动物造模,药效评价,结核病 z-bio 2024-08-19 427

　　The non-human primate tuberculosis model is the closest animal model of tuberculosis to human tuberculosis. The most commonly used method of model preparation is to directly inject tuberculous bacteria into the right lower lobe or middle lobe of the lung through fiberoptic bronchoscopy. The clinical testing of experimental monkeys after infection includes: physical examinations, such as measurement of body weight and temperature; Blood tests, including blood cell analysis, erythrocyte sedimentation rate, C-reactive protein, etc; Isolation of bacteria from a series of lung lavage fluid and gastric extracts; Chest X-ray and CT examination; Tuberculin reaction. Cellular immune analysis in monkey blood, lung lavage fluid, and tissues includes cell proliferation experiments, flow cytometry, Elispot, T cell analysis, ELISA, etc.

　　1. Transtracheal infection model The monkeys were infected with H37Rv through the trachea, and under the condition of using extremely high bacterial amount (10 000~100 000 cfu), a multi lobar tuberculosis model with acute rapid progress and high mortality was made. Under the condition of using moderate amount of bacteria (1000 cfu), tuberculosis models with clinical symptoms, chronic activities, naked eye pulmonary lesions, meningitis, etc. were made. When using low bacterial load (10-100 cfu), it can become a latent tuberculosis infection model.

　　2. The method of directly injecting tuberculosis bacteria into the lower right or middle lobe of monkey lungs through fiberoptic bronchoscopy to prepare an infection model is currently the most commonly used model for tuberculosis infection in monkeys. The infection dose of tuberculosis bacteria ranges from tens of cfu to thousands of cfu, and the strains used for infection include H37Rv and Erdman strains. J. L. Flynn infected 17 macaques with H37Rv at a dose of 15-25 cfu through fiberoptic bronchoscopy. All monkeys were assessed for infection using the old tuberculin test, lymphocyte proliferation test, and PPD. After euthanasia, the experimental monkeys underwent histopathological examination. Four weeks after infection, the lymphocyte proliferation test was positive, six weeks later the tuberculin skin test was positive, and all tests were positive at eight weeks. At a certain time, the majority of monkey lung lavage fluid and gastric extract cultures were positive for tuberculosis bacteria. According to the condition of the disease, the macaques were divided into three groups. One group developed symptoms rapidly and showed symptoms 4 weeks after infection, and was euthanized at 12 weeks; Nine macaques were in the active/chronic group, and significant progressive lung infiltration was observed on chest X-rays 4-6 weeks after infection, with clinical symptoms but progressing slowly; Seven macaques were in the latent infection group, and those with latent infection had no clinical symptoms, only small lung infiltration in the early stages. The pathology of the lungs is similar to that of humans. All monkeys have caseous granulomas, mainly in the lymph nodes of the right porta hepatis. There are few monkeys with latent infections of granulomas, and most of them are fibrotic. David M. Lewinsohn infected four rhesus monkeys with 1000 cfu of H37Rv through fiberoptic bronchoscopy, and monitored the entire development of the disease through CT scanning. All infected monkeys developed progressive pulmonary tuberculosis and were euthanized at 12 weeks.