Antibody epitope peptides as potential inducers of IgG antibodies against CD98 oncoprotein

Itoh, Ken; Ohshima, Motohiro; Sonobe, Momoyo; Saito, Misa; Yoshida, Akira; Hayashi, Hideki; Inoue, Ken‐ichiro; Masuko, Takashi

doi:10.1111/j.1349-7006.2008.00998.x

Cited by 10 publications

(14 citation statements)

References 28 publications

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“…Reactivity of GV5 with ME180 or HSC-3 seemed relatively heterogeneous; however, tumor formation or tumor growth was almost completely inhibited by GV5, suggesting that CSCs are mainly composed of GV5-reactive CD44R1 high cells but not of GV5-unreactive CD44R1 low cells. In this context, our recent data have provided evidence that the expression of CD44R1 and its association with xCT cysteine-glutamate antiporter, a light chain subunit of CD98 oncoprotein [10], [23], [30]–[32], [42], [43], block the reactive oxygen species (ROS)-induced stress signaling that results in growth arrest, cell differentiation and senescence, and thereby promote the proliferation of cancer cells and the formation of lethal gastrointestinal tumors [44]. Given that CD44R1-expressing CSCs play a central role in resistance to cancer therapy, it should be postulated that therapeutic mAb could target the CD44R1 high cell population in cancer.…”

Section: Resultsmentioning

confidence: 99%

Anti-Tumor Effect against Human Cancer Xenografts by a Fully Human Monoclonal Antibody to a Variant 8-Epitope of CD44R1 Expressed on Cancer Stem Cells

et al. 2012

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BackgroundCD44 is a major cellular receptor for hyaluronic acids. The stem structure of CD44 encoded by ten normal exons can be enlarged by ten variant exons (v1-v10) by alternative splicing. We have succeeded in preparing MV5 fully human IgM and its class-switched GV5 IgG monoclonal antibody (mAb) recognizing the extracellular domain of a CD44R1 isoform that contains the inserted region coded by variant (v8, v9 and v10) exons and is expressed on the surface of various human epithelial cancer cells.Methods and Principal FindingsWe demonstrated the growth inhibition of human cancer xenografts by a GV5 IgG mAb reshaped from an MV5 IgM. The epitope recognized by MV5 and GV5 was identified to a v8-coding region by the analysis of mAb binding to various recombinant CD44 proteins by enzyme-linked immunosorbent assay. GV5 showed preferential reactivity against various malignant human cells versus normal human cells assessed by flow cytometry and immunohistological analysis. When ME180 human uterine cervix carcinoma cells were subcutaneously inoculated to athymic mice with GV5, significant inhibition of tumor formation was observed. Furthermore, intraperitoneal injections of GV5markedly inhibited the growth of visible established tumors from HSC-3 human larynx carcinoma cells that had been subcutaneously transplanted one week before the first treatment with GV5. From in vitro experiments, antibody-dependent cellular cytotoxicity and internalization of CD44R1 seemed to be possible mechanisms for in vivo anti-tumor activity by GV5.ConclusionsCD44R1 is an excellent molecular target for mAb therapy of cancer, possibly superior to molecules targeted by existing therapeutic mAb, such as Trastuzumab and Cetuximab recognizing human epidermal growth factor receptor family.

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Section: Resultsmentioning

confidence: 99%

Anti-Tumor Effect against Human Cancer Xenografts by a Fully Human Monoclonal Antibody to a Variant 8-Epitope of CD44R1 Expressed on Cancer Stem Cells

et al. 2012

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“…6a). ( 72 ) Interestingly, the nearest VH and VL germ lines of these clones were identical to those of HBJ127 (Fig. 6b); however, amino acid sequences of VH and VL varied in these clones and in HBJ127, particularly in the framework regions (Fig.…”

