Isolation, ultrastructure and antigenicity of Mycoplasma gallisepticum membranes

The cell membrane of Mycoplasma gallisepticum was isolated by lysing the cells with digitonin. Chemical and density-gradient analyses and electron microscopy showed the isolated membranes to be relatively free of cytoplasmic contaminants. The density of the membranes exceeded that of other mycoplasma membranes, indicating a higher protein content. Small vesicular extensions seen in the sectioned membranes were interpreted as empty blebs.The isolated membranes, but not the cytoplasmic fraction, elicited in chickens the production of growth-inhibiting, agglutinating and haemagglutination-inhibition antibodies to M. gallisepticum in titres resembling those obtained by injection of whole cells. The peak of the serological response varied with the serological test employed. The rapid slide-agglutination test became positive as early as 3 days after the first injection of only 50 mug. of membrane protein. The haemagglutination-inhibition antibody titre reached its peak at about 10 days after the first injection, while that of the growth-inhibiting antibodies was reached only at about 25 days. The addition of adjuvant to the membrane antigen did not improve the production of the growth-inhibiting antibodies in chickens, but it produced some improvement in rabbits. Our results support the thesis that the chief immunogens of M. gallisepticum reside in the cell membrane of this organism.