The Experts below are selected from a list of 429 Experts worldwide ranked by ideXlab platform

Masato Nakamura - One of the best experts on this subject based on the ideXlab platform.

  • A Versatile Technique for the In Vivo Imaging of Human Tumor Xenografts Using Near-Infrared Fluorochrome- Conjugated Macromolecule Probes
    2016
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR

  • a versatile technique for the in vivo imaging of human tumor xenografts using near infrared fluorochrome Conjugated Macromolecule probes
    PLOS ONE, 2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR-Conjugated anti-HLA probe but not by the orange-red fluorescent signal derived from the tdTomato reporter. This result demonstrated the superiority of the NIR probes over the tdTomato reporter protein at enhancing tissue penetration. In another xenograft model, patient-derived xenografts (PDX) of LC11-JCK (human non-small cell lung cancer) were successfully visualized using the NIR-Conjugated Macromolecule probe without any genetic modification. These results suggested that NIR-Conjugated Macromolecule, preferably, anti-HLA antibody probe is a valuable tool for the detection of human tumors in experimental metastasis models using whole-body imaging.

  • Validation of in vivo imaging of human tumors with the NIR-Conjugated Macromolecule probes.
    2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    (A) Bright-field images and fluorescence images of the T-HCT 116 cells (which express tdTomato) and HCT 116 cells in vitro. Fluorescent signal from the orange-red fluorescent protein tdTomato and the NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively. The absence or presence of the NIR-αHLA antibody is indicated as NIR-αHLA (–) or (+), respectively. (B) In vivo fluorescence images of T-HCT 116 and HCT 116 tumor-bearing BRG nude mice. The NIR fluorescence intensity 2 days after iv injection of the NIR-αHLA probe can be observed. Fluorescent signal from tdTomato and NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively, using the Kodak In-Vivo Imaging System FX. The absence or presence of the NIR-αHLA probe is indicated as NIR-αHLA (–) or (+), respectively. The red and yellow arrows indicate engraftment sites of T-HCT 116 cells and HCT 116 cells, respectively. (C) Fluorescent signal of the NIR-Conjugated Macromolecule probes co-localized with tdTomato in T-HCT 116 cells in tumor-bearing BRG mice. The fluorescent signals at 535/600 nm and 745/800 nm were overlaid (composite) using Living Image software 4.1.3. Li; liver, Sp; spleen. (D) Immunohistochemical staining of dissected tumors; anti-RFP (RFP; left), anti-mouse IgG2a (MIgG2a; center), and anti-CD31 (CD31; right); Enlarged view of boxed area shown below. Arrowheads indicate same position the on serial section. Scale bar, 200 µm.

Hiroshi Suemizu - One of the best experts on this subject based on the ideXlab platform.

  • A Versatile Technique for the In Vivo Imaging of Human Tumor Xenografts Using Near-Infrared Fluorochrome- Conjugated Macromolecule Probes
    2016
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR

  • a versatile technique for the in vivo imaging of human tumor xenografts using near infrared fluorochrome Conjugated Macromolecule probes
    PLOS ONE, 2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR-Conjugated anti-HLA probe but not by the orange-red fluorescent signal derived from the tdTomato reporter. This result demonstrated the superiority of the NIR probes over the tdTomato reporter protein at enhancing tissue penetration. In another xenograft model, patient-derived xenografts (PDX) of LC11-JCK (human non-small cell lung cancer) were successfully visualized using the NIR-Conjugated Macromolecule probe without any genetic modification. These results suggested that NIR-Conjugated Macromolecule, preferably, anti-HLA antibody probe is a valuable tool for the detection of human tumors in experimental metastasis models using whole-body imaging.

