sars2 spike primary mouse antibody  (Native Antigen Inc)

Journal: Frontiers in Immunology

Article Title: Dual role of ACE2 in regulating inflammation triggered by Omicron S1 and other SARS-CoV-2 Spike variants

doi: 10.3389/fimmu.2025.1667880

Figure Lengend Snippet: Omicron S1 shows reduced immune cells recruitment and expansion compared with the ancestral variant. Recombinant S1WT, S1 Omicron or vehicle (-) were injected in the hindbrain ventricle (HBV) of Tg(lyz:DsRED) (A) , Tg(mfap4:Tomato) (B) , Tg(nfkb:eGFP) (C) of 2-dpf larvae. Neutrophil (A) and macrophage (B) recruitment and number and Nfkb reporter activity (C) were analyzed at 6, 12, and/or 24 hpi by fluorescence microscopy. Representative photos for each treatment are shown from 24 hpi. Scale bar 500 μm. Each dot represents one individual, and the means ± SEM for each group is also shown. P values were calculated using one-way analysis of variance (ANOVA) and Tukey multiple range test. ns, not significant; * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 and **** p < 0.0001. auf, arbitrary units of fluorescence.

Article Snippet: Recombinant His-tagged Spike S1 wild-type (#40591-V08B1), S1 Omicron (#40592-V08H121) or Spike S1/S2 TRIMER wild-type (#40589-V08H8), all from Sino Biological at a concentration of 0.25 mg/ml supplemented with phenol red were injected into the hindbrain (1 nl) of 48 hpf zebrafish larvae.

Techniques: Variant Assay, Recombinant, Injection, Activity Assay, Fluorescence, Microscopy

Journal: Frontiers in Immunology

Article Title: Dual role of ACE2 in regulating inflammation triggered by Omicron S1 and other SARS-CoV-2 Spike variants

doi: 10.3389/fimmu.2025.1667880

Figure Lengend Snippet: Omicron is more proinflammatory than the ancestral variant. Recombinant S1WT, S1 Omicron or vehicle (-) were injected in the hindbrain ventricle (HBV) of WT (A–F) 2-dpf larvae. The transcript levels of the indicated genes (A–E) were analyzed at 12 hpi by RT-qPCR in larval head and rest of the body and caspase-1 activity was determined at 24 hpi using a fluorogenic substrate (F) . Graphs shown are representative of three independent experiments; technical replicates are displayed in each graph. The means ± SEM for each group is shown. P values were calculated using one-way analysis of variance (ANOVA) and Tukey multiple range test. n=45 in (A–E) , n=35 in (F) . ns, not significant; * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 and **** p < 0.0001. auf, arbitrary units of fluorescence.

Article Snippet: Recombinant His-tagged Spike S1 wild-type (#40591-V08B1), S1 Omicron (#40592-V08H121) or Spike S1/S2 TRIMER wild-type (#40589-V08H8), all from Sino Biological at a concentration of 0.25 mg/ml supplemented with phenol red were injected into the hindbrain (1 nl) of 48 hpf zebrafish larvae.

Techniques: Variant Assay, Recombinant, Injection, Quantitative RT-PCR, Activity Assay, Fluorescence

Journal: Frontiers in Immunology

Article Title: Dual role of ACE2 in regulating inflammation triggered by Omicron S1 and other SARS-CoV-2 Spike variants

doi: 10.3389/fimmu.2025.1667880

Figure Lengend Snippet: Omicron causes higher neutrophil cell death than the ancestral variant. Recombinant S1WT, S1 Omicron or vehicle (-) were injected in the hindbrain ventricle (HBV) of Tg(lyz:DsRED) (A, B) of 2-dpf larvae. Tunel positive neutrophil number (double positive) was counted at 6 hpi in the head and tail of the larvae (A, B) . Representative photos for each treatment are shown. Scale bar 500 μm. Each dot represents one individual, and the means ± SEM for each group is also shown. P values were calculated using one-way analysis of variance (ANOVA) and Tukey multiple range test. ns, not significant; * P ≤ 0.05, ** P ≤ 0.01 and **** p < 0.0001.

