Ọdịiche Lambda: Ọgwụ mgbochi yana ọ na-efe efe karịa?

Nyocha data ọha

Data sitere na usoro SARS-CoV-2 yana ụbọchị ewepụtara ihe nlele ahụ site na usoro dị na Chile sitere na saịtị Consorcio Genomas CoV2 dị na https://auspice.cov2.cl/ncov/chile-global. Enwetara data ịgba ọgwụ mgbochi ọrịa sitere na data ọha sitere na Ministry of Science, Technology, Knowledge and Innovation dị na https://github.com/MinCiencia/Datos-COVID19 (Ngwaahịa 83).

Nnwale ọrịa

A kwadebere nje virus pseudotyped na-ebu protein dị iche iche SARS-CoV-2 dị ka anyị kọwara na mbụ¹². Na nkenke, emepụtara pseudotypes SARS-CoV-1 dabere na HIV-2 na sel HEK293T site na ibugharị pNL4.3-ΔEnv-Luc yana pCDNA-SARS-CoV-2 Spike coding vector kwekọrọ na 1: 1 molar ratio. Plasmids na-edobe spike kachasị nke codon enweghị amino acid 19 ikpeazụ nke njedebe C-terminal (SΔ19) mara iji zere njigide na reticulum endoplasmic.¹² enwetara site na njikọ mkpụrụ ndụ ihe nketa ma ọ bụ mutagenesis nke na-eduzi saịtị (GeneScript) ma nwee mgbanwe ndị a: lineage A (usoro ntụaka), lineage B (D614G), lineage B.1.1.7 (Δ69-70, Δ144, N501Y, A570D, D614G, P681H, T716I, S982A, D1118H), usoro ọmụmụ P.1 (L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y) 1027 (T37 75- 76, L246Q, F252S, D452G, T490N). A na-ekpochapụ nkwadebe pseudotype ọ bụla site na centrifugation na 614 rpm na ụlọ okpomọkụ, jiri HIV-859 Gag p3,000 Quantikine ELISA Kit (R & D Systems), nke a na-etinye na 1% fetal bovine serum (Sigma-Aldrich) ma chekwaa na -24 Celsius C ruo mgbe. jiri. A na-eji ụdị nje pseudotyped dị iche iche (dị ka ọkwa nke nje HIV-50 p80 siri kpebisie ike) iji bute mkpụrụ ndụ HEK-ACE1 na awa 24 ka e mesịrị, a tụrụ ọrụ luciferase nke ọkụ na-eji Luciferase Assay Reagent (Promega) na Glomax 2 Microplate. luminometer (Promega).

Nnwale nke ịnọpụ iche

A na-eme nyocha mkpochapụ nje nke Pseudotyped nke ọma dịka anyị kọwara na mbụ¹². N'ụzọ dị nkenke, a kwadebere usoro nke ihe nlele plasma (1: 4 ruo 1: 8748) na DMEM na 10% fetal bovine serum ma tinye ya na 5 ng nke p24 nke nje ọ bụla pseudotyped n'oge 1h na 37 Celsius C na mgbe ahụ, 1 × 10.⁴ sel HEK-ACE2 ka etinyere n'olulu mmiri ọ bụla. A na-eji mkpụrụ ndụ HEK293T (anaghị ekwupụta ACE2) nke ejiri nje pseudotyped (ahịrị A) mee ka njikwa na-adịghị mma. A na-ekpuchi mkpụrụ ndụ mgbe awa 48 gachara, a na-atụkwa ọrụ firefly luciferase site na iji Luciferase Assay Reagent (Promega) na Glomax 96 Microplate luminometer (Promega). A gbakọrọ pasentị nnọpụiche maka ntugharị ọ bụla yana gbakọọ ID50 nke nlele ọ bụla site na iji ụdị GraphPad Prism 9.0.1.

