Mbsa43

Manufactured by Thermo Fisher Scientific

Sourced in United States

The MBSA43 is a laboratory instrument designed for the analysis of biological samples. It is capable of performing various analytical measurements, but a detailed description of its core function cannot be provided in an unbiased and factual manner without the risk of extrapolation or interpretation.

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Lab products found in correlation

Facsaria 2, by BD (2 mentions) Rpa t4, by BD (2 mentions) Live dead cell viability assay, by Thermo Fisher Scientific (1 mentions) Lsrfortessa, by BD (1 mentions) Percoll, by Merck Group (1 mentions) Fluorescein isothiocyanate (fitc), by BD (1 mentions) Percp efluor 710, by Thermo Fisher Scientific (1 mentions) Dnase 1, by Worthington (1 mentions) Collagenase 4, by Worthington (1 mentions) Mih18, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rhil 2, by R&D Systems (1 mentions) Clone p3, by Thermo Fisher Scientific (1 mentions) Facsdiva, by BD (1 mentions) Cd56 buv737 ncam16.2, by BD (1 mentions) Eh12.2h7, by BioLegend (1 mentions) Cd27 bv650, by BioLegend (1 mentions) Cd107a apch7 h4a3, by BD (1 mentions) Mopc 21, by BioLegend (1 mentions) Facscalibur flow cytometer, by BD (1 mentions)

Spelling variants of this product

TIGIT (MBSA43, (5 citations) Anti-TIGIT– clone MBSA43 (2 citations) Anti‐TIGIT antibody (MBSA43, (2 citations) Anti-TIGIT (MBSA43, (2 citations)

5 protocols using mbsa43

Multiparametric Analysis of Tumor-Infiltrating T Cells

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Peripheral blood mononuclear cells were isolated from whole blood by Ficoll-Paque gradient centrifugation. Freshly resected tumor specimens were manually disrupted followed by digestion with collagenase IV (2.5 mg/mL) and DNase I (0.2 mg/mL) (Worthington Biochemical Corporation) for 1 hour. Tumor homogenates were separated on discontinuous 70%–30% Percoll (Sigma-Aldrich) gradients. Flow cytometric analysis was performed with antibodies targeting CD4 (BD Biosciences clone RPA-T4, V450 conjugated), CD8 (BD Biosciences clone RPA-T8, V500 conjugated), TIGIT (eBioscience clone MBSA43, PerCP-eFluor® 710 conjugated), CD226 (eBioscience clone TX25, FITC conjugated), and PD-1 (BD Biosciences clone EH12.1, Alexa Fluor® 647 conjugated). Cell viability was assessed using Live/Dead Cell Viability Assays (Life Technologies). Samples were run on a BD LSRFortessa or BD FACSAria II, as previously described. FlowJo software (Tree Star Inc.) was used for analysis after gating on live cells, with doublet exclusion followed by gating on CD4 and CD8 T cells.

Lucca L.E., Lerner B.A., Park C., DeBartolo D., Harnett B., Kumar V.P., Ponath G., Raddassi K., Huttner A., Hafler D.A, & Pitt D. (2020). Differential expression of the T-cell inhibitor TIGIT in glioblastoma and MS. Neurology® Neuroimmunology & Neuroinflammation, 7(3), e712.

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Suppression Assay of Tumor-infiltrating T Cells

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(CFSE)-labeled responders CD4⁺ Naive⁺ T cells from healthy donors were cocultured with different effector to target (E/T) ratios with unlabeled CD127⁻CD25^lowCD4⁺ T cells sorted from TILs or PBMCs of patients with CRC or NSCLC, using FACS Aria II (BD Biosciences), in the presence of CD11c⁺CD1c⁺dentritic cells as antigen-presenting cells and anti-CD3 (OKT3) mAb. Proliferation of CFSE-labeled cells was assessed after 96 hr. Some suppression assays were also performed with tumor Treg cells that were preincubated with the following antibodies (at a final concentration of 20 μg/ml): anti-human PD-L1 (Biolegend clone 29E.2 A 3), anti-human PD-L2 (Biolegend clone MIH18), and anti-human Functional Grade as isotype control (eBioscience clone MBSA43).

De Simone M., Arrigoni A., Rossetti G., Gruarin P., Ranzani V., Politano C., Bonnal R.J., Provasi E., Sarnicola M.L., Panzeri I., Moro M., Crosti M., Mazzara S., Vaira V., Bosari S., Palleschi A., Santambrogio L., Bovo G., Zucchini N., Totis M., Gianotti L., Cesana G., Perego R.A., Maroni N., Pisani Ceretti A., Opocher E., De Francesco R., Geginat J., Stunnenberg H.G., Abrignani S, & Pagani M. (2016). Transcriptional Landscape of Human Tissue Lymphocytes Unveils Uniqueness of Tumor-Infiltrating T Regulatory Cells. Immunity, 45(5), 1135-1147.

