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Construction and characterization of an anti-CD20 mAb nanocomb with exceptionally excellent lymphoma-suppressing activity.

Li HF, Wu C, Chen T, Zhang G, Zhao H, Ke CH, Xu Z - Int J Nanomedicine (2015)

Bottom Line: Comparing with free mAbs, PPRT nanocomb possesses a comparable binding ability and reduced "off-rate" to surface CD20 of NHL cells.Pharmacokinetic assays revealed that PPRT nanocomb experienced a relatively reduced clearance from peripheral blood compared with free antibodies.With the cooperation of all the abovementioned superiorities, PPRT nanocomb exhibits exceptionally excellent in vivo antitumor activities in both disseminated and localized human NHL xenotransplant models.

View Article: PubMed Central - PubMed

Affiliation: International Joint Cancer Institute, Translation Medicine Institute, the Second Military Medical University, Shanghai, People's Republic of China ; Planning Division, Scientific Research Department, Eastern Hepatobiliary Surgery Hospital, the Second Military Medical University, Shanghai, People's Republic of China ; Tumor Immunology and Gene Therapy Center, Eastern Hepatobiliary Surgery Hospital, the Second Military Medical University, Shanghai, People's Republic of China.

ABSTRACT
The CD20-directed monoclonal antibody rituximab (RTX) established a new era in the treatment of non-Hodgkin lymphoma (NHL); however, suboptimal response and/or resistance to RTX still limit its clinical merits. Although four effector mechanisms are validated to participate in CD20-based immunotherapy, including complement-dependent cytotoxicity, antibody-dependent cell-mediated cytotoxicity, caspase-dependent apoptosis, and lysosome-mediated programmed cell death (PCD), they could hardly be synchronously activated by any anti-CD20 mAb or mAb derivative until now. Herein, a novel mAb nanocomb (polyethylenimine polymer-RTX-tositumomab [PPRT nanocomb]) was firstly constructed through mass arming two different anti-CD20 mAbs (RTX and tositumomab) to one polymer by nanotechnology. Comparing with free mAbs, PPRT nanocomb possesses a comparable binding ability and reduced "off-rate" to surface CD20 of NHL cells. When treated by PPRT nanocomb, the caspase-dependent apoptosis was remarkably enhanced except for concurrently eliciting complement-dependent cytotoxicity, antibody-dependent cell-mediated cytotoxicity, and lysosome-mediated PCD. Besides, "cross-cell link"-assisted homotypic adhesion by PPRT nanocomb further enhanced the susceptibility to PCD of lymphoma cells. Pharmacokinetic assays revealed that PPRT nanocomb experienced a relatively reduced clearance from peripheral blood compared with free antibodies. With the cooperation of all the abovementioned superiorities, PPRT nanocomb exhibits exceptionally excellent in vivo antitumor activities in both disseminated and localized human NHL xenotransplant models.

No MeSH data available.


Related in: MedlinePlus

Biorecognition of PPRT nanocomb on surface CD20 of Raji cells.Notes: (A) Binding activity of PPRT nanocomb and free mAbs to surface CD20 of Raji cells. Cells incubated with 10 μg/mL RTX-488, Tos-647, or PPRT nanocomb-488/647 were observed with a confocal microscope. Scale bar: 10 μm. (B and C) Dissociation of PPRT nanocomb and parental mAbs from Raji cells. (B) The histogram represents the fluorescence intensity distribution of Raji cells. The black histogram shows phosphate-buffered saline-treated cells. Red and blue histograms show the fluorescence intensity distribution after 0 and 24 hours, respectively. (C) Numerical data representing the percentage of remaining mAbs or PPRT nanocombs on cellular surface after different time intervals. Data are mean ± standard deviation (n=3).Abbreviations: PPRT, polyethylenimine polymer–RTX–Tos; PPRT-488/647, Alexa Fluor-488/647-labeled PPRT; RTX, rituximab; RTX-488, Alexa Fluor-488-labeled RTX; Tos, tositumomab; Tos-647, Alexa Fluor-647-labeled Tos.
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f2-ijn-10-4783: Biorecognition of PPRT nanocomb on surface CD20 of Raji cells.Notes: (A) Binding activity of PPRT nanocomb and free mAbs to surface CD20 of Raji cells. Cells incubated with 10 μg/mL RTX-488, Tos-647, or PPRT nanocomb-488/647 were observed with a confocal microscope. Scale bar: 10 μm. (B and C) Dissociation of PPRT nanocomb and parental mAbs from Raji cells. (B) The histogram represents the fluorescence intensity distribution of Raji cells. The black histogram shows phosphate-buffered saline-treated cells. Red and blue histograms show the fluorescence intensity distribution after 0 and 24 hours, respectively. (C) Numerical data representing the percentage of remaining mAbs or PPRT nanocombs on cellular surface after different time intervals. Data are mean ± standard deviation (n=3).Abbreviations: PPRT, polyethylenimine polymer–RTX–Tos; PPRT-488/647, Alexa Fluor-488/647-labeled PPRT; RTX, rituximab; RTX-488, Alexa Fluor-488-labeled RTX; Tos, tositumomab; Tos-647, Alexa Fluor-647-labeled Tos.

Mentions: Figure 2A demonstrates that the exposure of Raji cells to RTX-488 or Tos-647 led to the decoration of cytomembranes with, respectively, green and red fluorescence, while the exposure to PPRT nanocomb led to both fluorescence decorations. The results suggest that the outstanding biorecognition between nanocomb and CD20 was not affected during the course of PPRT nanocomb preparation.


