HIV EcoHIV 腹腔感染MORKO小鼠模型

来源文献：An infectious murine model for studying the systemic effects of opioids on early HIV pathogenesis in the gut.

PMID:25502600

DOI:10.1007/s11481-014-9574-9

Male or female C57Bl/6 and BALB/c mice were purchased from the Jackson Laboratory, while Mu Opioid Receptor Knock Out (MORKO) and Toll-Like Receptor 4 Knock Out (TLR4KO) mice were bred in house. Allanimals were between 10 and 16 weeks. Mice were implanted with either a placebo or 25 mg slowrelease morphine pellet at 24 h prior to infection with EcoHIV. Pellets were generously provided by NIH/NIDA. For long term experiments (>7 days), mice were implanted with either 25 mg or 75 mg morphine pellets using a protocol that would increase released morphine every 7 days to avoid tolerance. Escalating doses in the order used were 1× 25 mg, 1× 75 mg, 2× 25 mg, and 2× 75 mg based on a study that showed 2× 25 mg morphine pellets resulted in higher morphine release than 1× 75 mg pellet, likely due to increased surface area with having two pellets. The first dose given (implant at day 0, exposed 0–7 days) was 25mg slow-release morphine pellet as in short term experiments. The second week (implant at day 7, exposed 7–14 days), mice were pelleted with a 75mg slow-release morphine pellet. The third week (implant at day 14, exposed 14–21 days) mice were pelleted with 2× 25 mg pellets and the final week (implant at day 21, exposed 21–28 days) with 2× 75 mg pellet.Mice were injected in the intraperitoneal cavity with 1 mL of either saline or 1×10^6 pg p24 units as measured by p24 ELISA following manufacturer’s instructions (ZeptoMetrix Corporation).

1、1. SIVmac251静脉感染恒河猴外周血病毒载量结果

8只SIVmac251静脉感染恒河猴均在感染后7d检出病毒载量阳性，之后迅速上升，在感染后14d达到高峰，峰值在107至108copies/mL之间，随后迅速下降（图1）。SIVmac251静脉感染后42d到140d感染猴的血浆载量比较稳定，1、2、3、4、9、11六只感染猴血浆病毒载量维持在105至107copies/mL之间；10号猴血浆病毒载量下降至105copies/mL以下；12号猴血浆病毒载量一直维持在较高水平，即107copies/mL以上。

图1 SIVmac251静脉感染中国恒河猴外周血病毒载量结果

Fig.1 Theviremia (viral RNA copies/ml plasma) in Chinese-original rhesus macaquesinfected intravenously with SIVmac251

1、 2. SIVmac251静脉感染恒河猴CD4+/CD8+比值结果

8只感染猴CD4+/CD8+的比例在SIVmac251静脉感染后持续下降，均于感染后14d出现倒置。2、4、10、11、12猴在感染后期一直维持在1以下的水平，其中，2、10、11三只猴维持在0.5上下波动。1、3、9三只感染猴在SIVmac251静脉感染后期CD4+/CD8+比值有些许反弹，在1.0上下波动。（图2）。

图2 SIVmac251静脉感染中国恒河猴外周血CD4+/CD8+T淋巴细胞比值

Fig.2 The changes of CD4+/CD8+ ratio of T lymphocytes in Chinese-original rhesus macaquesinfected intravenously with SIVmac251

3.SIVmac251静脉感染恒河猴CD4+ T细胞绝对数

8只感染猴CD4+细胞绝对数在SIVmac251静脉感染后迅速下降，随后发生回弹，在感染后56d下降至最低，之后维持在稳定水平（图3）。12号猴的CD4+细胞绝对数在感染后231天下降至50个/µL，出现艾滋病症状，继而死亡。4号猴的CD4+细胞绝对数在感染后259天下降至100个/µL以下；3号猴的CD4+细胞绝对数一直在800-1000个/µL上下浮动。

图3 SIVmac251静脉感染中国恒河猴外周血CD4+T淋巴细胞绝对数

Fig.3 The CD4+T cell counts (per l blood) in Chinese-origin rhesus macaques infectedintravenously with SIVmac251

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。