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European Journal of Medical Research

Erschienen in:

01.12.2011 | Research

Development of a novel monoclonal antibody to B7-H4: characterization and biological activity

verfasst von: Y. Qian, L. Shen, C. Xu, Z. Wu, N. H. Brockmeyer, P. Altmeyer, N. Wu, H. Yao

Erschienen in: European Journal of Medical Research | Ausgabe 7/2011

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Abstract

Objective

B7-H4, a member of the B7 family of immunoregulatory receptors, may participate in the negative regulation of cell-mediated immunity. Aberrant B7-H4 expression is detected in some tumors and it plays a role in the occurrence and development of tumors. The aim of this study was to elucidate the functional and structural properties of B7-H4.

Methods

We developed a monoclonal antibody (mAb) against the extracellular domain of B7-H4 through immunization of Balb/c mice with 3T3-mB7-H4 cells which expressed extrinsic B7-H4. A stable hybridoma cell line was established. Then, we analysised the characterization of the mAb through Enzyme linked immunosorbent assay (ELISA), Immunoprecipitation (IP), western blotting, Immunohistochemical (IHC), and tested the biological activity of the mAb.

Results

ELISA, IP, and western blotting analyses indicated that the mAb specifically recognized B7-H4. In addition, flow cytometry demonstrated that the mAb exhibits excellent reactivity when applied to leukemic cells. IHC staining revealed that the mAb stained in a predominantly diffuse plasmalemmal or cytoplasmic pattern when applied to certain tumor tissues. The preliminary results of the mAb's biological activity showed that the mAb could effectively inhibit the function of B7-H4 in the inhibition of T cell, while promotingg the growth of T cells and the secretion of Interleukin-2 (lL-2), Interleukin-4 (IL-4), Interleukin10 (IL-10) and Interferon-γ (IFN-γ).

Conclusion

This mAb will be a valuable tool for the further investigation of B7-H4 function.

Sica GL, Choi IH, Zhu G, Tamada K, Wang SD, Tamura H, Chapoval AI, Flies DB, Bajorath J, Chen L: B7-H4, a molecule of the B7 family, negatively regulates T cell immunity. Immunity 2003, 18: 849–861. 10.1016/S1074-7613(03)00152-3CrossRefPubMed

Chen L: Co-inhibitory molecules of the B7-CD28 family in the control of T cell immunity. Nature Rev Immunol 2004, 4: 336–347. 10.1038/nri1349CrossRef

B7x: a widely expressed B7 family member that inhibits T cell activation: Proc Natl Acad Sci USA. 2003, 100: 10388–10392. 10.1073/pnas.1434299100CrossRef

Prasad DVR, Richards S, Mai XM, Dong C: B7S1, a novel B7 family member that negatively regulates T cell activation. Immunity 2003, 18: 863–873. 10.1016/S1074-7613(03)00147-XCrossRefPubMed

Salceda S, Tang T, Kmet M, Munteanu A, Ghosh M, Macina R, Liu W, Pilkington G, Papkoff J: The immunomodulatory protein B7-H4 is overexpressed in breast and ovarian cancers and promotes epithelial cell transformation. Exp Cell Res 2005, 306: 128–141. 10.1016/j.yexcr.2005.01.018CrossRefPubMed

Tringler B, Zhuo S, Pilkington G, Torkko KC, Singh M, Lucia MS, Heinz DE, Papkoff J, Shroyer KR: B7-h4 is highly expressed in ductal and lobular breast cancer. Clin Cancer Res 2005, 11: 1842–1848. 10.1158/1078-0432.CCR-04-1658CrossRefPubMed

Choi IH, Zhu G, Sica GL, Strome SE, Cheville JC, Lau JS, Zhu Y, Flies DB, Tamada K, Chen L: Genomic organization and expression analysis of B7-H4, an immune inhibitory molecule of the B7 family. J Immunol 2003, 171: 4650–4654.CrossRefPubMed

Krambeck AE, Thompson RH, Dong H, Lohse CM, Park ES, Kuntz SM, Leibovich BC, Blute ML, Cheville JC, Kwon ED: B7-H4 expression in renal cell carcinoma and tumor vasculature: associations with cancer progression and survival. Proc Nati Acad Sci USA 2006, 103: 10391–10396. 10.1073/pnas.0600937103CrossRef

Zang X, Thompson RH, Al-Ahmadie HA, Serio AM, Reuter VE, Eastham JA, Scardino PT, Sharma P, Allison JP: B7-H3 and B7x are highly expressed in human prostate cancer and associated with disease spread and poor outcome. Proc Natl Acad Sci USA 2007, 104: 19458–19463. 10.1073/pnas.0709802104PubMedCentralCrossRefPubMed

10.

Sun Y, Wang Y, Zhao J, Gu M, Giscombe R, Lefvert AK, Wang X: B7-H3 and B7-H4 expression in non-small-cell lung cancer. Lung Cancer 2006, 53: 143–151. 10.1016/j.lungcan.2006.05.012CrossRefPubMed

11.

Kryczek I, Zou L, Rodriguez P, Zhu G, Wei S, Mottram P, Brumlik M, Cheng P, Curiel T, Myers L, Lackner A, Alvarez X, Ochoa A, Chen L, Zou W: B7-H4 expression identifies a novel suppressive macrophage population in human ovarian carcinoma. J Exp Me 2006, (203):871–881.CrossRef

12.

