Journal:

Article Title: Identification and characterization of macaque CD89 (immunoglobulin A Fc receptor)

doi: 10.1111/j.1365-2567.2004.01949.x

Figure Lengend Snippet: Alignment of CD89 derived amino acid sequences obtained by cloning and sequencing rhesus macaque (GenBank accession number AY386684) and cynomolgus macaque (GenBank accession number AY386690) cDNA from whole blood and comparison with the published human sequence (GenBank NM_002000). Amino acid differences are underlined. The first amino acid of the preprotein is numbered as residue 1. The mature peptide starts at residue 22. Arrows indicate distinct domains. The first two amino acids for EC1 are encoded at the end of the S2 exon. The signal peptide is encoded by both S1 and S2 sequences. Potential N-glycosylation sites, cysteines involved in disulfide bonds and arginine 209 critical for association with the FcRγ chain are bolded. Hu: Homo sapiens; Mamu: Macaca mulatta; Mafa: Macaca fascicularis.

Article Snippet: 1 × 10 6 cells were stained at 4° with either 20 µl of phycoerythrin-conjugated mouse anti-human CD89 (clone A59) or Simultest TM Control γ 1 /γ 2a (both from BD PharMingen, San Diego, CA) for 15 min, followed by three washes with PBS.

Techniques: Derivative Assay, Clone Assay, Sequencing

Journal:

Article Title: Identification and characterization of macaque CD89 (immunoglobulin A Fc receptor)

doi: 10.1111/j.1365-2567.2004.01949.x

Figure Lengend Snippet: Expression of recombinant macaque CD89 on HeLa cells. Cells were transfected with a plasmid containing full-length (a) rhesus macaque CD89 cDNA or (b) cynomolgus macaque CD89 cDNA and stained with anti-human CD89: thick line (rhesus MFI = 120·70, SD = 343·04; cynomolgus 30·35, SD = 345·04) or with isotype control mouse IgG: thin line (rhesus MFI = 8·65, SD = 7·40; cynomolgus MFI = 9·27, SD = 34·47). Untransfected HeLa cells stained with anti-human CD89: dotted line (MFI = 5·26, SD = 28·69). MFI = Mean fluorescence intensity, SD = standard deviation. Stable transfectants were used for detection of rhesus macaque CD89 and transient transfectants were used for detection of cynomolgus macaque CD89. 5000 events were counted per sample.

Article Snippet: 1 × 10 6 cells were stained at 4° with either 20 µl of phycoerythrin-conjugated mouse anti-human CD89 (clone A59) or Simultest TM Control γ 1 /γ 2a (both from BD PharMingen, San Diego, CA) for 15 min, followed by three washes with PBS.

Techniques: Expressing, Recombinant, Transfection, Plasmid Preparation, Staining, Fluorescence, Standard Deviation

Journal:

Article Title: Identification and characterization of macaque CD89 (immunoglobulin A Fc receptor)

doi: 10.1111/j.1365-2567.2004.01949.x

Figure Lengend Snippet: Alignment of the five rhesus macaque CD89 splice variants (GenBank accession number AY386684-AY386689) with full-length CD89 from the same species. Arrows indicate distinct domains. Mamu: Macaca mulatta.

Article Snippet: 1 × 10 6 cells were stained at 4° with either 20 µl of phycoerythrin-conjugated mouse anti-human CD89 (clone A59) or Simultest TM Control γ 1 /γ 2a (both from BD PharMingen, San Diego, CA) for 15 min, followed by three washes with PBS.

Techniques:

Journal:

Article Title: Identification and characterization of macaque CD89 (immunoglobulin A Fc receptor)

doi: 10.1111/j.1365-2567.2004.01949.x

Figure Lengend Snippet: Alignment of the three cynomolgus macaque CD89 splice variants (GenBank accession number AY386690-AY386693) with full-length CD89 from the same species. Arrows indicate distinct domains. Mafa: Macaca fascicularis.

