Kusanthula kwanthu pagulu
Zambiri pamizere ya SARS-CoV-2 ndi tsiku lomwe chitsanzocho chidatengedwa kuchokera kumayendedwe omwe amapezeka ku Chile adapezeka patsamba la Consorcio Genomas CoV2 lomwe likupezeka pa https://auspice.cov2.cl/ncov/chile-global. Deta ya katemera idapezedwa kuchokera kuzinthu zapagulu kuchokera ku Unduna wa Sayansi, Ukadaulo, Chidziwitso ndi Zatsopano zomwe zikupezeka pa https://github.com/MinCiencia/Datos-COVID19 (Chinthu 83).
Kusayesa kuyesa
Ma virus a pseudotyped okhala ndi mapuloteni osiyanasiyana a SARS-CoV-2 adakonzedwa monga tidafotokozera kale.12. Mwachidule, ma pseudotypes a HIV-1 a SARS-CoV-2 adapangidwa m'maselo a HEK293T popatsira pNL4.3-ΔEnv-Luc pamodzi ndi pCDNA-SARS-CoV-2 Spike coding vector yofananira mu 1: 1 molar ratio. Ma Plasmids amalemba spike yokhala ndi codon-wokometsedwa yopanda ma amino acid 19 omaliza a C-terminal end (SΔ19) omwe amadziwika kuti amapewa kusungidwa pa endoplasmic reticulum.12 adapezedwa ndi kaphatikizidwe ka majini kapena mutagenesis wotsogozedwa ndi tsamba (GeneScript) ndipo anali ndi masinthidwe awa: mzere A (zotsatizana), mzere B (D614G), mzere B.1.1.7 (Δ69-70, Δ144, N501Y, A570D, D614G, P681H, T716I, S982A, D1118H), mzere P.1 (L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027N, T37G75, T76G246) ndi T252G452 490- 614, L859Q, F3,000S, D1G, T24N). Kukonzekera kulikonse kwa pseudotype kunayeretsedwa ndi centrifugation pa 50 rpm kutentha kwa firiji, kuwerengedwera pogwiritsa ntchito HIV-80 Gag p1 Quantikine ELISA Kit (R&D Systems), yomwe ili mu 24% fetal bovine serum (Sigma-Aldrich) ndikusungidwa pa -2 ° C mpaka ntchito. Mitundu yosiyanasiyana ya ma virus a pseudotyped (monga momwe ma protein a HIV-48 p96) adagwiritsidwira ntchito kupatsira maselo a HEK-ACEXNUMX ndipo patatha maola XNUMX, ntchito ya firefly luciferase idayesedwa pogwiritsa ntchito Luciferase Assay Reagent (Promega) mu Glomax XNUMX Microplate. luminometer (Promega).
Zoyeserera zandale
Mayesero a pseudotyped virus neutralization adachitidwa makamaka monga tidafotokozera kale12. Mwachidule, serial dilutions of the plasma samples (1:4 mpaka 1:8748) anakonzedwa mu DMEM ndi 10% fetal bovine seramu ndi incubated ndi 5 ng wa p24 aliyense pseudotyped kachilombo pa 1h pa 37 °C ndiyeno, 1×10.4 ma cell a HEK-ACE2 adawonjezedwa pachitsime chilichonse. Maselo a HEK293T (osawonetsa ACE2) ophatikizidwa ndi kachilombo ka pseudotyped (mzera A) adagwiritsidwa ntchito ngati njira yoletsa. Maselo adasinthidwa maola 48 pambuyo pake, ndipo ntchito ya ziphaniphaniyi inayesedwa pogwiritsa ntchito Luciferase Assay Reagent (Promega) mu Glomax 96 Microplate luminometer (Promega). Peresenti ya kusalowerera ndale kwa dilution iliyonse idawerengedwa ndipo ID50 yachitsanzo chilichonse idawerengedwa pogwiritsa ntchito mtundu wa GraphPad Prism 9.0.1.
