Journal: bioRxiv

Article Title: Efficacy of Targeting SARS-CoV-2 by CAR-NK Cells

doi: 10.1101/2020.08.11.247320

Figure Lengend Snippet: ( a ) Immunophenotyping of CR3022-CAR. Antibodies against various immunomodulatory receptors including TIGIT, LAG-3, TIM-3, KLRG1, CTLA-4, PD-1, CD69, CD8A, NKG2C, CD94, DNAM-1, 2B4, NKG2D, NKP46, and CD16 were used to stain CR3022-CAR and NK-92MI. The expression of these receptors was determined by flow cytometry. ( b ) Diagram of CR3022-CAR binding to the RBD domain of SARS-CoV-2-S recombinant protein. CR3022-CAR binds to RBD of SARS-CoV-2-S protein which is then recognized by anti-His and its corresponding secondary antibody conjugated to a fluorophore. ( c ) Representative dot plots of CR3022-CAR binding to RBD of SARS-CoV-2. CR3022-CAR or NK-92MI cells were incubated with SARS-CoV-2-RBD or SARS-CoV-1-RBD recombinant protein. The binding efficiency was determined by flow cytometry.

Article Snippet: APC anti-human CD16 antibody (clone 3G8, BD Biosciences), BV711 anti-human CD314 (NKG2D) antibody (clone 1D11, BD Biosciences), and FITC anti-human CD107a antibody (clone H4A3, BD Biosciences) were purchased from BD Biosciences (San Jose, CA, USA).

Techniques: Staining, Expressing, Flow Cytometry, Binding Assay, Recombinant, Incubation

Journal: PLoS Pathogens

Article Title: Precision engineering for PRRSV resistance in pigs: Macrophages from genome edited pigs lacking CD163 SRCR5 domain are fully resistant to both PRRSV genotypes while maintaining biological function

doi: 10.1371/journal.ppat.1006206

Figure Lengend Snippet: Peripheral blood monocytes were isolated from the blood of the wild type (red), heterozygous (blue), and ΔSRCR5 (green) animals. Following cultivation in the presence of recombinant human CSF1 (rhCSF1) for seven days PMMs were analyzed by FACS. A) Co-staining with CD14-FITC and CD16-PE antibodies recognizing the native structure of the proteins (colored contour plots; red wild type, blue heterozygous, green ΔSRCR5) relative to isotype controls (grey). B) Co-staining with CD169-FITC and CD172a-PE antibodies recognizing the native structure of the proteins (colored contour plots) relative to isotype controls (grey). C) Co-staining with SWC9 (CD203a)-FITC and CD151-RPE antibodies recognizing the native structure of the proteins (colored contour plots) relative to isotype controls (grey). D) Staining against the native structure of surface expressed CD163 (colored) relative to an isotype control staining (grey).

Article Snippet: Cells were stained with antibodies targeting either mouse anti pig CD14 (AbD Serotec, MGA1273F, 1:50) and mouse anti pig CD16 (AbD Serotec, MCA2311PE, 1:200), mouse anti pig CD169 (AbD Serotec, MCA2316F, 1:50) and mouse anti pig CD172a (SoutherBiotech, 4525–09, 1:400), mouse anti human CD151 (AbD Serotec, MCA1856PE, 1:50) and mouse anti pig SWC9(CD203a) (AbD Serotec, MCA1973F, 1:50), mouse anti pig CD163 (AbD Serotec, MCA2311PE, 1:50), or mouse IgG1 or an IgG2b negative control (AbD Serotec, MCA928PE,MCA691F, or Sigma, F6397; same concentration as primary Ab).

Techniques: Isolation, Recombinant, Staining, Control