Article Snippet: 1 × 10 6 cells were stained at 4° with either 20 µl of phycoerythrin-conjugated mouse anti-human CD89 (clone A59) or Simultest TM Control γ 1 /γ 2a (both from BD PharMingen, San Diego, CA) for 15 min, followed by three washes with PBS.

Techniques:

Journal:

Article Title: Identification and characterization of macaque CD89 (immunoglobulin A Fc receptor)

doi: 10.1111/j.1365-2567.2004.01949.x

Figure Lengend Snippet: Schematic representation of the complete CD89 transcript and corresponding splice variants identified in rhesus and cynomolgus macaques. Mamu: Macaca mulatta; Mafa: Macaca fascicularis.

Article Snippet: 1 × 10 6 cells were stained at 4° with either 20 µl of phycoerythrin-conjugated mouse anti-human CD89 (clone A59) or Simultest TM Control γ 1 /γ 2a (both from BD PharMingen, San Diego, CA) for 15 min, followed by three washes with PBS.

Techniques:

Journal:

Article Title: Identification and characterization of macaque CD89 (immunoglobulin A Fc receptor)

doi: 10.1111/j.1365-2567.2004.01949.x

Figure Lengend Snippet: Two-colour dot-plots of whole blood leucocytes from a representative rhesus (a–d) and cynomolgus macaque (e–h). Forward scatter (FSC) versus side scatter (SSC) (a and e); CD89 versus CD20 (b and f); CD3 (c and g); and CD16 (d and h).

Article Snippet: 1 × 10 6 cells were stained at 4° with either 20 µl of phycoerythrin-conjugated mouse anti-human CD89 (clone A59) or Simultest TM Control γ 1 /γ 2a (both from BD PharMingen, San Diego, CA) for 15 min, followed by three washes with PBS.

Techniques:

Journal:

Article Title: Identification and characterization of macaque CD89 (immunoglobulin A Fc receptor)

doi: 10.1111/j.1365-2567.2004.01949.x

Figure Lengend Snippet: Two-colour dot-plots of whole-blood leucocytes from a representative rhesus (a–d) and cynomolgus macaque (e–h). (a and e) CD14 versus side scatter. CD14 versus CD89: (b and f) total leucocytes; (c and g) granulocytes; (d and h) monocytes. Gates used for granulocyte (R1) and monocyte (R2) populations of rhesus (a) and cynomolgus macaque (e) are shown.

Article Snippet: 1 × 10 6 cells were stained at 4° with either 20 µl of phycoerythrin-conjugated mouse anti-human CD89 (clone A59) or Simultest TM Control γ 1 /γ 2a (both from BD PharMingen, San Diego, CA) for 15 min, followed by three washes with PBS.

Techniques:

Journal: Cells

Article Title: Effective, Long-Term, Neutrophil Depletion Using a Murinized Anti-Ly-6G 1A8 Antibody

doi: 10.3390/cells11213406

Figure Lengend Snippet: Mouse-Ly-6G efficiently depleted neutrophils in C57Bl6/J, Balb/c, NXG, and SCID mice. ( a ) Experimental set-up; i.p. injections with the depletion antibodies were performed three times a week for four weeks. Blood was drawn via cheek puncture once a week before injection with the antibodies; ( b , c ) Longitudinal analysis of the number of CD45 + Siglec-F − CD115 − SSC high Gr-1 + CD11b + neutrophils in the peripheral blood ( n = five mice per group) per 5000 latex beads, showing; ( b ) Significant, almost complete, neutrophil depletion in all mouse strains tested (C57Bl6/J, Balb/c, NXG, and SCID) upon treatment with 100 μg mouse-Ly-6G antibody, and ( c ) Significantly better neutrophil depletion in 100 μg mouse-Ly-6G treated C57Bl6/J mice than when treated with 100 μg rat-Ly-6G antibody. Data is presented as mean with SEM. Statistics: one-way ANOVA with Bonferroni correction. Repeated measures one-way ANOVA with Bonferroni correction was used to compare groups in c, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001.

