14-5888-82
antibody from Invitrogen Antibodies
Targeting: PDCD1LG2
B7-DC, bA574F11.2, Btdc, CD273, PD-L2, PDL2
Antibody data
- Antibody Data
- Antigen structure
- References [17]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [8]
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- Product number
- 14-5888-82 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD273 (B7-DC) Monoclonal Antibody (MIH18), eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The MIH18 monoclonal antibody reacts with human B7-DC, also known as PD-L2. B7-DC, a recently identified member of the B7 family, has a predicted molecular weight of approximately 25 kDa and belongs to the Ig superfamily. B7-DC is primarily expressed by subpopulations of dendritic cells and monocytes/macrophages. Although B7-DC has structural and sequence similarities to the B7 family, it does not bind CD28/CTLA-4, rather it is a ligand for PD-1. The interactions between PD-1 and B7-DC/PD-L2 have been reported to be involved in costimulation or suppression of T cell proliferation depending on state of cellular activation. MIH18 is a useful tool for studies to elucidate the exact function of B7-DC/PD-L2 in APC/T cell interaction and to characterize the expression pattern of this molecule in humans. MIH18 is a blocking antibody. Applications Reported: The MIH18 antibody has been reported for use in flow cytometric analysis. Applications Tested: The MIH18 antibody has been tested by flow cytometric analysis of mature dendritic cells and normal human peripheral blood cells. This can be used at less than or equal to 0.5 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- MIH18
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- 4° C
Submitted references TLR9 Mediated Tumor-Stroma Interactions in Human Papilloma Virus (HPV)-Positive Head and Neck Squamous Cell Carcinoma Up-Regulate PD-L1 and PD-L2.
Osteosarcoma cell intrinsic PD-L2 signals promote invasion and metastasis via the RhoA-ROCK-LIMK2 and autophagy pathways.
Enhanced expression of PD-L1 in non-muscle-invasive bladder cancer after treatment with Bacillus Calmette-Guerin.
PD-1 axis expression in musculoskeletal tumors and antitumor effect of nivolumab in osteosarcoma model of humanized mouse.
Clinical Features of Nivolumab-Induced Thyroiditis: A Case Series Study.
Tolerogenic IDO(+) Dendritic Cells Are Induced by PD-1-Expressing Mast Cells.
Psoriasis is characterized by deficient negative immune regulation compared to transient delayed-type hypersensitivity reactions.
Antagonist antibodies to PD-1 and B7-H1 (PD-L1) in the treatment of advanced human cancer.
Tissue biomarkers in melanoma patients treated with TIL.
HIV-mediated phosphatidylinositol 3-kinase/serine-threonine kinase activation in APCs leads to programmed death-1 ligand upregulation and suppression of HIV-specific CD8 T cells.
Activated memory B cells may function as antigen-presenting cells in the joints of children with juvenile idiopathic arthritis.
Immunostaining of PD-1/PD-Ls in liver tissues of patients with hepatitis and hepatocellular carcinoma.
Mechanisms of immune suppression for CD8+ T cells by human corneal endothelial cells via membrane-bound TGFbeta.
Role played by the programmed death-1-programmed death ligand pathway during innate immunity against Mycobacterium tuberculosis.
PD-1 is a regulator of NY-ESO-1-specific CD8+ T cell expansion in melanoma patients.
B7-H3 is a potent inhibitor of human T-cell activation: No evidence for B7-H3 and TREML2 interaction.
Human corneal endothelial cells expressing programmed death-ligand 1 (PD-L1) suppress PD-1+ T helper 1 cells by a contact-dependent mechanism.
Baruah P, Bullenkamp J, Wilson POG, Lee M, Kaski JC, Dumitriu IE
Frontiers in immunology 2019;10:1644
Frontiers in immunology 2019;10:1644
Osteosarcoma cell intrinsic PD-L2 signals promote invasion and metastasis via the RhoA-ROCK-LIMK2 and autophagy pathways.
Ren T, Zheng B, Huang Y, Wang S, Bao X, Liu K, Guo W
Cell death & disease 2019 Mar 18;10(4):261
Cell death & disease 2019 Mar 18;10(4):261
Enhanced expression of PD-L1 in non-muscle-invasive bladder cancer after treatment with Bacillus Calmette-Guerin.
Hashizume A, Umemoto S, Yokose T, Nakamura Y, Yoshihara M, Shoji K, Wada S, Miyagi Y, Kishida T, Sasada T
Oncotarget 2018 Sep 25;9(75):34066-34078
Oncotarget 2018 Sep 25;9(75):34066-34078
PD-1 axis expression in musculoskeletal tumors and antitumor effect of nivolumab in osteosarcoma model of humanized mouse.
