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Characterization of ASKP1240, a fully human antibody targeting human CD40 with potent immunosuppressive effects.

Okimura K, Maeta K, Kobayashi N, Goto M, Kano N, Ishihara T, Ishikawa T, Tsumura H, Ueno A, Miyao Y, Sakuma S, Kinugasa F, Takahashi N, Miura T - Am. J. Transplant. (2014)

Bottom Line: In addition, ASKP1240 did not destabilize platelet thrombi under physiological high shear conditions while mouse anti-human CD154 mAb (mu5C8) did.And ASKP1240 itself did not activate platelet and endothelial cells.The immunosuppressive effect was well correlated with the CD40 receptor saturation.

View Article: PubMed Central - PubMed

Affiliation: Development Research Laboratories, Kyowa Hakko Kirin Co., Ltd., Shizuoka, Japan.

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Related in: MedlinePlus

Modulation of CD40 antigens on B cell lines analyzed by flow cytometry. Ramos cells were incubated with FITC-labeled ASKP1240 (A) or FITC-labeled G28.5 (B) in culture medium at 37°C. At the given time points, cells were washed with PBS containing NaN3 and then labeled with PE-labeled secondary antibodies. The fluorescence of FITC represents simultaneous measurements of cell surface and internalized immunocomplexes, while that of PE represents only cell surface immunocomplexes. The percentage of geoMFI at indicated time point relative to prevalue is plotted over time. FITC, fluorescein isothiocyanate; geoMFI, geometric mean fluorescence intensity; PBS, phosphate-buffered saline; PE, phycoerythrin.
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fig02: Modulation of CD40 antigens on B cell lines analyzed by flow cytometry. Ramos cells were incubated with FITC-labeled ASKP1240 (A) or FITC-labeled G28.5 (B) in culture medium at 37°C. At the given time points, cells were washed with PBS containing NaN3 and then labeled with PE-labeled secondary antibodies. The fluorescence of FITC represents simultaneous measurements of cell surface and internalized immunocomplexes, while that of PE represents only cell surface immunocomplexes. The percentage of geoMFI at indicated time point relative to prevalue is plotted over time. FITC, fluorescein isothiocyanate; geoMFI, geometric mean fluorescence intensity; PBS, phosphate-buffered saline; PE, phycoerythrin.

Mentions: Since CD40 ligation can lead to internalization of cell surface CD40 32, a determination was made as to whether ASKP1240 can elicit internalization of CD40. In this experiment, internalized antigen showed FITC fluorescence only, but residual cell surface antigen showed both PE and FITC fluorescence. Although binding of the anti-CD40 agonistic mAb, G28.5, leads to an immediate decrease in PE fluorescence, ASKP1240 affected neither FITC nor PE fluorescence levels (Figure 2).


Characterization of ASKP1240, a fully human antibody targeting human CD40 with potent immunosuppressive effects.

Okimura K, Maeta K, Kobayashi N, Goto M, Kano N, Ishihara T, Ishikawa T, Tsumura H, Ueno A, Miyao Y, Sakuma S, Kinugasa F, Takahashi N, Miura T - Am. J. Transplant. (2014)

Modulation of CD40 antigens on B cell lines analyzed by flow cytometry. Ramos cells were incubated with FITC-labeled ASKP1240 (A) or FITC-labeled G28.5 (B) in culture medium at 37°C. At the given time points, cells were washed with PBS containing NaN3 and then labeled with PE-labeled secondary antibodies. The fluorescence of FITC represents simultaneous measurements of cell surface and internalized immunocomplexes, while that of PE represents only cell surface immunocomplexes. The percentage of geoMFI at indicated time point relative to prevalue is plotted over time. FITC, fluorescein isothiocyanate; geoMFI, geometric mean fluorescence intensity; PBS, phosphate-buffered saline; PE, phycoerythrin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4225473&req=5

fig02: Modulation of CD40 antigens on B cell lines analyzed by flow cytometry. Ramos cells were incubated with FITC-labeled ASKP1240 (A) or FITC-labeled G28.5 (B) in culture medium at 37°C. At the given time points, cells were washed with PBS containing NaN3 and then labeled with PE-labeled secondary antibodies. The fluorescence of FITC represents simultaneous measurements of cell surface and internalized immunocomplexes, while that of PE represents only cell surface immunocomplexes. The percentage of geoMFI at indicated time point relative to prevalue is plotted over time. FITC, fluorescein isothiocyanate; geoMFI, geometric mean fluorescence intensity; PBS, phosphate-buffered saline; PE, phycoerythrin.
Mentions: Since CD40 ligation can lead to internalization of cell surface CD40 32, a determination was made as to whether ASKP1240 can elicit internalization of CD40. In this experiment, internalized antigen showed FITC fluorescence only, but residual cell surface antigen showed both PE and FITC fluorescence. Although binding of the anti-CD40 agonistic mAb, G28.5, leads to an immediate decrease in PE fluorescence, ASKP1240 affected neither FITC nor PE fluorescence levels (Figure 2).

Bottom Line: In addition, ASKP1240 did not destabilize platelet thrombi under physiological high shear conditions while mouse anti-human CD154 mAb (mu5C8) did.And ASKP1240 itself did not activate platelet and endothelial cells.The immunosuppressive effect was well correlated with the CD40 receptor saturation.

View Article: PubMed Central - PubMed

Affiliation: Development Research Laboratories, Kyowa Hakko Kirin Co., Ltd., Shizuoka, Japan.

Show MeSH
Related in: MedlinePlus