Article Snippet: Recombinant His-tagged Spike S1 wild-type (#40591-V08B1), S1 Omicron (#40592-V08H121) or Spike S1/S2 TRIMER wild-type (#40589-V08H8), all from Sino Biological at a concentration of 0.25 mg/ml supplemented with phenol red were injected into the hindbrain (1 nl) of 48 hpf zebrafish larvae.

Techniques: Variant Assay, Recombinant, Injection, TUNEL Assay

Journal: Frontiers in Immunology

Article Title: Dual role of ACE2 in regulating inflammation triggered by Omicron S1 and other SARS-CoV-2 Spike variants

doi: 10.3389/fimmu.2025.1667880

Figure Lengend Snippet: Trimeric ancestral variant induces a weaker immune response than its monomeric form. Recombinant S1WT (monomeric), S1/S2WT-T (trimeric) or vehicle (-) were injected in the hindbrain ventricle (HBV) of Tg(lyz:DsRED) (A) , Tg(mfap4:Tomato) (B) , Tg( nfkb :eGFP) (C) 2-day postfertilization larvae (dpf). Neutrophil (A) and macrophage (B) recruitment and number and Nfkb reporter activity (C) were analyzed at 6, 12, and/or 24 hpi by fluorescence microscopy. Representative photos for each treatment are shown from 24 hpi. Scale bar 500 μm. Each dot represents one individual, and the means ± SEM for each group is also shown. P values were calculated using one-way analysis of variance (ANOVA) and Tukey multiple range test. ns, not significant; * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 and **** p < 0.0001. auf, arbitrary units of fluorescence.

Article Snippet: Recombinant His-tagged Spike S1 wild-type (#40591-V08B1), S1 Omicron (#40592-V08H121) or Spike S1/S2 TRIMER wild-type (#40589-V08H8), all from Sino Biological at a concentration of 0.25 mg/ml supplemented with phenol red were injected into the hindbrain (1 nl) of 48 hpf zebrafish larvae.

Techniques: Variant Assay, Recombinant, Injection, Activity Assay, Fluorescence, Microscopy

Journal: Frontiers in Immunology

Article Title: Dual role of ACE2 in regulating inflammation triggered by Omicron S1 and other SARS-CoV-2 Spike variants

doi: 10.3389/fimmu.2025.1667880

Figure Lengend Snippet: Trimeric ancestral variant induces a weaker proinflammatory response than its monomeric form. Recombinant S1WT, S1/S2WT-T or vehicle (-) were injected in the hindbrain ventricle (HBV) of WT (A–E) 2-day postfertilization larvae (dpf). The transcript levels of the indicated genes (A–D) were analyzed at 12 hpi by RT-qPCR in larval head and rest of the body and caspase-1 activity was determined at 24 hpi using a fluorogenic substrate (E) . Graphs shown are representative of three independent experiments; technical replicates are displayed in each graph. The means ± SEM for each group is shown. P values were calculated using one-way analysis of variance (ANOVA) and Tukey multiple range test. n=40 in A-D, n=35 in E. ns, not significant; * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 and **** p < 0.0001. auf, arbitrary units of fluorescence.

Article Snippet: Recombinant His-tagged Spike S1 wild-type (#40591-V08B1), S1 Omicron (#40592-V08H121) or Spike S1/S2 TRIMER wild-type (#40589-V08H8), all from Sino Biological at a concentration of 0.25 mg/ml supplemented with phenol red were injected into the hindbrain (1 nl) of 48 hpf zebrafish larvae.