Zheng B, Ren T, Huang Y, Sun K, Wang S, Bao X, Liu K, Guo W
Journal of hematology & oncology 2018 Feb 6;11(1):16
Journal of hematology & oncology 2018 Feb 6;11(1):16
Clinical Features of Nivolumab-Induced Thyroiditis: A Case Series Study.
Yamauchi I, Sakane Y, Fukuda Y, Fujii T, Taura D, Hirata M, Hirota K, Ueda Y, Kanai Y, Yamashita Y, Kondo E, Sone M, Yasoda A, Inagaki N
Thyroid : official journal of the American Thyroid Association 2017 Jul;27(7):894-901
Thyroid : official journal of the American Thyroid Association 2017 Jul;27(7):894-901
Tolerogenic IDO(+) Dendritic Cells Are Induced by PD-1-Expressing Mast Cells.
Rodrigues CP, Ferreira AC, Pinho MP, de Moraes CJ, Bergami-Santos PC, Barbuto JA
Frontiers in immunology 2016;7:9
Frontiers in immunology 2016;7:9
Psoriasis is characterized by deficient negative immune regulation compared to transient delayed-type hypersensitivity reactions.
Gulati N, Suárez-Fariñas M, Correa da Rosa J, Krueger JG
F1000Research 2015;4:149
F1000Research 2015;4:149
Antagonist antibodies to PD-1 and B7-H1 (PD-L1) in the treatment of advanced human cancer.
Sznol M, Chen L
Clinical cancer research : an official journal of the American Association for Cancer Research 2013 Mar 1;19(5):1021-34
Clinical cancer research : an official journal of the American Association for Cancer Research 2013 Mar 1;19(5):1021-34
Tissue biomarkers in melanoma patients treated with TIL.
Knol AC, Nguyen JM, Pandolfino MC, Quéreux G, Brocard A, Peuvrel L, Saint-Jean M, Saiagh S, Khammari A, Dréno B
PloS one 2012;7(12):e48729
PloS one 2012;7(12):e48729
HIV-mediated phosphatidylinositol 3-kinase/serine-threonine kinase activation in APCs leads to programmed death-1 ligand upregulation and suppression of HIV-specific CD8 T cells.
Muthumani K, Shedlock DJ, Choo DK, Fagone P, Kawalekar OU, Goodman J, Bian CB, Ramanathan AA, Atman P, Tebas P, Chattergoon MA, Choo AY, Weiner DB
Journal of immunology (Baltimore, Md. : 1950) 2011 Sep 15;187(6):2932-43
Journal of immunology (Baltimore, Md. : 1950) 2011 Sep 15;187(6):2932-43
Activated memory B cells may function as antigen-presenting cells in the joints of children with juvenile idiopathic arthritis.
Morbach H, Wiegering V, Richl P, Schwarz T, Suffa N, Eichhorn EM, Eyrich M, Girschick HJ
Arthritis and rheumatism 2011 Nov;63(11):3458-66
Arthritis and rheumatism 2011 Nov;63(11):3458-66
Immunostaining of PD-1/PD-Ls in liver tissues of patients with hepatitis and hepatocellular carcinoma.
Wang BJ, Bao JJ, Wang JZ, Wang Y, Jiang M, Xing MY, Zhang WG, Qi JY, Roggendorf M, Lu MJ, Yang DL
World journal of gastroenterology 2011 Jul 28;17(28):3322-9
World journal of gastroenterology 2011 Jul 28;17(28):3322-9
Mechanisms of immune suppression for CD8+ T cells by human corneal endothelial cells via membrane-bound TGFbeta.
Yamada Y, Sugita S, Horie S, Yamagami S, Mochizuki M
Investigative ophthalmology & visual science 2010 May;51(5):2548-57
Investigative ophthalmology & visual science 2010 May;51(5):2548-57
Role played by the programmed death-1-programmed death ligand pathway during innate immunity against Mycobacterium tuberculosis.
Alvarez IB, Pasquinelli V, Jurado JO, Abbate E, Musella RM, de la Barrera SS, García VE
The Journal of infectious diseases 2010 Aug 15;202(4):524-32
The Journal of infectious diseases 2010 Aug 15;202(4):524-32
PD-1 is a regulator of NY-ESO-1-specific CD8+ T cell expansion in melanoma patients.
Fourcade J, Kudela P, Sun Z, Shen H, Land SR, Lenzner D, Guillaume P, Luescher IF, Sander C, Ferrone S, Kirkwood JM, Zarour HM
Journal of immunology (Baltimore, Md. : 1950) 2009 May 1;182(9):5240-9
Journal of immunology (Baltimore, Md. : 1950) 2009 May 1;182(9):5240-9
B7-H3 is a potent inhibitor of human T-cell activation: No evidence for B7-H3 and TREML2 interaction.