  • Validation of in vivo imaging of human tumors with the NIR-Conjugated Macromolecule probes.
    2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    (A) Bright-field images and fluorescence images of the T-HCT 116 cells (which express tdTomato) and HCT 116 cells in vitro. Fluorescent signal from the orange-red fluorescent protein tdTomato and the NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively. The absence or presence of the NIR-αHLA antibody is indicated as NIR-αHLA (–) or (+), respectively. (B) In vivo fluorescence images of T-HCT 116 and HCT 116 tumor-bearing BRG nude mice. The NIR fluorescence intensity 2 days after iv injection of the NIR-αHLA probe can be observed. Fluorescent signal from tdTomato and NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively, using the Kodak In-Vivo Imaging System FX. The absence or presence of the NIR-αHLA probe is indicated as NIR-αHLA (–) or (+), respectively. The red and yellow arrows indicate engraftment sites of T-HCT 116 cells and HCT 116 cells, respectively. (C) Fluorescent signal of the NIR-Conjugated Macromolecule probes co-localized with tdTomato in T-HCT 116 cells in tumor-bearing BRG mice. The fluorescent signals at 535/600 nm and 745/800 nm were overlaid (composite) using Living Image software 4.1.3. Li; liver, Sp; spleen. (D) Immunohistochemical staining of dissected tumors; anti-RFP (RFP; left), anti-mouse IgG2a (MIgG2a; center), and anti-CD31 (CD31; right); Enlarged view of boxed area shown below. Arrowheads indicate same position the on serial section. Scale bar, 200 µm.

Yuichiro Higuchi - One of the best experts on this subject based on the ideXlab platform.

  • A Versatile Technique for the In Vivo Imaging of Human Tumor Xenografts Using Near-Infrared Fluorochrome- Conjugated Macromolecule Probes
    2016
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR

  • a versatile technique for the in vivo imaging of human tumor xenografts using near infrared fluorochrome Conjugated Macromolecule probes
    PLOS ONE, 2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR-Conjugated anti-HLA probe but not by the orange-red fluorescent signal derived from the tdTomato reporter. This result demonstrated the superiority of the NIR probes over the tdTomato reporter protein at enhancing tissue penetration. In another xenograft model, patient-derived xenografts (PDX) of LC11-JCK (human non-small cell lung cancer) were successfully visualized using the NIR-Conjugated Macromolecule probe without any genetic modification. These results suggested that NIR-Conjugated Macromolecule, preferably, anti-HLA antibody probe is a valuable tool for the detection of human tumors in experimental metastasis models using whole-body imaging.

  • Validation of in vivo imaging of human tumors with the NIR-Conjugated Macromolecule probes.
    2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    (A) Bright-field images and fluorescence images of the T-HCT 116 cells (which express tdTomato) and HCT 116 cells in vitro. Fluorescent signal from the orange-red fluorescent protein tdTomato and the NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively. The absence or presence of the NIR-αHLA antibody is indicated as NIR-αHLA (–) or (+), respectively. (B) In vivo fluorescence images of T-HCT 116 and HCT 116 tumor-bearing BRG nude mice. The NIR fluorescence intensity 2 days after iv injection of the NIR-αHLA probe can be observed. Fluorescent signal from tdTomato and NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively, using the Kodak In-Vivo Imaging System FX. The absence or presence of the NIR-αHLA probe is indicated as NIR-αHLA (–) or (+), respectively. The red and yellow arrows indicate engraftment sites of T-HCT 116 cells and HCT 116 cells, respectively. (C) Fluorescent signal of the NIR-Conjugated Macromolecule probes co-localized with tdTomato in T-HCT 116 cells in tumor-bearing BRG mice. The fluorescent signals at 535/600 nm and 745/800 nm were overlaid (composite) using Living Image software 4.1.3. Li; liver, Sp; spleen. (D) Immunohistochemical staining of dissected tumors; anti-RFP (RFP; left), anti-mouse IgG2a (MIgG2a; center), and anti-CD31 (CD31; right); Enlarged view of boxed area shown below. Arrowheads indicate same position the on serial section. Scale bar, 200 µm.

Haruo Hashimoto - One of the best experts on this subject based on the ideXlab platform.