Techniques: Variant Assay, Recombinant, Injection, Quantitative RT-PCR, Activity Assay, Fluorescence

Journal: bioRxiv

Article Title: Design and development of a SARS and MERS Combination Vaccine

doi: 10.1101/2025.10.27.683653

Figure Lengend Snippet: ZRS=Zoogenic Recombinant SARS; blue=vaccines formulated with alum and MPLA; red=vaccines formulated with alum and CpG; orange=vaccines formulated with SWE. ZRS03=Zoogenic Recombinant SARS version 3 containing as antigens the S1 sub-unit of (SARS2-Wuhan + SARS2-variant [either alpha, beta or gamma] + SARS1-bat) (ZRS04, ZRS05 see ). Alum=aluminium hydroxide; CpG=CpG oligodeoxynucleotides; MPLA=Monophosphoryl-Lipid A; SWE=Squalene-in-Water-Emulsion.

Article Snippet: A SARS2 spike primary mouse antibody (CE5, Native Antigen Company, MAB12443) was included as a positive control, and normal mouse serum (Invitrogen, 10410) used as a negative control.

Techniques: Recombinant, Vaccines, Variant Assay, Emulsion

Journal: bioRxiv

Article Title: Design and development of a SARS and MERS Combination Vaccine

doi: 10.1101/2025.10.27.683653

Figure Lengend Snippet: ZRS=Zoogenic Recombinant SARS; blue=vaccines formulated with alum and MPLA; red=vaccines formulated with alum and CpG; ZRS03=Zoogenic Recombinant SARS version 3 containing as antigens the S1 sub-unit of (SARS2-Wuhan + SARS2-variant [either alpha, beta or gamma] + SARS1-bat) (see ). Antigens tested see . Alum=aluminium hydroxide; CpG=CpG oligodeoxynucleotides; MPLA=Monophosphoryl-Lipid A.

Article Snippet: A SARS2 spike primary mouse antibody (CE5, Native Antigen Company, MAB12443) was included as a positive control, and normal mouse serum (Invitrogen, 10410) used as a negative control.

Techniques: Recombinant, Vaccines, Variant Assay

Journal: bioRxiv

Article Title: Design and development of a SARS and MERS Combination Vaccine

doi: 10.1101/2025.10.27.683653

Figure Lengend Snippet: ZRS=Zoogenic Recombinant SARS; blue=vaccines formulated with alum and MPLA; red=vaccines formulated with alum and CpG; ZRS03=Zoogenic Recombinant SARS version 3 containing as antigens the S1 sub-unit of (SARS2-Wuhan + SARS2-variant [either alpha, beta or gamma] + SARS1-bat) (see ). Alum=aluminium hydroxide; CpG=CpG oligodeoxynucleotides; MPLA=Monophosphoryl-Lipid A.

Article Snippet: A SARS2 spike primary mouse antibody (CE5, Native Antigen Company, MAB12443) was included as a positive control, and normal mouse serum (Invitrogen, 10410) used as a negative control.

Techniques: Recombinant, Vaccines, Variant Assay

Journal: bioRxiv

Article Title: Design and development of a SARS and MERS Combination Vaccine

doi: 10.1101/2025.10.27.683653

Figure Lengend Snippet: ZRS=Zoogenic Recombinant SARS; blue=vaccines formulated with alum and MPLA; MP; red=vaccines formulated with alum and CpG; ZRS03=Zoogenic Recombinant SARS version 3 containing as antigens the S1 sub-unit of (SARS2-Wuhan + SARS2-variant [either alpha, beta or gamma] + SARS1-bat). (see ). Alum=aluminium hydroxide; CpG = CpG oligodeoxynucleotides; MPLA=Monophosphoryl-Lipid A.

Article Snippet: A SARS2 spike primary mouse antibody (CE5, Native Antigen Company, MAB12443) was included as a positive control, and normal mouse serum (Invitrogen, 10410) used as a negative control.

