来源文献：Frequency of Human CD45+ Target Cells is a Key Determinant of Intravaginal HIV-1 Infection in Humanized Mice

PMID：29127409

DOI：10.1038/s41598-017-15630-z

NOD-Rag1tm1Mom Il2rgtm1Wjl (NRG) mice was obtained from Jackson Laboratories.Umbilical cord blood was obtained with parental consent from healthy newborns in the Department of Obstetrics and Gynecology at the McMaster University Medical Centre. Te umbilical cord blood was processed using the RosetteSep Human Cord Blood Progenitor Enrichment Kit from Stem Cell Technologies (Vancouver, BC, Canada), to purify and enrich CD34+ cells. Purifed CD34+ cells were cryopreserved until required. Four day old NRG pups were sub-lethally irradiated with a single-dose γ-ray (300 cGy) and then administered an intrahepatic injection with 1×10^6 to 2×10^6 CD34-enriched hematopoietic stem cells. Te degree of human immune cell reconstitution in the NRG mice was evaluated by fow cytometry 90–120 days following the intrahepatic injection by quantifying the expression of human leukocyte markers, hCD45, hCD3, hCD4, as well as mouse CD45 in peripheral blood mononuclear cells (PBMCs) to determine the percent reconstitution in each Hu-mouse. Hu-mice in the progesterone high/diestrus stage of the estrous cycle were anesthetized by IP injection of anesthetic (150 mg ketamine per kg/10 mg xylazine per mL). Mice were placed in a supine position, the vaginal tract was gently swabbed and an IVAG inoculation with a 25 µl dose of 10^3 (low) to 10^5(high) TCID50/mL of NL4.3-Bal-Env HIV-1 was delivered, by gently inserting the viral inoculum using a pipette.

诊断：病毒在肺和阴道中复制。

所有感染动物必须在ABSL-3室中进行。

1.该模型制备比较成熟，国内外都有此模型，构建和评价指标一致，所以该模型应用广泛，具有统一的操作规程。

2. 该模型制备的关键：1）小鼠必须是重度免疫缺陷的，而且饲养环境必须洁净；2）HIV操作需要生物安全。

3.此模型具有完善的人的免疫系统，HIV感染后，具有艾滋病病人相似的临床症状，所以具有很高的比较医学应用价值。