Section: Acquisition Of Mab With Superior Quality To Original Mab By mentioning

confidence: 86%

Towards therapeutic antibodies to membrane oncoproteins by a robust strategy using rats immunized with transfectants expressing target molecules fused to green fluorescent protein

et al. 2010

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Cell‐surface molecules containing growth factor receptors, adhesion molecules and transporter proteins are often over‐expressed in various cancer cells, and could be regarded as suitable targets for therapeutic monoclonal antibodies (mAb). Anti‐cancer therapeutic mAb are claimed to bind these cell‐surface molecules on viable cancer cells: therefore, it is necessary to produce mAb recognizing epitopes on the extracellular domains of native but not denatured proteins. We have experienced difficulty in obtaining mAb bound to viable cancer cells using synthetic peptides or recombinant proteins produced in bacteria as immunogens, although these immunogens are relatively easy to prepare. In this context, we have concluded that viable cancer cells or cells transfected with cDNA encoding target proteins are suitable immunogens for the production of anti‐cancer therapeutic mAb. Furthermore, we selected rats as the immunized animals, because of their excellent capacity to generate diverse antibodies. Because many target candidates are multi‐pass (type IV) membrane proteins, such as 7‐pass G protein‐coupled receptors and 12‐pass transporter proteins belonging to the solute carrier family, and their possible immunogenic extracellular regions are very small, production of specific mAb was extremely difficult. In this review, we summarize the successful preparation and characterization of rat mAb immunized against the extracellular domain of type I, type II and type IV membrane oncoproteins fused to green fluorescent protein as an approach using reverse genetics, and also introduce the discovery of cell‐death‐inducing antibodies as an approach using forward genetics and a strategy to produce reshaped antibodies using mimotope peptides as the immunogen. (Cancer Sci 2011; 102: 25–35)

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“…Each well was added with 100 µl TMB/H 2 O 2 (Boster, China) and incubated at 37°C in dark for 30 minutes, and then 100 µl 2 M H 2 SO 4 was added into wells as a stop solution. The optical density (OD) was read at 450 nm, and all of the tests were conducted in triplicate [25], [33].…”

Section: Methodsmentioning

confidence: 99%

Novel Antibody against a Glutamic Acid-Rich Human Fibrinogen-Like Protein 2-Derived Peptide near Ser91 Inhibits hfgl2 Prothrombinase Activity

Wang

Long

et al. 2014

PLoS ONE

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Fibrinogen-like protein 2 (fgl2) is highly expressed in microvascular endothelial cells in diseases associated with microcirculatory disturbances and plays a crucial role in microthrombosis. Previous studies have demonstrated that the Ser89 residue is a critical site for mouse fgl2 prothrombinase activity. The aim of this study was to investigate the prothrombinase inhibitory ability of antibodies against an hfgl2-derived peptide. The peptide was termed NPG-12 because it is located at the N-terminus of membrane-bound hfgl2, contains 12 amino acid residues (corresponding to residues 76 to 87), and is rich in Glu. This peptide was selected as an antigenic determinant to produce antibodies in immunized rabbits using the DNAStar and HomoloGene software program. Abundant hfgl2 expression was induced in human umbilical vein endothelial cells through treatment with TNF-α. The generated anti-NPG-12 antibodies specifically recognize fgl2, as determined by ELISA, Western Blot and immunostaining. Moreover, one-stage clotting and thrombin generation tests provide evidence that the antibodies can reduce the hfgl2 prothrombinase activity without affecting the platelet-poor plasma prothrombin time (PT) or the activated partial thromboplastin time (APTT). In addition, the antibodies exerted undetectable influence on the proliferation or activation of bulk T cell populations. In conclusion, the selected peptide sequence NPG-12 may be a critical domain for hfgl2 prothrombinase activity, and the development of inhibitors against this sequence may be promising for research or management of hfgl2-associated microcirculatory disturbances.

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Antibody epitope peptides as potential inducers of IgG antibodies against CD98 oncoprotein

Cited by 10 publications

References 28 publications

Anti-Tumor Effect against Human Cancer Xenografts by a Fully Human Monoclonal Antibody to a Variant 8-Epitope of CD44R1 Expressed on Cancer Stem Cells

Anti-Tumor Effect against Human Cancer Xenografts by a Fully Human Monoclonal Antibody to a Variant 8-Epitope of CD44R1 Expressed on Cancer Stem Cells

Towards therapeutic antibodies to membrane oncoproteins by a robust strategy using rats immunized with transfectants expressing target molecules fused to green fluorescent protein

Novel Antibody against a Glutamic Acid-Rich Human Fibrinogen-Like Protein 2-Derived Peptide near Ser91 Inhibits hfgl2 Prothrombinase Activity

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