Journal: bioRxiv

Article Title: Molecular basis of broad neutralization against SARS-CoV-2 variants including Omicron by a human antibody

doi: 10.1101/2022.01.19.476892

Figure Lengend Snippet: (A) Daily prevalence of the variants and their distributions in all collected SARS-CoV-2 sequences in the world deposited to GISAID in 2021. The left y-axis indicates the percentage of each variant collected everyday, which is displayed by areas in different colors. Orange: Beta, Gray: Delta, Yellow: Mu, Light blue: C.1.2, Green: Omicron, Dark blue: Others. The right y-axis indicates the total number of collected sequences each day, which is displayed by the purple line. (B) The prevalence of C.1.2 variant was separated to shown due to its rarity among the reported sequences. (C) The landscape of key mutations in spike proteins of SARS-CoV-2 variants used in this study. The full-length mutated spike genes were synthesized to construct pseudoviruses. The WT, Beta, Delta, Mu, C.1.2, and Omicron variants were shown from top to bottom. (D) The neutralization of WT SARS-CoV-2 and variants by plasma samples of 19 convalescent patients infected with the WT virus and recovered from the first wave of COVID-19 pandemic. The GMT of nAbs, fold change, and statistical analysis were calculated and noted on the top of each column. The data are shown in Geometric mean ± SD. “-” indicates decreased neutralization activity. Fold changes in GMTs were compared between each variant and WT or between two variants. Statistical analysis was performed with a paired t test using GraphPad Prism 9 software. **: P < 0.01, ****: P < 0.0001. The horizontal dashed line indicates the limit of detection (1:20 dilution) for the neutralizing assay. The non-neutralizing data below the limit were set to 20 for visualization. (E) The neutralization of each plasma sample against WT and variants was represented in ID 50 value. (F) The neutralization (IC 50 ) of 12 representative nAbs of Class 1 to 4 against WT SARS-CoV-2 and variants. The cutoff value of neutralization was set as 50 μg/mL. (G) The binding affinity (K D ) of 12 representative nAbs to RBD proteins of WT SARS-CoV-2 and variants by SPR. The neutralizing potency or binding affinity is highlighted in different colors. Red: high, Yellow: moderate, Green: weak, Gray: non-neutralizing or not binding (n.b.). The data are means of two independent experiments.

Article Snippet: The RBD proteins of SARS-CoV-2 Omicron (40592-V08H121) and SARS-CoV (40150-V08B2) were purchased from Sino Biological.

Techniques: Variant Assay, Synthesized, Construct, Neutralization, Infection, Activity Assay, Software, Neutralizing Assay, Binding Assay

Journal: bioRxiv

Article Title: Molecular basis of broad neutralization against SARS-CoV-2 variants including Omicron by a human antibody

doi: 10.1101/2022.01.19.476892

Figure Lengend Snippet: (A) Overall structure of ACE2 (PDB: 7DMU) and 12 representative nAbs in complex with SARS-CoV-2 RBD. Footprints of four classes of representative nAbs were represented on the RBD in different colors. The mutated residues appeared in their epitopes were shown in red and labeled beside. The structural analysis of Omicron escaping from nAbs of Class 1 (B) , Class 2 (C) , Class 3 (D) , and Class 4 (E) . Cartoon diagrams showing the detailed interface between nAbs and RBD. Hydrogen bond and salt bridge interactions were represented by dashed and black lines, respectively. “H:” indicated antibody heavy chain. “L:” indicated antibody light chain.

Article Snippet: The RBD proteins of SARS-CoV-2 Omicron (40592-V08H121) and SARS-CoV (40150-V08B2) were purchased from Sino Biological.

