pcmv3 hdpp4 cha  (Sino Biological)

Journal: bioRxiv

Article Title: Dysregulation of lung epithelial cell homeostasis and immunity contributes to Middle East Respiratory Syndrome coronavirus disease severity

doi: 10.1101/2024.10.03.616483

Figure Lengend Snippet: (A) Study design. 19-23 week old female C57BL/6 hDPP4 mice were infected with PBS (“mock”, N = 14), 5×10 4 PFU mouse adapted MERS ma1 (“low dose”, N = 14) or 5×10 6 PFU MERS ma1 (“high dose”, N = 16). On 2, 4, and 7dpi, lung tissue from 4 mice per group was harvested for virological, pathological, and multi-omics measures. (B) Percent starting weight. The boxes encompass the 25th to 75th percentile, the line is at the median, while the whiskers represent the range. Asterisks denote statistical significance as determined by Two-way ANOVA with a Tukey’s multiple comparison test. * = 0.01, **** = < 0.0001. (C) Percent Survival and significance as determined by Mantel-Cox test (***P = 0.0001). (D) Virus lung titer per lung lobe by plaque assay. Asterisks denote statistical significance by Two-way ANOVA with a Tukey’s multiple comparison test. * = 0.01. (E) Lung hemorrhage scored on a scale of 0–4, where 0 is a normal pink healthy lung and 4 is a completely dark red lung. Asterisks denote statistical significance as determined by Two-way ANOVA with a Tukey’s multiple comparison test. * = 0.01, ** = 0.001, **** < 0.0001.

Article Snippet: To determine the importance of functional T-cells and B-cells on MERS-CoV pathogenesis, we bred C57BL/6 hDPP4 mice with C57BL/6 RAG1 −/− (Jackson Labs Strain #002216) mice resulting in C57BL/6 hDPP4 RAG1 −/− offspring.

Techniques: Infection, Biomarker Discovery, Comparison, Virus, Plaque Assay

Journal: bioRxiv

Article Title: Dysregulation of lung epithelial cell homeostasis and immunity contributes to Middle East Respiratory Syndrome coronavirus disease severity

doi: 10.1101/2024.10.03.616483

Figure Lengend Snippet: (A) Study design or MERS-CoV infection of mice deficient in functional T and B-cells. (B) Percent starting weight of 20-week old male and female WT C57BL/6 hDPP4 mice or C57BL/6 hDPP4 RAG1 −/− mice infected with 5×10 4 PFU MERS ma1 (WT, N = 11; RAG1 −/− N = 10) or 5×10 6 PFU MERS ma1 WT, N = 10; RAG1 −/− N = 12). Asterisks denote statistical significance as determined by Two-way ANOVA with a Tukey’s multiple comparison test. (C) Percent Survival. (D) Virus lung titer on 7dpi by plaque assay. Asterisks denote statistical significance as determined by Kruskal-Wallis test with a Dunn’s multiple comparison test. (E) Gross pathology “lung discoloration score” on 7dpi.

Article Snippet: To determine the importance of functional T-cells and B-cells on MERS-CoV pathogenesis, we bred C57BL/6 hDPP4 mice with C57BL/6 RAG1 −/− (Jackson Labs Strain #002216) mice resulting in C57BL/6 hDPP4 RAG1 −/− offspring.

Techniques: Infection, Functional Assay, Comparison, Virus, Plaque Assay

Journal: npj Viruses

Article Title: Transmission dynamics of MERS-CoV in a transgenic human DPP4 mouse model

doi: 10.1038/s44298-024-00048-y

Figure Lengend Snippet: Detection of hDPP4 expression in hDPP4 mice using immunohistochemistry in ( a ) nasal mucosa; ( b ) trachea; and ( c ) type I and II pneumocytes, bronchiolar and endothelial cells in lung tissue. d Comparison of hDPP4 expression in lung and kidney tissue obtained from wildtype and hDPP4 mice using flow cytometry. N = 3, bars represent median. e Survival curves of mice after inoculation with 5 × 10 5 TCID 50 MERS-CoV. N = 4 (Wildtype) or 5 (hDPP4). ** = p < 0.01. f Relative weight loss in mice after MERS-CoV inoculation. The lines represent median±range. Mice were euthanized upon reaching >20% of body weight loss (dotted line). g Viral load (gRNA) in oropharyngeal swabs obtained from mice after inoculation with 5 × 10 5 TCID 50 MERS-CoV. h Infectious MERS-CoV titers in lung, brain, and kidney tissue of hDPP4 mice. Dotted line detection limit.

Article Snippet: hDPP4 mice were developed by ingenious Targeting Laboratory.

Techniques: Expressing, Immunohistochemistry, Comparison, Flow Cytometry

Journal: npj Viruses

Article Title: Transmission dynamics of MERS-CoV in a transgenic human DPP4 mouse model

doi: 10.1038/s44298-024-00048-y

Figure Lengend Snippet: a Survival curves of mice inoculated with 3.8 × 10 2 TCID 50 MERS-CoV via aerosols. N = 6. b Viral load (gRNA) in oropharyngeal swabs obtained from mice inoculated with 3.8 × 10 2 TCID 50 MERS-CoV via aerosols. Bars represent median. b Viral load (gRNA) and c (infectious virus) in lung and brain tissue of hDPP4 mice inoculated with 3.8 × 10 2 TCID 50 MERS-CoV via aerosols.

Article Snippet: hDPP4 mice were developed by ingenious Targeting Laboratory.

