeutic potential for GBS and related conditions.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.

ABSTRACT
Plasma exchange and intravenous immunoglobulin are effective in treating Guillain-Barré syndrome (GBS) probably because the former removes IgG autoantibodies and complement and the latter inhibits complement activation subsequent to the autoantibody binding to peripheral nerve antigens. IgG degrading enzyme of Streptococcus pyogenes (IdeS) can cleave the pathogenic autoantibodies into F(ab')2 and Fc. The purpose of this study is to show whether IdeS has novel therapeutic potential for GBS. Sera with anti-ganglioside IgG antibodies from 15 patients with GBS or Miller Fisher syndrome were used. We tested whether IdeS cleaved the anti-ganglioside IgG antibodies and inhibited deposition of activated complement component on ELISA plates. IdeS efficiently cleaved IgG and blocked complement activation mediated by anti-GM1, anti-GD1a and anti-GQ1b IgG antibodies. IdeS has therapeutic potential for GBS and related conditions.

No MeSH data available.


Related in: MedlinePlus

(A) Schema of IdeS treatment for binding of autoantibodies. Anti-ganglioside antibodies in the diluted patients’ sera bind to the ganglioside coating on the microtiter plates. IdeS cleaves IgG antibodies into F(ab’)2 and Fc fragments. Peroxidase-conjugated anti-human IgG (Fc) does not bind to the F(ab’)2 residue and does not work as the luminescent substrate in IdeS treated plates. (B) Concentration dependence of IdeS treatment. The representative change of GM1-IgG binding depends on its concentration. The clearance of IgG (Fc) accumulated to the highest levels at a concentration of 10 μg/ml or more of IdeS. (C) Time to react after adding IdeS. The binding of GM1-IgG halved after around 10 minutes and was almost eliminated after one hour. (D) The binding of IgG was detected by ELISA with both anti-Fc and anti-F(ab’)2 antibodies. Fc deposition was degraded by IdeS (10 μg/ml), whereas F(ab’)2 deposition remained unaltered. IdeS inhibited the Fc deposition of not only anti-GM1 IgG (n = 5) but also anti-GD1a IgG (n = 5) and anti-GQ1b IgG (n = 5) antibodies. (E) The binding of IgM was detected by ELISA. IdeS did not affect the binding of anti-GM1 IgM antibodies (n = 5).
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f1: (A) Schema of IdeS treatment for binding of autoantibodies. Anti-ganglioside antibodies in the diluted patients’ sera bind to the ganglioside coating on the microtiter plates. IdeS cleaves IgG antibodies into F(ab’)2 and Fc fragments. Peroxidase-conjugated anti-human IgG (Fc) does not bind to the F(ab’)2 residue and does not work as the luminescent substrate in IdeS treated plates. (B) Concentration dependence of IdeS treatment. The representative change of GM1-IgG binding depends on its concentration. The clearance of IgG (Fc) accumulated to the highest levels at a concentration of 10 μg/ml or more of IdeS. (C) Time to react after adding IdeS. The binding of GM1-IgG halved after around 10 minutes and was almost eliminated after one hour. (D) The binding of IgG was detected by ELISA with both anti-Fc and anti-F(ab’)2 antibodies. Fc deposition was degraded by IdeS (10 μg/ml), whereas F(ab’)2 deposition remained unaltered. IdeS inhibited the Fc deposition of not only anti-GM1 IgG (n = 5) but also anti-GD1a IgG (n = 5) and anti-GQ1b IgG (n = 5) antibodies. (E) The binding of IgM was detected by ELISA. IdeS did not affect the binding of anti-GM1 IgM antibodies (n = 5).

Mentions: The established assays demonstrated the binding of autoantibodies to each ganglioside and the deposition of active complement component. The detection of Fc domain was obscured due to cleavage by IdeS and subsequent rinsing out. Nevertheless F(ab’)2 remained to bind stably to the ganglioside coating on the microtiter plates (Fig. 1A and 2A). The clearance of Fc depended on the concentration of IdeS (Fig. 1B) as well as the time after the addition of IdeS to the serum (Fig. 1C). The cleaving effect emerged in a few minutes and reached the maximum in one hour. IdeS cleaved all the anti-GM1, anti-GD1a and anti-GQ1b IgG antibodies (Fig. 1D). Fc deposition was degraded by IdeS (10 μg/ml), whereas F(ab’)2 deposition remained unaltered. Thus, the subsequent complement deposition mediated by anti-ganglioside IgG autoantibodies was inhibited by IdeS, resulting in the blocking of the C3 deposition (Fig. 2B). The blocking effect depended on the concentration of IdeS as well as the clearance of Fc (Fig. 2C). In contrast, IdeS did not affect the binding of anti-GM1 IgM antibodies (Fig. 1E).


Streptococcal IdeS: therapeutic potential for Guillain-Barré syndrome.