Techniques: Labeling

Journal: bioRxiv

Article Title: Molecular basis of broad neutralization against SARS-CoV-2 variants including Omicron by a human antibody

doi: 10.1101/2022.01.19.476892

Figure Lengend Snippet: (A) The neutralization (IC 50 ) of 9 nAbs isolated from individuals vaccinated with WT SARS-CoV-2 vaccine against 13 tested pseudoviruses. The cutoff value of neutralization was set as 50 μg/mL. The neutralizing breadth was calculated as the percentage of viruses neutralized by each nAb. Geometric mean potency was calculated by the neutralizing value of less than 50 μg/mL. (B) The binding affinity (K D ) of 9 nAbs to RBD proteins of WT SARS-CoV-2 and variants by SPR. The neutralizing and/or binding activities of 9 nAbs to WT, Delta, and Kappa have been reported in our previous study, which are re-tested here for a head-to-head comparison with other variants. (C) The neutralizing activities of 9 nAbs against SARS-CoV. VacW-209 is marked in red, the other non-neutralizing mAbs are marked in black. (D) The binding affinity of VacW-209 to SARS-CoV RBD by SPR. (E) Competition ELISA of VacW-209 with human ACE2 for binding to WT RBD. P2C-1F11 was a known competitive antibody as a positive control. EY6A was a known non-competitive antibody as a negative control. (F) Competition ELISA of VacW-209 with 15 representatives nAbs of four classes and with itself for binding to WT RBD. The neutralizing activities of VacW-209 combined with REGN10987 or S309 against WT, Beta, Delta, and Omicron were represented in IC 50 values (G) and curves (H) . The data represented here are means of at least two independent experiments. The neutralizing potency or binding affinity is highlighted in different colors. Red: high, Yellow: moderate, Green: weak, Gray: non-neutralizing or not binding (n.b.). The curve of neutralization, binding affinity, or competition was one out of similar results. A cutoff value of 50% is indicated by a horizontal dashed line in neutralization and competition.

Article Snippet: The RBD proteins of SARS-CoV-2 Omicron (40592-V08H121) and SARS-CoV (40150-V08B2) were purchased from Sino Biological.

Techniques: Neutralization, Isolation, Binding Assay, Enzyme-linked Immunosorbent Assay, Positive Control, Negative Control

Journal: bioRxiv

Article Title: Molecular basis of broad neutralization against SARS-CoV-2 variants including Omicron by a human antibody

doi: 10.1101/2022.01.19.476892

Figure Lengend Snippet: Single-particle cryo-EM images processing workflow and the global and local resolution estimation for the immune complex of SARS-CoV-2 WT-S2P:VacW-209.

Article Snippet: The RBD proteins of SARS-CoV-2 Omicron (40592-V08H121) and SARS-CoV (40150-V08B2) were purchased from Sino Biological.

Techniques: Single Particle, Cryo-EM Sample Prep

Journal: bioRxiv

Article Title: Molecular basis of broad neutralization against SARS-CoV-2 variants including Omicron by a human antibody

doi: 10.1101/2022.01.19.476892

Figure Lengend Snippet: (A) Comparison of the binding mode of VacW-209 to representative Class 1-4 nAbs. Class 1: CB6 (7C01), Class 2: BD-368-2 (7CHH), Class 3: REGN10987 (6XDG) and S309 (7R6W), and Class 4: H014 (7CAH). (B-F) Structural compassion of VacW-209 bound to SARS-CoV-2 WT-RBD (B), Delta-RBD (C), Mu-RBD (D), C.1.2-RBD (E), and Omicron-RBD (F). (G-K) Structure comparison of RBDs of Alpha (G) (7LWV), Beta (H) (7VX1), Gamma (I) (7M8K), Kappa (J) (7VXE) variants, and SARS-CoV (K) (7JN5). RBDs are shown as gray surface representations and key mutations on RBDs are labeled. The modeled VacW-209 footprints are shown on (G-K) based on the epitope information revealed in SARS-CoV-2 WT-S. (L) RBD sequences of SARS-CoV-2 WT and its 12 variants as well as SARS-CoV with highlighted footprint of VacW-209 (slate blue: heavy chain, dodger blue: light chain, dark blue: both chains). (M) VacW-209-like nAbs and their binding modes on RBD. VacW-209, C118 (7RKS), C022 (7RKU), S2X35 (7R6W), and S2X259 (7M7W) are shown as sticks and colored in blue, orange, cyan, red, and green, respectively. (N) RBD sequence of SARS-CoV-2 WT and Omicron variant with highlighted footprints of VacW-209, C118, C022, S2X35, and S2X259. Amino acids substitutions revealed on Omicron variant are boxed.