Article Snippet: Experiments were conducted using C57Bl/6J (C57Bl/6JRj), Balb/c (Balb/cByJRj), NXG (NOD.Prkdc scid Il2rg tm1 /Rj), SCID (NOD.CB17-Prkdc scid/scid /Rj), and human FcαRI (CD89) transgenic SCID mice (all housed and bred at Janvier Labs, Paris, France) [ ], or C57Bl/6J FcRγ −/− (C57Bl/6JRj FcRγ-chain knockout) mice (housed and bred at the University of Utrecht) [ ].

Techniques: Injection

Journal: Cells

Article Title: Effective, Long-Term, Neutrophil Depletion Using a Murinized Anti-Ly-6G 1A8 Antibody

doi: 10.3390/cells11213406

Figure Lengend Snippet: Mouse-Ly-6G efficiently depletes neutrophils in the blood and tumor of IMR32 tumor-bearing SCID mice ( a ) Experimental set-up; 2.5 × 10 6 IMR32 cells were s.c. injected in SCID mice in a 1:2 mix with Vitrogel Hydrogel Matrix. Tumors were established for 28 days, after which mice were randomized over the different treatment groups, and treatment was started. I.p. injections with PBS, IgA ch14.18, and mouse-Ly-6G were performed three times a week for six weeks. The SIRPα-D1 fusion protein was injected i.p. every nine days. Blood was drawn via cheek puncture once a week before injection with the antibodies; ( b ) Longitudinal analysis of the percentage of SSC high Ly-6C low Ly-6G + CD11b + neutrophils in the peripheral blood ( n = 2–5 mice per group) of the CD45+ leukocyte population, showing complete neutrophil depletion upon treatment with 100 μg mouse-Ly-6G antibody. Of note; blood at day 70 was collected from the orbit and processed in a similar fashion as the tumor material, meaning cells were fixed at a later time point, possibly resulting in some neutrophil cell death; ( c ) Intratumoral analysis of the percentage of SSC high Ly-6C low Ly-6G + CD11b + neutrophils of the live (TO-PRO-3 negative) cells, CD45+ leukocytes, and CD11b+ myeloid populations. Data showed increased tumor infiltration when the mice were treated with IgA ch14.18/SIRPα-D1 fusion, which was completely ablated when 100 μg mouse Ly-6G was added. Data is presented as mean with SEM. Statistics: one-way ANOVA with Bonferroni correction. Repeated measures one-way ANOVA with Bonferroni correction was used to compare groups in b, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001.

Article Snippet: Experiments were conducted using C57Bl/6J (C57Bl/6JRj), Balb/c (Balb/cByJRj), NXG (NOD.Prkdc scid Il2rg tm1 /Rj), SCID (NOD.CB17-Prkdc scid/scid /Rj), and human FcαRI (CD89) transgenic SCID mice (all housed and bred at Janvier Labs, Paris, France) [ ], or C57Bl/6J FcRγ −/− (C57Bl/6JRj FcRγ-chain knockout) mice (housed and bred at the University of Utrecht) [ ].

Techniques: Injection

Journal: Journal of Immunology Research

Article Title: Overcoming the Constraints of Anti-HIV/CD89 Bispecific Antibodies That Limit Viral Inhibition

doi: 10.1155/2016/9425172

Figure Lengend Snippet: Binding affinity of F240 × 14A8 Bi-Fab with neutrophil through CD89. Neutrophils were washed twice with PBS and resuspended at a concentration of 1 × 10 7 c/mL in HBSS containing 2.5% FBS. 100 µ L of neutrophils was mixed with 100 µ L of serial diluted Bi-Fab antibodies with Bi-Fab A in (a) and Bi-Fab B in (b). Cells were incubated with FITC-labeled goat anti-human Ig Kappa for 30 minutes. Neutrophils were fixed in 1% paraformaldehyde. Results are representative of four different neutrophil donors.

Article Snippet: The 14A8 is a human anti-CD89 antibody that was generated in Medarex-Mouse human Ig transgenic mice [ ].

Techniques: Binding Assay, Concentration Assay, Incubation, Labeling