Sequences of Synthesis of Γ-1 Macroglobulin and Γ-2 Globulin Antibodies During Primary and Secondary Responses to Proteins, Salmonella Antigens, and Phage

Abstract

The nature of the antibodies produced by the rabbit during the primary and secondary responses to T2 phage, proteins, and the O and H antigens of Salmonella typhosa has been determined. Immune sera have been fractionated by zone electrophoresis, sucrose density ultracentrifugation, and anion exchange chromatography. The resulting fractions have been assayed by phage neutralization or hemagglutination (antisera to proteins) or bacterial agglutination. In confirmation and extension of earlier work from this laboratory, the primary response to these antigens, with the exception of the O antigen of the Salmonella, included the early synthesis of 19S, γ-1 globulin antibody, and the later synthesis of 7S, γ-2 globulin antibody. The primary response to the O antigen consisted of the synthesis of only a macroglobulin agglutinin. The secondary response to the proteins, including the H antigen of the Salmonella, comprised the early synthesis of large amounts of the 7S γ-2 globulin antibody to the same level attained during the primary response. The secondary response to the phage consisted in the synthesis of 7S, γ-2 globulin antibody alone. Treatment of the macroglobulin phage-neutralizing antibody with mercaptoethanol resulted in complete loss of its neutralizing activity. A working hypothesis to explain these observations was presented. A salient feature of this hypothesis was the suggestion that different cells synthesized the two distinct molecular forms of antibody. The significance of the sequential synthesis of the two forms of antibody is not known. It was proposed that the system for synthesis of macroglobulin antibody is an auxiliary system for antibody synthesis, perhaps the first to develop phylogenetically and ontogenetically. It is felt that the present observations indicate a clear-cut qualitative distinction between the primary and secondary responses to immunization whereby these responses might be identified in various experimental situations. It is also felt that these findings with the primary and secondary responses to various antigens in the rabbit may be of widespread occurrence in nature among a variety of species.

Metabolism of Human Gamma Macroglobulins*

“…At the same time, the 18 S antibodies would be predominantly located in the intravascular space while the smaller 6.6 S antibodies distribute themselves more widely. Since macroglobulin antibodies are formed first in response to antigen administration (7,9), intravascular retention of antibodies may have some significance in the early phase of the immune response. Later, with formation of 6.6 S antibodies, the large antibodies may no longer be needed or may even be deleterious, so that their rapid removal would be of value.…”

“…Antibodies such as the isohemagglutinins, typhoid 0 antibodies, and cold agglutinins are predominantly in the y,-macroglobulin group. Infants respond principally with macroglobulin antibodies (3)(4)(5), and in adult animals and man the response to many (perhaps all) antigens normally passes through an early stage in which antibody activity is entirely limited to the 'yi-macroglobulins (6)(7)(8)(9).…”

The Enhancement of 19s Antibody Production by Particulate Antigen

“…The phenomenon could depend upon a different organ localization of the particulate antigen although there is no clear evidence of preferential synthesis of a particular immunoglobulin type in any one tissue. The transient macroglobnlin antibody response to a single injection of most antigens is maintained only by further administration of antigen (Bauer, Mathies, and Stavitsky, 1963;Uhr and Finkelstein, 1963;Svehag and Mandel, 1964 b). The ability of acryl-bound antigen to enhance 19S production may be due to prolonged persistence of the antigen in macrophages as a result of its association with metabolically inert particles or perhaps to an increased uptake into these cells.…”

Expression of T-cell differentiation antigens on effector cells in cell-mediated cytotoxicity in vitro. Evidence for functional heterogeneity related to the surface phenotype of T cells.

“…We wished to determine whether the surface antigen phenotype of killer cells generated at an Mice TABLE VIII early stage of immunization would differ from that of hyperimmune donors, i.e . whether there might be some qualitative change in CMC-active T cells with progressive immunization, of the sort that characterizes the progression of antibody responses (29) . We knew from our own experience that CMC activity of PC as a whole is augmented by repeated immunization .…”