Journal: Viruses

Article Title: Clinical Utility of Quantitative HBV Core Antibodies for Solving Diagnostic Dilemmas

doi: 10.3390/v15020373

Figure Lengend Snippet: Methods and units used for anti-HBc antibodies quantitation.

Article Snippet: CMIA for total anti-HBc and anti-HBc IgM , Abbott Diagnostics, IL, USA , S (sample)/Co (cut-off) of RLU/mL , [ ] .

Techniques: Quantitation Assay, Sandwich ELISA

Journal: PLOS Pathogens

Article Title: Characterisation of the antibody-mediated selective pressure driving intra-host evolution of SARS-CoV-2 in prolonged infection

doi: 10.1371/journal.ppat.1012624

Figure Lengend Snippet: Peripheral blood samples obtained from the patient on the days indicated were tested by the quantitative Abbott Anti-RBD-CMIA (a) and the GenScript cPass sVNT (b) . Antibody levels detected by Anti-RBD-CMIA and sVNT are given as arbitrary units (AU)/mL, cut-off ≥50 AU/mL, and signal reduction (% inhibition, cut-off >30%) as compared to the negative control. The GenScript cPass sVNT was performed with technical duplicates at a serum dilution of 1:20. The cut-off of the sVNT is indicated by the horizontal dotted line. (c) Qualitative results of the Mikrogen line blot testing for Anti-N IgG, as well as Anti-S1 and Anti-RBD IgM and IgG, are indicated as follows: open squares: non-reactive, grey squares: reactive below cut-off, black squares: reactive above cut-off. Administration of bamlanivimab (B), first vaccination (V) and therapy with sotrovimab (S) is indicated by vertical dotted lines.

Article Snippet: IgG antibodies against the nucleocapsid (N) protein were not detected in CMIA (Abbott) until day 381 ( ).

Techniques: Inhibition, Negative Control

Journal: Analytical Biochemistry

Article Title: A review post-vaccination SARS-CoV-2 serological test: Method and antibody titer response

doi: 10.1016/j.ab.2022.114902

Figure Lengend Snippet: Several methods of postvaccine SARS-Cov-2 serological tests have been reported.

Article Snippet: CMIA , 4.3 AU/mL , Pfizer , USA , [ ] .

Techniques: Enzyme-linked Immunosorbent Assay, Immunofluorescence, Modification

Journal: Nature Communications

Article Title: Hepatitis B virus rigs the cellular metabolome to avoid innate immune recognition

doi: 10.1038/s41467-020-20316-8

Figure Lengend Snippet: a PHHs were infected with or without HBV virions (MOI = 30) for 9 days and harvested for metabolomics analysis. A heat map shows changes of glycolysis or oxidative phosphorylation metabolites. b Quantification of indicated intermediates of glycolysis and TCA cycle. c Simplified scheme of the glucose metabolic pathway. d , e PHHs were infected with or without HBV virions (MOI = 30) for 7 days. The supernatants were pretreated with or without anti-HBs, an HBV neutralizing antibody. Then, cells were cultured with high glucose (25 mM) or low glucose (5 mM) for 36 h, and treated with or without poly(I:C) (1 μg/mL) for 12 h prior to qPCR analyses. f , g Experiments were performed similar to those in d , e , except that cells were treated with or without HTDNA (1 μg/mL) or LPS (50 ng/mL). h C57BL/6 mice received HI with 10 µg of plasmid pAAV-HBV1.2 for 7 days. Mice were fasted overnight and then treated with high glucose (1.5 g/kg) or low glucose (0.2 g/kg) and were treated with PBS or poly(I:C) (10 μg/g) for 2 days (left panel). Mouse liver samples were collected and subjected to qPCR analyses (right panel). i IRNAR1 expression in IFNAR1 WT (WT) and IFNAR1−/− cells was measured using RT-PCR and western blot analyses. j IFNAR1 WT (WT) and IFNAR1−/− cells were cultured with high glucose or low glucose and transfected with pHBV-1.3 for 48 h. HBsAg and HBeAg (middle panel) secreted in culture supernatants were quantified by chemiluminescence immunoassay (CMIA), or HBV genomes in culture supernatants were determined by qPCR. k IFNAR expression in liver ( n = 3) was measured using RT-PCR and western blot analyses. l Experiments were performed similar to those in h , except HBsAg, HBeAg and HBV DNA were analyzed. Data b , d – g , j represent the means ± SD (Student’s t test), data h , j , l represent the means ± SEM (one-way ANOVA) (** P < 0.01; * P < 0.05, n.s., not significant). S:CO signal to cutoff ratio, HG high glucose, LG low glucose, HI hydrodynamic injection. See also Supplementary Figs. and .

Article Snippet: The levels of secreted HBsAg and HBeAg were determined using the Architect System and HBsAg and HBeAg CMIA kits (Abbott Diagnostics, 06C3622 and 06C3237) from culture supernatants and mouse sera, according to the manufacturer’s instructions.

