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A novel PET imaging using ⁶⁴Cu-labeled monoclonal antibody against mesothelin commonly expressed on cancer cells.

Kobayashi K, Sasaki T, Takenaka F, Yakushiji H, Fujii Y, Kishi Y, Kita S, Shen L, Kumon H, Matsuura E - J Immunol Res (2015)

Bottom Line: Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum.Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN.In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan ; Department of Urology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan.

ABSTRACT
Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum. Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN. In this study, we applied the 11-25 mAb to in vivo imaging to detect MSLN-expressing tumors. In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells. We showed the accumulation of Alexa Fluor 750-labeled 11-25 mAb in MSLN-expressing tumor xenografts in athymic nude mice. Then, 11-25 mAb was labeled with (64)Cu via a chelating agent DOTA and was used in both in vitro cell binding assay and in vivo positron emission tomography (PET) imaging in the tumor-bearing mice. We confirmed that (64)Cu-labeled 11-25 mAb highly accumulated in MSLN-expressing tumors as compared to MSLN-negative ones. The (64)Cu-labeled 11-25 mAb is potentially useful as a PET probe capable of being used for wide range of tumors, rather than (18)F-FDG that occasionally provides nonspecific accumulation into the inflammatory lesions.

No MeSH data available.


Related in: MedlinePlus

Analysis of MSLN protein expression in cancer cell lines by Western blot (a and b) and by flow cytometry using 11-25 mAb (b). β-actin served as a loading control. Expression of MSLN mRNA in cultured BxPC-3 (open circles), CFPAC-1 (closed triangles), PANC-1 (open squares), and A-431 (x-marks) cells (d and e).
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fig1: Analysis of MSLN protein expression in cancer cell lines by Western blot (a and b) and by flow cytometry using 11-25 mAb (b). β-actin served as a loading control. Expression of MSLN mRNA in cultured BxPC-3 (open circles), CFPAC-1 (closed triangles), PANC-1 (open squares), and A-431 (x-marks) cells (d and e).

Mentions: To determine the expression of the MSLN protein on the pancreatic carcinoma cells, Western blot analysis was performed. Figure 1(a) shows that BxPC-3 and CFPAC-1 cells expressed the MSLN protein but PANC-1 and A-431 cells did not. A-431 is human epidermal carcinoma cell line and is known as MSLN-negative [31]. Figure 1(b) shows the expression of MSLN relative to that of β-actin. Figure 1(c) shows the results of flow cytometric analysis of the four cancer cells with anti-MSLN mAb, 11-25. The antibody reacted with 50.9% of BxPC-3 cells, 31.4% of CFPAC-1 cells, 0.6% of PANC-1 cells, and 4.4% of A-431 cells. Next, the expression of MSLN mRNA on the four cell lines was investigated by semi-quantitative PCR (Figures 1(d) and 1(e)). BxPC-3 and CFPAC-1 showed MSLN mRNA expression but PANC-1 and A-431 did not.


A novel PET imaging using ⁶⁴Cu-labeled monoclonal antibody against mesothelin commonly expressed on cancer cells.

Kobayashi K, Sasaki T, Takenaka F, Yakushiji H, Fujii Y, Kishi Y, Kita S, Shen L, Kumon H, Matsuura E - J Immunol Res (2015)

Analysis of MSLN protein expression in cancer cell lines by Western blot (a and b) and by flow cytometry using 11-25 mAb (b). β-actin served as a loading control. Expression of MSLN mRNA in cultured BxPC-3 (open circles), CFPAC-1 (closed triangles), PANC-1 (open squares), and A-431 (x-marks) cells (d and e).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4390102&req=5

fig1: Analysis of MSLN protein expression in cancer cell lines by Western blot (a and b) and by flow cytometry using 11-25 mAb (b). β-actin served as a loading control. Expression of MSLN mRNA in cultured BxPC-3 (open circles), CFPAC-1 (closed triangles), PANC-1 (open squares), and A-431 (x-marks) cells (d and e).
Mentions: To determine the expression of the MSLN protein on the pancreatic carcinoma cells, Western blot analysis was performed. Figure 1(a) shows that BxPC-3 and CFPAC-1 cells expressed the MSLN protein but PANC-1 and A-431 cells did not. A-431 is human epidermal carcinoma cell line and is known as MSLN-negative [31]. Figure 1(b) shows the expression of MSLN relative to that of β-actin. Figure 1(c) shows the results of flow cytometric analysis of the four cancer cells with anti-MSLN mAb, 11-25. The antibody reacted with 50.9% of BxPC-3 cells, 31.4% of CFPAC-1 cells, 0.6% of PANC-1 cells, and 4.4% of A-431 cells. Next, the expression of MSLN mRNA on the four cell lines was investigated by semi-quantitative PCR (Figures 1(d) and 1(e)). BxPC-3 and CFPAC-1 showed MSLN mRNA expression but PANC-1 and A-431 did not.

Bottom Line: Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum.Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN.In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan ; Department of Urology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan.

ABSTRACT
Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum. Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN. In this study, we applied the 11-25 mAb to in vivo imaging to detect MSLN-expressing tumors. In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells. We showed the accumulation of Alexa Fluor 750-labeled 11-25 mAb in MSLN-expressing tumor xenografts in athymic nude mice. Then, 11-25 mAb was labeled with (64)Cu via a chelating agent DOTA and was used in both in vitro cell binding assay and in vivo positron emission tomography (PET) imaging in the tumor-bearing mice. We confirmed that (64)Cu-labeled 11-25 mAb highly accumulated in MSLN-expressing tumors as compared to MSLN-negative ones. The (64)Cu-labeled 11-25 mAb is potentially useful as a PET probe capable of being used for wide range of tumors, rather than (18)F-FDG that occasionally provides nonspecific accumulation into the inflammatory lesions.

No MeSH data available.


Related in: MedlinePlus