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Chlamydial hemagglutinin: interaction of ether-ethanol extracted fraction with sensitive erythrocytes
Chlamydial particles and soluble hemagglutinin were separated by differential centrifugation from the supernatant of L-cells and the allantoic fluid of chick embryo infected with C. psittaci 6BC and C. trachomatis TW-3. Concentrated hemagglutinin was fractionated with ether-ethanol; specimens were compared using sensitive erythrocytes of adult white Leghorn chickens. The ether-ethanol extract had a 40- to 80-fold higher hemagglutinin titer than the crude hemagglutinin or the ether-ethanol insoluble fraction. The extracted hemagglutinin also showed a higher complement-fixing activity than the other two fractions. Extracted hemagglutinin was stable for 3 months at 4 degrees and --70 degrees C when sonicated immediately before hemagglutinin; it agglutinated to a similar titer at 37 degrees, 25 degrees and 4 degrees C, showing the reaction to be temperature independent; it agglutinated to a similar titer within a pH range of 7.0 to 8.0 McIlvaine citrate buffer-saline solution and Dulbecco's phosphate buffer solution without Ca++ and Mg++ at pH 7.0 were both suitable for hemagglutinin-titration. Hemagglutination failed to take place in non-electrolyte solutions.
Chlamydial particles and soluble hemagglutinin were separated by differential centrifugation from the supernatant of L-cells and the allantoic fluid of chick embryo infected with C. psittaci 6BC and C. trachomatis TW-3. Concentrated hemagglutinin was fractionated with ether-ethanol; specimens were compared using sensitive erythrocytes of adult white Leghorn chickens. The ether-ethanol extract had a 40- to 80-fold higher hemagglutinin titer than the crude hemagglutinin or the ether-ethanol insoluble fraction. The extracted hemagglutinin also showed a higher complement-fixing activity than the other two fractions. Extracted hemagglutinin was stable for 3 months at 4 degrees and --70 degrees C when sonicated immediately before hemagglutinin; it agglutinated to a similar titer at 37 degrees, 25 degrees and 4 degrees C, showing the reaction to be temperature independent; it agglutinated to a similar titer within a pH range of 7.0 to 8.0 McIlvaine citrate buffer-saline solution and Dulbecco's phosphate buffer solution without Ca++ and Mg++ at pH 7.0 were both suitable for hemagglutinin-titration. Hemagglutination failed to take place in non-electrolyte solutions.