Techniques: Recombinant, Vaccines, Variant Assay

Journal: bioRxiv

Article Title: Design and development of a SARS and MERS Combination Vaccine

doi: 10.1101/2025.10.27.683653

Figure Lengend Snippet: ZRS=Zoogenic Recombinant SARS; orange=vaccines formulated with SWE; ZRS04=Zoogenic Recombinant SARS version 4 containing as antigens the S1 sub-unit of (SARS2-Wuhan + SARS1-bat) (see ). Antigens tested see . SWE=Squalene-in-Water-Emulsion.

Article Snippet: A SARS2 spike primary mouse antibody (CE5, Native Antigen Company, MAB12443) was included as a positive control, and normal mouse serum (Invitrogen, 10410) used as a negative control.

Techniques: Recombinant, Vaccines, Emulsion

Journal: bioRxiv

Article Title: Design and development of a SARS and MERS Combination Vaccine

doi: 10.1101/2025.10.27.683653

Figure Lengend Snippet: a. Neutralisation of ACE2 receptor SARS-CoV2 interaction; b. SARS-CoV2 in vitro viral neutralisation. ZRS = Zoogenic Recombinant SARS; orange = vaccines formulated with SWE; ZRS04 = Zoogenic Recombinant SARS version 4 containing as antigens the S1 sub-unit of (SARS2-Wuhan + SARS1-bat) (see ). SWE = Squalene-in-Water-Emulsion.

Article Snippet: A SARS2 spike primary mouse antibody (CE5, Native Antigen Company, MAB12443) was included as a positive control, and normal mouse serum (Invitrogen, 10410) used as a negative control.

Techniques: In Vitro, Recombinant, Vaccines, Emulsion

Journal: bioRxiv

Article Title: Design and development of a SARS and MERS Combination Vaccine

doi: 10.1101/2025.10.27.683653

Figure Lengend Snippet: ZRS = Zoogenic Recombinant SARS; orange = vaccines formulated with SWE; ZRS04 = Zoogenic Recombinant SARS version 4 containing as antigens the S1 sub-unit of (SARS2-Wuhan + SARS1-bat) (see ). SWE = Squalene-in-Water-Emulsion.

Article Snippet: A SARS2 spike primary mouse antibody (CE5, Native Antigen Company, MAB12443) was included as a positive control, and normal mouse serum (Invitrogen, 10410) used as a negative control.

Techniques: Recombinant, Vaccines, Emulsion

Journal: bioRxiv

Article Title: Design and development of a SARS and MERS Combination Vaccine

doi: 10.1101/2025.10.27.683653

Figure Lengend Snippet: ZRS=Zoogenic Recombinant SARS; orange=vaccines formulated with SWE; ZRS05=Zoogenic Recombinant SARS version 5 containing as antigens the S1 sub-unit of (SARS2-Wuhan + SARS1-bat) and MERS (see ). ISO=isotonic formulation; Antigens tested see . SWE=Squalene-in-Water-Emulsion.

Article Snippet: A SARS2 spike primary mouse antibody (CE5, Native Antigen Company, MAB12443) was included as a positive control, and normal mouse serum (Invitrogen, 10410) used as a negative control.

Techniques: Recombinant, Vaccines, Formulation, Emulsion

Journal: International Journal of Molecular Sciences

Article Title: Artificial Intelligence Reveals Nature: Functional Parallels Between a Designed and a Natural Peptide

doi: 10.3390/ijms262110607

Figure Lengend Snippet: RBD-ACE2 interaction inhibition assays. ( a ) Schematic illustration of ELISA experiment to evaluate the ability of the A13 or Nat1 to prevent the interaction of SARS-CoV-2 S1 protein RBD to human receptor ACE2. First the plates were incubated with immobilized RBD. They were then inoculated with peptides prior to the addition of recombinant human receptor ACE2. ( b ) Inhibition of RBD-ACE2 binding via A13 or Nat1. The level of RBD-ACE2 binding was measured by chemiluminescence. The negative control with no peptide representing RBD-ACE2 interaction was set to 1.00 (3.2 × 10 4 RLU) and other values were normalized to this number. To the same extent, at the concentration of 0.1 µg/mL, both peptides reduced ACE2 binding compared to the control. At 1.0 µg/mL, inhibition was further enhanced. The negative control peptide did not exhibit a notable inhibition. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001).

