閆麗夢博士第一份報告中關於動物傳代實驗是如何描述的

撰稿:喜馬拉雅的肉夾饃;審核:喜馬拉雅的饃夾肉;校對:Maarago

路德社4/3/2021路德時評(路博艾談):HBO王牌脫口秀主持人Bill Maher節目都開始談論的話題太重磅了意味著什麼? 訪談開始點出了Dr. Lawrence [email protected]上午1:03 · 2021年4月4日·Twitter Web App發佈的推文——

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

這則推文列出兩個中共超限生物戰的專家——鄧宏魁秦川

那麼Dr. Lawrence Sellin到底是什麼人呢?這則推文又意味著什麼呢?據路德社4/3/2021路德時評(路博艾談):HBO王牌脫口秀主持人Bill Maher節目都開始談論的話題太重磅了意味著什麼?時間點25:52

[Larence Sellin博士是美國前軍情部門專家,他的推是零FOLLOW,從來不FOLLOW任何人,這個推就是純粹傳遞資訊的,他發的是什麼東西?今天發的這個東西絕對重磅,他說COVID19病毒很可能是通過提前適應了讓人類感染被一系列的“人性化”的動物模型通過連續傳代感染,然後具備傳染人的能力,從這裡開始——超限生化武器/冠狀病毒的來源。然後附上鄧宏魁和秦川的照片,這個人絕對是軍情部門的,他是生化武器專家,第一次提到動物傳達,我們在119第一次提到動物傳代,閆博士告訴大家通過動物傳代打磨,然後她在第一份報告裡頭,閆博士的第一份報告裡面就講了一整頁,第19頁第五部分大家去看。]

下面我們按圖索驥看一下閆麗夢博士的報告原文(注:以下英文取自閆麗夢博士的報告原文,中文部分摘自GNEWS此前發佈過的閆麗夢博士第一份報告譯文)——

Step 5: Optimize the virus for fitness and improve its hACE2-binding affinity in vivo (2.5-3 months) 步驟5:優化病毒的適應性,提⾼病毒在體內的hACE2結合親和⼒(2.5-3個⽉)

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.

從步驟4中回收的病毒需要經過經典的實驗—實驗室動物的連續傳代—進⾏進⼀步調整101。這最後⼀步將驗證病毒的適應性,並確保其⾯向受體的適應性,以適應其預定的宿主,根據上述分析,宿主應該是⼈類。重要的是,RBM和弗林酶切位點,被分別引⼊到刺突蛋⽩中,現在將作為⼀個功能單元⼀起優化。在各種可⽤的冠狀病毒動物模型(如⼩⿏、倉⿏、雪貂和猴⼦)中,hACE2轉基因⼩⿏(hACE2-mice)應該是這⾥最合適、最⽅便的選擇。這種動物模型在SARS-CoV的研究過程中已經建⽴起來了,並且在傑克遜實驗室已經有多年的使⽤經驗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.

連續傳代的程式是簡單直接的。簡⽽⾔之,從步驟4中選擇的病毒株,即SARS-CoV-2的前體,將被⿐內接種到⼀組⿇醉的hACE2-⼩⿏中。感染後2-3天左右,肺部的病毒通常會放⼤到⼀個峰值滴度。然後,⼩⿏將死去,其肺部最後均質化。通常,攜帶病毒量最⾼的⿏肺上清液將被⽤於提取候選病毒,以備下⼀輪傳代。經過⼤約10~15輪的傳代,病毒株的hACE2結合親和⼒、感染效率和致死率都會得到充分的提⾼,病毒基因組也會趨於穩定101。最後,經過⼀系列的表徵實驗(如病毒動⼒學化驗、抗體反應化驗、症狀觀察和病理檢查等),得到最終產品SARS-CoV-2,從⽽結束了整個研製過程。從此,這種病毒病原體可以被擴增(很可能使⽤VeroE6細胞),並進⾏常規⽣產。

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,根據對SARS-CoV的研究,hACE2⼩⿏雖然適合SARS-CoV-2的適應性,但並不是反映病毒在⼈類中的傳播性和相關臨床症狀的良好模型。我們認為,在COVID-19爆發前,那些科學家可能沒有使⽤合適的動物模型(如敘利亞倉⿏)來測試SARS-CoV-2的傳播性。如果他們使⽤適當的動物模型進⾏實驗,SARS-CoV-2的⾼傳染性就會⾮常明顯,因此SARS-CoV-2不會在疫情爆發之初被描述為“不會造成⼈傳⼈”。

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.

我們還假定,針對提⾼傳播能⼒和致死率的⼴泛實驗室適應措施可能會讓病毒變得更加強⼤。因此,SARS-CoV-2在⽬前對⼈類的適應過程中,可能已經失去了在傳播性和致死性上弱化的能⼒。這⼀假設與SARS-CoV-2儘管在⼤流⾏,但到⽬前為⽌還沒有明顯的減弱跡象,也與最近出現的主要變異體只表現出更強的傳播性的觀察相⼀致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.

連續傳代是⼀個快速⽽密集的過程,在這個過程中,病毒的適應性被加速。雖然意在模仿⾃然進化,但連續傳代在時間和規模上都受到更多限制。因此,與⾃然進化相⽐,預計連續傳代過程中隨機突變的情況會更少。這對於保守的病毒蛋⽩來說尤其如此,例如E蛋⽩。E蛋⽩在病毒複製中⾄關重要,是毒性的決定因素,對它進⾏⼯程化處理可能會使SARS-CoV-2減毒109-111。因此,在最初的組裝階段,這些科學家可能決定保持E蛋⽩的氨基酸序列與ZC45/ZXC21的序列不變。由於E蛋⽩的保守性和連續傳代的限制,實際上沒有發⽣氨基酸突變,導致SARS-CoV-2和ZC45/ZXC21之間E蛋⽩的序列⼀致性100%相同。同樣的情況也可能發⽣在分⼦克隆的標記(RBM兩側的限制性位點)上。連續傳代,應該使SARS-CoV-2基因組部分⾃然化,但可能沒有消除所有⼈⼯操縱的痕跡。

*******閆麗夢博士第一份報告中英文內容引用完畢*******

綜述:我們大部分人都不是中共病毒專家,所以即使把閆麗夢博士的報告放在我們面前,我們也無法徹底讀懂它,但是閆麗夢博士的報告正在喚醒這個世界,美國軍情生化武器專家Larence Sellin博士已經通過推文公開認可了閆麗夢博士關於中共病毒是[超限生化武器]的定義和定性,並且Larence Sellin博士已經在推文中列出中共的超限生化武器專家名單,我相信這份名單要甚于當年美軍在伊拉克公佈的伊拉克前政要人員撲克牌,我相信凡是上這個榜單的所有中共專家他們都無法逃避最終的審判!

(文章內容僅代表作者個人觀點)

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