Studies on Antibody Formation by Peritoneal Exudate Cells in Vitro

Abstract

Cells from peritoneal exudates of rabbits sacrificed 3 days after an intraperitoneal injection of sterile mineral oil were grown in tissue cultures in medium 199 (75 per cent); normal rabbit serum (25 per cent). Antibody produced by the cells was assayed by an hemagglutination technique in which the antigens used were adsorbed to formalinized tanned sheep erythrocytes. These sensitized cells agglutinate in the presence of antibody specific to the adsorbed antigen. It has been demonstrated that: Peritoneal exudate cells produced hemagglutinating antibody to bovine gamma globulin (BGG) in a replicating tissue culture system for approximately 3 weeks when taken from animals given either primary or secondary injections of BGG. The mean hemagglutinating titer was 30 for the primary and 32 for the secondary systems. Since the other cell types did not persist, it is felt that monocytes were responsible for these results. Monocytes taken from normal rabbits and exposed to either BGG or egg albumen (EA) in vitro produced titers of 28 for about 2 weeks. Monocytes taken from rabbits given hyperimmunizing injections of BGG produced titers of 147 for about 1 week. Endotoxin from Salmonella typhosa caused the monocytes to form antibody as if they had been taken from hyperimmunized rabbits. This was true both when the antigen was given in vivo together with the endotoxin as well as when the cells were exposed to antigen in vitro. The titers were 223 and 97, respectively. Neither freshly harvested nor cultured monocytes were phagocytic for carbon particles or bacteria in vitro. Monocytes in tissue culture appeared to assume the morphology of fibroblasts, but did not stain with the characteristics of fibroblasts. The morphologic changes and staining characteristics of monocytes in tissue culture have been described. The implications of these findings have been discussed and an attempt made to integrate them into general biological theory.

Antibody Formation in Vitro

“…The role of the macrophage in antibody production has been described by Burnet (17) as an entirely passive one; by removing excess antigen the macrophages merely protect antibody-producing cells from the repeated exposure to antigen which might exert a deleterious effect on these cells. The opposite view is held by a number of other workers who consider the macrophage as an active producer of antibody (4,18). Still others believe that the maerophage solubilizes antigens and thereby merely enhances the stimulus to antibody-producing cells (19).…”

Aspects of the Pathogenesis of Virus Diseases

“…It is difficult to avoid the conclusions that macrophages are involved in the initiation of the' immune response to viruses because of their primary importance in the phagocytosis of foreign materials. It has been suggested that macrophageg turn into antibody-producing cells (54), and that macrophages themselves produce antibody (92). It has also been suggested that macrophages prepare antigen for antibodyproducing cells (58,129,134), and the evidence for this has sometimes been good.…”

Transfer Agent of Immunity

“…McKenna and Stevens [12] described that antibody production against T2 bacteriophages or bovine serum albumin were observed in cultures of mononuclear cells from rabbit peritoneal exudates. Jacherts On the other hand, Burnet [3] described the role of macrophages in antibody formation as an entirely passive one; by removing excess antigen the macrophages protect antibody forming cells from the repeated exposure to antigen.…”