hdpp4 mice  (inGenious Targeting Laboratory)

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: Nature Communications

Article Title: Isolation and characterization of a pangolin-borne HKU4-related coronavirus that potentially infects human-DPP4-transgenic mice

doi: 10.1038/s41467-024-45453-2

Figure Lengend Snippet: a Kinetics of replication of pangolin-CoV-HKU4-P251T in human Huh7, Caco-2, Calu-3, and BEAS-2B cell lines. Viral RNA levels of virus in cell culture supernatant were detected at 0, 12, 24, 48, and 72 HPI, with three independent biological replicates per time point and three technical replicates per sample. The viral RNA levels were normalized relative to initial phase of infection (0 HPI). Data are presented as mean ± SD (shown as error bars). ANOVA was used for multiple comparisons. b Fluorescence in situ hybridization locating ORF1ab gene of Pangolin-CoV-HKU4-P251T in each cell lines at 48 HPI. Nuclei, DAPI (blue); ORF1ab probe, Quasar 570 (red). Representative microscopy fields are shown. Original magnification ×200. See also Supplementary Fig. . c Western blotting analyses of expressions of hDPP4 and hACE2 in various cell lines. Α-Tubulin was used as the loading control. d Cytopathic effect (CPE) of virus-infected Huh7 cells at 5 DPI. Original magnification×100. The uninfected Huh7 cells cultivated in parallel are used as controls. Each imaging experiment was independently performed at least three times with similar results, and representative images are shown ( b – d ). e Viral titers of infected Huh7 cells at different time points post-infection. n = 3 biologically independent experiments per time point. Data are presented as mean ± SD (shown as error bars). ANOVA was used for multiple comparisons. f The growth curve of passages 1 and 15 viruses in Huh7 cells. Cells were infected with the virus at an MOI of 0.01. At the specified time intervals, the supernatants were collected and the viral titers were measured as instructed. n = 3 biologically independent experiments per time point. Data are displayed as a line representing the mean, with individual data points shown as dots. Source data are provided as a Source Data file.

Article Snippet: Five weeks old hDPP4 female mice (Cat. NO. NM-HU-190042) were purchased from Shanghai Model Organisms Center, Inc. ( https://www.modelorg.com/ ).

Techniques: Virus, Cell Culture, Infection, Fluorescence, In Situ Hybridization, Microscopy, Western Blot, Control, Imaging

Journal: Nature Communications

Article Title: Isolation and characterization of a pangolin-borne HKU4-related coronavirus that potentially infects human-DPP4-transgenic mice

doi: 10.1038/s41467-024-45453-2

Figure Lengend Snippet: a The copy number of viral RNA in lungs of pangolin-CoV-HKU4-P251T-infected hDPP4 (orange) and WT (blue) mice was determined using qRT-PCR. n = 3 biologically independent animals per genotype for each time point. Data are presented as mean ± SD with scatter plot at each time point. Dashed lines denote the detection limit. Whenever the sample measurement was below the detection limit, the result was assigned a value equal to the minimum detection limit to facilitate statistical analysis of the data. ANOVA was used for multiple comparisons. b Relative sgmRNA quantification in lungs of virus-infected hDPP4 (orange) and WT (blue) mice. n = 3 biologically independent animals per genotype for each time point. Data are presented as mean ± SD with scatter plot at each time point. ANOVA was used for multiple comparisons. c The expression of viral nucleocapsid protein in lungs of infected hDPP4-mice. Representative IHC staining images of viral nucleocapsid protein expression in lungs of infected hDPP4-mice at 3, 6, and 12 DPI. The hDPP4-mice inoculated with cell medium are used as control. Images are representative of three experimental animals. Original magnification ×200 (upper panel). Red frames indicate regions shown in high magnification (lower panel, ×400). See also Supplementary Fig. and . Source data are provided as a Source Data file.

Article Snippet: Five weeks old hDPP4 female mice (Cat. NO. NM-HU-190042) were purchased from Shanghai Model Organisms Center, Inc. ( https://www.modelorg.com/ ).

Techniques: Infection, Quantitative RT-PCR, Virus, Expressing, Immunohistochemistry, Control