Article Snippet: The ability of the peptides to block the interaction between the RBD and ACE2 was assessed using a commercial ACE2 inhibition assay kit (BPS Bioscience, San Diego, CA, United States, CAT# 79931).

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Incubation, Recombinant, Binding Assay, Negative Control, Concentration Assay, Control, Standard Deviation, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Artificial Intelligence Reveals Nature: Functional Parallels Between a Designed and a Natural Peptide

doi: 10.3390/ijms262110607

Figure Lengend Snippet: NanoLuc bioreporter assays. ( a ) A schematic illustration for the NanoLuc bioreporter assay to determine SARS-CoV-2 S1 RBD—ACE2 inhibition by A13 or Nat1. ( b ) NanoLuc bioreporter assay indicates that both peptides reduce luciferase complementation to the same extent. The signal value for no peptide control was set to 1.00 (6.30 × 10 5 RLU) and all other values were normalized to it. In cell lysates, A13 and Nat1 both reduced luminescence, indicating inhibition of RBD-ACE2 binding. The negative control peptide had no significant effect. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).

Article Snippet: The ability of the peptides to block the interaction between the RBD and ACE2 was assessed using a commercial ACE2 inhibition assay kit (BPS Bioscience, San Diego, CA, United States, CAT# 79931).

Techniques: Inhibition, Luciferase, Control, Binding Assay, Negative Control, Standard Deviation, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Artificial Intelligence Reveals Nature: Functional Parallels Between a Designed and a Natural Peptide

doi: 10.3390/ijms262110607

Figure Lengend Snippet: Pseudovirus infectivity assays. ( a ) A schematic illustration for pseudovirus infectivity assay to determine the inhibition ability of the peptides to prevent psuedovirus to enter the cell. ( b ) Pseudovirus infectivity assays indicates that both peptides exhibit similar effects on pseudovirus infected cells. The signal value for pseudovirus infection with no peptide was set to 1.00 and corresponding values were normalized to it. Cells were incubated with pseudovirus in the absence/presence of A13 or Nat1, and infection was measured using a luciferase reporter. Both peptides reduce pseudovirus entry into ACE2-expressing cells. Neither peptide shows any visible effects on the non-peptide sample. Data represents the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).

Article Snippet: The ability of the peptides to block the interaction between the RBD and ACE2 was assessed using a commercial ACE2 inhibition assay kit (BPS Bioscience, San Diego, CA, United States, CAT# 79931).

Techniques: Infection, Inhibition, Incubation, Luciferase, Expressing, Standard Deviation, Two Tailed Test

Journal: Journal of Virology

Article Title: Spike proteins of coronaviruses activate mast cells for degranulation via stimulating Src/PI3K/AKT/Ca 2+ intracellular signaling cascade

