16-5889-82

antibody from Invitrogen Antibodies

Targeting: ICOSLG B7-H2, B7h, B7H2, B7RP-1, B7RP1, CD275, GL50, ICOS-L, ICOSL, KIAA0653

Immunohistochemistry (Recommended by provider)

Flow cytometry (Supportive data in Antibodypedia)

Blocking/Neutralizing (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

16-5889-82

Provider

Invitrogen Antibodies

Product name

CD275 (B7-H2) Monoclonal Antibody (MIH12), Functional Grade, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The MIH12 monoclonal antibody reacts with human B7RP-1, also known as B7h, B7-H2, GL50 and ICOS Ligand. B7RP-1, a member of the B7 family, has a predicted molecular weight of approximately 40 kDa and belongs to the Ig superfamily. Human B7RP-1 is expressed by activated monocytes/macrophages. B7RP-1 binds to the ICOS molecule (AILIM, CRP-1) expressed by activated T cells. The interaction of ICOS/B7RP-1 plays an important role in the T cell costimulation pathway. Applications Reported: This MIH12 antibody has been reported for use in flow cytometric analysis, and immunohistology staining of frozen tissue sections. The MIH12 monoclonal antibody has also been demonstrated to block the co-stimulatory function of B7RP-1 including the inhibition of T-cell proliferation and IL-10 production. Applications Tested: The MIH12 antibody has been tested by flow cytometric analysis of normal human peripheral blood monocyte-derived dendritic cells. This can be used at less than or equal to 1 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. Storage and handling: Use in a sterile environment. Filtration: 0.2 µm post-manufacturing filtered. Purity: Greater than 90%, as determined by SDS-PAGE. Endotoxin Level: Less than 0.001 ng/µg antibody, as determined by LAL assay. Aggregation: Less than 10%, as determined by HPLC.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

MIH12

Vial size

100 µg

Concentration

1 mg/mL

Storage

4° C

References

Submitted references

Senescence-induced endothelial phenotypes underpin immune-mediated senescence surveillance.

Yin K, Patten D, Gough S, de Barros Gonçalves S, Chan A, Olan I, Cassidy L, Poblocka M, Zhu H, Lun A, Schuijs M, Young A, Martinez-Jimenez C, Halim TYF, Shetty S, Narita M, Hoare M
Genes & development 2022 May 1;36(9-10):533-549

---

Cytomegalovirus restricts ICOSL expression on antigen-presenting cells disabling T cell co-stimulation and contributing to immune evasion.

Angulo G, Zeleznjak J, Martínez-Vicente P, Puñet-Ortiz J, Hengel H, Messerle M, Oxenius A, Jonjic S, Krmpotić A, Engel P, Angulo A
eLife 2021 Jan 18;10

---

Dysregulated NF-κB-Dependent ICOSL Expression in Human Dendritic Cell Vaccines Impairs T-cell Responses in Patients with Melanoma.

Maurer DM, Adamik J, Santos PM, Shi J, Shurin MR, Kirkwood JM, Storkus WJ, Butterfield LH
Cancer immunology research 2020 Dec;8(12):1554-1567

---

Comprehensive characterization of chorionic villi-derived mesenchymal stromal cells from human placenta.

Ventura Ferreira MS, Bienert M, Müller K, Rath B, Goecke T, Opländer C, Braunschweig T, Mela P, Brümmendorf TH, Beier F, Neuss S
Stem cell research & therapy 2018 Feb 5;9(1):28

---

Polyfunctional Melan-A-specific tumor-reactive CD8(+) T cells elicited by dacarbazine treatment before peptide-vaccination depends on AKT activation sustained by ICOS.

Franzese O, Palermo B, Di Donna C, Sperduti I, Ferraresi V, Stabile H, Gismondi A, Santoni A, Nisticò P
Oncoimmunology 2016 May;5(5):e1114203

---

Development of a Novel Functional Monoclonal Antibody to Human CD275: Characterization and Biological Activity.

Hu X, Liu C, An J, Shen Y, Hu Y, Jiang J, Wu J, Zhang X
Monoclonal antibodies in immunodiagnosis and immunotherapy 2016 Feb;35(1):18-24

---

Pulmonary sarcoidosis is associated with high-level inducible co-stimulator (ICOS) expression on lung regulatory T cells--possible implications for the ICOS/ICOS-ligand axis in disease course and resolution.

Sakthivel P, Grunewald J, Eklund A, Bruder D, Wahlström J
Clinical and experimental immunology 2016 Feb;183(2):294-306

---

Tumor-infiltrating plasmacytoid dendritic cells promote immunosuppression by Tr1 cells in human liver tumors.

Pedroza-Gonzalez A, Zhou G, Vargas-Mendez E, Boor PP, Mancham S, Verhoef C, Polak WG, Grünhagen D, Pan Q, Janssen H, Garcia-Romo GS, Biermann K, Tjwa ET, IJzermans JN, Kwekkeboom J, Sprengers D
Oncoimmunology 2015 Jun;4(6):e1008355

---

Endothelial microparticles interact with and support the proliferation of T cells.