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NK Cell Stimulation and TIGIT Blocking

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Cells were cultured in RPMI media containing 10% FBS (Thermo Fisher Scientific) and 1% penicillin/streptomycin/amphotericin (Thermo Fisher Scientific) (RP10). NK cells were incubated in 24-well plates at a concentration of 2x10⁶ cells/ml in for 3 days at 37°C with 5% CO2, with addition of rhIL-2 (R&D Systems, 300 IU/ml). After 3 days, NK cells were washed with fresh media and plated in 96-well plates (80,000-100,000 cells/well). Blocking mIgG1κ anti-hTIGIT antibody (eBioscience, clone MBSA43, 10 μg/ml) or a mIgG1κ isotype control (eBioscience, clone P3.6.2.8.1, 10 μg/ml) were added. Cells were incubated at 4°C for 20 minutes, washed with PBS + 2% FBS and resuspended in media.

VENDRAME E., SEILER C., RANGANATH T., ZHAO N.Q., VERGARA R., ALARY M., LABBÉ A.C., GUÉDOU F., POUDRIER J., HOLMES S., ROGER M, & BLISH C.A. (2020). TIGIT is upregulated by HIV-1 infection and marks a highly functional adaptive and mature subset of natural killer cells. AIDS (London, England), 34(6), 801-813.

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Assessing Immune Checkpoint Markers on PBMC from PLWH

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Fresh PBMC isolated from PLWH/ART were rested overnight in RF10 at 37°C 5% CO₂. The next day, 0.5 × 10⁶ PBMC were added to a 96‐well round bottom plate at a 1:1 ratio with murine P815 cells (ATCC, Manassas, USA) and stimulated with 40 ng mL⁻¹ of monoclonal antibodies against either CD3 (OKT3; Biolegend), CD16 (3G8; Biolegend) or an isotype control (MOPC‐21; Biolegend). 5 μg mL⁻¹ of monoclonal antibodies against either CD160 (BY55; Biolegend), 2B4 (C1.7; Biolegend), Tim‐3 (F38‐2E2; Biolegend), TIGIT (MBSA43; eBioscience, San Diego, USA), PD‐1 (EH12.2H7; Biolegend), NKG2C (134522; R&D Systems) or an isotype control (MOPC‐21; Biolegend) were then added to CD3 or CD16 stimulated conditions. CD107a APCH7 (H4A3; BD Biosciences) was added to each well, and the plate was briefly centrifuged before incubation at 37°C with 5% CO₂ for 5 h. After incubation, wells were washed, and cells stained with UV viability dye (ThermoFisher) then a cocktail containing Vδ1 FITC (TS8.2; Invitrogen, Carlsbad, USA), CD3 BV510 (SK7; Biolegend), CD27 BV650 (0323; Biolegend), CD69 PE Dazzle (FN50; Biolegend) and CD56 BUV737 (NCAM16.2; BD Biosciences). After staining, cells were washed and resuspended in PBS containing 2% FCS before acquisition on a BD LSR Fortessa using BD FACS Diva.

Field K.R., Wragg K.M., Kent S.J., Lee W.S, & Juno J.A. (2024). γδ T cells mediate robust anti‐HIV functions during antiretroviral therapy regardless of immune checkpoint expression. Clinical & Translational Immunology, 13(2), e1486.

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Flow Cytometric Analysis of Immune Cells

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Cell surface staining was performed at 4 C for 30 minutes and was analyzed using a FACSCalibur flow cytometer (BD Biosciences) and CellQuest Software (BD Biosciences). A minimum of 1 Â 10 4 cells were examined. FcR Blocking Reagent (human cell and mouse cell, Miltenyi Biotec) was used according to the manufacturer's instructions. Antibodies to mouse CD3e (145-2C11, BD Biosciences), CD107a (1D4B, BD Biosciences), IFNg (XMG1.2, BioLegend), TNF (MP6-XT22, BioLegend), CD155 (4.24.1, BioLegend), CD8a (53-6.7, BioLegend) and antibodies to human CD3 (HIT3a, BioLegend), CD45 (2D1, BioLegend), CD4 (RPA-T4, BD Biosciences), PD-1 (EH12, BD Biosciences), TIGIT (MBSA43, eBioscience), and CD8 (RPA-T8, BD Biosciences) were used in this study. For TIL experiments, the cells were stimulated with phorbol 12-myristate 13-acetate (30 ng/mL; Sigma) and 1 mmol/L ionomycin (Sigma) in the presence of monensin (2.5 mg/mL; eBioscience) for 4 hours, after which they were stained for surface markers and then fixed and permeabilized with eBioscience FoxP3 fixation buffer according to the manufacturer's instructions. Fixed cells were stained with antibodies to IFNg (XMG1.2; BioLegend) and TNF (MP6-XT22, BioLegend), and Rat IgG1 antibody was used as an isotype control (RTK2071, BioLegend).

Shen X., Fu W., Wei Y., Zhu J., Yu Y., Lei C., Zhao J, & Hu S. (2021). TIGIT-Fc Promotes Antitumor Immunity. Cancer immunology research, 9(9).

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