doi: 10.1128/jvi.00078-25

Figure Lengend Snippet: Binding of Spike/RBD proteins to receptors triggers MC degranulation. ( A ) Expression of ACE2, DPP4, and APN in LUVA and HMC-1 cells was detected by immunostaining with specific antibodies and analyzed with flow cytometry. ( B ) LUVA, LAD2, and HMC-1 cells (1 × 10 6 cells for each) were treated with HCoV-229E and HCoV-NL63 (M.O.I = 1) or medium (mock) for 24 or 48 h. Viral replication was quantified by detecting the expression of nucleocapsid gene and normalizing with the gapdh gene. ( C ) HMC-1 and LUVA cells were exposed to Spike/RBD proteins (5 µg/mL) for 1 h at 4°C. The co-localization of these proteins with their respective receptors of APN, ACE2, and DPP4 was detected with confocal microscopy. Scale bar: 10 µm. ( D through G ) MC degranulation. MC degranulation in Spike/RBD protein-treated ( D ), HCoV-229E and HCoV-NL63 (M.O.I = 1) virus-infected LUVA, LAD2, or HMC-1 cells ( E ), Spike/RBD mutant protein-treated LUVA cells ( F ), and pre- and post-fusion S2 subunits from SARS-CoV-2-treated LUVA and HMC-1 cells (G) were detected by quantifying the β-hexosaminidase release. The compound 48/80 (C48/80) was used as the control. Data are presented as mean ± SD. One representative result from three independent repeats is shown. *** P < 0.001 is considered significant differences.

Article Snippet: The recombinant HCoV-NL63 Spike/RBD (receptor-binding domains) (40600-V08H), MERS-CoV Spike/RBD protein (40071-V08B1), SARS-CoV-2 Spike/RBD (40592-V08B), HCoV-229E Spike/RBD (40601-V08H), HCoV-HKU1 Spike/RBD (40021-V08H), HCoV-OC43 (40607-V08H1), and SARS-CoV Spike/RBD (40150-V08B2) were purchased from Sino Biological, Beijing, China.

Techniques: Binding Assay, Expressing, Immunostaining, Flow Cytometry, Confocal Microscopy, Virus, Infection, Mutagenesis, Control

Journal: Journal of Virology

Article Title: Spike proteins of coronaviruses activate mast cells for degranulation via stimulating Src/PI3K/AKT/Ca 2+ intracellular signaling cascade

doi: 10.1128/jvi.00078-25

Figure Lengend Snippet: Transcriptome analysis reveals Spike/RBD proteins-induced MC activation. LAD2 cells were exposed to Spike/RBD proteins (5 µg/mL) or virions of HCoV-229E and HCoV-NL63 (MOI = 1) for 24 h. Total RNAs were extracted from cells, and the transcriptome analysis was conducted. Data from three independent repeats were summarized. ( A ) Volcano plot of DEGs comparing Spike/RBD proteins treated or HCoV-229E and HCoV-NL63 infected cells to that of mock-infection or medium-treatment. The symbols of top upregulated or downregulated genes are shown. ( B ) Summary of DEGs. The consistently upregulated and downregulated genes in LAD2 cells from these treatments with spike/RBD proteins and viral particles. ( C ) GO functional enrichment analysis of DEGs. The color bar indicates the minus logarithm of Q values, and bubble size indicates the absolute gene counts enriched in a GO term. ( D-F ) Expression of inflammatory factors. Total RNAs from LAD2 cells or LUVA cells were isolated. The expression of IL-1β, IL-6, TNF-α, or IL-8 in virus-infected LAD2 cells ( D ) or Spike/RBD protein-treated LUVA cells ( E, F ) were detected by real-time (RT-)PCR. Data are presented as mean ± SD. One representative result from three independent repeats is shown. * P < 0.05, ** P < 0.01 and *** P < 0.001 are considered significant differences.

Article Snippet: The recombinant HCoV-NL63 Spike/RBD (receptor-binding domains) (40600-V08H), MERS-CoV Spike/RBD protein (40071-V08B1), SARS-CoV-2 Spike/RBD (40592-V08B), HCoV-229E Spike/RBD (40601-V08H), HCoV-HKU1 Spike/RBD (40021-V08H), HCoV-OC43 (40607-V08H1), and SARS-CoV Spike/RBD (40150-V08B2) were purchased from Sino Biological, Beijing, China.