Journal: Nature Communications

Article Title: Isolation and characterization of a pangolin-borne HKU4-related coronavirus that potentially infects human-DPP4-transgenic mice

doi: 10.1038/s41467-024-45453-2

Figure Lengend Snippet: a Percentage of body weight changes of infected hDPP4-mice and WT-mice. n = 17 biologically independent animals per genotype. Data are reported as mean ± SD (shown as error bars). The solid dots and lines indicate the weight changes of hDPP4-mice. The hollow dots and dashed lines indicate the weight changes of WT-mice. Orange, live virus-infected mice; blue, heat-inactivated viruses-infected mice; dark gray, cell culture supernatant-infected mice. ANOVA was used for multiple comparisons. b Pathological features of virus-infected hDPP4-mice. H&E-stained lung sections at 3, 6, and 12 DPI are displayed. The hDPP4-mice inoculated with cell medium are used as control. Images are representative of three experimental animals. Original magnification ×200. See also Supplementary Fig. . c Immunohistochemical analysis of lung tissue stained with MAC2 antibody for macrophage detection. The hDPP4-mice inoculated with cell medium are used as control. Images are representative of three experimental animals. Original magnification ×400. See also Supplementary Fig. . d Cytokine response in the lung of hDPP4 (orange) and WT (blue) mice following virus infection. mRNA levels were monitored by qPCR at 1, 3, 6, and 12 DPI. The relative expression of mRNA at 3, 6, and 12 DPI was measured by comparative 2 −∆∆CT method related to 1 DPI and the mean ± SD (shown as error bars) of fold change of three independent biological replicates for each gene at each time point are presented. Student’s T -test (two-tailed) was used for comparisons of hDPP4- and WT-mice. Source data are provided as a Source Data file.

Article Snippet: Five weeks old hDPP4 female mice (Cat. NO. NM-HU-190042) were purchased from Shanghai Model Organisms Center, Inc. ( https://www.modelorg.com/ ).

Techniques: Infection, Virus, Cell Culture, Staining, Control, Immunohistochemical staining, Expressing, Two Tailed Test

Journal: Vaccines

Article Title: Comparing the Immunogenicity and Protective Effects of Three MERS-CoV Inactivation Methods in Mice

doi: 10.3390/vaccines10111843

Figure Lengend Snippet: The three inactivated MERS-CoV induced a protective immune response against MERS-CoV infection in the hDPP4-transgenic mouse model. ( A ) Changes in body weight and survival rate were monitored for two weeks after MERS-CoV infection. The survival symbols of H 2 O 2 -MERS-CoV and BEI-MERS-CoV overlapped with FA-MERS-CoV. ( B ) MERS-CoV specific neutralizing antibodies (nAbs) and ( C , D ) S1 protein-specific serum IgG/IgG2c/IgG1 levels were measured for two weeks after the third immunization. ( E ) MERS-CoV titers in lung and brain tissues were observed by plaque assay on dpi 4 and 8 after the MERS virus challenge. Data are represented as the mean ± S.E.M. *** p < 0.001 compared with alum only. ### p < 0.001, #### p < 0.0001 compared with alum only lung. † p < 0.05, †††† p < 0.00001 compared with alum only brain.

Article Snippet: The 6-week-old female hDPP4 (C57BL/6 with human DPP4) transgenic mice were provided by Macrogen (Seoul, Korea) and were maintained in an ABSL-3 facility at the KDCA.

Techniques: Infection, Transgenic Assay, Plaque Assay, Virus

Journal: Nature Communications

Article Title: Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV

doi: 10.1038/s41467-019-13940-6

Figure Lengend Snippet: a Percent starting weight of 9–12-week-old male and female Ces1c −/− hDPP4 mice prophylactically administered subcutaneous vehicle or remdesivir (RDV, 25 mg/kg) BID the day prior to infection with either 5E + 04 (vehicle n = 14, RDV n = 14) or 5E + 05 (vehicle n = 14, RDV n = 15) plaque-forming units (pfu) MERS M35C4. Asterisks indicate statistically significant differences ( P < 0.05) as determined by two-way ANOVA and Tukey’s multiple comparison test. b Percent survival of each cohort and survival analysis by Mantel–Cox test ( P < 0.05, N per group noted in a ). c 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. On 4 dpi, N = 4/group, and on 6 dpi the remaining animals are plotted. Asterisks indicate statistically significant differences ( P < 0.05) as determined by two-way ANOVA and Tukey’s multiple comparison test. d MERS-CoV lung titer on 4 ( N = 4) and 6 dpi (all remaining animals). Asterisks indicate statistically significant differences ( P < 0.05) as determined by two-way ANOVA and Sidek’s multiple comparison test. For a , c , d , the boxes encompass the 25th to 75th percentile, the line is at the median, while the whiskers represent the range. e Hematoxylin (nuclei, blue) and immunostaining for MERS-CoV antigen (brown) in lung tissue sections from 4 dpi. All photos were taken with the same magnification. The black bar indicates 100 µM scale. Images from representative mice for each group are shown.

Article Snippet: Fig. 2 Prophylactic RDV reduces MERS-CoV replication and disease. a Percent starting weight of 9–12-week-old male and female Ces1c −/− hDPP4 mice prophylactically administered subcutaneous vehicle or remdesivir (RDV, 25 mg/kg) BID the day prior to infection with either 5E + 04 (vehicle n = 14, RDV n = 14) or 5E + 05 (vehicle n = 14, RDV n = 15) plaque-forming units (pfu) MERS M35C4.