Article Snippet: The RBD proteins of SARS-CoV-2 Omicron (40592-V08H121) and SARS-CoV (40150-V08B2) were purchased from Sino Biological.

Techniques: Binding Assay, Labeling, Sequencing, Variant Assay

Journal: bioRxiv

Article Title: Molecular basis of broad neutralization against SARS-CoV-2 variants including Omicron by a human antibody

doi: 10.1101/2022.01.19.476892

Figure Lengend Snippet: (A-E) Binding modes (upper) and footprints (lower) of VacW-209 (A), C118 (B) (7RKS), C022 (C) (7RKU), S2X35 (D) (7R6W), and S2X259 (E) (7M7W). RBDs are shown as gray surface and nAbs are presented as colored cartoon. The footprints of nAbs and mutations involved in nAbs interactions are labeled in the lower panels. (F) The neutralization of VacW-209-like nAbs against SARS-CoV-2 WT, Beta, Delta, Omicron, and SARS-CoV pseudoviruses. (G) The binding affinity of VacW-209- like nAbs to SARS-CoV-2 WT and Omicron RBDs by SPR. The data represented here was mean of at least two independent experiments.

Article Snippet: The RBD proteins of SARS-CoV-2 Omicron (40592-V08H121) and SARS-CoV (40150-V08B2) were purchased from Sino Biological.

Techniques: Binding Assay, Labeling, Neutralization

Journal: iScience

Article Title: Controllable self-replicating RNA vaccine delivered intradermally elicits predominantly cellular immunity

doi: 10.1016/j.isci.2023.106335

Figure Lengend Snippet: c-srRNA vaccine elicits cellular immunity (A–C) Cellular immunity assessed by enzyme-linked immunospot (ELISpot) assays. A single dose or two doses of a placebo (PBO) or 5 μg, or 25 μg of EXG-5003 RNAs, prepared as naked RNAs without LNP or other transfection reagents and in lactated Ringer’s solution, were administered intradermally to BALB/c mice. To assess cellular immunity, an ELISPOT assay, which quantitates the number of cytokine-secreting cells, was performed on splenocytes isolated on Day 14 and Day 42. The splenocytes were stimulated for 24 h with pools of peptides: 9mer peptides for RBD of SARS-CoV-2 (original strain) or 15mer peptides for RBD of SARS-CoV-2 (original strain). Data are presented as mean (SD). (D) Cellular immunity was assessed by intracellular cytokine staining (ICS) and flow cytometry assays. The FACS-ICS assays were performed on splenocytes isolated on Day 42. MFI, mean fluorescence intensity; p, placebo; v, EXG-5003. Data are presented as mean ± SEM. Two-tailed unpaired t-test: ns (p > 0.05), ∗ (p ≤ 0.05), ∗∗ (p ≤ 0.01), ∗∗∗ (p ≤ 0.001), ∗∗∗∗ (p ≤ 0.0001). (E) Cellular immunity assessed by in vivo tumor cell elimination assays. A single EXG-5003 vaccination suppresses the growth of 4T1-LUC tumor cells carrying SARS-CoV-2 spike protein (4T1-LUC-Spike cells). Data are presented as mean (SD).

Article Snippet: On day 14 (second treatment), the mice were treated with an intradermal injection of a placebo (PBO: buffer only), 25 μg EXG-5003 (c-srRNA1 encoding RBD [original strain]), 25 μg of EXG-5003o (c-srRNA3 encoding RBD [omicron variant B.1.1.529 BA.1]), or 10 μg RBD protein (Sino Biological SARS-CoV-2) + adjuvant (AddaVax).