Techniques: Activation Assay, Infection, Functional Assay, Expressing, Isolation, Virus, Quantitative RT-PCR

Journal: Journal of Virology

Article Title: Spike proteins of coronaviruses activate mast cells for degranulation via stimulating Src/PI3K/AKT/Ca 2+ intracellular signaling cascade

doi: 10.1128/jvi.00078-25

Figure Lengend Snippet: Spike/RBD proteins induce activation of the cellular Src/PI3K/AKT signaling pathway. ( A ) LUVA cells were exposed to Spike/RBD proteins (5 µg/mL) for 2 h. Src-pY416, Src, P-PI3K(P85), PI3K, P-AKT, and AKT levels were assessed by immunoblot analysis. ( B ) LUVA cells were exposed to Spike/RBD proteins (5 µg/mL) in the presence or not of LY294002 (100 µM) for 2 h. ( C ) LUVA or HMC-1 cells were treated with HCoV-229E and HCoV-NL63 (M.O.I = 1) for 2 h. MC degranulation was detected by quantifying the β-hexosaminidase release. Data are presented as mean ± SD. One representative result from three independent repeats is shown. *** P < 0.001 is considered significant differences.

Article Snippet: The recombinant HCoV-NL63 Spike/RBD (receptor-binding domains) (40600-V08H), MERS-CoV Spike/RBD protein (40071-V08B1), SARS-CoV-2 Spike/RBD (40592-V08B), HCoV-229E Spike/RBD (40601-V08H), HCoV-HKU1 Spike/RBD (40021-V08H), HCoV-OC43 (40607-V08H1), and SARS-CoV Spike/RBD (40150-V08B2) were purchased from Sino Biological, Beijing, China.

Techniques: Activation Assay, Western Blot

Journal: Journal of Virology

Article Title: Spike proteins of coronaviruses activate mast cells for degranulation via stimulating Src/PI3K/AKT/Ca 2+ intracellular signaling cascade

doi: 10.1128/jvi.00078-25

Figure Lengend Snippet: Binding of Spike/RBD proteins to receptors triggers MC degranulation. ( A ) Expression of ACE2, DPP4, and APN in LUVA and HMC-1 cells was detected by immunostaining with specific antibodies and analyzed with flow cytometry. ( B ) LUVA, LAD2, and HMC-1 cells (1 × 10 6 cells for each) were treated with HCoV-229E and HCoV-NL63 (M.O.I = 1) or medium (mock) for 24 or 48 h. Viral replication was quantified by detecting the expression of nucleocapsid gene and normalizing with the gapdh gene. ( C ) HMC-1 and LUVA cells were exposed to Spike/RBD proteins (5 µg/mL) for 1 h at 4°C. The co-localization of these proteins with their respective receptors of APN, ACE2, and DPP4 was detected with confocal microscopy. Scale bar: 10 µm. ( D through G ) MC degranulation. MC degranulation in Spike/RBD protein-treated ( D ), HCoV-229E and HCoV-NL63 (M.O.I = 1) virus-infected LUVA, LAD2, or HMC-1 cells ( E ), Spike/RBD mutant protein-treated LUVA cells ( F ), and pre- and post-fusion S2 subunits from SARS-CoV-2-treated LUVA and HMC-1 cells (G) were detected by quantifying the β-hexosaminidase release. The compound 48/80 (C48/80) was used as the control. Data are presented as mean ± SD. One representative result from three independent repeats is shown. *** P < 0.001 is considered significant differences.

Article Snippet: The recombinant HCoV-NL63 Spike/RBD (receptor-binding domains) (40600-V08H), MERS-CoV Spike/RBD protein (40071-V08B1), SARS-CoV-2 Spike/RBD (40592-V08B), HCoV-229E Spike/RBD (40601-V08H), HCoV-HKU1 Spike/RBD (40021-V08H), HCoV-OC43 (40607-V08H1), and SARS-CoV Spike/RBD (40150-V08B2) were purchased from Sino Biological, Beijing, China.