Kusanthula kusanthula
Kusanthula kwachiwerengero kunachitika pogwiritsa ntchito pulogalamu ya GraphPad Prism 9.1.2. Kuyerekeza kwamagulu angapo kwa neutralizing antibody titers (NAbTs) motsutsana ndi gulu la ma virus a pseudotyped a SARS-CoV-2 komanso kufananitsa mayankho a NAbs pakugonana ndi utsi kunachitika pogwiritsa ntchito mayeso osainidwa a Wilcoxon. Kusintha kwa Factor kunawerengedwa ngati kusiyana kwa geometric mean titer mu ID50 poyerekeza ndi kachilombo ka Wild mtundu wa pseudotyped. Kusanthula kwamalumikizidwe pakati pa NAbTs ndi zaka kapena BMI kunachitika pogwiritsa ntchito mayeso a Spearman. Mayeso anjira imodzi a ANOVA ndi Tukey oyerekeza angapo adachitidwa kuti afufuze za matenda. Mtengo wa p ≤0.05 unkaganiziridwa ngati wofunikira kwambiri.
Kuvomerezeka kakhalidwe
Ndondomeko yophunzirira inavomerezedwa ndi Ethics Committee ya Faculty of Medicine ku Universidad de Chile (Projects N ° 0361-2021 ndi N ° 096-2020) ndi Clínica Santa Maria (Project N ° 132604-21). Onse opereka ndalama adasaina chilolezo chodziwitsidwa, ndipo zitsanzo zawo sizinatchulidwe.
Zotsatira zakusintha kwa ma spike mu mtundu wa Lambda pakusintha ndi kusokoneza mayankho a ma antibodies
Kusanthula kwamayendedwe 3695 ochokera ku Chile osungidwa ku GISAID monga pa Juni 24th 2021 ikuwonetsa kutsogola koonekeratu kwa mitundu ya SARS-CoV-2 ya Gamma ndi Lambda munthawi yowerengera ya trimester yomaliza, palimodzi, pa 79% yazotsatira zonse.
Chosangalatsa ndichakuti nthawi imeneyi idadziwika ndi ntchito yayikulu yotemera pomwe 65.6% ya anthu omwe akuyembekezeredwa (azaka 18 ndi okalamba) adalandira katemera wathunthu monga pa June 27.th 2021
Poganizira kuti 78.2% ya anthu omwe adalandira katemera wa kachilombo ka CoronaVac wotsekedwa adalandira katemera wa CoronaVac kuchokera ku Sinovac Biotech, tinayesetsa kufufuza momwe kusintha kwamtundu wa Lambda kumakhudzira mphamvu ya ma antibodies obwera chifukwa cha katemerayu.
Pazimenezi, tidapanga kachilombo ka HIV-1 kochokera ku SARS-CoV-2 pseudotyped puloteni yochokera ku Wuhan-1 reference lineage (Wild Type; mzere A), masinthidwe a D614G (mzera B), ndi Alpha (mzera B). .1.1.7), Gamma (mzera wa P.1) ndi Lambda (mzera wa C.37).
Pokonzekera ma virus, tidawona nthawi zonse kuti ma cell omwe ali ndi kachilombo ka pseudotyped omwe amanyamula chiwombankhanga cha Lambda amatulutsa ma bioluminescence apamwamba kwambiri poyerekeza ndi D614G mutant kapena mitundu ya Alpha ndi Gamma zomwe zikuwonetsa kuchulukirachulukira komwe kumayendetsedwa ndi mapuloteni a Lambda.
Chithunzi 1.Infectivity mkhalapakati ndi osiyana spike mapuloteni.
(A) Kuyimira kwadongosolo kwa SARS-CoV-2 spike protein ndi zosiyana zomwe zimagwiritsidwa ntchito mu kafukufukuyu. Mizere ikuwonetsedwa m'makolo. RBD, receptor-binding domain, CM; cytoplasmic mchira.
(B) Kutanthauzira kwa pseudotypes a mzere uliwonse pogwiritsa ntchito kuchuluka kwa HIV-1 p24. Ntchito ya firefly luciferase idayesedwa ngati mayunitsi a luminescence (RLU) pa maola 48 atadwala. Avereji ndi SD zidawerengedwa kuchokera ku kuyesa koyimilira katatu.