Techniques: Infection, Immunostaining

Journal: Nature Communications

Article Title: Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV

doi: 10.1038/s41467-019-13940-6

Figure Lengend Snippet: a Percent starting weight (Left) of 12–14-week-old female Ces1c −/− hDPP4 mice infected with 5E + 04 pfu MERS M35C4 and treated BID with either vehicle ( n = 9) or remdesivir (RDV, 25 mg/kg, n = 9) subcutaneously beginning −1 dpi. Asterisks indicate statistically significant differences ( P < 0.05) as determined by two-way ANOVA and Tukey’s multiple comparison test. (Middle) MERS-CoV lung titer on 2 ( N = 3) and 6 dpi (all remaining animals). Asterisks indicate statistically significant differences ( P < 0.05) as determined by Mann–Whitney test. (Right) WBP was used to assess pulmonary function in mice. PenH is a surrogate measure of airway resistance or bronchoconstriction. Asterisks indicate statistical differences by two-way ANOVA with Sidek’s multiple comparison test. b Percent starting weight (left), virus lung titer (middle), and pulmonary function metric PenH (right) of cohorts of mice similar in age and sex and infected similarly with MERS-CoV as in b but treated with vehicle ( n = 9), LPV/RTV + IFNb low (1× human equivalent) ( n = 9), LPV/RTV + IFNb high (25× human equivalent) ( n = 9), or IFNb high only ( n = 9). Oral vehicle or lopinavir/ritonavir (160/40 mg/kg) were administered orally once daily beginning the −1 dpi. IFNb treatment was initiated 2 h prior to infection and every other day thereafter. To control for dosing effects, vehicle-treated mice received both LPV/RTV vehicle and subcutaneous PBS to mirror IFNb injections. Likewise, IFNb only group received oral vehicle to mirror that seen in orally dosed groups. Similar statistical tests performed on a were performed on b . For the box and whisker plots, the boxes encompass the 25th to 75th percentile, the line is at the median, while the whiskers represent the range.

Article Snippet: Fig. 2 Prophylactic RDV reduces MERS-CoV replication and disease. a Percent starting weight of 9–12-week-old male and female Ces1c −/− hDPP4 mice prophylactically administered subcutaneous vehicle or remdesivir (RDV, 25 mg/kg) BID the day prior to infection with either 5E + 04 (vehicle n = 14, RDV n = 14) or 5E + 05 (vehicle n = 14, RDV n = 15) plaque-forming units (pfu) MERS M35C4.

Techniques: Infection, MANN-WHITNEY, Whisker Assay

Journal: Nature Communications

Article Title: Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV

doi: 10.1038/s41467-019-13940-6

Figure Lengend Snippet: Percent starting weight of 10–12-week-old female Ces1c −/− hDPP4 mice infected with 5E + 04 pfu MERS M35C4 and treated with a subcutaneous vehicle for RDV ( N = 13) or remdesivir (RDV, 25 mg/kg, N = 14) BID beginning 1 dpi or b vehicle for LPV/RTV-IFNb ( N = 15), LPV/RTV-IFNb low ( N = 16) or LPV/RTV-IFNb high ( N = 16) beginning 1 dpi. Oral vehicle or lopinavir/ritonavir (160/40 mg/kg) was administered orally once daily. IFNb low (1x human equivalent dose of 1.6 MIU/kg) and high (25x human equivalent dose of 40 MIU/kg) or PBS vehicle were administered via subcutaneous injection every other day. Asterisks indicate statistical differences by two-way ANOVA with Tukey’s multiple comparison test. c Lung hemorrhage 6 dpi for all animals in a , b scored on a scale of 0–4, where 0 is a normal pink healthy lung and 4 is a diffusely discolored dark red lung. d MERS-CoV lung titer 6 dpi in mice as described in a , b . Asterisks indicate statistical significance ( N group described in a and b , P < 0.05) by one-way ANOVA with Kruskal–Wallis test for ( c , d ). Data for a – d are compiled from two independent experiments. For the box and whisker plots, the boxes encompass the 25th to 75th percentile, the line is at the median, while the whiskers represent the range. e Representative photomicrographs of MERS-CoV antigen (brown) and hematoxylin stained nuclei (blue) in mouse lung tissue sections from 6 dpi. The black bar is 100 µM.

Article Snippet: Fig. 2 Prophylactic RDV reduces MERS-CoV replication and disease. a Percent starting weight of 9–12-week-old male and female Ces1c −/− hDPP4 mice prophylactically administered subcutaneous vehicle or remdesivir (RDV, 25 mg/kg) BID the day prior to infection with either 5E + 04 (vehicle n = 14, RDV n = 14) or 5E + 05 (vehicle n = 14, RDV n = 15) plaque-forming units (pfu) MERS M35C4.

Techniques: Infection, Injection, Whisker Assay, Staining