Differentiation of hemagglutinins in tissues infected withChlamydia

Crude, soluble, chlamydial hemagglutinin was prepared from allantoic fluid harvested from embryonated chick eggs and the supernatant fluid of mouse L cells infected with eitherChalamydia psittaci strain 6BC orChlamydia trachomatis strain TW-3. Control nonhemagglutinating specimens of uninfected allantoic fluid and mouse L cells were also prepared. The six preparations were separated by ether-ethanol extraction into lipid-rich and lipid-depleted fractions. Complement-fixing activity was found in the lipid-rich (but not in the lipid-depleted) fraction of infected preparations. In contrast, lipid-rich fractions of infected and uninfected preparations had similar agglutinating activity when sensitive erythrocytes of white Leghorn chickens were used. The lipid-rich fraction of infected and uninfected preparations was separated by thin-layer chromatography (TLC) into seven components with similarR_f values, hemagglutinating patterns, and chemical composition (lipid, protein, and carbohydrate). The highest hemagglutination titers of normal and infected preparations were found in a TLC fraction with similarR_f values and contained lipid, protein, and carbohydrate. This TLC fraction fromC. psittaci 6BC preparations was used in hemagglutination-inhibition studies. The results indicated that chlamydial hemagglutinin extracted by ether-ethanol and separated by TLC contained, in addition to specific hemagglutinin, nonspecific tissue-lipid hemagglutinin(s) identical to that found in normal preparations.