Fig. 4.

GSH prevents antibody-mediated agglutination of RBCs and antibody-sensitized microbeads. (A) Effect of GSH on antibody-mediated RBC agglutination. Mouse RBCs were exposed to the indicated dilutions of anti-mouse RBC antibody in the presence or absence of 5 mM GSH for 90 min. The formation of agglutinated RBCs was photographed. (B) Effect of GSH on antibody-initiated and complement-mediated RBC lysis. Mouse RBCs were exposed to the anti-mouse antibody (1 μg/ml) in the presence or absence of 5 mM GSH or NAC for 1.5 h. Afterward, 5% human serum were added as a source of complement. After reaction for 4.5 h, the supernatants were collected and measured for the absorbance at 405 nm. (C-G) The influence of GSH on the Easy-Titer IgG Assay. (C) Effect of GSH on antibody-elicited agglutination of the microbeads. The assay beads were incubated with the indicated concentrations of mouse anti-Thy-1 antibody that was pretreated with 3 mM GSH for 1 h. After reaction for 5 min, the agglutination of the microbeads was photographed. Note the obvious difference in the size of the aggregates between control and GSH-treated wells. (D) The standard curve generated from the OD obtained from a series of diluted mouse IgG in the presence or absence of 3 mM GSH. (E) The influence of GSH on the Easy-Titer IgG Assay. The same amount of anti-Thy-1 antibody in the presence or absence of 3 mM GSH was measured for their concentrations using the Easy-Titer IgG Assay Kit. Data shown are mean ±SE (n =4), ** P<0.01. (F) Effect of different concentrations of GSH on Easy-Titer IgG assay. The IgG assay was done in the presence of the indicated concentrations of GSH. Data shown are mean ±SE (n =4), * P<0.05; ** P<0.01. (G) Influence of NAC on Easy-Titer IgG assay. The IgG assay was done in the presence of 3 mM NAC. Data shown are mean ±SE (n =4). ** P<0.01.