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MT95-4, a fully humanized antibody raised against aminopeptidase N, reduces tumor progression in a mouse model.

Akita S, Hattori N, Masuda T, Horimasu Y, Nakashima T, Iwamoto H, Fujitaka K, Miyake M, Kohno N - Cancer Sci. (2015)

Bottom Line: To develop a novel monoclonal antibody-based cancer therapy targeting APN/CD13, we established a fully humanized anti-APN/CD13 monoclonal antibody, MT95-4.We found that expression of human APN/CD13 in murine melanoma cells increased the size of subcutaneous tumors, extent of lung metastasis and degree of angiogenesis in the subcutaneous tumors; these tumor-promoting and angiogenesis-promoting characteristics were reduced by the i.p. administration of MT95-4.These results suggested that the antitumor and anti-angiogenic effects of MT95-4 were dependent on APN/CD13 expression in tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Internal Medicine, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

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Effects of human APN/CD13 expression in murine B16-F1 melanoma cells. (a) Expression levels of human APN/CD13 mRNA in control-B16 and APN-B16 cells were evaluated by quantitative real-time PCR. (b) Expression levels of APN/CD13 on the surface of control-B16 and APN-B16 cells were assessed by flow cytometry. Data (a) represent the mean (± SEM) of triplicate samples. $$$P < 0.001. (c) Comparison of the numbers of lung surface nodules in the tail vein metastasis model between control-B16 and APN-B16 cells. The numbers of lung surface nodules in mice were counted 21 days after injection of control-B16 and APN-B16 cells. Each bar represents the mean number of nodules for eight mice per group. $P < 0.05. (d) Comparison of tumor volumes in a subcutaneous tumor model for tumors derived from control-B16 and APN-B16 cells. The sizes of subcutaneous tumors were measured twice a week for 2 weeks after inoculation of control-B16 and APN-B16 cells. The data represent the mean (± SEM) for eight mice per group. $P < 0.05. (e) Evaluation of angiogenesis in subcutaneous tumors derived from control-B16 and APN-B16 cells. The area containing CD31-positive vessels was measured. Data represent the means (± SEM) from eight mice per group. $$P < 0.01. (f) Immunohistochemical staining for CD31 in a subcutaneous tumor. Scale bar, 100 μm.
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fig02: Effects of human APN/CD13 expression in murine B16-F1 melanoma cells. (a) Expression levels of human APN/CD13 mRNA in control-B16 and APN-B16 cells were evaluated by quantitative real-time PCR. (b) Expression levels of APN/CD13 on the surface of control-B16 and APN-B16 cells were assessed by flow cytometry. Data (a) represent the mean (± SEM) of triplicate samples. $$$P < 0.001. (c) Comparison of the numbers of lung surface nodules in the tail vein metastasis model between control-B16 and APN-B16 cells. The numbers of lung surface nodules in mice were counted 21 days after injection of control-B16 and APN-B16 cells. Each bar represents the mean number of nodules for eight mice per group. $P < 0.05. (d) Comparison of tumor volumes in a subcutaneous tumor model for tumors derived from control-B16 and APN-B16 cells. The sizes of subcutaneous tumors were measured twice a week for 2 weeks after inoculation of control-B16 and APN-B16 cells. The data represent the mean (± SEM) for eight mice per group. $P < 0.05. (e) Evaluation of angiogenesis in subcutaneous tumors derived from control-B16 and APN-B16 cells. The area containing CD31-positive vessels was measured. Data represent the means (± SEM) from eight mice per group. $$P < 0.01. (f) Immunohistochemical staining for CD31 in a subcutaneous tumor. Scale bar, 100 μm.

Mentions: We established two B16-F1-derived cell lines that differed in the expression levels of human APN/CD13 (control-B16 and APN-B16 cells). Quantitative real-time PCR and flow cytometric analysis revealed that APN-B16 cells expressed high levels of human APN/CD13, while control-B16 cells did not (Fig.2a,b). When the in vitro cell proliferation rates of the two cell lines were compared, no differences were observed (data not shown).


