| 制作方法Making method |
Construction of the BLTS humanized mouse model. In Step I, NSG mice are myoablated via γ irradiation (200 rads) using a Cesium-137 irradiator. In Step II, fetal tissues (spleen, thymus, and liver) are processed into 1 mm2 pieces and transplanted as a “sandwich” under the kidney capsule in irradiated NSG mice, following the administration of antibiotic and analogesic therapy and the induction of general anesthesia. In Step III, autologous CD34+ hematopoietic stem cells are isolated from the fetal liver via magnetic selection and transplanted at 2 × 10^5
cells per mouse via retro-orbital injection following kidney capsule transplantation of the lymphoid tissues. In Step IV, transplanted NSG mice were maintained under specific pathogen free conditions, and the human spleen and thymus organoids — along with other lymphoid tissues and associated immune cells — were allowed to develop over a period of 10
weeks, resulting in the BLTS humanized mouse model. 293T supernatant without HIV was used as mock control. Humanized mice with stable human leukocyte reconstitution were anesthetized and inoculated with mock or HIV-1 (~1 × 10^5 infectious units) i.v. (via retroorbital route) or via intravaginal route. ART in HIV-infected human immune system–humanized mice. Chronic HIV-infected humanized mice were treated with ART (daily i.p. injections emtricitabine [FTC, Cayman Chemicals, catalog 16879, 200 mg/ kg body weight], tenofovir disoproxil fumarate [TDF, Cayman Chemicals, catalog 16922, 200 mg/kg body weight], and raltegravir [RAL, Cayman Chemicals, catalog 16071, 80 mg/kg body weight) beginning at 8 weeks after infection for a duration of 4 weeks. |
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