Techniques: Binding Assay, Expressing, Immunostaining, Flow Cytometry, Confocal Microscopy, Virus, Infection, Mutagenesis, Control

Journal: Journal of Virology

Article Title: Spike proteins of coronaviruses activate mast cells for degranulation via stimulating Src/PI3K/AKT/Ca 2+ intracellular signaling cascade

doi: 10.1128/jvi.00078-25

Figure Lengend Snippet: Transcriptome analysis reveals Spike/RBD proteins-induced MC activation. LAD2 cells were exposed to Spike/RBD proteins (5 µg/mL) or virions of HCoV-229E and HCoV-NL63 (MOI = 1) for 24 h. Total RNAs were extracted from cells, and the transcriptome analysis was conducted. Data from three independent repeats were summarized. ( A ) Volcano plot of DEGs comparing Spike/RBD proteins treated or HCoV-229E and HCoV-NL63 infected cells to that of mock-infection or medium-treatment. The symbols of top upregulated or downregulated genes are shown. ( B ) Summary of DEGs. The consistently upregulated and downregulated genes in LAD2 cells from these treatments with spike/RBD proteins and viral particles. ( C ) GO functional enrichment analysis of DEGs. The color bar indicates the minus logarithm of Q values, and bubble size indicates the absolute gene counts enriched in a GO term. ( D-F ) Expression of inflammatory factors. Total RNAs from LAD2 cells or LUVA cells were isolated. The expression of IL-1β, IL-6, TNF-α, or IL-8 in virus-infected LAD2 cells ( D ) or Spike/RBD protein-treated LUVA cells ( E, F ) were detected by real-time (RT-)PCR. Data are presented as mean ± SD. One representative result from three independent repeats is shown. * P < 0.05, ** P < 0.01 and *** P < 0.001 are considered significant differences.

Article Snippet: The recombinant HCoV-NL63 Spike/RBD (receptor-binding domains) (40600-V08H), MERS-CoV Spike/RBD protein (40071-V08B1), SARS-CoV-2 Spike/RBD (40592-V08B), HCoV-229E Spike/RBD (40601-V08H), HCoV-HKU1 Spike/RBD (40021-V08H), HCoV-OC43 (40607-V08H1), and SARS-CoV Spike/RBD (40150-V08B2) were purchased from Sino Biological, Beijing, China.

Techniques: Activation Assay, Infection, Functional Assay, Expressing, Isolation, Virus, Quantitative RT-PCR

Journal: Journal of Virology

Article Title: Spike proteins of coronaviruses activate mast cells for degranulation via stimulating Src/PI3K/AKT/Ca 2+ intracellular signaling cascade

doi: 10.1128/jvi.00078-25

Figure Lengend Snippet: Spike/RBD proteins induce activation of the cellular Src/PI3K/AKT signaling pathway. ( A ) LUVA cells were exposed to Spike/RBD proteins (5 µg/mL) for 2 h. Src-pY416, Src, P-PI3K(P85), PI3K, P-AKT, and AKT levels were assessed by immunoblot analysis. ( B ) LUVA cells were exposed to Spike/RBD proteins (5 µg/mL) in the presence or not of LY294002 (100 µM) for 2 h. ( C ) LUVA or HMC-1 cells were treated with HCoV-229E and HCoV-NL63 (M.O.I = 1) for 2 h. MC degranulation was detected by quantifying the β-hexosaminidase release. Data are presented as mean ± SD. One representative result from three independent repeats is shown. *** P < 0.001 is considered significant differences.

Article Snippet: The recombinant HCoV-NL63 Spike/RBD (receptor-binding domains) (40600-V08H), MERS-CoV Spike/RBD protein (40071-V08B1), SARS-CoV-2 Spike/RBD (40592-V08B), HCoV-229E Spike/RBD (40601-V08H), HCoV-HKU1 Spike/RBD (40021-V08H), HCoV-OC43 (40607-V08H1), and SARS-CoV Spike/RBD (40150-V08B2) were purchased from Sino Biological, Beijing, China.

Techniques: Activation Assay, Western Blot