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A previous report from this laboratory demonstrated that radioactive poliovirus could diffuse through agar to combine with electrophoretically separated colostrum and serum antibodies ( 1 ) . The technique as used was relatively insensitive, requiring concentrated, purified immunoglobulins and large volumes of poliovirus. A more sensitive technique, combining immunoelectrophoresis and radioautography with radioactive antigen as one of the reactants, has been described for demonstrating antibody activity. Thus antibodies which bind insulin, thyroglobulins and ragweed have been detected in the IgG,t IgA and IgM classes of immunoglobulins by this method(2-6). The adaptation and use of this radioimmunoelectrophoresis technique to the study of antibody directed against poliovirus is described in this paper.Material and methods. Antigen. P2 labelled type I poliovirus was prepared by growing virus in HeLa cell cultures with citrate-Eagle's medium, 5 % dialysed calf serum and sodium phosphate P2 (0.066 mc/ml). The labelled virus was purified by ultracentrifugation and DEAE column chromatography ( 7 ) . The preparation used for diffusion contained approximately 10T.O TIDS0 and 100,-000 cpm per 0.1 ml.Serum. Serum specimens were obtained from adult human males and females who had not been immunized within the past 2 years. Serum polio neutralizing antibodies were demonstrated by a standard one hour tube-neutralization method, using 100 TIDBo of poliovirus, in HeLa cell cultures(8). The *This research was supponted by U.S.P.H.S. Research Grant AI-05009-03 from Nat. Inst. M t h .t The World Health Organization (Bull. 447, World Health Organ., v30, 1%4) has recommended the following nomenclature for the classes ob immumglobulins: G Immunoglobulin (IgG) for 7s (y2) -globulin ; A Immunoglobulin (IgA) for yla-globulin and M Immunoglobulin (IgM) for y,,,-globulin. Sinai Hospital, New York Citysera were fractionated by a combination of DEAE and Sephadex G-2Q0 column chromatography (9). The type of immunoglobulin in the eluates was determined by radial immunodiffusion( 10). The neutralization titer of each fraction was determined and correlated with the type of immunoglobulin present in the fraction.Radioimmunoelectrophoresis was performed as described by Yagi (2) and Morse(3) using agar consisting of 1% Difco Special Agar Noble in a 25% solution of 0.1 M Verona1 buffer (pH 8.6) on microscope slides. After electrophoresis of the serum, rabbit antiserum directed against human serum was added to the antiserum troughs. As soon as the antiserum had diffused into the agar, the radioactive poliovirus preparation was added to the trough. The plates were incubated 24 hours at room temperature and then washed for 48 hours with a 1% solution of NaCl. The slides were dried and then placed in contact with Kodak Royal Pan film for one week. Exposure was carried out in total darkness. After removal from contact with the film, the slides were stained with amido black. In some experiments the antiserum was allowed to react with the electrophoretic...

Summary We have measured by radioimmunodiffusion (RID) the specific IgA and IgG polioantibody in the sera of 36 men and 36 women. We have found in 8 women the IgA polioantibody titer was equal to or greater than the IgG polioantibody titer. In 6 women, the IgA polioantibody was 50% of the IgG titer. No man was found to have an IgA polioantibody titer equal to 50% of his IgG polioantibody titer. In fact, 33 of the 36 men had an IgA polioantibody titer which was less than 20% of their IgG polioantibody titer. The other 3 men had an IgA polioantibody titer which was 25% of their IgG titer. These data show that the adult female population contains a significant number of women in whom the IgA polioantibody is equal to or greater than the IgG polioantibody. No such individuals were found among the adult male population. Thus we have demonstrated for the first time that there is a significant difference in the immunoglobulin pattern of the polioantibody in the serum of the adult male and the adult female.

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