Construction and characterization of an anti-CD20 mAb nanocomb with exceptionally excellent lymphoma-suppressing activity.

Li HF, Wu C, Chen T, Zhang G, Zhao H, Ke CH, Xu Z - Int J Nanomedicine (2015)

Biorecognition of PPRT nanocomb on surface CD20 of Raji cells.Notes: (A) Binding activity of PPRT nanocomb and free mAbs to surface CD20 of Raji cells. Cells incubated with 10 μg/mL RTX-488, Tos-647, or PPRT nanocomb-488/647 were observed with a confocal microscope. Scale bar: 10 μm. (B and C) Dissociation of PPRT nanocomb and parental mAbs from Raji cells. (B) The histogram represents the fluorescence intensity distribution of Raji cells. The black histogram shows phosphate-buffered saline-treated cells. Red and blue histograms show the fluorescence intensity distribution after 0 and 24 hours, respectively. (C) Numerical data representing the percentage of remaining mAbs or PPRT nanocombs on cellular surface after different time intervals. Data are mean ± standard deviation (n=3).Abbreviations: PPRT, polyethylenimine polymer–RTX–Tos; PPRT-488/647, Alexa Fluor-488/647-labeled PPRT; RTX, rituximab; RTX-488, Alexa Fluor-488-labeled RTX; Tos, tositumomab; Tos-647, Alexa Fluor-647-labeled Tos.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525799&req=5

f2-ijn-10-4783: Biorecognition of PPRT nanocomb on surface CD20 of Raji cells.Notes: (A) Binding activity of PPRT nanocomb and free mAbs to surface CD20 of Raji cells. Cells incubated with 10 μg/mL RTX-488, Tos-647, or PPRT nanocomb-488/647 were observed with a confocal microscope. Scale bar: 10 μm. (B and C) Dissociation of PPRT nanocomb and parental mAbs from Raji cells. (B) The histogram represents the fluorescence intensity distribution of Raji cells. The black histogram shows phosphate-buffered saline-treated cells. Red and blue histograms show the fluorescence intensity distribution after 0 and 24 hours, respectively. (C) Numerical data representing the percentage of remaining mAbs or PPRT nanocombs on cellular surface after different time intervals. Data are mean ± standard deviation (n=3).Abbreviations: PPRT, polyethylenimine polymer–RTX–Tos; PPRT-488/647, Alexa Fluor-488/647-labeled PPRT; RTX, rituximab; RTX-488, Alexa Fluor-488-labeled RTX; Tos, tositumomab; Tos-647, Alexa Fluor-647-labeled Tos.
Mentions: Figure 2A demonstrates that the exposure of Raji cells to RTX-488 or Tos-647 led to the decoration of cytomembranes with, respectively, green and red fluorescence, while the exposure to PPRT nanocomb led to both fluorescence decorations. The results suggest that the outstanding biorecognition between nanocomb and CD20 was not affected during the course of PPRT nanocomb preparation.

Bottom Line: Comparing with free mAbs, PPRT nanocomb possesses a comparable binding ability and reduced "off-rate" to surface CD20 of NHL cells.Pharmacokinetic assays revealed that PPRT nanocomb experienced a relatively reduced clearance from peripheral blood compared with free antibodies.With the cooperation of all the abovementioned superiorities, PPRT nanocomb exhibits exceptionally excellent in vivo antitumor activities in both disseminated and localized human NHL xenotransplant models.

View Article: PubMed Central - PubMed

Affiliation: International Joint Cancer Institute, Translation Medicine Institute, the Second Military Medical University, Shanghai, People's Republic of China ; Planning Division, Scientific Research Department, Eastern Hepatobiliary Surgery Hospital, the Second Military Medical University, Shanghai, People's Republic of China ; Tumor Immunology and Gene Therapy Center, Eastern Hepatobiliary Surgery Hospital, the Second Military Medical University, Shanghai, People's Republic of China.

ABSTRACT
The CD20-directed monoclonal antibody rituximab (RTX) established a new era in the treatment of non-Hodgkin lymphoma (NHL); however, suboptimal response and/or resistance to RTX still limit its clinical merits. Although four effector mechanisms are validated to participate in CD20-based immunotherapy, including complement-dependent cytotoxicity, antibody-dependent cell-mediated cytotoxicity, caspase-dependent apoptosis, and lysosome-mediated programmed cell death (PCD), they could hardly be synchronously activated by any anti-CD20 mAb or mAb derivative until now. Herein, a novel mAb nanocomb (polyethylenimine polymer-RTX-tositumomab [PPRT nanocomb]) was firstly constructed through mass arming two different anti-CD20 mAbs (RTX and tositumomab) to one polymer by nanotechnology. Comparing with free mAbs, PPRT nanocomb possesses a comparable binding ability and reduced "off-rate" to surface CD20 of NHL cells. When treated by PPRT nanocomb, the caspase-dependent apoptosis was remarkably enhanced except for concurrently eliciting complement-dependent cytotoxicity, antibody-dependent cell-mediated cytotoxicity, and lysosome-mediated PCD. Besides, "cross-cell link"-assisted homotypic adhesion by PPRT nanocomb further enhanced the susceptibility to PCD of lymphoma cells. Pharmacokinetic assays revealed that PPRT nanocomb experienced a relatively reduced clearance from peripheral blood compared with free antibodies. With the cooperation of all the abovementioned superiorities, PPRT nanocomb exhibits exceptionally excellent in vivo antitumor activities in both disseminated and localized human NHL xenotransplant models.

No MeSH data available.


Related in: MedlinePlus