Wheway J, Latham SL, Combes V, Grau GE
Journal of immunology (Baltimore, Md. : 1950) 2014 Oct 1;193(7):3378-87

---

Expression and regulation of human CD275 on endothelial cells in healthy and inflamed mucosal tissues.

Youngnak-Piboonratanakit P, Tsushima F, Otsuki N, Igarashi H, Omura K, Azuma M
Scandinavian journal of immunology 2006 Mar;63(3):191-8

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Staining of normal human peripheral blood cells with Anti-Human CD275 (B7-H2) PE.Appropriate isotype controls were used (open histogram).Cells in the lymphocyte population were used for analysis.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 11. HCMV, HSV-1, and HSV-2 also limit cell surface expression of ICOSL on antigen-presenting cells (APCs). ( A ) Primary monocyte-derived macrophages (upper panels) and PMA-treated THP-1 cells (bottom panels) were mock-infected or infected for 72 hr with HCMV-GFP at an moi of 10 and analyzed by flow cytometry for cell-surface expression of human ICOSL or CD70 using specific mAbs against each of these receptors. Gray histograms represent the expression of mock-infected cells, green histograms represent the expression on HCMV-infected (GFP+) cells, and blue histograms represent the expression on uninfected (GFP-) cells from the same culture. ( B ) PMA-treated THP-1 cells were mock-infected (time 0) or infected with HCMV-GFP as in A and analyzed by flow cytometry for surface expression of ICOSL at the different time points after infection indicated. ( C ) Same as in A, except that an moi of 20 was used, and THP-1 cells were also exposed for 72 hr to the same amount of HCMV-GFP UV-inactivated (red histogram). ( D ) Equal amounts of lysates from PMA-treated THP-1 cells mock-infected (lane 1) or infected for 72 hr at an moi of 20 with HCMV-GFP (lanes 2 and 3), and when indicated, treated with 250 muM leupeptin and 20 nM of bafilomycin A1 (lane 3), were lysed and analyzed by western blot with antibodies against ICOSL and actin, followed by anti-rabbit IgG-HRP (ICOSL) or anti-mouse IgG-HRP (actin). ( E ) PMA-treated THP-1 cells were mock-infected or infected with HSV-1-GFP, HS

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 2 a Cumulative population-doubling (cPD) levels versus passage number for the four different sources of MSC. Black represents CV-MSC ( n = 7), dark gray UC-MSC ( n = 4), medium gray AT-MSC ( n = 5), and light gray BM-MSC ( n = 6). b IHC-based senescence-associated beta-galactosidase (SA-beta-gal) staining of CV-MSC in early (i, passage 4) and late (ii, passage 9) passages, AT-MSC in passage 6 (iii), BM-MSC in passage 6 (iv), and UC-MSC in passage 2 (v) and passage 4 (vi). Scale = 200 mum. c IHC of CV-MSC (i, ii) and BM-MSC (iii, iv) stained for osteopontin (i, iii) and fibronectin (ii, iv). Scale = 1 mm. d Collagen area (%) after collagen contraction assay for CV-MSC ( n = 4), BM-MSC ( n = 3), UC-MSC ( n = 4), and AT-MSC ( n = 3). Cells in passage 3 were used. Results expressed as mean +- SD, percentage of the total collagen area of the collagen gels without cells. e Surface marker expression of CV-MSC in early passages ( n = 5). Results expressed as mean +- SD (%). f Representative immunofluorescence of early passaged CV-MSC (i, iii) and BM-MSC (iii, iv) stained for SM22alpha (i, iii) and alpha-SMA (ii, iv). Scale = 50 mum. AT adipose tissue, BM bone marrow, CV chorionic villi, MSC mesenchymal stromal cells, UC umbilical cord

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Increased proportion of non-classical blood monocytes with higher inducible co-stimulator ligand (ICOS-L) expression in sarcoidosis patients. (a) Representative fluorescence activated cell sorter (FACS) dot-plot gated on blood monocyte populations in peripheral blood mononuclear cells (PBMCs) showing the frequencies of classical, intermediate and non-classical (proinflammatory) monocytes in sarcoidosis patients ( n = 11) and healthy controls ( n = 13). (b) Box-plots represent the % CD14 high CD16 low monocytes (classical) in sarcoidosis patients and healthy controls (upper left) and the intensities of ICOS-L [as mean fluorescence intensity (MFI)] on classical monocytes in patients and controls (upper right). Box-plots represent the % CD14 low CD16 high monocytes (non-classical) in sarcoidosis patients and healthy controls (lower left) and the intensities of ICOS-L (as MFI) on non-classical monocytes in patients and controls (lower right). P -values were calculated using the Mann-Whitney U -test.