ALTERATION OF IgG
نویسندگان
چکیده
Streptococcus pyogenes type 12, isolated from the pharynx of a patient with acute poststreptococcal glomerulonephritis, was subcultured in Todd-Hewitt broth and incubated at 37°C for 24 hr. The organisms were removed by centrifugation and washed with Dulbecco’s solution, and serial dilutions were plated on blood-agar plates; 1 ml of suspension containing 3 x 104 organisms was used for treatment of human IgG under different conditions. Human IgG was precipitated from pooled human serum with 50 per cent. ammonium sulphate and purified by gel-filtration on Sephadex G 200 (Flodin and Killander, 1962) followed by Chromatography on diethylaminoethyl (DEAE) cellulose (Tomasi and Kunkel, 1964). The IgG thus isolated was centrifuged at 15,000 r.p.m. and the supernatant fluid was tested for purity by immuno-electrophoresis and immunodiffusion in agar gel. The IgG was dialysed extensively against distilled water and was lyophilised. Eleven tests were set up. Test I. 100 mg of IgG was dissolved in 20 ml of sterile Todd-Hewitt broth and centrifuged at 15,000 r.p.m. The supernatant fluid was centrifuged at 15,000 r.p.m. and passed through a Millipore filter; 3 x 104 organisms were added, and the mixture was incubated at 37°C for 24 hr. Test II. The same procedure as in test I was performed, but incubation was at 4°C. Test III. 100 mg of IgG was dissolved in sterile Todd-Hewitt broth as in test I, but no
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