Simon I, Zhuo S, Corral L, Diamandis EP, Sarno MJ, Wolfert RL, Kim NW: B7-H4 Is a Novel MembraneBound Protein and a Candidate Serum and Tissue Biomarker for Ovarian Cancer. Cancer Res 2006, 66: 1570–1575. 10.1158/0008-5472.CAN-04-3550CrossRefPubMed

13.

Thompson RH, Zang X, Lohse CM, Leibovich BC, Slovin SF, Reuter VE, Cheville JC, Blute ML, Russo P, Kwon ED, Allison JP: Serum-soluble B7x is elevated in renal cell carcinoma patients and is associated with advanced stage. Cancer Res 2008, 68: 6054–6058. 10.1158/0008-5472.CAN-08-0869PubMedCentralCrossRefPubMed

14.

Mugler KC, Singh M, Tringler B, Torkko KC, Liu W, Papkoff J, Shroyer KR: B7-h4 expression in a range of breast pathology: correlation with tumor T-cell infiltration. Appl Immunohistochem Mol Morphol 2007,15(4):363–370. 10.1097/01.pai.0000213159.79557.71CrossRefPubMed

15.

Qian Y, Shen L, Cheng LF, Wu ZG, Yao HP: B7-H4 expression in various tumors determined using a novel developed monoclonal antibody. Clin Exp Med 2010. [printed in head]

16.

Yao HP, Luo YL, Feng L, Cheng LF, Lu Y, Li W, Wang MH: Agonistic monoclonal antibodies potentiate tumorigenic and invasive activities of splicing variant of the RON receptor tyrosine kinase. Cancer Biol Ther 2006, 5: 1179–1186. 10.4161/cbt.5.9.3073CrossRefPubMed

17.

Nelson PN, Reynolds GM, Waldron EE, Ward E, Giannopoulos K, Murray PG: Monoclonal antibodies. Molecular Pathology 2000, 53: 111–117. 10.1136/mp.53.3.111PubMedCentralCrossRefPubMed

18.

Qian Y, Yao HP, Cheng LF, Shen L, Gu LJ, Tao L, Zhang LH: Expression and identification of recombinant mouse gene B7-H4. Journal of Zhejiang University (medical sciences) 2009, 38: 117–124.

19.

Allred DC, Harvey JM, Berardo M, Clark GM: Prognostic and predictive factors in breast cancer by immunohistochemical analysis. Mod Pathol 1998, 11: 155–168.PubMed

20.

Park GB, Song H, Kim YS, Sung M, Ryu JW, Lee HK, Cho DH, Kim D, Lee WJ, Hur Dy: Cell cycle arrest induced by engagement of B7-H4 on Epstein-Barr viruspositive B-cell lymphoma cell lines. Immunology 2009,128(3):360–368. 10.1111/j.1365-2567.2009.03111.xPubMedCentralCrossRefPubMed

21.

Guin S, Yao HP, Wang MH: RON receptor tyrosine kinase as a target for delivery of chemodrugs by antibody directed pathway for cancer cell cytotoxicity. Mol Pharm 2010,7(2):386–397. 10.1021/mp900168vCrossRefPubMed

22.

Tringler B, Liu W, Corral L, Torkko KC, Enomoto T, Davidson S, Lucia MS, Heinz DE, Papkoff J, Shroyer KR: B7-H4 overexpression in ovarian tumors. Gynecol Onco 2006, (100):44–52.

23.

Miyatake T, Tringler B, Liu W, Liu SH, Papkoff J, Enomoto T, Torkko KC, Dehn DL, Swisher A, Shroyer KR: B7-H4 (DD-O110) is overexpressed in high risk uterine endometrioid adenocarcinomas and inversely correlated with tumor T-cell infiltration. Gynecol Oncol 2007, 106: 119–127. 10.1016/j.ygyno.2007.03.039CrossRefPubMed

24.

Cheng L, Jiang J, Gao R, Wei S, Nan F, Li S, Kong B: B7-H4 expression promotes tumorigenesis in ovarian cancer. Int J Gynecol Cancer 2009, 19: 1481–1486. 10.1111/IGC.0b013e3181ad0fa2CrossRefPubMed

25.

Jiang J, Zhu Y, Wu C, Shen Y, Wei W, Chen L, Zheng X, Sun J, Lu B, Zhang X: Tumor expression of B7-H4 predicts poor survival of patients suffering from gastric cancer. Cancer Immunol Immunother 2010,59(11):1707–1714. 10.1007/s00262-010-0900-7CrossRefPubMed

26.

Azzoni C, Bottarelli L, Cecchini S, Lagrasta C, Pizzi S, D'Adda T, Tamburini E, Rindi G, Bordi C: Involvement of HER-2/neu and metastasis-related proteins in the development of ileal neuroendocrine tumors. Virchows Arch 2011, in press.

27.

Zynek-Litwin M, Kuzniar J, Nocen-Rychlewska B, Kusztal M, Klinger M: An inconspicuous insect bite as the cause of sepsis in a patient treated with infliximab due to rheumatoid arthritis. Adv Med Sci 2011, in press.

Titel: Development of a novel monoclonal antibody to B7-H4: characterization and biological activity
verfasst von: Y. Qian
L. Shen
C. Xu
Z. Wu
N. H. Brockmeyer
P. Altmeyer
N. Wu
H. Yao
Publikationsdatum: 01.12.2011
Verlag: BioMed Central
Erschienen in: European Journal of Medical Research / Ausgabe 7/2011
Elektronische ISSN: 2047-783X
DOI: https://doi.org/10.1186/2047-783X-16-7-295

Springer Medizin

Abstract

Objective

Methods

Results

Conclusion

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