闫丽梦博士第一份报告中关于动物传代实验是如何描述的

撰稿:喜马拉雅的肉夹馍;审核:喜马拉雅的馍夹肉;校对:Maarago

路德社在4/3/2021路德时评(路博艾谈):HBO王牌脱口秀主持人Bill Maher节目都开始谈论的话题太重磅了意味着什么? 访谈开始点出了Dr. Lawrence [email protected]上午1:03 · 2021年4月4日·Twitter Web App发布的推文——

The #COVID19 virus was likely pre-adapted for human infection by serial passage through “humanized” animal models To explore such capabilities, begin here #COVID #Corona #coronavirus #DRASTIC #coronavirus #CCPVirus #UnrestrictedBiowarfare #UnrestrictedBioweapon #OriginOfCOVID19

这则推文列出两个中共超限生物战的专家——邓宏魁秦川

那么Dr. Lawrence Sellin到底是什么人呢?这则推文又意味着什么呢?据路德社4/3/2021路德时评(路博艾谈):HBO王牌脱口秀主持人Bill Maher节目都开始谈论的话题太重磅了意味着什么?时间点25:52

[Larence Sellin博士是美国前军情部门专家,他的推是零FOLLOW,从来不FOLLOW任何人,这个推就是纯粹传递信息的,他发的是什么东西?今天发的这个东西绝对重磅,他说COVID19病毒很可能是通过提前适应了让人类感染被一系列的“人性化”的动物模型通过连续传代感染,然后具备传染人的能力,从这里开始——超限生物武器/冠状病毒的来源。然后附上邓宏魁和秦川的照片,这个人绝对是军情部门的,他是生物武器专家,第一次提到动物传达,我们在119第一次提到动物传代,闫博士告诉大家通过动物传代打磨,然后她在第一份报告里头,闫博士的第一份报告里面就讲了一整页,第19页第五部分大家去看。]

下面我们按图索骥看一下闫丽梦博士的报告原文(注:以下英文取自闫丽梦博士的报告原文,中文部分摘自GNEWS此前发布过的闫丽梦博士第一份报告译文)——

Step 5: Optimize the virus for fitness and improve its hACE2-binding affinity in vivo (2.5-3 months) 步骤5:优化病毒的适应性,提⾼病毒在体内的hACE2结合亲和⼒(2.5-3个⽉)

Virus recovered from step 4 needs to be further adapted undergoing the classic experiment – serial passage in laboratory animals101. This final step would validate the virus’ fitness and ensure its receptororiented adaptation toward its intended host, which, according to the analyses above, should be human. Importantly, the RBM and the furin-cleavage site, which were introduced into the Spike protein separately, would now be optimized together as one functional unit. Among various available animal models (e.g. mice, hamsters, ferrets, and monkeys) for coronaviruses, hACE2 transgenic mice (hACE2-mice) should be the most proper and convenient choice here. This animal model has been established during the study of SARS-CoV and has been available in the Jackson Laboratory for many years102-104.

从步骤4中回收的病毒需要经过经典的实验—实验室动物的连续传代—进⾏进⼀步调整101。这最后⼀步将验证病毒的适应性,并确保其⾯向受体的适应性,以适应其预定的宿主,根据上述分析,宿主应该是⼈类。重要的是,RBM和弗林酶切位点,被分别引⼊到刺突蛋⽩中,现在将作为⼀个功能单元⼀起优化。在各种可⽤的冠状病毒动物模型(如⼩⿏、仓⿏、雪貂和猴⼦)中,hACE2转基因⼩⿏(hACE2-mice)应该是这⾥最合适、最⽅便的选择。这种动物模型在SARS-CoV的研究过程中已经建⽴起来了,并且在杰克逊实验室已经有多年的使⽤经验102-104

The procedure of serial passage is straightforward. Briefly, the selected viral strain from step 4, a precursor of SARS-CoV-2, would be intranasally inoculated into a group of anaesthetized hACE2-mice. Around 2-3 days post infection, the virus in lungs would usually amplify to a peak titer. The mice would then be sacrificed and the lungs homogenized. Usually, the mouse-lung supernatant, which carries the highest viral load, would be used to extract the candidate virus for the next round of passage. After approximately 10~15 rounds of passage, the hACE2-binding affinity, the infection efficiency, and the lethality of the viral strain would be sufficiently enhanced and the viral genome stabilized101. Finally, after a series of characterization experiments (e.g. viral kinetics assay, antibodies response assay, symptom observation and pathology examination), the final product, SARS-CoV-2, would be obtained, concluding the whole creation process. From this point on, this viral pathogen could be amplified (most probably using Vero E6 cells) and produced routinely.

