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Constitutive interaction between 4-1BB and 4-1BBL on murine LPS-activated bone marrow dendritic cells masks detection of 4-1BBL by TKS-1 but not 19H3 antibody.
J Immunol Methods. 2017 11; 450:81-89.JI

Abstract

4-1BB is a TNFR family member associated with NF-κB mediated survival signaling. 4-1BB is widely expressed on activated cells of the immune system, including activated T cells, NK cells and dendritic cells. Its ligand, 4-1BBL, is transiently expressed on activated antigen presenting cells and at low levels on activated T cells. Although 4-1BBL-deficient mice clearly demonstrate a role for 4-1BBL in CD8 T cell responses to viruses such as influenza, 4-1BBL can be difficult to detect following infection of mice. Here we provide evidence for a constitutive interaction between endogenous 4-1BB and 4-1BBL on LPS activated bone marrow-derived murine dendritic cells that can mask its detection, with implications for measurement of 4-1BBL expression. The masking of 4-1BBL by its receptor results in loss of reactivity to the anti-4-1BBL antibody TKS-1, whereas the 19H3 antibody binds to 4-1BBL in the presence or absence of 4-1BB. Moreover, 4-1BB/4-1BBL interaction can occur in trans between 4-1BB+/+ and 4-1BB-/- dendritic cells in culture. These data suggest that 19H3 is the preferable antibody to use to detect 4-1BBL in the presence of its receptor.

Authors+Show Affiliations

Department of Immunology, University of Toronto, Toronto, ON M5S1A8, Canada.Department of Immunology, University of Toronto, Toronto, ON M5S1A8, Canada.Department of Immunology, University of Toronto, Toronto, ON M5S1A8, Canada.Department of Immunology, University of Toronto, Toronto, ON M5S1A8, Canada. Electronic address: tania.watts@utoronto.ca.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

28789924

Citation

Mbanwi, Achire N., et al. "Constitutive Interaction Between 4-1BB and 4-1BBL On Murine LPS-activated Bone Marrow Dendritic Cells Masks Detection of 4-1BBL By TKS-1 but Not 19H3 Antibody." Journal of Immunological Methods, vol. 450, 2017, pp. 81-89.
Mbanwi AN, Lin GHY, Wang KC, et al. Constitutive interaction between 4-1BB and 4-1BBL on murine LPS-activated bone marrow dendritic cells masks detection of 4-1BBL by TKS-1 but not 19H3 antibody. J Immunol Methods. 2017;450:81-89.
Mbanwi, A. N., Lin, G. H. Y., Wang, K. C., & Watts, T. H. (2017). Constitutive interaction between 4-1BB and 4-1BBL on murine LPS-activated bone marrow dendritic cells masks detection of 4-1BBL by TKS-1 but not 19H3 antibody. Journal of Immunological Methods, 450, 81-89. https://doi.org/10.1016/j.jim.2017.08.001
Mbanwi AN, et al. Constitutive Interaction Between 4-1BB and 4-1BBL On Murine LPS-activated Bone Marrow Dendritic Cells Masks Detection of 4-1BBL By TKS-1 but Not 19H3 Antibody. J Immunol Methods. 2017;450:81-89. PubMed PMID: 28789924.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Constitutive interaction between 4-1BB and 4-1BBL on murine LPS-activated bone marrow dendritic cells masks detection of 4-1BBL by TKS-1 but not 19H3 antibody. AU - Mbanwi,Achire N, AU - Lin,Gloria H Y, AU - Wang,Kuan Chung, AU - Watts,Tania H, Y1 - 2017/08/05/ PY - 2017/01/18/received PY - 2017/06/16/revised PY - 2017/08/03/accepted PY - 2017/8/10/pubmed PY - 2017/10/3/medline PY - 2017/8/10/entrez KW - 19H3 KW - 4-1BB KW - 4-1BBL KW - Antibody blocking KW - Dendritic cells KW - TKS-1 SP - 81 EP - 89 JF - Journal of immunological methods JO - J Immunol Methods VL - 450 N2 - 4-1BB is a TNFR family member associated with NF-κB mediated survival signaling. 4-1BB is widely expressed on activated cells of the immune system, including activated T cells, NK cells and dendritic cells. Its ligand, 4-1BBL, is transiently expressed on activated antigen presenting cells and at low levels on activated T cells. Although 4-1BBL-deficient mice clearly demonstrate a role for 4-1BBL in CD8 T cell responses to viruses such as influenza, 4-1BBL can be difficult to detect following infection of mice. Here we provide evidence for a constitutive interaction between endogenous 4-1BB and 4-1BBL on LPS activated bone marrow-derived murine dendritic cells that can mask its detection, with implications for measurement of 4-1BBL expression. The masking of 4-1BBL by its receptor results in loss of reactivity to the anti-4-1BBL antibody TKS-1, whereas the 19H3 antibody binds to 4-1BBL in the presence or absence of 4-1BB. Moreover, 4-1BB/4-1BBL interaction can occur in trans between 4-1BB+/+ and 4-1BB-/- dendritic cells in culture. These data suggest that 19H3 is the preferable antibody to use to detect 4-1BBL in the presence of its receptor. SN - 1872-7905 UR - https://www.unboundmedicine.com/medline/citation/28789924/Constitutive_interaction_between_4_1BB_and_4_1BBL_on_murine_LPS_activated_bone_marrow_dendritic_cells_masks_detection_of_4_1BBL_by_TKS_1_but_not_19H3_antibody_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-1759(17)30033-9 DB - PRIME DP - Unbound Medicine ER -