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J Bacteriol. 1968 May; 95(5): 1572-1579
Copyright © 1968 American Society for Microbiology. All Rights Reserved.

Detection by Hemagglutination of Antibodies to Group A and Group E Streptococci by the Use of O-Stearoyl Derivatives of Their Cell Wall Carbohydrate-grouping Antigens

^a Department of Microbiology, Northwestern University Medical School, Chicago, Illinois 60611
^b Max-Planck Institut für Immunbiologie, Freiburg, Germany

ABSTRACT

The streptococcal group A and group E cell wall polysaccharide antigens were extracted with trichloroacetic acid from the cell or cell wall and esterified with stearic acid. The stearoyl derivatives contained 5 to 8% (by weight) of the ester. Sheep or human red blood cells were sensitized with the esterified antigens and were shown to agglutinate in the presence of specific rabbit antisera. Sera from (i) children hospitalized with group A streptococcal respiratory disease and (ii) swine possessing group E streptococcal lymphadenitis were shown to possess antibody titers significantly higher than the controls. The use of the two esterified antigens as controls for each other established the specificity of the reaction in each case. The general shape of the antigen-antibody precipitin curves was not changed when the stearoyl antigens were used; however, the quantitative aspects differed markedly. Oligosaccharides which inhibit the normal antigen-antibody precipitin reaction did not inhibit the hemagglutination reaction. The adsorption of antisera with whole streptococcal cells reduced the hemagglutination titer in relation to the quantity of cells employed. Data are given on the (i) optimal concentration of stearoyl antigen for sensitization, (ii) time of adsorption of antigen to red cells, (iii) use of albumin as diluting fluid, and (iv) condition of red cells. Properties of the esterified antigens and the mechanism of the agglutination reaction are discussed. The results indicate that polysaccharide antigens of other bacteria may be esterified and employed in a similar manner.

¹ Research Career Awardee (K6-GM-16284) of the National Institute of General Medical Sciences, National Institutes of Health.

² Present address: Sloan-Kettering Institute for Cancer Research, New York, N.Y. 10021.