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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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Function of a fully humanized monoclonal antibody, MT95-4, in vitro. (a) Evaluation of the specificity of MT95-4 by western blotting analysis using cell lysates from HT1080 cells. (b) Effects of MT95-4 on the aminopeptidase activity of HT1080 cells. The levels of AMC released from Ala-MCA were measured. Data represent the mean (± SEM) of triplicate samples. $P < 0.05 versus the control. NS, not significant. (c) Effects of MT95-4 on the invasion of HT1080 cells in a Matrigel invasion assay. Data represent the mean (± SEM) of five wells. $$$P < 0.001 versus the control.
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fig01: Function of a fully humanized monoclonal antibody, MT95-4, in vitro. (a) Evaluation of the specificity of MT95-4 by western blotting analysis using cell lysates from HT1080 cells. (b) Effects of MT95-4 on the aminopeptidase activity of HT1080 cells. The levels of AMC released from Ala-MCA were measured. Data represent the mean (± SEM) of triplicate samples. $P < 0.05 versus the control. NS, not significant. (c) Effects of MT95-4 on the invasion of HT1080 cells in a Matrigel invasion assay. Data represent the mean (± SEM) of five wells. $$$P < 0.001 versus the control.

Mentions: By immunizing KM mice with HT1080 fibrosarcoma cells abundantly expressing APN/CD13, we established a fully humanized monoclonal antibody, MT95-4, which had inhibitory effects against aminopeptidase activity. To verify the specificity of MT95-4, western blotting analysis was performed using HT1080 cell lysates. As shown in Figure1a, MT95-4 detected a band at 150 kDa corresponding to the predicted size of APN/CD13, which coincided with the band from a commercially available anti-APN/CD13 antibody.


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)

Function of a fully humanized monoclonal antibody, MT95-4, in vitro. (a) Evaluation of the specificity of MT95-4 by western blotting analysis using cell lysates from HT1080 cells. (b) Effects of MT95-4 on the aminopeptidase activity of HT1080 cells. The levels of AMC released from Ala-MCA were measured. Data represent the mean (± SEM) of triplicate samples. $P < 0.05 versus the control. NS, not significant. (c) Effects of MT95-4 on the invasion of HT1080 cells in a Matrigel invasion assay. Data represent the mean (± SEM) of five wells. $$$P < 0.001 versus the control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4520645&req=5

fig01: Function of a fully humanized monoclonal antibody, MT95-4, in vitro. (a) Evaluation of the specificity of MT95-4 by western blotting analysis using cell lysates from HT1080 cells. (b) Effects of MT95-4 on the aminopeptidase activity of HT1080 cells. The levels of AMC released from Ala-MCA were measured. Data represent the mean (± SEM) of triplicate samples. $P < 0.05 versus the control. NS, not significant. (c) Effects of MT95-4 on the invasion of HT1080 cells in a Matrigel invasion assay. Data represent the mean (± SEM) of five wells. $$$P < 0.001 versus the control.
Mentions: By immunizing KM mice with HT1080 fibrosarcoma cells abundantly expressing APN/CD13, we established a fully humanized monoclonal antibody, MT95-4, which had inhibitory effects against aminopeptidase activity. To verify the specificity of MT95-4, western blotting analysis was performed using HT1080 cell lysates. As shown in Figure1a, MT95-4 detected a band at 150 kDa corresponding to the predicted size of APN/CD13, which coincided with the band from a commercially available anti-APN/CD13 antibody.

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