Techniques: Virus

Journal: npj Viruses

Article Title: Transmission dynamics of MERS-CoV in a transgenic human DPP4 mouse model

doi: 10.1038/s44298-024-00048-y

Figure Lengend Snippet: hDPP4 mice were exposed to 5 × 10 4 TCID 50 MERS-CoV ( N = 18) or 5 × 10 6 TCID 50 MERS-CoV ( N = 20). a Survival curves of hDPP4 mice exposed to fomites containing MERS-CoV. Viral gRNA ( b ) or sgRNA ( c ) in lung and brain tissue of hDPP4 mice that reached endpoint criteria. d Infectious MERS-CoV detected in lung and brain tissue of hDPP4 mice that reached endpoint criteria. e Serology titers in sera of survivors obtained 28 dpe. ELISA assays were performed using MERS-CoV S1 protein. Dotted line = limit of detection. ** = p < 0.01.

Article Snippet: hDPP4 mice were developed by ingenious Targeting Laboratory.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: npj Viruses

Article Title: Transmission dynamics of MERS-CoV in a transgenic human DPP4 mouse model

doi: 10.1038/s44298-024-00048-y

Figure Lengend Snippet: hDPP4 mice were inoculated intranasally with 10 4 TCID 50 MERS-CoV ( N = 20) or 10 5 TCID 50 MERS-CoV ( N = 38). a Survival curves of mice directly exposed to donor mice infected with MERS-CoV. Viral gRNA ( b ) or sgRNA ( c ) in lung and brain tissue of hDPP4 mice that reached endpoint criteria. d Infectious MERS-CoV detected in lung and brain tissue of hDPP4 mice that reached endpoint criteria. e Serology titers in sera of survivors obtained 28 dpe. ELISA assays were performed using MERS-CoV S1 protein. Dotted line = limit of detection.

Article Snippet: hDPP4 mice were developed by ingenious Targeting Laboratory.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: npj Viruses

Article Title: Transmission dynamics of MERS-CoV in a transgenic human DPP4 mouse model

doi: 10.1038/s44298-024-00048-y

Figure Lengend Snippet: hDPP4 mice were inoculated intranasally with 10 4 TCID 50 MERS-CoV ( N = 5) or 10 5 TCID 50 MERS-CoV ( N = 54). a Survival curves of mice exposed to donor mice infected with MERS-CoV. Viral gRNA ( b ) or sgRNA ( c ) in lung and brain tissue of hDPP4 mice that reached endpoint criteria. d Infectious MERS-CoV detected in lung and brain tissue of hDPP4 mice that reached endpoint criteria. e Serology titers in sera of survivors obtained 28 dpe. ELISA assays were performed using MERS-CoV S1 protein. Dotted line = limit of detection.

Article Snippet: hDPP4 mice were developed by ingenious Targeting Laboratory.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: Virus Research

Article Title: A dual-targeting approach using a human bispecific antibody against the receptor-binding domain of the Middle East Respiratory Syndrome Coronavirus

doi: 10.1016/j.virusres.2024.199383

Figure Lengend Snippet: Neutralizing properties of selected antibodies against MERS-CoV pseudotyped virus in vitro. (A) Schematic representation of the neutralizing activity analysis using a pseudotyped virus assay. (B) Verification of hDPP4 expression in 293T/hDPP4 cell lines using immunoblot analysis. (C) Multiplicity of infection (MOI)-dependent infection by the MERS-CoV pseudotyped virus in 293T/hDPP4 cells. (D–G) Concentration-dependent neutralization of MERS-CoV pseudotyped virus infection in 293T/hDPP4 cells by the control IgG and selected mAbs: (D) K111.1, (E) K111.2, (F) K111.3, and (G) K111.4.

Article Snippet: Briefly, 50 ng of purified Fc-tagged hDPP4 (Acro Biosystems, Newark, DE, USA) was coated onto each well of a 96-well plate and incubated for 2 h at room temperature (RT).

Techniques: Virus, In Vitro, Activity Assay, Expressing, Western Blot, Infection, Concentration Assay, Neutralization, Control

Journal: Virus Research

Article Title: A dual-targeting approach using a human bispecific antibody against the receptor-binding domain of the Middle East Respiratory Syndrome Coronavirus

doi: 10.1016/j.virusres.2024.199383

Figure Lengend Snippet: Biochemical characterization and neutralization efficacy analysis of bsAbs for MERS-CoV RBD. (A) SPR sensorgrams showing the binding of bsAb K207.C on an RBD-immobilized sensor chip that was pre-saturated with K111.3 (left) and the binding of K207.C to an RBD-immobilized sensor chip that was pre-saturated with K111.1 (right). (B) SPR sensorgrams demonstrating the competitive binding of K111.1 (left) and K111.3 (right) on an RBD-immobilized sensor chip pre-saturated with bsAb K207.C. (C) Results from the neutralization activity assay, showing the efficacy of parental mAb K111.3, bsAb K207.C, and control IgG against MERS-CoV pseudotyped virus infections in 293T/hDPP4 cells, displayed in a dose−response curve. (D) Bar graph representing the neutralization potency of parental mAb K111.3, bsAb K207.C, and control IgG in disrupting the interaction of MERS-CoV RBD with the hDPP4 protein.

Article Snippet: Briefly, 50 ng of purified Fc-tagged hDPP4 (Acro Biosystems, Newark, DE, USA) was coated onto each well of a 96-well plate and incubated for 2 h at room temperature (RT).

Techniques: Neutralization, Binding Assay, Activity Assay, Control, Virus