动物造模,药效评价,SLE小鼠 z-bio 2024-08-01 385

　　[Modeling mechanism] Using activated lymphocytes from inbred mice as antigens to immunize homologous mice can induce the host to produce self anti dsDNA antibodies, anti nuclear antibodies, etc. Antigen antibody complexes deposit in renal tissue, causing immune complex glomerulonephritis, similar to lupus nephritis in humans.

　　[Modeling Method] Aseptic spleen and mesenteric lymph nodes were taken to prepare lymphocytes. The cell concentration was adjusted to 2 × 1000000 cells/ml using 1640 culture medium and divided into three groups. ConA (final concentration 10 μ g/ml), lipopolysaccharides (LPs, final concentration 10 μ g/ml), and IL-2 (final concentration 1000U/ml) were added to each group. The cells were cultured at 37 ℃ and 5% CO2 for 72 hours, and subcutaneously injected at a rate of 2 × 10000000 cells/mouse, once a week, for a total of three times.

　　Active cell nuclei and their chromatin were extracted from ConA activated spleen cells, while quiescent cell nuclei and their chromatin were extracted from non activated spleen cells. Inject subcutaneously with 2 × 10000000 nuclei per cell and 50 μ g chromatin, once per week, for a total of 3 times. Blood samples were collected from the posterior orbital venous plexus after the last immunization, and serum was isolated. Detect the generation of IgG class anti dsDNA antibodies. Take frozen sections of mouse kidneys and use direct immunofluorescence to detect the deposition of IgG immune complexes in the glomeruli.

　　High titer IgG anti dsDNA antibodies were detected in mice immunized with ConA activated T lymphocytes, LPS activated B lymphocytes, and IL-2 activated LAK cells. IgG class anti dsDNA antibodies were detected in both mice immunized with active cell nuclei and mice immunized with active chromatin. Mice with quiescent nuclear immunity and quiescent chromatin immunity did not exhibit IgG class anti dsDNA antibodies.

　　For mice with positive IgG anti dsDNA antibodies, direct immunofluorescence can be used to detect the deposition of IgG immune complexes in the kidneys of mice, which appear as patchy structures. Mice with negative IgG anti dsDNA antibodies did not show renal deposition of IgG immune complexes.

　　This model can be used for the etiology and pathogenesis of SLE, immune regulation of autoimmune diseases, immune complex nephritis, and pharmacological studies of SLE drugs.