Techniques: Enzyme-linked Immunospot, Transfection, Isolation, Staining, Flow Cytometry, Fluorescence, Two Tailed Test, In Vivo

Journal: iScience

Article Title: Controllable self-replicating RNA vaccine delivered intradermally elicits predominantly cellular immunity

doi: 10.1016/j.isci.2023.106335

Figure Lengend Snippet: c-srRNA vaccines can prime the immune system for the subsequent induction of neutralizing antibodies against a variant antigen upon exposure to the variant antigen (A) A schematic diagram of experimental procedures. EXG-5003 RNAs were prepared as naked RNAs, without LNP or other transfection reagents, in lactated Ringer’s solution. (B–D) Groups of CD-1 outbred mice (N = 10) received one of three formulations by intradermal injection on Day 0 and Day 14 (black arrows): (B) a placebo (buffer only), (C) 5 μg of EXG-5003 RNA, or (D) 5 μg of EXG-5003 RNA in combination with an RNase inhibitor (3 units of RNasin Plus; Promega, Madison, WI). Subsequently, all mice received a recombinant RBD protein (Ala319-Phe541, with a C-terminal 6-His tag, NCBI accession number: YP_009724390.1, R&D Systems, Minneapolis, MN) by intradermal injection on Day 49 (open arrows). To assess humoral immunity, an enzyme-linked immunosorbent assay (ELISA), which quantitates the amount of immunoglobulin G (IgG) specific to a recombinant RBD protein (represented as the geometric mean of endpoint titer in triplicate), was performed on serum obtained from the mice on Day −3, Day 14, Day 28, Day 46, Day 56, and Day 63. Data are presented as mean (SD). (E) A schematic diagram of experimental procedures. On day −40, blood was drawn from female BALB/c mice for a plaque reduction neutralization test (PRNT). On day −36, these mice received an intradermal injection of EXG-5003 RNA, which was prepared as a naked RNA, without an LNP or transfection reagent. On day −22 (14 days after EXG-5003 vaccination), half of the mice were sacrificed to obtain splenocytes for ELISpot assays. On day 0, the remaining mice were intradermally injected with the spike protein of the SARS-CoV-2 delta variant (B.1.617.2: R&D Systems) mixed with adjuvant—AddaVax (Invivogen). On day 7 (7 days after the spike protein injection), blood was drawn for the PRNT assay. (F) The induction of cellular immunity against the RBD protein by a single intradermal administration of EXG-5003 RNA. The ELISpot assay shows the number of IFN-γ spot-forming cells (SFCs) in 1 × 10ˆ6 splenocytes from immunized mice restimulated by culturing in the presence or absence of a pool of 15mer peptides for RBD of SARS-CoV-2 (original strain). The number of SFC obtained in the presence of peptides was plotted on the graph after subtracting the number of SFC obtained in the absence of peptides (background). The average and standard deviation (error bars) of five mice (n = 5) are shown for each group. (G) The titer of serum antibodies that can neutralize (50%) the SARS-CoV-2 virus (delta variant B.1.617.2), measured by the PRNT assay. Exposure to the spike protein of the SARS-CoV-2 virus (delta variant B.1.617.2) induced neutralization antibodies against the delta variant of the SARS-CoV-2 virus only in mice vaccinated with EXG-5003, which encodes the RBD of SARS-CoV-2 (original strain). Data are presented as mean (SD).

Article Snippet: On day 14 (second treatment), the mice were treated with an intradermal injection of a placebo (PBO: buffer only), 25 μg EXG-5003 (c-srRNA1 encoding RBD [original strain]), 25 μg of EXG-5003o (c-srRNA3 encoding RBD [omicron variant B.1.1.529 BA.1]), or 10 μg RBD protein (Sino Biological SARS-CoV-2) + adjuvant (AddaVax).