Takahashi R, Yuki N - Sci Rep (2015)

(A) Schema of IdeS treatment for binding of autoantibodies. Anti-ganglioside antibodies in the diluted patients’ sera bind to the ganglioside coating on the microtiter plates. IdeS cleaves IgG antibodies into F(ab’)2 and Fc fragments. Peroxidase-conjugated anti-human IgG (Fc) does not bind to the F(ab’)2 residue and does not work as the luminescent substrate in IdeS treated plates. (B) Concentration dependence of IdeS treatment. The representative change of GM1-IgG binding depends on its concentration. The clearance of IgG (Fc) accumulated to the highest levels at a concentration of 10 μg/ml or more of IdeS. (C) Time to react after adding IdeS. The binding of GM1-IgG halved after around 10 minutes and was almost eliminated after one hour. (D) The binding of IgG was detected by ELISA with both anti-Fc and anti-F(ab’)2 antibodies. Fc deposition was degraded by IdeS (10 μg/ml), whereas F(ab’)2 deposition remained unaltered. IdeS inhibited the Fc deposition of not only anti-GM1 IgG (n = 5) but also anti-GD1a IgG (n = 5) and anti-GQ1b IgG (n = 5) antibodies. (E) The binding of IgM was detected by ELISA. IdeS did not affect the binding of anti-GM1 IgM antibodies (n = 5).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4508529&req=5

f1: (A) Schema of IdeS treatment for binding of autoantibodies. Anti-ganglioside antibodies in the diluted patients’ sera bind to the ganglioside coating on the microtiter plates. IdeS cleaves IgG antibodies into F(ab’)2 and Fc fragments. Peroxidase-conjugated anti-human IgG (Fc) does not bind to the F(ab’)2 residue and does not work as the luminescent substrate in IdeS treated plates. (B) Concentration dependence of IdeS treatment. The representative change of GM1-IgG binding depends on its concentration. The clearance of IgG (Fc) accumulated to the highest levels at a concentration of 10 μg/ml or more of IdeS. (C) Time to react after adding IdeS. The binding of GM1-IgG halved after around 10 minutes and was almost eliminated after one hour. (D) The binding of IgG was detected by ELISA with both anti-Fc and anti-F(ab’)2 antibodies. Fc deposition was degraded by IdeS (10 μg/ml), whereas F(ab’)2 deposition remained unaltered. IdeS inhibited the Fc deposition of not only anti-GM1 IgG (n = 5) but also anti-GD1a IgG (n = 5) and anti-GQ1b IgG (n = 5) antibodies. (E) The binding of IgM was detected by ELISA. IdeS did not affect the binding of anti-GM1 IgM antibodies (n = 5).
Mentions: The established assays demonstrated the binding of autoantibodies to each ganglioside and the deposition of active complement component. The detection of Fc domain was obscured due to cleavage by IdeS and subsequent rinsing out. Nevertheless F(ab’)2 remained to bind stably to the ganglioside coating on the microtiter plates (Fig. 1A and 2A). The clearance of Fc depended on the concentration of IdeS (Fig. 1B) as well as the time after the addition of IdeS to the serum (Fig. 1C). The cleaving effect emerged in a few minutes and reached the maximum in one hour. IdeS cleaved all the anti-GM1, anti-GD1a and anti-GQ1b IgG antibodies (Fig. 1D). Fc deposition was degraded by IdeS (10 μg/ml), whereas F(ab’)2 deposition remained unaltered. Thus, the subsequent complement deposition mediated by anti-ganglioside IgG autoantibodies was inhibited by IdeS, resulting in the blocking of the C3 deposition (Fig. 2B). The blocking effect depended on the concentration of IdeS as well as the clearance of Fc (Fig. 2C). In contrast, IdeS did not affect the binding of anti-GM1 IgM antibodies (Fig. 1E).

Bottom Line: We tested whether IdeS cleaved the anti-ganglioside IgG antibodies and inhibited deposition of activated complement component on ELISA plates.IdeS efficiently cleaved IgG and blocked complement activation mediated by anti-GM1, anti-GD1a and anti-GQ1b IgG antibodies.IdeS has therapeutic potential for GBS and related conditions.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.

ABSTRACT
Plasma exchange and intravenous immunoglobulin are effective in treating Guillain-Barré syndrome (GBS) probably because the former removes IgG autoantibodies and complement and the latter inhibits complement activation subsequent to the autoantibody binding to peripheral nerve antigens. IgG degrading enzyme of Streptococcus pyogenes (IdeS) can cleave the pathogenic autoantibodies into F(ab')2 and Fc. The purpose of this study is to show whether IdeS has novel therapeutic potential for GBS. Sera with anti-ganglioside IgG antibodies from 15 patients with GBS or Miller Fisher syndrome were used. We tested whether IdeS cleaved the anti-ganglioside IgG antibodies and inhibited deposition of activated complement component on ELISA plates. IdeS efficiently cleaved IgG and blocked complement activation mediated by anti-GM1, anti-GD1a and anti-GQ1b IgG antibodies. IdeS has therapeutic potential for GBS and related conditions.

No MeSH data available.


Related in: MedlinePlus