[Antibody formation in the mouse by the ascites technic].
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Biomedical subjects
Publications and source records attributed to A Negro Ponzi.
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Human cord blood and adult red blood cells (rbc) and pigeon rbc were compared for sensitivity to rubella virus hemagglutination by a new method, consisting of the direct counting of free and aggregate red blood cells by using an hemocytometer chamber. With this method, more reliable than the "pattern test", hemagglutination is quantitated by the "aggregation index" (A.I.). Pigeon rbc were more sensitive than human rbc; cord blood and trypsinized adult human rbc showed the same sensitivity. However the kinetics of hemagglutination, evaluated by A.I. after different incubation times, were essentially the same with the three rbc types.
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Rubella virus hemagglutinating (HA) antigen shows different titers against non treated, trypsin-modified and cord blood human erythrocytes. Trypsin-modified erythrocytes give titers eleven fold higher, and cord blood red cells sixteen fold higher than those seen with the non treated cells. These differences in sensitivity agree with a different ability to bind HA antigen: with an excess of antigen cord blood erythrocytes bind two, and trypsin-modified erythrocytes three times as much antigen was bound by non treated cells. The amount of bound antigen shows little or no dependence on 4 degrees or 37 degrees C temperatures.
Rubella virus hemagglutination non-specific inhibitors, in sera fractionated on Bio-Gel A 5 m, elute in IgM containing fractions. In addition, in heparin-MnCl2 treated sera, the same inhibitors display a different chromatographic behaviour on Bio-Gel A 5m, being eluted earlier than IgM. Therefore, in order to avoid false-positive results due to the presence of non-specific inhibitors in IgM containing fractions it is necessary to verify the disappearance of hemagglutination-inhibition activity after 2-ME reduction.
High flow rates permit drastic reduction of time required in human serum fractionation by gel filtration giving relatively pure IgM. Fractionation of human serum on Bio-Gel A5m, Superose 6B and Sephacryl S-400, packed in 0.9 X 30 cm columns, at flow rate of 24 cm/h gives IgM fractions employable in diagnosis, in one hour time only.
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