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1.
Biotechnol Lett ; 34(7): 1183-91, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22421972

ABSTRACT

The use of a bi-specific antibody (BsAb) is an attractive and specific approach to cancer therapy. We have constructed a fully human recombinant single chain Fv BsAb against CD19 and CD3 that was an effective treatment in an animal model of non-Hodgkin's lymphoma (NHL). The CD19/CD3 BsAb was expressed in CHO cells and purified by Ni-column chromatography. Flow cytometry revealed that the CD19/CD3 BsAb specifically bound to both CD19 and CD3-positive cells. In vitro, the CD19/CD3 BsAb could stimulate T cell proliferation and induce the lysis of cultured Raji cells in the presence of unstimulated T lymphocytes. In vivo, the CD19/CD3 BsAb efficiently inhibited tumour growth in SCID mice of NHL, and the survival time of the mice was significantly prolonged. Therefore, our CD19/CD3 BsAb is a useful tool that could be a suitable candidate for treatment of NHL.


Subject(s)
Antibodies, Bispecific/immunology , Antigens, CD19/immunology , CD3 Complex/immunology , Lymphoma, Non-Hodgkin/immunology , T-Lymphocytes, Cytotoxic/immunology , Animals , Antibodies, Bispecific/genetics , Antibodies, Bispecific/therapeutic use , CHO Cells , Cell Proliferation , Chromatography, Affinity , Cricetinae , Disease Models, Animal , Humans , Immunotherapy/methods , Mice , Mice, SCID , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/therapeutic use , Treatment Outcome
2.
Cancer Immunol Immunother ; 61(10): 1735-43, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22406982

ABSTRACT

The CD40 receptor is a member of the tumour necrosis factor receptor family and is widely expressed on various cell types. The antitumour activity of CD40 agonist antibody has been observed in B-cell-derived malignancies, but its activity on ovarian cancer remains unclear. However, in this paper, we first confirmed that the anti-CD40 agonist antibody could inhibit the growth of ovarian cancer cells and induce apoptosis. This study investigated the expression of CD40 by ovarian carcinoma tissues and cell lines, at the same time, we evaluated the effect of a recombinant soluble human CD40L (rshCD40L) and an anti-CD40 agonist antibody on cell growth and apoptosis. Flow cytometry and immunohistochemistry assay demonstrated that CD40 was expressed on ovarian carcinoma cell lines and primary ovarian carcinoma cells derived from ascites, as well as on ovarian carcinoma tissues. The growth inhibition of rshCD40L and the anti-CD40 agonist antibody on ovarian carcinoma cells was examined by MTT assay, and the proportion of apoptotic tumour cells was analysed by flow cytometry and Hoechst staining. Our study showed that CD40 was expressed on all ovarian carcinoma cell lines and was examined in 86.2% (162/188) of ovarian cancer tissue samples, but not in normal ovarian tissues (n = 20). Treatment with rshCD40L or anti-CD40 agonist antibody significantly inhibited ovarian carcinoma cell growth and induced apoptosis. Theses results suggest that CD40 is expressed on ovarian carcinoma cells, moreover, that rshCD40L and anti-CD40 agonist antibody have therapeutic potential to inhibit human ovarian cancer growth.


Subject(s)
Antibodies, Monoclonal, Humanized/therapeutic use , CD40 Antigens/agonists , CD40 Antigens/biosynthesis , Carcinoma/drug therapy , Ovarian Neoplasms/drug therapy , Adult , Aged , Antibodies, Monoclonal, Humanized/immunology , Apoptosis/drug effects , Apoptosis/immunology , CD40 Antigens/immunology , Carcinoma/immunology , Cell Line, Tumor , Female , Humans , Middle Aged , Ovarian Neoplasms/immunology , Young Adult
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