Kenako, tidagwiritsa ntchito mavairasi ongoyerekeza omwe tawatchula pamwambapa kuti tiyesere kusalowerera ndale pogwiritsa ntchito zitsanzo 79 za plasma kuchokera kwa ogwira ntchito yazaumoyo ochokera ku Universidad de Chile ndi Clínica Santa María ku Santiago, Chile.
Sitinaphatikizepo zitsanzo 4 chifukwa sitinathe kuwerengera ID50 titer. Kuchokera pazitsanzo zomwe zafufuzidwa, 73% imagwirizana ndi akazi, zaka zapakati pa zaka 34 (IQR 29 - 43) ndi index max index (BMI) ya 25 (IQR 22.7 - 27). A 20.5% mwa omwe adatenga nawo gawo adalengeza kuti ndi osuta pomwe nthawi ya katemera idapitilira. Zitsanzo zidapezedwa mkati mwa masiku 95 (IQR 76 - 96) pambuyo pa mlingo wachiwiri wa katemera wa CoronaVac.
Tidawona kuti kusalowerera ndale kwa kachilombo ka pseudotyped komwe kamakhala ndi mapuloteni amtundu wa Wild spike kudapangitsa kuti 50% inhibitory dilution (ID)50) Kutanthauza kuti 191.46 (154.9 - 227.95, 95% CI), 153.92) ) ndi 115.68 (192.16 - 95, 124.73% CI) za ma virus ongoyerekeza omwe amanyamula puloteni ya spike kuchokera ku D86.2G mutant kapena mitundu ya Alpha, Gamma ndi Lambda, motsatana.
Tidawonanso kuti ma geometric amatanthauza titer ya ID50 titers atsika ndi 3.05 (2.57 - 3.61, 95% CI) pa kachilombo ka pseudotyped kamene kamanyamula Lambda spike, 2.33 (1.95 - 2.80, 95% CI) kwa Gamma spike, 2.03 (1.71 - 2.41% 95%) kwa Alpha spike ndi 1.37 (1.20 - 1.55, 95% CI) kwa D614G spike poyerekeza ndi Wild mtundu spike.
Palibe kulumikizana pakati pa kugonana, zaka, index-mass index (BMI) kapena mawonekedwe a utsi ndi ma antibody titers omwe adawonedwa mu gulu lathu lophunzirira.
Chithunzi 2.Kuyesa kusalowerera ndale pogwiritsa ntchito zitsanzo za plasma zochokera ku katemera wa CoronaVac
(A) Kusintha kwa 50% neutralization titer (ID50) mu zitsanzo za plasma kuchokera kwa 75 olandira katemera wa CoronaVac motsutsana ndi D614G (mzera B), Alpha (mzera B.1.1.7),
Mitundu ya Gamma (mzera wa P.1) ndi Lambda (mzera wa C.37) poyerekeza ndi kachilombo koyambitsa matenda. Zotsatira zikuwonetsedwa ngati kusiyana kwa ma neutralization titers a zitsanzo zofananira. P zofananira za ID50 amawerengedwa ndi mayeso osainidwa a Wilcoxon.
(B) Magawo a bokosi amawonetsa mtundu wapakati ndi interquartile (IQR) wa ID50 pa pseudotyped virus iliyonse. Kusintha kwa zinthu kumawonetsedwa ngati kusiyana kwa geometric mean titer mu ID50 poyerekeza ndi a Wild mtundu pseudotyped kachilombo. Kusanthula kwachiwerengero kunachitika pogwiritsa ntchito mayeso a Wilcoxon ofanana ndi awiriawiri omwe adasaina.
Pamodzi, zambiri zathu zidawulula kuti puloteni yokulirapo ya mtundu womwe wangodziwika kumene wa Lambda, womwe ukufalikira kwambiri kumayiko aku Chile ndi South America, umakhala ndi masinthidwe opatsa kufalikira komanso kuthekera kothawa ma antibodies oyambitsidwa ndi CoronaVac.