Techniques: Variant Assay, Transfection, Injection, Recombinant, Enzyme-linked Immunosorbent Assay, Plaque Reduction Neutralization Test, Enzyme-linked Immunospot, Standard Deviation, Neutralization

Journal: iScience

Article Title: Controllable self-replicating RNA vaccine delivered intradermally elicits predominantly cellular immunity

doi: 10.1016/j.isci.2023.106335

Figure Lengend Snippet: Improvement of c-srRNA and its use as a booster vaccine to induce antibodies against an antigen, following earlier administration of the antigen protein (A) Comparison of srRNA and c-srRNAs for T cell-inducibility. A schematic diagram of experimental procedures. On day 0, mice were intradermally injected with either placebo (PBO, buffer only), srRNA0, c-srRNA1, or c-srRNA3. The srRNA0, c-srRNA1, and c-srRNA3 encode the same RBD of SARS-CoV-2 (original strain). On day 14, mice were sacrificed and splenocytes were isolated for ELISpot assays. (B) The ELISpot assay shows the number of IFN-γ spot-forming cells (SFCs) in 1 × 10ˆ6 splenocytes from immunized mice restimulated by culturing in the presence or absence of a pool of 15mer peptides for RBD of SARS-CoV-2 (original strain). The number of SFC obtained in the presence of peptides was plotted on the graph after subtracting the number of SFC obtained in the absence of peptides (background). The average and standard deviation (error bars) are shown for each group. Two-tailed unpaired t test: ns (p > 0.05), ∗ (p ≤ 0.05), ∗∗ (p ≤ 0.01), ∗∗∗∗ (p ≤ 0.0001). (C) Use of c-srRNA vaccine as a booster to induce antibodies against an antigen, following earlier administration of the antigen protein. A schematic diagram of experimental procedures. On day 0 (first treatment), female C57BL/6 mice were treated with an intradermal injection of 10 μg RBD protein (Sino Biological SARS-CoV-2 [original strain]) + adjuvant (AddaVax). On day 14 (second treatment), the mice were treated with an intradermal injection of a placebo (PBO: buffer only), 25 μg EXG-5003 (c-srRNA1 encoding RBD [original strain]), 25 μg of EXG-5003o (c-srRNA3 encoding RBD [omicron variant B.1.1.529 BA.1]), or 10 μg RBD protein (Sino Biological SARS-CoV-2) + adjuvant (AddaVax). On day 28, the mice were sacrificed, and splenocytes and serum were collected for ELISpot and ELISA assays. (D) The induction of cellular immunity is shown by the frequency of IFN-γ spot-forming cells (SFC) in 1 × 10ˆ6 splenocytes restimulated by culturing in the presence or absence of a pool of 15mer peptides for RBD of SARS-CoV-2 (original strain) (RBD-PBO, RBD-EXG-5003, RBD-RBD) or 15mer peptides for RBD of SARS-CoV-2 (omicron variant) (RBD-EXG-5003o). The frequency obtained in the presence of peptides is plotted in the graph after subtracting the frequency obtained in the absence of peptides (background). Data are represented as mean (SD). (E) The levels of serum antibodies against the RBD protein of the SARS-CoV-2 (original strain), measured by an ELISA assay. The levels of antibodies are represented by the OD450 measurement. The average and standard deviation (error bars) of five mice (n = 5) are shown for each group. The data from Day −1 (before the first treatment) and the data from Day 28 (after the second treatment) are shown for each group. Data are represented as mean (SD).

Article Snippet: On day 14 (second treatment), the mice were treated with an intradermal injection of a placebo (PBO: buffer only), 25 μg EXG-5003 (c-srRNA1 encoding RBD [original strain]), 25 μg of EXG-5003o (c-srRNA3 encoding RBD [omicron variant B.1.1.529 BA.1]), or 10 μg RBD protein (Sino Biological SARS-CoV-2) + adjuvant (AddaVax).