连续传代的程序是简单直接的。简⽽⾔之,从步骤4中选择的病毒株,即SARS-CoV-2的前体,将被⿐内接种到⼀组⿇醉的hACE2-⼩⿏中。感染后2-3天左右,肺部的病毒通常会放⼤到⼀个峰值滴度。然后,⼩⿏将死去,其肺部最后均质化。通常,携带病毒量最⾼的⿏肺上清液将被⽤于提取候选病毒,以备下⼀轮传代。经过⼤约10~15轮的传代,病毒株的hACE2结合亲和⼒、感染效率和致死率都会得到充分的提⾼,病毒基因组也会趋于稳定101。最后,经过⼀系列的表征实验(如病毒动⼒学化验、抗体反应化验、症状观察和病理检查等),得到最终产品SARS-CoV-2,从⽽结束了整个研制过程。从此,这种病毒病原体可以被扩增(很可能使⽤VeroE6细胞),并进⾏常规⽣产。

It is noteworthy that, based on the work done on SARS-CoV, the hACE2-mice, although suitable for SARS-CoV-2 adaptation, is not a good model to reflect the virus’ transmissibility and associated clinical symptoms in humans. We believe that those scientists might not have used a proper animal model (such as the golden Syrian hamster) for testing the transmissibility of SARS-CoV-2 before the outbreak of COVID-19. If they had done this experiment with a proper animal model, the highly contagious nature of SARS-CoV-2 would be extremely evident and consequently SARS-CoV-2 would not have been described as “not causing human-to-human transmission” at the start of the outbreak.

值得注意的是,根据对SARS-CoV的研究,hACE2⼩⿏虽然适合SARS-CoV-2的适应性,但并不是反映病毒在⼈类中的传播性和相关临床症状的良好模型。我们认为,在COVID-19爆发前,那些科学家可能没有使⽤合适的动物模型(如叙利亚仓⿏)来测试SARS-CoV-2的传播性。如果他们使⽤适当的动物模型进⾏实验,SARS-CoV-2的⾼传染性就会⾮常明显,因此SARS-CoV-2不会在疫情爆发之初被描述为“不会造成⼈传⼈”。

We also speculate that the extensive laboratory-adaptation, which is oriented toward enhanced transmissibility and lethality, may have driven the virus too far. As a result, SARS-CoV-2 might have lost the capacity to attenuate on both transmissibility and lethality during its current adaptation in the human population. This hypothesis is consistent with the lack of apparent attenuation of SARS-CoV-2 so far despite its great prevalence and with the observation that a recently emerged, predominant variant only shows improved transmissibility105-108.

我们还假定,针对提⾼传播能⼒和致死率的⼴泛实验室适应措施可能会让病毒变得更加强⼤。因此,SARS-CoV-2在⽬前对⼈类的适应过程中,可能已经失去了在传播性和致死性上弱化的能⼒。这⼀假设与SARS-CoV-2尽管在⼤流⾏,但到⽬前为⽌还没有明显的减弱迹象,也与最近出现的主要变异体只表现出更强的传播性的观察相⼀致105-108

Serial passage is a quick and intensive process, where the adaptation of the virus is accelerated. Although intended to mimic natural evolution, serial passage is much more limited in both time and scale. As a result, less random mutations would be expected in serial passage than in natural evolution. This is particularly true for conserved viral proteins, such as the E protein. Critical in viral replication, the E protein is a determinant of virulence and engineering of it may render SARS-CoV-2 attenuated109-111 Therefore, at the initial assembly stage, these scientists might have decided to keep the amino acid sequence of the E protein unchanged from that of ZC45/ZXC21. Due to the conserved nature of the E protein and the limitations of serial passage, no amino acid mutation actually occurred, resulting in a 100% sequence identity on the E protein between SARS-CoV-2 and ZC45/ZXC21. The same could have happened to the marks of molecular cloning (restriction sites flanking the RBM). Serial passage, which should have partially naturalized the SARS-CoV-2 genome, might not have removed all signs of artificial manipulation.

连续传代是⼀个快速⽽密集的过程,在这个过程中,病毒的适应性被加速。虽然意在模仿⾃然进化,但连续传代在时间和规模上都受到更多限制。因此,与⾃然进化相⽐,预计连续传代过程中随机突变的情况会更少。这对于保守的病毒蛋⽩来说尤其如此,例如E蛋⽩。E蛋⽩在病毒复制中⾄关重要,是毒性的决定因素,对它进⾏⼯程化处理可能会使SARS-CoV-2减毒109-111。因此,在最初的组装阶段,这些科学家可能决定保持E蛋⽩的氨基酸序列与ZC45/ZXC21的序列不变。由于E蛋⽩的保守性和连续传代的限制,实际上没有发⽣氨基酸突变,导致SARS-CoV-2和ZC45/ZXC21之间E蛋⽩的序列⼀致性100%相同。同样的情况也可能发⽣在分⼦克隆的标记(RBM两侧的限制性位点)上。连续传代,应该使SARS-CoV-2基因组部分⾃然化,但可能没有消除所有⼈⼯操纵的痕迹。

*******闫丽梦博士第一份报告中英文内容引用完毕*******

综述:我们大部分人都不是中共病毒专家,所以即使把闫丽梦博士的报告放在我们面前,我们也无法彻底读懂它,但是闫丽梦博士的报告正在唤醒这个世界,美国军情生物武器专家Larence Sellin博士已经通过推文公开认可了闫丽梦博士关于中共病毒是[超限生物武器]的定义和定性,并且Larence Sellin博士已经在推文中列出中共的超限生物武器专家名单,我相信这份名单要甚于当年美军在伊拉克公布的伊拉克前政要人员扑克牌,我相信凡是上这个榜单的所有中共专家他们都无法逃避最终的审判!

(文章内容仅代表作者个人观点)

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