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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an enveloped virus responsible for the COVID-19 pandemic. Despite recent advances in the structural elucidation of SARS-CoV-2 proteins and the complexes of the spike (S) proteins with the cellular receptor ACE2 or neutralizing antibodies, detailed architecture of the intact virus remains to be unveiled. Here we report the molecular assembly of the authentic SARS-CoV-2 virus using cryo-electron tomography (cryo-ET) and subtomogram averaging (STA). Native structures of the S proteins in both pre- and postfusion conformations were determined to average resolutions of 8.7-11 [A]. Compositions of the N-linked glycans from the native spikes were analyzed by mass-spectrometry, which revealed highly similar overall processing states of the native glycans to that of the recombinant glycoprotein glycans. The native conformation of the ribonucleoproteins (RNP) and its higher-order assemblies were revealed. Overall, these characterizations have revealed the architecture of the SARS-CoV-2 virus in unprecedented detail, and shed lights on how the virus packs its [~]30 kb long single-segmented RNA in the [~]80 nm diameter lumen.
RESUMEN
Objective To investigate the associations of IFNL3/IFNL4 single-nucleotide polymorphisms(SNPs)with the efficacy of highly active antiretroviral therapy(HAART)in patients with HIV-1 infection.Methods Sixty-three adult patients with HIV-1 infection receiving HAART for at least 1 year in the First Affiliated Hospital, Zhejiang University School of Medicine were enrolled.HIV-1 RNA loads in plasma and HIV-1 DNA loads in peripheral blood mononuclear cells(PBMCs),and blood SNPs were detected by quantitative polymerase chain reaction(qPCR).Plasma inflammatory cytokines were examined by magnetic beads method,and the CD4 +T and CD8 +T lymphocyte counts in peripheral blood were measured by flow cytometry.According to response to HAART,the patients were classified as low HIV-1 RNA group(viral load <100 copies/mL)and high HIV-1 RNA group(viral load≥100 copies/mL);according to CD4+T lymphocyte counts,the patients were defined as low CD4+T cell group(<250 cells/μL), and high CD4+T cell group(≥250 cells/μL);according to HIV-1 DNA levels,the patients were divided into low(<100 copies/106cells)and high(≥100 copies/106cells)HIV-1 DNA groups.Results Three candidate SNPs rs368234815,rs8099917 and rs4803223 had significantly different distribution between low and high HIV-1 RNA groups(χ2=0.043,0.047 and 0.032,all P<0.05).The levels of interleukin(IL)-10 were declined in the low HIV-1 RNA group(U=4.00,P<0.05);the levels of IL-13 were decreased in the high HIV-1 RNA group and the high HIV-1 DNA group(U=0.00 and 2.00,both P<0.05);the levels of IL-21 were reduced in the high HIV-1 RNA group and in the low CD4 +T cell group(U=3.00 and 2.00, both P<0.05),the levels of IL-28A were decreased in the high HIV-1 RNA group,the high HIV-1 DNA group,and the low CD4 +T cell group(U=3.00, 0.50 and 3.00,P<0.05 or <0.01).In addition, rs368234815 was associated with IL-21 level(H=6.690,P<0.05),the IL-21 level in rs368234815 ΔG/ΔG [131.88(2.66,174.00)]was higher than that in TT/TT[6.79(2.81,26.48)](P<0.05);rs4803223 was correlated with IFN-γlevel(H=6.690, P<0.05),the IFN-γlevel in GG subtype[62.26(19.45, 96.49)]was higher than that in GA subtype[6.98(2.19, 99.14)](P<0.05).Conclusion The polymorphisms of IFNL3/IFNL4 rs368234815, rs8099917 and rs4803223 are associated with efficacy of HAART and immune-associated cytokines levels in patients with HIV-1 infection.