• 中国精品科技期刊
  • 中国高校百佳科技期刊
  • 中国中文核心期刊
  • 中国科学引文数据库核心期刊
Advanced Search
CHEN Yali, LI Anqi, ZHANG Ningyue, ZHANG Liang. Purification of monoclonal anti-B7-H4 antibody and the blockade of B7-H4-mediated tumor immune evasion by the antibody[J]. Journal of China Pharmaceutical University, 2017, 48(4): 461-468. DOI: 10.11665/j.issn.1000-5048.20170411
Citation: CHEN Yali, LI Anqi, ZHANG Ningyue, ZHANG Liang. Purification of monoclonal anti-B7-H4 antibody and the blockade of B7-H4-mediated tumor immune evasion by the antibody[J]. Journal of China Pharmaceutical University, 2017, 48(4): 461-468. DOI: 10.11665/j.issn.1000-5048.20170411

Purification of monoclonal anti-B7-H4 antibody and the blockade of B7-H4-mediated tumor immune evasion by the antibody

More Information
  • The purified 3C8 was obtained by two step column purification, including Protein G affinity purification and DEAE anion exchange purification. The purity of purified 3C8 was about 93% when analyzed by reverse column. SDS-PAGE showed that the purity of 3C8 was increased greatly by two step purification. By flowcytometry we found that 3C8 specifically binded with B7-H4/293T cells and did not bind with Mock/293T cells, moreover 3C8 did not bind with other B7 family members transgene cells. In confocal experiment 3C8 could specifically stained B7-H4/293T cells. In Western blot only B7-H4/293T cells showed positive band while Mock/293T cells showed negative result. The result of immunohistochemistry showed that B7-H4 was highly expressed in prostate cancer and renal cell carcinoma, while para-cancer tissues did not express B7-H4. The T cell proliferation experiment showed that B7-H4-Ig could bind to activate T cells and inhibit T cell proliferation, while 3C8 could block the binding of B7-H4-Ig and reverse the T cell proliferation inhibition effect of B7-H4-Ig by CFSE and CCK8 assay. The cytokine IFN-γ and IL-2 secreted by activating T cells was decreased by B7-H4-Ig and 3C8 could reverse the effect of B7-H4-Ig.
  • [1]
    Bretscher PA.A two-step,two-signal model for the primary activation of precursor helper T cells[J].Proc Natl Acad Sci U S A,1999,96(1):185-190.
    [2]
    Schildberg FA,Klein SR,Freeman GJ,et al.Coinhibitory pathways in the B7-CD28 ligand-receptor family[J].Immunity,2016,44(5):955-972.
    [3]
    Swaika A,Hammond WA,Joseph RW.Current state of anti-PD-L1 and anti-PD-1 agents in cancer therapy[J].Mol Immunol,2015,67(2 Pt A):4-17.
    [4]
    Sica GL,Choi IH,Zhu G,et al.B7-H4,a molecule of the B7 family,negatively regulates T cell immunity[J].Immunity,2003,18(6):849-861.
    [5]
    Prasad DV,Richards S,Mai XM,et al.B7S1,a novel B7 family member that negatively regulates T cell activation[J].Immunity,2003,18(6):863-873.
    [6]
    Zang X,Loke P,Kim J,et al.B7x:a widely expressed B7 family member that inhibits T cell activation[J].Proc Natl Acad Sci U S A,2003,100(18):10388-10392.
    [7]
    Awadallah NS,Shroyer KR,Langer DA,et al.Detection of B7-H4 and p53 in pancreatic cancer:potential role as a cytological diagnostic adjunct[J].Pancreas,2008,36(2):200-206.
    [8]
    Miyatake T,Tringler B,Liu W,et al.B7-H4(DD-O110)is overexpressed in high risk uterine endometrioid adenocarcinomas and inversely correlated with tumor T-cell infiltration[J].Gynecol Oncol,2007,106(1):119-127.
    [9]
    Podojil JR,Miller SD.Potential targeting of B7-H4 for the treatment of cancer[J].Immunol Rev,2017,276(1):40-51.
    [10]
    Ni L,Dong C.New B7 family checkpoints in human cancers[J].Mol Cancer Ther,2017,16(7):1203-1211.
    [11]
    MacGregor HL,Ohashi PS.Molecular pathways:evaluating the potential for B7-H4 as an immunoregulatory target[J].Clin Cancer Res,2017,23(12):2934-2941.
    [12]
    Jeon H,Vigdorovich X,Garrett-Thomson SC,et al.Structure and cancer immunotherapy of the B7 family member B7x[J].Cell Rep,2014,9(3):1089-1098.
    [13]
    Dangaj D,Lanitis E,Zhao A,et al.Novel recombinant human b7-h4 antibodies overcome tumoral immune escape to potentiate T-cell antitumor responses[J].Cancer Res,2013,73(15):4820-4829.
    [14]
    Dangaj D,Scholler N.Isolation and validation of Anti-B7-H4 scFvs from an ovarian cancer scFv yeast-display library[J].Methods Mol Biol,2015,1319:37-49.
    [15]
    Leong SR,Liang WC,Wu Y,et al.An anti-B7-H4 antibody-drug conjugate for the treatment of breast cancer[J].Mol Pharm,2015,12(6):1717-1729.
  • Related Articles

