动物造模,衣原体附睾炎模型 z-bio 2021-11-01 327

　　(1) Replication method C.trachomatis MoPn Nigg Ⅱ strain was cultured and proliferated on HeLa-229 cells, and the infected monolayer cells were harvested. After 30 seconds of ultrasonic ultrasound, the supernatant was collected by centrifugation at 4°C at 500×g for 10 minutes. Centrifuge at 30000×g at 4°C for 45min, and dissolve the precipitate with SPG [0.2mol/L glucose, 20mmol/L phosphate (pH 7.2), 5mmol/L glutamic acid], and freeze at -70°C for use. Take 7-8 weeks old C3H/HeN(H-2k) mice anesthetized, place the abdomen up under a 4x dissecting microscope, pull out the foreskin of the penis, and place 2μl of CT inoculum containing 1000000IFU in the urethral opening to make The vaccination flows into the urethra. CT can be cultured from the bladder in the first 4 weeks after vaccination, CT can be isolated from the testis and epididymis in the first 2 weeks, and CT can also be isolated from the urethra at the 7th week.

　　(2) Characteristics of the model After experiments, it was found that C3H/HeN mice were vaccinated with CT in the urethra, and its ID50 was 5×10000IFU/ml. Inoculation of ID50 20 times the amount of bacteria from the urethra can cause acute and chronic epididymitis in mice. The method for making the model does not require surgical inoculation of the testis and epididymis, and is directly inoculated from the urethral opening, which is similar to the natural infection route of humans, and is simple to operate and easy to master. The symptoms and manifestations of model animals are similar to those of human infection. It can be used to study the pathogenic mechanism of CT, as well as preventive and therapeutic measures, and can be used to study the host's cellular and humoral immune mechanism to CT.

　　(3) Comparing the severity of CT infection in medical model animals is closely related to the route of inoculation. Mice were vaccinated with CT in the urethra. Acute inflammation of the urethra appeared after 1 week, and mixed acute and chronic inflammation appeared at 3 weeks, and started at 5 weeks. Manifested as chronic inflammation. A similar inflammatory reaction can also occur in the bladder, and the epididymis also has an acute inflammatory reaction in 2 weeks. Injecting CT directly from the vas deferens can cause more serious lesions, such as epididymal swelling and testicular atrophy. Mild to moderate mononuclear cell infiltration, interstitial fibrosis, and multifocal granulomas can still occur at 90 days.