MT95-4, a fully humanized antibody raised against aminopeptidase N, reduces tumor progression in a mouse model.

Akita S, Hattori N, Masuda T, Horimasu Y, Nakashima T, Iwamoto H, Fujitaka K, Miyake M, Kohno N - Cancer Sci. (2015)

Effects of human APN/CD13 expression in murine B16-F1 melanoma cells. (a) Expression levels of human APN/CD13 mRNA in control-B16 and APN-B16 cells were evaluated by quantitative real-time PCR. (b) Expression levels of APN/CD13 on the surface of control-B16 and APN-B16 cells were assessed by flow cytometry. Data (a) represent the mean (± SEM) of triplicate samples. $$$P < 0.001. (c) Comparison of the numbers of lung surface nodules in the tail vein metastasis model between control-B16 and APN-B16 cells. The numbers of lung surface nodules in mice were counted 21 days after injection of control-B16 and APN-B16 cells. Each bar represents the mean number of nodules for eight mice per group. $P < 0.05. (d) Comparison of tumor volumes in a subcutaneous tumor model for tumors derived from control-B16 and APN-B16 cells. The sizes of subcutaneous tumors were measured twice a week for 2 weeks after inoculation of control-B16 and APN-B16 cells. The data represent the mean (± SEM) for eight mice per group. $P < 0.05. (e) Evaluation of angiogenesis in subcutaneous tumors derived from control-B16 and APN-B16 cells. The area containing CD31-positive vessels was measured. Data represent the means (± SEM) from eight mice per group. $$P < 0.01. (f) Immunohistochemical staining for CD31 in a subcutaneous tumor. Scale bar, 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig02: Effects of human APN/CD13 expression in murine B16-F1 melanoma cells. (a) Expression levels of human APN/CD13 mRNA in control-B16 and APN-B16 cells were evaluated by quantitative real-time PCR. (b) Expression levels of APN/CD13 on the surface of control-B16 and APN-B16 cells were assessed by flow cytometry. Data (a) represent the mean (± SEM) of triplicate samples. $$$P < 0.001. (c) Comparison of the numbers of lung surface nodules in the tail vein metastasis model between control-B16 and APN-B16 cells. The numbers of lung surface nodules in mice were counted 21 days after injection of control-B16 and APN-B16 cells. Each bar represents the mean number of nodules for eight mice per group. $P < 0.05. (d) Comparison of tumor volumes in a subcutaneous tumor model for tumors derived from control-B16 and APN-B16 cells. The sizes of subcutaneous tumors were measured twice a week for 2 weeks after inoculation of control-B16 and APN-B16 cells. The data represent the mean (± SEM) for eight mice per group. $P < 0.05. (e) Evaluation of angiogenesis in subcutaneous tumors derived from control-B16 and APN-B16 cells. The area containing CD31-positive vessels was measured. Data represent the means (± SEM) from eight mice per group. $$P < 0.01. (f) Immunohistochemical staining for CD31 in a subcutaneous tumor. Scale bar, 100 μm.
Mentions: We established two B16-F1-derived cell lines that differed in the expression levels of human APN/CD13 (control-B16 and APN-B16 cells). Quantitative real-time PCR and flow cytometric analysis revealed that APN-B16 cells expressed high levels of human APN/CD13, while control-B16 cells did not (Fig.2a,b). When the in vitro cell proliferation rates of the two cell lines were compared, no differences were observed (data not shown).

Bottom Line: To develop a novel monoclonal antibody-based cancer therapy targeting APN/CD13, we established a fully humanized anti-APN/CD13 monoclonal antibody, MT95-4.We found that expression of human APN/CD13 in murine melanoma cells increased the size of subcutaneous tumors, extent of lung metastasis and degree of angiogenesis in the subcutaneous tumors; these tumor-promoting and angiogenesis-promoting characteristics were reduced by the i.p. administration of MT95-4.These results suggested that the antitumor and anti-angiogenic effects of MT95-4 were dependent on APN/CD13 expression in tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Internal Medicine, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

Show MeSH
Related in: MedlinePlus