    [1]LIU Xiaohan, TAN Yunying, LI Qiang, CHEN Xu, FU Junjie, YIN Jian. Synthesis and antitumor activity evaluation of glycoconjugates derived from natural product harmine[J]. Journal of China Pharmaceutical University, 2023, 54(6): 729-742. DOI: 10.11665/j.issn.1000-5048.2023041101
    [2]SUN Dongyu, GONG Jingxu, LI Xuwen, HAN Guanying, GUO Yuewei. Studies on methyl xestopongoate analogues:design, synthesis and antitumor activities[J]. Journal of China Pharmaceutical University, 2018, 49(4): 413-421. DOI: 10.11665/j.issn.1000-5048.20180405
    [3]QIAO Yixue, MOU Yi, HUANG Zhangjian, AI Yong, KANG Fenghua, LAI Yisheng, ZHANG Yihua. Synthesis and antitumor activities of novel CDDO-Me analogues[J]. Journal of China Pharmaceutical University, 2015, 46(3): 289-293. DOI: 10.11665/j.issn.1000-5048.20150305
    [4]GU Xiaoke, TANG Xiaobo, HUANG Zhangjian, PENG Hui, ZHANG Yihua. Synthesis and antitumor activity of NO-releasing alkoxylbiphenyl derivatives[J]. Journal of China Pharmaceutical University, 2014, 45(6): 657-661. DOI: 10.11665/j.issn.1000-5048.20140606
    [5]HU Kun, ZHOU Anfei, JIANG Hefei, XU Yuanyuan, HUANG Qianhui, YANG Jie, CHEN Xin, REN Jie. Synthesis and antitumor activities of podophyllotoxin derivatives[J]. Journal of China Pharmaceutical University, 2014, 45(1): 33-38. DOI: 10.11665/j.issn.1000-5048.20140105
    [6]LIU Dexiu, PAN Huaying, SHI Hui, HUANG Xiaoying, LOU Hongxiang. Synthesis and antitumor activity of a novel bisbibenzyl derivative[J]. Journal of China Pharmaceutical University, 2013, 44(6): 515-519. DOI: 10.11665/j.issn.1000-5048.20130605
    [7]HU Kun, XU Hua-jin, XIN Wen-qun, CHEN Xin, REN Jie. Synthesis and antitumor activity of nitrogen mustard derivatives of formononetin[J]. Journal of China Pharmaceutical University, 2012, 43(2): 113-119.
    [8]ZHA Xiao-ming, ZHANG Fei-ran, SHAN Jia-qi, CHEN Yan-ke, ZHANG Yi-hua, LIU Jun, SUN Hong-bin. Synthesis and in vitro antitumor activities of novel soladulcidine derivatives[J]. Journal of China Pharmaceutical University, 2010, 41(6): 493-498.
    [9]Synthesis and antitumor activity of ent-kaurene diterpenoids[J]. Journal of China Pharmaceutical University, 2010, 41(1): 20-25.
    [10]ZHOU Mei, MA Lin, HAO Xiao-jiang, YANG Xiao-sheng. Antitumor activities of chemical constituents of Cephalotaxus fortunei in Guizhou province[J]. Journal of China Pharmaceutical University, 2009, 40(3): 209-212.

Catalog

    Article views (636) PDF downloads (1389) Cited by()

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return