Immunotherapy with ciamexon in the non obese diabetic (NOD) mouse.

Ciamexon (CMX), a new immunomodulatory compound acting mainly on B-lymphocytes was given orally to 42 NOD mice divided into three sex and litter matched groups (A: 0.3 mg/mouse/day CMX, B: 1.5 mg/mouse/day CMX, C: control) from 7 weeks of age. Animals were followed up for evaluation of diabetes incidence up to 32 weeks of age. There was a tendency for a delayed onset of hyperglycemia in mice of group B up to 26 weeks of age; however no significant difference in the cumulative incidence of diabetes at 32 weeks of age was observed (A: 57.5%, B: 38.5%, C: 38.5%). No differences were found in the number of infiltrated islets in animals culled at 10 weeks of age treated with CMX from 4 weeks of age. We conclude that CMX does not modify the course of insulitis and diabetes incidence in NOD mice although though the appearance of glycosuria was delayed by this treatment.

Production of polyclonal antibodies in ascitic fluid of mice: time and dose relationships.

Further studies of the humoral immune response have been undertaken using ovine immunoglobulin G as the immunogen and BALB/C mice as the recipients. Mice immunized by the intraperitoneal route weekly for five weeks with an emulsion of the immunogen in Freund's complete adjuvant and given an intraperitoneal injection of pristane on day 14 produce relatively large volumes of ascitic fluid. A simple fluoroimmunoassay is used to determine specific antibody titres. The mice do not lose condition, remain active and eat normally throughout, although all show extensive granuloma formation within the peritoneal cavity. Tapping is performed before ascitic fluid production is excessive and each study is complete within 49 days, with a maximum of 5 taps. Only a single immunization site is required, since no higher antibody titres are obtained using combined intraperitoneal plus intramuscular plus subcutaneous immunization routes. A dose response study has shown that relatively large amounts (100 micrograms) of ovine immunoglobulin G must be injected to evoke a maximum response. Studies are in progress of various adjuvants and potential modulators of the immune response and of ways in which the immunogenicity of macromolecules can be changed. This approach can also be used to obtain antibodies for diagnostic purposes more rapidly than in rabbits and sheep.

Production of polyclonal antibodies in ascitic fluid of mice: technique and applications.

Balb/c mice immunised intraperitoneally with insulin developed significantly higher antibody concentrations in their serum than mice immunised subcutaneously. The antibody response was dose related, 50 ug amounts being more effective than either 20 ug or 5 ug amounts. In contrast, the antibody titres in the ascitic fluid of mice immunised with growth hormone were significantly higher after 5 ug when compared to 25 and 100 ug amounts. Repeated intraperitoneal injections of an emulsion of sheep IgG or human growth hormone in complete Freund's adjuvant, together with a single intraperitoneal injection of Pristane, induced ascites formation in most mice within 5 weeks. Up to 90 mL could be obtained from a single mouse by abdominal tapping, and the antibodies derived from the ascitic fluid proved suitable for application in a radioimmunoassay.