Techniques: Injection, Isolation, Enzyme-linked Immunospot, Standard Deviation, Two Tailed Test, Variant Assay, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Controllable self-replicating RNA vaccine delivered intradermally elicits predominantly cellular immunity

doi: 10.1016/j.isci.2023.106335

Figure Lengend Snippet: Generation of EXG-5008, encoding RBD and nucleoproteins of SARS-CoV-2 and MERS (A) A schematic diagram of EXG-5008, encoding a fusion protein of the signal peptide of CD5, the RBD and nucleoprotein of SARS-CoV-2, and the RBD and nucleoprotein of MERS-CoV. (B and C) The frequency of IFN-γ spot-forming cells (B) and the frequency of IL-4 spot-forming cells (C) in 1 × 10ˆ6 splenocytes obtained from female C57BL/6 mice that were immunized by a single intradermal injection of 100 μL solution containing either placebo (PBO: buffer only), 25 μg of EXG-5006a, or 25 μg of EXG-5008. The splenocytes were restimulated by culturing in the presence or absence of pools of peptides: 15mer peptides for RBD of SARS-CoV-2 (original strain) ( 1 ); 15mer peptides for nucleoprotein of SARS-CoV-2 (original strain) ( 2 ); 15mer peptides for nucleoprotein of MERS-CoV ( 3 ); 15mer peptides for the spike of MERS-CoV ( 4 ). The cellular immunity was analyzed by ELISpot assays. The frequency obtained in the presence of peptides is plotted in the graph after subtracting the frequency obtained in the absence of peptides (background). The average and standard deviation (error bars) of five mice (n = 5) for PBO, four mice (n = 4) for EXG-5006, and five mice (n = 5) for EXG-5008, are shown for each group. Splenocytes were isolated 14 days after vaccination. (D) A model showing how the c-srRNA booster vaccine works. Primary series of vaccines or prior infections expose naive B cells to viral surface proteins and turn them into memory B cells in a manner dependent on CD4 + helper T cells. Skin delivery of a c-srRNA booster vaccine encoding a fusion antigen of the viral surface proteins and internal proteins, whose sequences are more evolutionarily conserved than the surface proteins, is primarily incorporated into skin antigen-presenting cells such as Langerhans cells and dendritic cells. Within the antigen-presenting cells, c-srRNA replicates and produces the fusion antigen. The antigen-presenting cells digest the antigen into peptides and present these peptides to T cells. The peptides presented through this pathway stimulate MHC-I-restricted CD8 + cytotoxic T cells as well as MHC-II-restricted CD4 + helper T cells. CD8 + cytotoxic T cells eliminate infected cells (B) CD4 + helper T cells stimulate the memory B cells and enhance or restore the production of neutralizing antibody, which prevents infection.

Article Snippet: On day 14 (second treatment), the mice were treated with an intradermal injection of a placebo (PBO: buffer only), 25 μg EXG-5003 (c-srRNA1 encoding RBD [original strain]), 25 μg of EXG-5003o (c-srRNA3 encoding RBD [omicron variant B.1.1.529 BA.1]), or 10 μg RBD protein (Sino Biological SARS-CoV-2) + adjuvant (AddaVax).

Techniques: Injection, Enzyme-linked Immunospot, Standard Deviation, Isolation, Infection

Journal: iScience

Article Title: Controllable self-replicating RNA vaccine delivered intradermally elicits predominantly cellular immunity

doi: 10.1016/j.isci.2023.106335

Figure Lengend Snippet:

Article Snippet: On day 14 (second treatment), the mice were treated with an intradermal injection of a placebo (PBO: buffer only), 25 μg EXG-5003 (c-srRNA1 encoding RBD [original strain]), 25 μg of EXG-5003o (c-srRNA3 encoding RBD [omicron variant B.1.1.529 BA.1]), or 10 μg RBD protein (Sino Biological SARS-CoV-2) + adjuvant (AddaVax).