Nnyocha nchịkọta akụkọ

Emere nyocha ndekọ aha site na iji ụdị ngwanrọ GraphPad Prism 9.1.2. Ọtụtụ ntụnyere otu maka iwepu ihe mgbochi mgbochi (NAbTs) megide otu panel nke nje SARS-CoV-2 pseudotyped yana ntụnyere nzaghachi NAbs site na mmekọahụ na ọnọdụ anwụrụ ọkụ ka emere site na iji nyocha akara Wilcoxon jikọtara ọnụ. A gbakọrọ mgbanwe ihe dị ka ọdịiche nke ihe nrịbama geometric dị na ID ahụ₅₀ dị ka atụnyere nke ụdị anụ ọhịa pseudotyped virus. A na-eme nyocha mmekọrịta n'etiti NABT na afọ ma ọ bụ BMI site na iji ule Spearman. Otu ụzọ ANOVA na Tukey ka emere ọtụtụ ntụle maka nyocha ọnụọgụgụ nke ọrịa. A tụlere p uru ≤0.05 dị ka ihe dị mkpa n'ọnụ ọgụgụ.

Nkwenye usoro

Ndị Kọmitii Ethics nke ngalaba ọgwụ na Universidad de Chile kwadoro usoro ọmụmụ a (Projects N° 0361-2021 na N° 096-2020) na Clínica Santa Maria (Project N°132604-21). Ndị nyere onyinye niile bịanyere aka n'akwụkwọ nkwenye ahụ, ma anabataghị ihe nlele ha.

Mmetụta mmụgharị spike dị na Lambda dị iche iche na ọrịa nje na iwepu azịza mgbochi ọrịa.

Nyocha nke usoro 3695 sitere na Chile nke edobere na GISAID dịka kwa June 24^th 2021 na-egosi ikike doro anya nke ụdị SARS-CoV-2 dị iche iche Gamma na Lambda n'oge nyocha nke ọnwa atọ ikpeazụ, ọnụ, maka 79% nke usoro niile.

N'ụzọ na-akpali mmasị, e jiriwo mgbasa ozi ịgba ọgwụ mgbochi mee mara oge a nke 65.6% nke ndị a na-achọsi ike (ndị gbara afọ 18 na ndị okenye) nwetara atụmatụ ịgba ọgwụ mgbochi ọrịa zuru oke ka ọ dị na June 27.^th 2021

Nyere na 78.2% nke ndị ejiri atụmatụ zuru oke nweta ọgwụ mgbochi nje CoronaVac sitere na Sinovac Biotech, anyị chọrọ nyocha mmetụta nke mmụgharị spike dị na ụdị Lambda na ikike iwepụ ọgwụ mgbochi nke ọgwụ mgbochi a kpatara.

Maka nke a, anyị butere nje virus SARS-CoV-1 pseudotyped nke sitere na HIV-2 na-ebu protein spike sitere na usoro nrụtụ aka Wuhan-1 (ụdị ọhịa; lineage A), ngbanwe D614G (usoro ọmụmụ B), na Alfa (usoro ọmụmụ B). .1.1.7), Gamma (agbụrụ P.1) na Lambda (agbụrụ C.37) dị iche iche.

N'oge nkwadebe nje, anyị na-achọpụtakarị na mkpụrụ ndụ ndị bu nje pseudotyped na-ebu Lambda spike na-emepụta ụkpụrụ bioluminescence dị elu dị elu ma e jiri ya tụnyere D614G mutant ma ọ bụ Alfa na Gamma variants na-egosi mmụba na-efe efe nke protein Lambda spike na-ebute.

Nyocha 1.Ọrịa nje na-ebute site na protein dị iche iche spike.

(A) Ngosipụta atụmatụ nke SARS-CoV-2 spike protein na ụdị dị iche iche ejiri mee ihe n'ọmụmụ ihe a. A na-egosi usoro ọmụmụ na mbo. RBD, ngalaba nnabata-njikọ, CM; cytoplasmic ọdụ.

(B) Titration nke pseudotypes nke usoro ọmụmụ ọ bụla na-eji nje HIV-1 p24 nhata. A tụrụ ọrụ luciferase firefly dị ka nkeji luminescence (RLU) na awa 48 ka ọrịa butere ya. A gbakọrọ nkezi na SD site na nnwale ugboro atọ nke nnọchite anya.

