Recombinant Mouse Antibody (20.1) is capable of binding to BTN3A1. This agonist antibody, 20.1, binds to the extracellular domain of BTN3A molecules and induces Vγ9Vδ2 activation with similar kinetics and potency as with addition of PiPs.
Figure 1 HLA haplo-identical HeLa-L and HeLa-M cells vary in their ability to present phosphoantigens to Vγ9Vδ2 T cells.
Activation of Vγ9/Vδ2 T cells detected by ELISA for IFN-γ using a concentration gradient of HMB-PP, agonist antibody CD277 20.1 and T cells expanded from two separate donors. Activation was induced by pretreatment of HeLa cells using anti-BTN3A monoclonal antibody 20.1 for 30 min. Graphs show mean and SD from n = 2 and are representative of two independent measurements.
Rhodes, D. A., Chen, H. C., Williamson, J. C., Hill, A., Yuan, J., Smith, S., ... & Eberl, M. (2018). regulation of human γδ T cells by BTn3a1 Protein stability and aTP-Binding cassette Transporters. Frontiers in immunology, 9, 662.
Figure 2 Transcripts for the three BTN3A isoforms are unchanged in HeLa-L and HeLa-M.
Analysis of activation markers MHC-I (w6/32), MHC-II (L243), and BTN3A (CD277 20.1) by flow cytometry in HeLa cells treated with or without IFN-γ. Results representative of triplicate experiments. Red lines indicate IFN-γ treated and black lines untreated samples. Solid lines are HeLa-M and dotted lines HeLa-L.
Rhodes, D. A., Chen, H. C., Williamson, J. C., Hill, A., Yuan, J., Smith, S., ... & Eberl, M. (2018). regulation of human γδ T cells by BTn3a1 Protein stability and aTP-Binding cassette Transporters. Frontiers in immunology, 9, 662.
Figure 3 Western blot of HeLa-L cells transiently transfected with expression vectors for BTN3A1, BTN3A2, and BTN3A3.
Monoclonal anti-BTN3A (CD277 20.1) antibody and polyclonal sera 056 and B6, directed to B30.2 domains of BTN3A1 and BTN3A3, respectively, were used. Anti-GFP and calnexin (CNX) antibodies provided loading controls for expression of transgene and endogenous proteins, respectively.
Rhodes, D. A., Chen, H. C., Williamson, J. C., Hill, A., Yuan, J., Smith, S., ... & Eberl, M. (2018). regulation of human γδ T cells by BTn3a1 Protein stability and aTP-Binding cassette Transporters. Frontiers in immunology, 9, 662.
Figure 4 CD107a expression on the human Vγ9Vδ2 T cell line GUI following incubation with either 20.1 full-lengthantibody (open circles), 20.1 Fab fragments (filled triangles), or 20.1 scFv (filled circles).
The data are presented as thepercentages of CD107a+ γδT cells and are representative of more than three experiments.
Palakodeti, A., Sandstrom, A., Sundaresan, L., Harly, C., Nedellec, S., Olive, D., ... & Adams, E. J. (2013). The Molecular Basis for Modulation of Human V;# 947; 9V [delta] 2 T Cell Responses by CD277/Butyrophilin-3 (BTN3A)-specific Antibodies. J. Biol. Chem., 287(39); 09, 2012).
Girard, Pauline, et al. "Dysfunctional BTN3A together with deregulated immune checkpoints and type I/II IFN dictate defective interplay between pDCs and γδ T cells in melanoma patients, which impacts clinical outcomes." Clinical & Translational Immunology 10.11 (2021): e1329. https://doi.org/10.1002/cti2.1329
This study investigates the dysfunctional BTN3A and its interaction with deregulated immune checkpoints and type I/II interferons in the context of melanoma. The research focuses on the roles of plasmacytoid dendritic cells (pDCs) and gamma-delta (γδ) T cells, examining their phenotypic and functional interactions in both the blood and tumor microenvironment of melanoma patients. The study highlights severe alterations in the bidirectional cross-talk between pDCs and γδ T cells in melanoma, noting that while healthy pDCs can drive potent activation of γδ T cells, those from melanoma patients exhibit impaired functionality.
Creative Biolabs provided critical reagents for this study, specifically anti-BTN3A agonist antibody (clone 20.1, Cat# PABL-414) and anti-BTN3A blocking antibody (clone 103.2, Cat# PABL-415). These antibodies were used to assess the functionality of BTN3A on γδ T cells and pDCs within PBMCs and tumor infiltrates. The use of these antibodies was essential in revealing the potential functional impairment of BTN3A in melanoma patients, thereby providing insights into the therapeutic targeting of BTN3A to improve immune responses in cancer therapy.
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CAT | Product Name | Application | Type |
---|---|---|---|
MOB-127 | Recombinant Anti-human BTN3A1 Antibody | WB, Dot, FuncS | IgG |
MHH-127 | Recombinant Human Anti-human BTN3A1 Antibody | WB, IHC, FuncS | IgG |
PABL-415 | Mouse Anti-BTN3A1 Recombinant Antibody (clone 103.2) | FuncS | Mouse IgG |
VS-0723-WK16 | Human Anti-BTN3A1 Recombinant Antibody (VS-0723-WK16) | FC | Human IgG1 |
VS-0723-WK287 | Rabbit Anti-BTN3A1 Recombinant Antibody (VS-0723-WK287) | ELISA, FC | Rabbit IgG |
To accurately reference this product in your publication, please use the following citation information:
(Creative Biolabs Cat# PABL-414, RRID: AB_3111654)
For Research Use Only. Not For Clinical Use.
For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.
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