Techniques: Recombinant, Modification, Derivative Assay, Variant Assay, Plasmid Preparation, Enzyme-linked Immunospot, Double-Color Enzymatic ELISPOT, Enzyme-linked Immunosorbent Assay, Plaque Reduction Neutralization Test, Software, Imaging

Journal: Scientific Reports

Article Title: Functional antibody responses targeting the Spike protein of SARS-CoV-2 Omicron XBB.1.5 in elderly nursing home residents following Wuhan-Hu-1-based mRNA booster vaccination

doi: 10.1038/s41598-024-62874-7

Figure Lengend Snippet: Neutralizing antibody titres (reciprocal IC 50 ) against the spike protein of Wuhan-Hu-1 ( A ), Omicron BA.4/5 ( B ), and Omicron XBB.1.5 ( C ) in elderly nursing home residents before and after one (pre-3D and post-3D, respectively) or two (pre-4D and post-4D, respectively) COVID-19 vaccine booster doses. The number of plasma samples analysed at the different sampling times is shown. Bars indicate medians and IQRs. P values for differences are shown.

Article Snippet: Briefly, purified, recombinant S proteins, corresponding to the SARS-CoV-2 Spike S1 + S2 trimer protein of either the D614G or Omicron XBB.1.5 variants (SinoBiological, Eschborn, Germany), were plated overnight in Thermo NUNC MaxiSorp 96-well plates at 3 μg/ml in borate buffered saline at 4ºC.

Techniques: Sampling

Journal: Scientific Reports

Article Title: Functional antibody responses targeting the Spike protein of SARS-CoV-2 Omicron XBB.1.5 in elderly nursing home residents following Wuhan-Hu-1-based mRNA booster vaccination

doi: 10.1038/s41598-024-62874-7

Figure Lengend Snippet: Neutralizing antibody titres (reciprocal IC 50 ) against the spike protein of Wuhan-Hu-1, Omicron BA.4/5 , and Omicron XBB.1.5 in elderly nursing home residents before and after one (pre-3D and post-3D, respectively) or two (pre-4D and post-4D, respectively) COVID-19 vaccine booster doses, according to their SARS-CoV-2 infection status (experienced/Vac-ex or naïve/Vac-n). The number of plasma samples analysed at the different sampling times is shown. Bars indicate medians and IQRs. P values for differences are shown.

Article Snippet: Briefly, purified, recombinant S proteins, corresponding to the SARS-CoV-2 Spike S1 + S2 trimer protein of either the D614G or Omicron XBB.1.5 variants (SinoBiological, Eschborn, Germany), were plated overnight in Thermo NUNC MaxiSorp 96-well plates at 3 μg/ml in borate buffered saline at 4ºC.

Techniques: Infection, Sampling

Journal: Scientific Reports

Article Title: Functional antibody responses targeting the Spike protein of SARS-CoV-2 Omicron XBB.1.5 in elderly nursing home residents following Wuhan-Hu-1-based mRNA booster vaccination

doi: 10.1038/s41598-024-62874-7

Figure Lengend Snippet: Comparison of antibody-dependent NK cell-mediated responses against SARS-CoV-2 Wuhan-Hu-1 and Omicron XBB.1.5 spike variants in elderly nursing home residents before and after one (pre-3D and post-3D, respectively) or two (pre-4D and post-4D, respectively) COVID-19 vaccine booster doses. Frequencies of LAMP1- (lysosomal-associated membrane protein 1) and IFN (interferon) γ-producing NK cells are shown in panels A and B, respectively. The number of participants analysed at the different sampling times is shown. Bars indicate medians and IQRs. P values for differences are shown.

Article Snippet: Briefly, purified, recombinant S proteins, corresponding to the SARS-CoV-2 Spike S1 + S2 trimer protein of either the D614G or Omicron XBB.1.5 variants (SinoBiological, Eschborn, Germany), were plated overnight in Thermo NUNC MaxiSorp 96-well plates at 3 μg/ml in borate buffered saline at 4ºC.