Leitner J, Klauser C, Pickl WF, Stöckl J, Majdic O, Bardet AF, Kreil DP, Dong C, Yamazaki T, Zlabinger G, Pfistershammer K, Steinberger P
European journal of immunology 2009 Jul;39(7):1754-64
European journal of immunology 2009 Jul;39(7):1754-64
Human corneal endothelial cells expressing programmed death-ligand 1 (PD-L1) suppress PD-1+ T helper 1 cells by a contact-dependent mechanism.
Sugita S, Usui Y, Horie S, Futagami Y, Yamada Y, Ma J, Kezuka T, Hamada H, Usui T, Mochizuki M, Yamagami S
Investigative ophthalmology & visual science 2009 Jan;50(1):263-72
Investigative ophthalmology & visual science 2009 Jan;50(1):263-72
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Supportive validation
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- Experimental details
- Staining of human in vitro derived dendritic cells with 0.25 µg of Mouse IgG1 K Isotype Control Purified (Product # 14-4714-82) (open histogram) or 0.25 µg of Anti-Human CD273 (B7-DC) Purified followed by Anti-Mouse IgG Biotin (Product # 13-4013-85) and Streptavidin PE (Product # 12-4317-87) (filled histogram).Total viable cells were used for analysis.
Supportive validation
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- Fig. 2 Effect of PD-L2 expression on the proliferation of osteosarcoma cells. a PD-L2 expression in osteosarcoma cell lines was evaluated by western blotting and real-time PCR. b The osteosarcoma cell lines (red) exhibited differing degrees of PD-L2 expression compared with the isotype control (black) by flow cytometry. c KHOS and U2OS cells were stably transfected with shPD-L2 or shNC lentivirus, following evaluation of PD-L2 expression by western blotting and real-time PCR. d Cell proliferation of KHOS and U2OS cells in response to PD-L2 knockdown determined by cell colony formation assay. e Cell viability of KHOS and U2OS cells after PD-L2 knockdown assayed by CCK-8. All experiments were repeated three times. Data are presented as the mean +- SD. * P < 0.05, ** P < 0.01, *** P < 0.001
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- Figure 3 HPV-positive HNSCCs increase PD-L1 and PD-L2 on fibroblasts. PD-L1 and PD-L2 expression on primary BJ human fibroblasts, HPV-positive (SCC154), and HPV-negative (SCC099) HNSCC cell lines (HNSCCs) was detected by flow cytometry. Illustrative histograms show PD-L1 and PD-L2 expression on fibroblasts (black histograms), HPV-positive (red histograms), or HPV-negative (blue histograms) HNSCCs (A) . Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 5) in fibroblasts, HPV-positive, and HPV-negative HNSCCs (B) . Fibroblasts were cultured alone or co-cultured in direct contact (direct) with HPV-positive (SCC154) or HPV-negative (SCC099) HNSCCs. Fibroblasts were identified in co-cultures by lack of EpCAM expression. Illustrative histograms show PD-L1 and PD-L2 expression on fibroblasts cultured alone (black histograms) or co-cultured directly with HPV-positive SCC154 ( C ; red histograms) or HPV-negative SCC099 ( D ; blue histograms). Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 5) on fibroblasts cultured alone (w/o) or co-cultured directly with HNSCC cells (E) . HPV-positive (SCC154) or HPV-negative (SCC099) HNSCCs were cultured alone or co-cultured in direct contact (direct) with fibroblasts. HNSCCs were identified in co-cultures by EpCAM expression. Illustrative histograms show PD-L1 and PD-L2 expression on HPV-positive SCC154 cultured alone (black histograms) or co-cultured directly with fibroblasts ( F ; red histograms). Graphs show PD-L1 and PD-
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- Figure 4 PD-L1 and PD-L2 expression in co-cultures of macrophages and HPV-positive or HPV-negative HNSCCs. Macrophages were cultured alone (w/o) or co-cultured in direct contact (direct) with HPV-positive (SCC154) or HPV-negative (SCC099) HNSCC cell lines (HNSCCs). Illustrative histograms show PD-L1 and PD-L2 expression on macrophages cultured alone ( A ; black histograms). Macrophages were identified in co-cultures by lack of EpCAM expression. Illustrative histograms show PD-L1 and PD-L2 expression on macrophages cultured alone (black histograms) or co-cultured directly with HPV-positive SCC154 ( B ; red histograms) or HPV-negative SCC099 ( C ; blue histograms). Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 4) on macrophages cultured alone (w/o) or co-cultured directly with HNSCCs (D) . HPV-positive (SCC154) or HPV-negative (SCC099) HNSCCs were cultured alone or co-cultured in direct contact (direct) with macrophages. HNSCCs were identified in co-cultures by EpCAM expression. Illustrative histograms show PD-L1 and PD-L2 expression on HPV-positive SCC154 cultured alone (black histograms) or co-cultured directly with macrophages ( E ; red histograms). Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 3) on HPV-positive SCC154 cultured alone or co-cultured directly with macrophages (F) . Illustrative histograms show PD-L1 and PD-L2 expression on HPV-negative SCC099 cultured alone (black histograms) or co-cultured directly with macrophages ( G ; blue hist
