Production in Mice of Ascitic Fluid Containing Antibodies Induced by Staphylococcus- or Salmonella- Adjuvant Mixtures

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Abstract
Summary: Substantial quantities of ascitic fluid containing antibodies were obtained from virtually 100% of mice injected with antigenic materials derived from either Staphylococcus aureus 18, Staphylococcus aureus 2 or Salmonella enteriditis mixed with incomplete Freund's adjuvant. High yields of ascitic fluid were elicited with 2–3 intraperitoneal inoculations of 0.4–0.5 ml of antigen-adjuvant mixtures administered at 3–7 day intervals. “Booster” doses caused a further increase in yield. Production of ascitic fluid, however, was not continuous for all the injected mice and for a 4-month period actual yield was 12–15 ml/mouse for persistent producers and 4–6 ml/mouse for the total number inoculated. Increased ascitic fluid yield was also obtained by concomitant employment of antigen-adjuvant mixtures and enzymes, hyaluronidase or trypsin. Ascites was elicited in four different strains of mice with production of substantial amounts of ascitic fluid comparable for all strains except A/HeN where yield was low. Heavier mice, weighing in excess of 20 g were found to be less susceptible to the lethal effects of inoculations with antigen-adjuvant mixtures. Studies of gross and microscopic pathology of ascitic mice revealed the presence of large numbers of plasma cells in peritoneal granulomas and in peripheral lymph nodes. Employing S. aureus 18-paraffin oil adjuvant mixtures, agglutinin titers showed an early rise (1:500–1:1000) which persisted at constant high levels for a period of 4–5 months. Comparable titers were found for the limited number of serums tested. Changes made in treatment of antigenic materials or in immunization schedules did not appreciably affect agglutinin titer. S. enteriditis protective antibodies were demonstrated in both immune mouse serum and immune mouse ascitic fluid. “Immune” ascitic fluid (0.005 ml) or immune serum (0.01 ml) afforded 50% protection to mice against S. enteriditis challenge. Injections of immunizing mixtures of S. enteriditis and paraffin oil by different routes produced comparable titers of agglutinating antibodies in the ascitic fluid. Total and differential counts of leukocytes in ascitic fluid typically revealed early in ascites 54,000 leukocytes/cu mm of which only 40% were mononuclears, whereas fluid obtained in late ascites contained 16,500 cells of which more than 90% were mononuclears. Eighty-eight per cent of the ascitic fluids measured for specific gravity fell in the range of 1.010 to 1.040. The γ-globulin average level for ascitic fluid was 0.86 g/100 ml and constituted on the average about 22% of the total protein. Protein levels for ascitic fluid were about 70% that for serum. Ascitic fluid was demonstrated to be a potential source of protective and agglutinating antibodies, viable leukocytes and γ-globulin. Induction of ascites in mice was accompanied by plasma cell proliferation in abdominal lymph nodes and granulomas.

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