Techniques: Comparison, Membrane, Sampling

Journal: Scientific Reports

Article Title: Functional antibody responses targeting the Spike protein of SARS-CoV-2 Omicron XBB.1.5 in elderly nursing home residents following Wuhan-Hu-1-based mRNA booster vaccination

doi: 10.1038/s41598-024-62874-7

Figure Lengend Snippet: Impact of COVID-19 booster vaccination on antibody-dependent NK cell-mediated responses against SARS-CoV-2 Wuhan-Hu-1 and Omicron XBB.1.5 spike variants in elderly nursing home residents. The frequencies of LAMP1 (lysosomal-associated membrane protein 1)-producing NK cells against Wuhan-Hu-1 and Omicron XBB.1.5 are shown in ( A ) and ( B ), respectively. The frequencies of IFN (interferon) γ-producing NK cells against Wuhan-Hu-1 and Omicron XBB.1.5 are shown in ( C ) and ( D ), respectively. The number of participants analysed at the different sampling times is shown. Bars indicate medians and IQRs. P values for differences are shown.

Article Snippet: Briefly, purified, recombinant S proteins, corresponding to the SARS-CoV-2 Spike S1 + S2 trimer protein of either the D614G or Omicron XBB.1.5 variants (SinoBiological, Eschborn, Germany), were plated overnight in Thermo NUNC MaxiSorp 96-well plates at 3 μg/ml in borate buffered saline at 4ºC.

Techniques: Membrane, Sampling

Journal: Scientific Reports

Article Title: Functional antibody responses targeting the Spike protein of SARS-CoV-2 Omicron XBB.1.5 in elderly nursing home residents following Wuhan-Hu-1-based mRNA booster vaccination

doi: 10.1038/s41598-024-62874-7

Figure Lengend Snippet: Antibody-dependent NK cell-mediated responses against SARS-CoV-2 Wuhan-Hu-1 and Omicron XBB.1.5 spike variants in elderly nursing home residents according to their SARS-CoV-2 infection status (experienced/Vac-ex or naïve/Vac-n). The frequencies of LAMP1- (lysosomal-associated membrane protein 1)-producing NK cells against Wuhan-Hu-1 and Omicron XBB.1.5 at the different testing times are shown in ( A ) and ( B ), respectively. The frequencies of IFN (interferon)γ-producing NK cells against Wuhan-Hu-1 and Omicron XBB.1.5 are shown in ( C ) and ( D ), respectively. The number of participants analysed at the different sampling times is shown. Bars indicate medians and IQRs. P values for differences are shown.

Article Snippet: Briefly, purified, recombinant S proteins, corresponding to the SARS-CoV-2 Spike S1 + S2 trimer protein of either the D614G or Omicron XBB.1.5 variants (SinoBiological, Eschborn, Germany), were plated overnight in Thermo NUNC MaxiSorp 96-well plates at 3 μg/ml in borate buffered saline at 4ºC.

Techniques: Infection, Membrane, Sampling

Journal: Scientific Reports

Article Title: Functional antibody responses targeting the Spike protein of SARS-CoV-2 Omicron XBB.1.5 in elderly nursing home residents following Wuhan-Hu-1-based mRNA booster vaccination

doi: 10.1038/s41598-024-62874-7

Figure Lengend Snippet: Correlation between Neutralizing antibody (NtAb) levels and frequencies of LAMP1- (lysosomal-associated membrane protein 1)-producing or (interferon)γ-producing NK cells against Wuhan-Hu-1 ( A and B , respectively) and Omicron XBB.1.5 ( C and D , respectively). Rho and P values by the Spearman Rank test are shown.

Article Snippet: Briefly, purified, recombinant S proteins, corresponding to the SARS-CoV-2 Spike S1 + S2 trimer protein of either the D614G or Omicron XBB.1.5 variants (SinoBiological, Eschborn, Germany), were plated overnight in Thermo NUNC MaxiSorp 96-well plates at 3 μg/ml in borate buffered saline at 4ºC.

Techniques: Membrane