Journal: Emerging Infectious Diseases

Article Title: Livestock Susceptibility to Infection with Middle East Respiratory Syndrome Coronavirus

doi: 10.3201/eid2302.161239

Figure Lengend Snippet: Presence of MERS-CoV receptor DPP4 (IHC) and of mucosubstances (PAS) in upper and lower respiratory tract tissues from sheep, pigs, llamas, and horses. A) In the nose, DPP4 (red cytoplasmic or membrane staining) was present on the lining epithelium of pigs, llamas, and horses but not sheep. PAS staining (magenta) demonstrated more mucous cells in the lining epithelium of sheep and horses and a layer of mucus on the lining epithelium of the horses. B) DPP4 (red cytoplasmic or membrane staining) was present on the lining epithelium of the trachea, bronchus/bronchioles, and alveoli in the pigs, llamas and horses but not in the sheep. Original magnification ×400 for all images. DPP4, dipeptidyl peptidase-4; IHC, immunohistochemistry; MERS-CoV, Middle East respiratory syndrome coronavirus; PAS, periodic acid–Schiff; term., terminal.

Article Snippet: In brief, we performed DPP4 IHC staining by using 5 μg/mL of polyclonal goat IgG anti-human DPP4 antibody (R&D Systems, Abingdon, UK) and peroxidase-labeled rabbit anti-goat IgG (1:200; DAKO; Agilent Technologies Company, Santa Clara, CA, USA) as a secondary antibody.

Techniques: Membrane, Staining, Immunohistochemistry

Journal: Oncotarget

Article Title: Synergistic cytotoxic activity of cannabinoids from cannabis sativa against cutaneous T-cell lymphoma (CTCL) in-vitro and ex-vivo

doi: 10.18632/oncotarget.27528

Figure Lengend Snippet: PBL were isolated from blood samples of Sézary patients ( n = 6), and were treated with S4 (5 μg/mL), S5 (6 μg/mL), S4 (5 μg/mL) +S5 (6 μg/mL) and control (methanol; Vehicle treatment) for 48 h. Cells were harvested and analyzed by FACS following CD4-APC, CD26-alexa 405, Annexin V-FITC and PI staining. Apoptotic-induced cells (Annexin positive cells) were determined in the CD4 + CD26 - cell population and in non-CD4 + CD26 - cells of treated cells minus control. ( A ) The percent of apoptotic-induced CD4 + CD26 - cells is presented for single treatment compared to combined treatment; ( n = 4); *** denote significant difference between means (one way ANOVA; p < 0.001). ( B ) The percent of apoptotic-induced cells following the combined treatment was compared between CD4 + CD26 - cells and non-CD4 + CD26 - cells of SPBL; ( n = 6); ** denote significant difference between means (paired student T test; 0.001 < P < 0.05).

Article Snippet: Cells were suspended in 100 μL binding buffer with 1 μL Annexin V-FITC (4830-01-1, eBioscience) + 2 μL CD26 Alexa Fluor 405-conjugated antibody (FAB1180V-100UG, R&D SYSTEMS) + 10 μL CD4-APC-conjugated antibody (FAB3791A-100, R&D SYSTEMS) and incubated for 15 min at room temperature.

Techniques: Isolation, Control, Staining

Journal: Cytotherapy

Article Title: Differentiation of immortalized human multi-lineage progenitor to alveolar type 2-like cells: angiotensin-converting enzyme 2 expression and binding of severe acute respiratory syndrome coronavirus 2 spike and spike 1 proteins

doi: 10.1016/j.jcyt.2021.07.017

Figure Lengend Snippet: Summary of confocal analysis of control E12 MLPCs, E12 AT2-like cells and SAEpis.