Ọzọ, anyị jiri nje pseudotyped a kpọtụrụ aha n'elu mee nyocha nkwụsịtụ site na iji ihe nlele plasma 79 sitere na ndị ọrụ ahụike ahụike sitere na Universidad de Chile na Clínica Santa María na Santiago, Chile.

Anyị wepụrụ sample 4 ka anyị enweghị ike ịgbakọ titer ID50. Site na ihe atụ ndị a tụlere, 73% kwekọrọ na ụmụ nwanyị, etiti afọ 34 (IQR 29-43) na ọnụọgụ anụ ahụ (BMI) nke 25 (IQR 22.7-27). Pasent 20.5 nke ndị sonyere kwuru na ha bụ ndị na-ese anwụrụ ọkụ mgbe oge ịgba ọgwụ mgbochi ahụ dịruru. Enwetara ihe nlele n'ime etiti ụbọchị 95 (IQR 76-96) ka ọgwụ mgbochi CoronaVac nke abụọ gasịrị.

Anyị hụrụ na mwepụ nke nje pseudotyped bu protein spike ụdị anụ ọhịa butere 50% inhibitory dilution (ID).₅₀) pụtara na 191.46 (154.9 - 227.95 ci), 95), 153.92 - 115.68 - 192.16, 95, 124.73% CI ) na 86.2 (163.2 - 95, 104.57% CI) maka nje pseudotyped na-ebu protein spike sitere na mutant D75.02G ma ọ bụ Alfa, Gamma na Lambda variants, n'otu n'otu.

Anyị hụkwara na geometric pụtara titer nke ID ahụ₅₀ titers belatara site na ihe nke 3.05 (2.57 - 3.61, 95% CI) maka nje pseudotyped na-ebu spike Lambda, 2.33 (1.95 - 2.80, 95% CI) maka spike Gamma, 2.03 (1.71 - 2.41), 95. maka Alfa spike na 1.37 (1.20 - 1.55, 95% CI) maka D614G spike ma e jiri ya tụnyere ụdị anụ ọhịa.

Ọnweghị njikọ dị n'etiti mmekọahụ, afọ, index of the body- mass index (BMI) ma ọ bụ ọkwa anwụrụ ọkụ na mkpochapụ ndị na-emegide mmadụ ka ahụrụ n'òtù ọmụmụ anyị.

Nyocha 2.Nyochaa ịnọpụ iche site na iji sample plasma sitere na ọgwụ mgbochi CoronaVac

(A) Mgbanwe na nkwụghachi ụgwọ 50% neutralization titer (ID₅₀N'ihe nlele plasma sitere na ndị nnata 75 nke ọgwụ CoronaVac megide D614G (usoro ọmụmụ B), Alpha (usoro ọmụmụ B.1.1.7),

Gamma (usoro ọmụmụ P.1) na Lambda (usoro ọmụmụ C.37) dị iche iche ma e jiri ya tụnyere nje ụdị anụ ọhịa. E gosipụtara nsonaazụ ya dị ka ọdịiche dị na titers neutralization nke ihe atụ dakọtara. P ụkpụrụ maka ntụnyere nke ID₅₀ A na-agbakọ ya na nyocha ọkwa ọkwa Wilcoxon.

(B) Mpempe igbe gosipụtara n'etiti etiti na etiti anọ (IQR) nke ID₅₀ maka nje ọ bụla pseudotyped. Egosiri mgbanwe ihe kpatara dị ka ihe dị iche nke ihe nrịbama geometric dị na NJ₅₀ dị ka atụnyere ndị nke anụ ọhịa ụdị pseudotyped virus. Emere nyocha ndekọ aha site na iji Wilcoxon matched-pairs test-ọkwa nlebanyere.

Ọnụ, data anyị kpughere na protein spike nke ụdị mmasị Lambda a ma ama, na-ekesa nke ukwuu na Chile na mba ndịda America, na-ebu mmụgharị na-enye mmụba na-efe efe na ikike ịgbanarị ọgwụ mgbochi nke CoronaVac kpatara.