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- Figure 5 Conditioned medium from HPV-positive HNSCCs up-regulates PD-L1 and PD-L2 on fibroblasts. Fibroblasts were cultured alone or co-cultured in direct contact with HPV-positive SCC154 (direct) or with conditioned medium from HPV-positive SCC154 (CM). Illustrative histograms show PD-L1 and PD-L2 expression on fibroblasts cultured alone (black histograms) or co-cultured with conditioned medium from HPV-positive SCC154 ( A ; red histograms). Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 13) on fibroblasts cultured alone (w/o), co-cultured directly with HPV-positive SCC154 (direct) or with conditioned medium from HPV-positive SCC154 (CM) (B) . HPV-positive (SCC154) HNSCCs were cultured alone or co-cultured in direct contact with fibroblasts (Fibro direct) or with conditioned medium from fibroblasts (Fibro CM). Illustrative histograms show PD-L1 and PD-L2 expression on HPV-positive SCC154 cultured alone (black histograms) or co-cultured with conditioned medium from fibroblasts ( C ; red histograms). Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 4) on HPV-positive SCC154 cultured alone (w/o), co-cultured directly with fibroblasts (Fibro direct) or with conditioned medium from fibroblasts (Fibro CM) (D) . Macrophages were cultured alone (w/o) or co-cultured in direct contact with HPV-positive SCC154 (direct) or with conditioned medium from HPV-positive SCC154 (CM). Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 3) on macrophages for the in
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- Figure 6 Blockade of IFN-gamma, TNF-alpha, or CD81 does not affect PD-L1 and PD-L2 up-regulation by HPV-positive HNSCCs. Fibroblasts were cultured alone (w/o) or co-cultured in direct contact with HPV-positive SCC154 (SCC154 direct) or with conditioned medium from HPV-positive SCC154 (SCC154 CM) as indicated. Graphs (A) show IFN-gamma and TNF-alpha levels in culture supernatants (mean +- SEM; n = 4). The dashed red line indicates the lowest value (15.6 pg/ml) of the dynamic range for the ELISA assays used. Neutralizing antibodies anti-IFN-gamma (B,C) , anti-TNF-alpha (D,E) , or anti-CD81 (F,G) were added to the cultures as indicated. Illustrative histograms show PD-L1 and PD-L2 expression on fibroblasts cultured alone (black histograms), co-cultured directly with HPV-positive SCC154 or with conditioned medium from HPV-positive SCC154 alone (red histograms) or in the presence of blocking antibodies (green histograms). Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 3) on fibroblasts for the indicated treatments. ns, not significant (one-way ANOVA with Bonferroni correction for multiple comparisons) Numbers adjacent to plots represent MFI values; dashed histograms show control staining with isotype-matched antibodies. MFI, mean fluorescence intensity.
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- Figure 7 The TLR9 antagonists ODN TTAGGG and chloroquine inhibit PD-1 ligands up-regulation on fibroblasts co-cultured with HPV-positive HNSCCs. Fibroblasts were cultured alone (w/o) or co-cultured in direct contact with HPV-positive SCC154 (SCC154 direct) or with conditioned medium from HPV-positive SCC154 (SCC154 CM) in the presence or absence of the TLR9 antagonists ODN TTAGGG (ODN) or chloroquine (CHQ). Illustrative histograms show PD-L1 and PD-L2 expression on fibroblasts cultured alone (black histograms), co-cultured directly with HPV-positive SCC154 (red histograms) or co-cultured directly with HPV-positive SCC154 in the presence of ODN (A) or CHQ (H) (green histograms). Illustrative histograms show PD-L1 and PD-L2 expression on fibroblasts cultured alone (black histograms), cultured with conditioned medium from HPV-positive SCC154 (red histograms) or with conditioned medium from HPV-positive SCC154 in the presence of ODN (B) or CHQ (I) (green histograms). Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 6) on fibroblasts for the indicated treatments (C,J) . Illustrative histograms show PD-L1 and PD-L2 expression on fibroblasts cultured alone (black histograms) or in the presence of ODN (D) or CHQ (K) (green histograms). Graphs show PD-L1 and PD-L2 expression (mean +- SEM; n = 4) on fibroblasts for the indicated treatments (E,L) . HPV-positive SCC154 were cultured alone or co-cultured in direct contact (direct) with fibroblasts in the presence