Anti cd4 buv737

Manufactured by BD

Anti-CD4-BUV737 is a laboratory reagent used for the detection and analysis of CD4-positive cells in biological samples. It is a fluorochrome-conjugated monoclonal antibody that binds specifically to the CD4 receptor, allowing for the identification and quantification of CD4-expressing cells through flow cytometry.

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Anti-CD4 (255 citations) CD4 BUV737 (11 citations)

4 protocols using anti cd4 buv737

Detecting Mumps-Specific CD8+ T Cells

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Five immunogenic peptides were selected that were able to induce a T cell response. Subsequently, fluorochrome-labeled peptide-HLA class I (p:HLA-I-)dextramers each loaded with one of these five peptides were synthesized: B*0702-APIQGTNLL (FITC-labeled), B*0702-KPRTSTPVTEF (PE-labeled), B*0702-IPNARANL (APC-labeled), A*0101-YSDPNNHEVY (PE-labeled), and A*0101-VTDSNLIY (APC-labeled) (Custom made, all Immudex). In order to detect ex vivo MuV-specific CD8⁺ T cells, 4 × 10⁶ PBMCs from three HLA-B*0702- and three HLA-A*0101-positive mumps patients and from two HLA-B7- and two HLA-A1-positive young adults that recently received a third dose of the MMR vaccine, were stained with a combination of the different p:HLA-I-dextramers and incubated for 10 min at room temperature. Subsequently, cells were stained with anti-CD3- APC/R700, anti-CD4-BUV737 (564,305 ,BD), anti-CD8-BV786, anti-CD45RO-BUV395 (564,291, BD) and anti-CCR7 for 15 min at 4 °C. On average 175,000–350,000 CD8⁺ T cells were acquired per sample on an LSRFortessaX20.

Kaaijk P., Emmelot M.E., Meiring H.D., van Els C.A, & de Wit J. (2021). Novel mumps virus epitopes reveal robust cytotoxic T cell responses after natural infection but not after vaccination. Scientific Reports, 11, 13664.

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T Cell Proliferation Assay with MSCs

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Collected cells were washed in washing buffer (PBS containing 2% (vol/vol) FBS and 2 mM EDTA (Sigma, St. Louis, MO) and stained with: anti-CD3 (BV605, Biolegend, Dedham, MA), anti-CD4 (BUV737, BD, Franklin Lakes, NJ), anti-CD8 (V500, BD, Franklin Lakes, NJ), anti-CD38 (BUV395, BD, Franklin Lakes, NJ), anti-CD25 (PE/Cy7, Biolegend, Dedham, MA) for 20 minutes on ice. Cells were washed twice in washing buffer and cell pellets were then resuspended in 100 uL of fixation buffer (Cytofix, BD, Franklin Lakes, NJ) for 20 minutes on ice. Cells were then washed twice in washing buffer and resuspended in 200 uL of washing buffer. All flow cytometry was performed using an LSRII (BD, Franklin Lakes, NJ). Flow cytometry analysis was performed in FlowJo (Tree Star, Ashland, OR; version 8.7.3). Proliferative cells were gated based on their overall or individual number of cell divisions and raw flow cytometry values (P_raw) for proliferation were normalized to the positive stimulated control that did not have MSCs (P₊) (Eq. 1).

P r o l i f e r a t i v e F o l d C h a n g e = \frac{P_{r a w}}{P_{+}}

Li M., Khong D., Chin L.Y., Singleton A, & Parekkadan B. (2018). Therapeutic Delivery Specifications Identified Through Compartmental Analysis of a Mesenchymal Stromal Cell-Immune Reaction. Scientific Reports, 8, 6816.

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Comprehensive Immunophenotyping of T Cell Subsets

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Flow cytometric analysis was performed on an LSR Fortessa (BD Biosciences) and data analyzed using FlowJo Version 10.7.0 (Treestar). The following anti-human antibodies were used: anti-CD3-PE-Cy7 (BD Biosciences, clone HIT3a), anti-CD4-V500 (BD Biosciences, clone RPA-T4), anti-CD25-APC (Biolegend, clone BC96), anti-CD152-PE (CTLA-4) (BD Biosciences, clone BNI3), LIVE/DEAD TM Fixable Near-IR dead cell stain kit (Thermo Fisher Scientific, L10119). Intracellular staining was performed using the eBioscience TM Foxp3/Transcription factor staining buffer set (Thermo Fisher Scientific, 00-5523-00) and anti-FOXP3-APC (Thermo Fisher Scientific, clone 236A/E7) and anti-GFP-FITC (Rockland, 600-402-215).
The following anti-mouse antibodies were used: anti-CD3-BV421 (BD Biosciences, clone 17A2), anti-CD4-BUV737 (BD Biosciences, clone GK1.5), anti-CD4-PerCP-Cy5.5 (BD Biosciences, clone RM4-5), anti-CD152-PE (CTLA-4) (BD Biosciences, clone UC10-4F10-11), anti-FOXP3-APC (Thermo Fisher Scientific, FJK-16s), Fixable Viability Dye eFluor™ 780 (Thermo Fisher Scientific).

Fox T.A., Houghton B.C., Petersone L., Waters E., Edner N.M., McKenna A., Preham O., Hinze C., Williams C., de Albuquerque A.S., Kennedy A., Pesenacker A.M., Genovese P., Walker L.S.K., Burns S.O., Sansom D.M., Booth C, & Morris E.C. (2022). Therapeutic gene editing of T cells to correct CTLA-4 insufficiency. Science translational medicine, 14(668).

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Comprehensive Immune Cell Phenotyping

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Cells were stained according to the manufacturer's instructions. Briefly, cells were stained with anti-CD69 BUV395 (BD, clone FN50) on the surface and washed prior to stimulation. After the stimulation, cells were surface stained with anti-CD103 BV711 (Biolegend, Ber-ACT8), anti-CD45RA BUV563 (BD, clone HI100), anti-CD27 BV650 (Biolegend, clone 0323), anti-PD-1 PE-Cy7 (BD, clone EH12.2H7), and anti-CD39 BV510 (Biolegend, clone A1). Cells were then fixed and permeabilized with the Foxp3 Staining Buffer Set (Thermofisher), followed by intracellular staining of the cells with anti-CD3 BV605 (eBioscience, clone OKT3) or BUV661 (BD, clone UCHT1), anti-CD4 BUV737 (BD, clone SK3), anti-CD8 BUV805 (BD, clone SK1), anti-CD40L PeDazzle594 (Biolegend, clone 24-31), anti-CD137 AF647 or PeCy7 (Thermofisher Scientific, clone 4B4-1), anti-TNF PeCy7 or FITC (BD, Mab11), anti-IFNγ eF450 or BV785 or Pe (Biolegend, clone 4S.B3), anti-IL17a BV605 (Biolegend, clone BL168), and anti-IL-2 BB700 or APC (BD, clone MQ1-17H12). Samples were analyzed in 0.5% FCS PBS and acquired on a BD FACSymphony. Data analysis was performed using FlowJo (TreeStar, Version 10.0.7).

Oja A.E., Morgana F., Hagen R., Ghandour C.A., Nolte M.A., Rooijakkers S.H.M., Lier R.A.W.v., Bardoel B.W., & Hombrink P. (2022). Polyfunctional pathogen-specific CD4⁺ T cells reside in the lungs and tumors of NSCLC patients.

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