Structure and Specificity of Guinea Pig 7s Antibodies

119

A previous report described the development of the immune response in rabbits to streptococcal group-specific carbohydrates following intravenous immunization with streptococcal vaccines (1). I n a continuation of these studies, certain rabbits were noted to develop an unusually high level of streptococcal group-specific antibodies. Examination of the zone electrophoresis patterns of these immune sera revealed a remarkably sharp and narrow band within the ,y-globulin region. This report is concerned with the immunologic, chemical, and physical properties of the specific 3,o-globulin isolated from these sera. The findings suggest a selective manufacture in certain rabbits of specific immune 3,G-globulin which exhibits a restricted range of electrophoretic properties compared to that of the normal complement of q-globulin. Materials and MethodsStreptococd.--Group A-variant strain A486 was obtained from Dr. R. C. Lancefield of the Rockefeller University.RabMts.--7-1b. New Zealand red rabbits were supplied by Baumgartd Bunny Farm, Marion, Iowa.Streptococcal Vaccine.--The vaccine was prepared from Group A-variant streptococci, strain A486. The bacteria were grown in 1000 ml of DiSco Todd-Hewitt broth, collected by centrifugation, and washed three times with saline. The collected bacteria were resuspended in 10 ml of saline, adjusted to pH 2.0, which contained 1 mg/ml of pepsin, and were incubated for 2 hr at 37°C. The treated bacteria were washed two times with saline and finally resuspended in 50 ml of formalized saline. The vaccine was stored at 4°C.

Section: Discussionsupporting

confidence: 92%

Characteristics of Streptococcal Group-Specific Antibody Isolated From Hyperimmune Rabbits

Osterland

Miller

Karakawa

et al. 1966

119

“…Our cells which switch from 19S to 7S production apparently continue to produce only 1 type of antibody-combining site per cell (13). Presumably this is due to changes in the synthesis and/or assembly of the subunits, but until uniformity of opinion on the location of the combining site has been achieved (26)(27)(28), it would be premature to attempt an interpretation of our results in terms of current concepts of globulin structure.…”

Section: Discussionmentioning

confidence: 79%

“…Immunoglobulins consist of L (light) chains and H (heavy) chains (26,28), and H chains from macroglobulins differ in a number of respects from those of 7S ~,-globulins (26,27). Our cells which switch from 19S to 7S production apparently continue to produce only 1 type of antibody-combining site per cell (13).…”

Section: Discussionmentioning

confidence: 99%

Single Cell Studies on 19s Antibody Production

Nossal

Szenberg

Ada

et al. 1964

274

“…There is general agreement that the L chains are contained in the S fragment (3,29,30) and also mounting evidence that both L chains (3, 31) and a piece of the H chain present in the S fragment (30,32) are involved in the acquisition of antibody specificity (33).…”

Section: Discussionmentioning

confidence: 96%

“…In this study, the electrophoretic mobilities of guinea pig antihapten antibodies have been investigated and compared because previous studies on structure and specificity made with similar purified antibody preparations have demonstrated reproducible differences in their urea starch gel electrophoretic patterns, after reduction and alkylation (3). A number of specifically purified guinea pig 7S antibodies were compared in agar gel electrophoresis, and distinct and characteristic differences in mobility have been observed which have been shown to be related to their immunological specificities.…”

Section: (From the Department Of Pathology New York University Schoomentioning

confidence: 99%

Differences in the Electrophoretic Mobilities of Guinea Pig 7s Antibodies of Different Specificities

Nussenzweig

Benacerraf

1964

Guinea pig 7S antibodies have been found to consist of two main populations, q'x and ~,~, with different electrophoretic mobilities (1). A broad electrophoretic heterogeneity can be shown, however, to exist within each of these two populations, as in 7S immune globulins of other mammalian species. In contrast, myeloma proteins formed in malignant plasmocytes, which probably represent the proliferation of clones of cells, are notable for their more homogeneous physicochemical properties (2). These proteins which have many of the characteristics of individual antibodies migrate as different but distinct peaks in zone eiectrophoresis. It is reasonable to assign the differences in electrophoretic mobility between normal immune globulins and myeloma proteins to the cell populations involved in their synthesis and to consider that differences in electrophoretic mobility should also be detected among antibodies in relation to their specificity, if the proper antigenic stimulation were used.In this study, the electrophoretic mobilities of guinea pig antihapten antibodies have been investigated and compared because previous studies on structure and specificity made with similar purified antibody preparations have demonstrated reproducible differences in their urea starch gel electrophoretic patterns, after reduction and alkylation (3). A number of specifically purified guinea pig 7S antibodies were compared in agar gel electrophoresis, and distinct and characteristic differences in mobility have been observed which have been shown to be related to their immunological specificities. Papain digestion (4) of these antibody preparations revealed that the electrophoretic mobility of one of the resulting fragments, which contains the antibody combining site (S fragment, reference 5), could be correlated with the mobility of the intact molecule.