Techniques: Infection, Phospho-proteomics, Cell Culture, Plasmid Preparation, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Chemiluminescence Immunoassay, Injection

Journal: Nature Communications

Article Title: Hepatitis B virus rigs the cellular metabolome to avoid innate immune recognition

doi: 10.1038/s41467-020-20316-8

Figure Lengend Snippet: a PHHs were infected with HBV (MOI = 30) for 8 days and treated with or without UK5099 (10 μM) for 12 h. HBsAg (left panel) and HBeAg (middle panel) secreted in culture supernatants were quantified using CMIA, or the levels of HBV genomes in culture supernatants were determined using qPCR (right panels). b PHHs were infected with or without HBV virions (MOI = 30) for 8 days, and transfected with or without poly(I:C) (1 μg/mL), treated with or without UK5099 (10 μM) for 12 h, and subjected to qPCR analyses for IFN-β. c , d Experiments were performed as in a and b , except that PHHs were treated with or without DCA (10 mM) for 12 h. e , f Experiments were performed as in a , b , except that PHHs were exposed to normoxia (20% O 2 ) or hypoxia (1% O 2 ) for 12 h. g , h Experiments were performed as in a , b , except that PHHs were cultured in mediums containing glucose (25 mM) or galactose (25 mM) for 12 h. All data represent the means ± SD (Student’s t test) (** P < 0.01; * P < 0.05, n.s., not significant). S:CO signal to cutoff ratio. See also Supplementary Figs. .

Article Snippet: The levels of secreted HBsAg and HBeAg were determined using the Architect System and HBsAg and HBeAg CMIA kits (Abbott Diagnostics, 06C3622 and 06C3237) from culture supernatants and mouse sera, according to the manufacturer’s instructions.

Techniques: Infection, Transfection, Cell Culture

Journal: Nature Communications

Article Title: Hepatitis B virus rigs the cellular metabolome to avoid innate immune recognition

doi: 10.1038/s41467-020-20316-8

Figure Lengend Snippet: a HepG2-NTCP cells were infected with or without HBV virions (MOI = 300) for 3 days, and transfected the indicated plasmids or shRNAs for 48 h. HBsAg (left panel) and HBeAg (right panel) secreted in culture supernatants were quantified using CMIA. b , c Experiments were performed as in a , except the levels of HBV genomes in culture supernatants were determined using qPCR ( b ), or lactate secretion in supernatants was analyzed ( c ). d HepG2-NTCP cells were infected with or without HBV virions (MOI = 300) for 3 days. Cells were transfected with the indicated plasmids or shRNAs for 36 h, and transfected with or without poly(I:C) (1 μg/mL) for 12 h, and subjected to qPCR (left panel) and ELISA (right panel) analyses. e – h Experiments were performed as in a – d , except sh-LDHAs were used. i HepG2-NTCP cells were infected with or without HBV virions (MOI = 300) for 3 days and treated with or without sodium oxamate for 24 h. HBsAg (left panel) and HBeAg (right panel) secreted in culture supernatants were quantified by CMIA. j , k Experiments were performed as in i , except the levels of HBV genomes in culture supernatants were determined using qPCR ( j ), or the lactate secretion in the supernatant was analyzed ( k ). l HepG2-NTCP cells were infected with or without HBV virions (MOI = 300) for 3 days and treated with or without sodium oxamate. Twelve hours later, cells were transfected with or without poly(I:C) (1 μg/mL) for 12 h, and were subjected to qPCR (left panel) and ELISA (right panel) analyses. m – o Experiments were performed as in i , j , l , except lactate was used. All data represent the means ± SD (Student’s t test) (** P < 0.01; * P < 0.05, n.s., not significant). S:CO signal to cutoff ratio, Oxa oxamate, Lac lactate. See also Supplementary Fig. .

Article Snippet: The levels of secreted HBsAg and HBeAg were determined using the Architect System and HBsAg and HBeAg CMIA kits (Abbott Diagnostics, 06C3622 and 06C3237) from culture supernatants and mouse sera, according to the manufacturer’s instructions.

Techniques: Infection, Transfection, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: Hepatitis B virus rigs the cellular metabolome to avoid innate immune recognition

doi: 10.1038/s41467-020-20316-8

Figure Lengend Snippet: a – d C57BL/6 mice ( n = 5 for each group) received HI with 10 µg of plasmid pAAV-HBV1.2. At 7 days after HI, the mice were treated with high glucose (1.5 g/kg) or low glucose (0.2 g/kg) with or without injection of sodium oxamate (750 mg/kg) for 2 days ( a ). Serum HBsAg and HBeAg were analyzed using CMIA ( b ). Serum HBV DNA was quantified by qPCR ( c ). HBV replicative intermediates in liver tissues were extracted and detected by Southern blotting ( d ). e – h Experiments were performed as in a – d , except sodium lactate (1 g/kg) was used. i – l Experiments were performed similar as in a – d , except sodium oxamate (750 mg/kg) and sodium lactate (1 g/kg) were used. m – p WT ( n = 5) and IFNAR −/− ( n = 5) mice received HI with 10 µg of plasmid pAAV-HBV1.2. At 7 days after HI, the mice were treated with high glucose (1.5 g/kg) or low glucose (0.2 g/kg) for 2 days ( m ). Serum HBsAg and HBeAg were analyzed using CMIA ( n ). Serum HBV DNA was quantified by qPCR ( o ). HBV replicative intermediates in liver tissues were extracted and detected by Southern blotting ( p ). q – t Experiments were performed as in m – p , except sodium oxamate (750 mg/kg) was used. u – x Experiments were performed as in m – p , except sodium lactate (1 g/kg) was used. Data in b , c , f , g , j , k , n , o , r , s v , w represent means ± SEM (one-way ANOVA) (** P < 0.01; * P < 0.05, n.s., not significant). See also Supplementary Fig. .

Article Snippet: The levels of secreted HBsAg and HBeAg were determined using the Architect System and HBsAg and HBeAg CMIA kits (Abbott Diagnostics, 06C3622 and 06C3237) from culture supernatants and mouse sera, according to the manufacturer’s instructions.

Techniques: Plasmid Preparation, Injection, Southern Blot