Article Snippet: Antibodies specific for cytokeratin 19 (CK19) (MAB3608), epithelial cell adhesion molecule (EpCAM) (MAB9601), SOX17 (MAB1924), CD26 (MAB1180) and ACE-2 labeled with Alexa Fluor 594 (FAB9332T) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Control

Journal: Cytotherapy

Article Title: Differentiation of immortalized human multi-lineage progenitor to alveolar type 2-like cells: angiotensin-converting enzyme 2 expression and binding of severe acute respiratory syndrome coronavirus 2 spike and spike 1 proteins

doi: 10.1016/j.jcyt.2021.07.017

Figure Lengend Snippet: Confocal analysis of undifferentiated E12 control MLPCs, differentiated E12 AT2-like cells and SAEpis. Cells were labeled with primary antibodies (TERT, EpCAM, CD26 and TM4SF1) and secondary antibodies labeled with Alexa Fluor 594. Positive cells are stained red. DAPI staining shows the nuclei as blue. Negative cells are indicated only by blue nuclei. The results are representative of at least six different culture experiments and determinations done on different days.

Article Snippet: Antibodies specific for cytokeratin 19 (CK19) (MAB3608), epithelial cell adhesion molecule (EpCAM) (MAB9601), SOX17 (MAB1924), CD26 (MAB1180) and ACE-2 labeled with Alexa Fluor 594 (FAB9332T) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Control, Labeling, Staining

Journal: Cytotherapy

Article Title: Differentiation of immortalized human multi-lineage progenitor to alveolar type 2-like cells: angiotensin-converting enzyme 2 expression and binding of severe acute respiratory syndrome coronavirus 2 spike and spike 1 proteins

doi: 10.1016/j.jcyt.2021.07.017

Figure Lengend Snippet: The qRT-PCR analysis of ACE-2, DPP4 (CD26), EpCAM, SPC, CK19 and TM4SF1 expression of control E12 MLPCs, E12 AT2-like cells and SAEpis. Data were normalized to GAPDH. The results represent at least four different culture experiments done on different days.

Article Snippet: Antibodies specific for cytokeratin 19 (CK19) (MAB3608), epithelial cell adhesion molecule (EpCAM) (MAB9601), SOX17 (MAB1924), CD26 (MAB1180) and ACE-2 labeled with Alexa Fluor 594 (FAB9332T) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Quantitative RT-PCR, Expressing, Control

Journal: PLoS ONE

Article Title: Multi-Organ Damage in Human Dipeptidyl Peptidase 4 Transgenic Mice Infected with Middle East Respiratory Syndrome-Coronavirus

doi: 10.1371/journal.pone.0145561

Figure Lengend Snippet: (A) Schematic diagram of the optimized hDPP4 expression vector cassette. The optimized hDPP4 was cloned into a pCAGGS plasmid in which hDPP4 expression was driven by the CAG promoter. hDPP4 expression was confirmed in vitro by transfection of Cos-7 cells with the pCAGGS-hDPP4 (B) or pCAGGS (C) plasmids and detected by direct immunofluorescence assay with FITC-labeled anti-human CD26–Fluorescein antibody. Confirmation of the binding between DPP4 and MERS-CoV RBD was achieved by transfecting Cos-7 cells with pCAGGS-hDPP4 (D) or pCAGGS (E) plasmid followed by indirect immunofluorescence assay with MERS-RBD-Fc protein and DyLight 549-conjugated goat anti-human IgG antibody. (F) Determination of the copy numbers of hDPP4 cDNA in four transgenic founder lines by qPCR. (G) Expression of hDPP4 mRNA in the indicated tissues of transgenic mice in two founder lines as determined by qRT-PCR. Results are mean±SEM ( n = 3). (H) Four lines of hDPP4 transgenic mice were infected with MERS-CoV and monitored for body weight changes. Results are mean±SEM ( n = 6). (I) Lung viral titer at day 5 postinfection was determined for four lines of hDPP4 transgenic mice. The data are expressed as mean±SEM ( n = 3). The dotted line indicates the limit of detection.

Article Snippet: Expression of the codon-optimized hDPP4 was achieved by transfection of the pCAGGS-hDPP4 plasmid into Cos-7 cells, followed by confirmation of expression using anti-human CD26 monoclonal antibody (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Plasmid Preparation, Clone Assay, In Vitro, Transfection, Immunofluorescence, Labeling, Binding Assay, Transgenic Assay, Quantitative RT-PCR, Infection