  • A Versatile Technique for the In Vivo Imaging of Human Tumor Xenografts Using Near-Infrared Fluorochrome- Conjugated Macromolecule Probes
    2016
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR

  • a versatile technique for the in vivo imaging of human tumor xenografts using near infrared fluorochrome Conjugated Macromolecule probes
    PLOS ONE, 2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR-Conjugated anti-HLA probe but not by the orange-red fluorescent signal derived from the tdTomato reporter. This result demonstrated the superiority of the NIR probes over the tdTomato reporter protein at enhancing tissue penetration. In another xenograft model, patient-derived xenografts (PDX) of LC11-JCK (human non-small cell lung cancer) were successfully visualized using the NIR-Conjugated Macromolecule probe without any genetic modification. These results suggested that NIR-Conjugated Macromolecule, preferably, anti-HLA antibody probe is a valuable tool for the detection of human tumors in experimental metastasis models using whole-body imaging.

  • Validation of in vivo imaging of human tumors with the NIR-Conjugated Macromolecule probes.
    2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    (A) Bright-field images and fluorescence images of the T-HCT 116 cells (which express tdTomato) and HCT 116 cells in vitro. Fluorescent signal from the orange-red fluorescent protein tdTomato and the NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively. The absence or presence of the NIR-αHLA antibody is indicated as NIR-αHLA (–) or (+), respectively. (B) In vivo fluorescence images of T-HCT 116 and HCT 116 tumor-bearing BRG nude mice. The NIR fluorescence intensity 2 days after iv injection of the NIR-αHLA probe can be observed. Fluorescent signal from tdTomato and NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively, using the Kodak In-Vivo Imaging System FX. The absence or presence of the NIR-αHLA probe is indicated as NIR-αHLA (–) or (+), respectively. The red and yellow arrows indicate engraftment sites of T-HCT 116 cells and HCT 116 cells, respectively. (C) Fluorescent signal of the NIR-Conjugated Macromolecule probes co-localized with tdTomato in T-HCT 116 cells in tumor-bearing BRG mice. The fluorescent signals at 535/600 nm and 745/800 nm were overlaid (composite) using Living Image software 4.1.3. Li; liver, Sp; spleen. (D) Immunohistochemical staining of dissected tumors; anti-RFP (RFP; left), anti-mouse IgG2a (MIgG2a; center), and anti-CD31 (CD31; right); Enlarged view of boxed area shown below. Arrowheads indicate same position the on serial section. Scale bar, 200 µm.

Tomoyuki Ogura - One of the best experts on this subject based on the ideXlab platform.

  • A Versatile Technique for the In Vivo Imaging of Human Tumor Xenografts Using Near-Infrared Fluorochrome- Conjugated Macromolecule Probes
    2016
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR

  • a versatile technique for the in vivo imaging of human tumor xenografts using near infrared fluorochrome Conjugated Macromolecule probes
    PLOS ONE, 2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    Here, we present a versatile method for detecting human tumor xenografts in vivo, based on the enhanced permeability and retention (EPR) effect, using near-infrared (NIR) fluorochrome-Conjugated Macromolecule probes. Bovine serum albumin (BSA) and two immunoglobulins—an anti-human leukocyte antigen (HLA) monoclonal antibody and isotype control IgG2a—were labeled with XenoLight CF770 fluorochrome and used as NIR-Conjugated Macromolecule probes to study whole-body imaging in a variety of xenotransplantation mouse models. NIR fluorescent signals were observed in subcutaneously transplanted BxPC-3 (human pancreatic cancer) cells and HCT 116 (colorectal cancer) cells within 24 h of NIR-Macromolecule probe injection, but the signal from the fluorochrome itself or from the NIR-Conjugated small molecule (glycine) injection was not observed. The accuracy of tumor targeting was confirmed by the localization of the NIR-Conjugated immunoglobulin within the T-HCT 116 xenograft (in which the orange-red fluorescent protein tdTomato was stably expressed by HCT 116 cells) in the subcutaneous transplantation model. However, there was no significant difference in the NIR signal intensity of the region of interest between the anti-HLA antibody group and the isotype control group in the subcutaneous transplantation model. Therefore, the antibody accumulation within the tumor in vivo is based on the EPR effect. The liver metastasis generated by an intrasplenic injection of T-HCT 116 cells was clearly visualized by the NIR-Conjugated anti-HLA probe but not by the orange-red fluorescent signal derived from the tdTomato reporter. This result demonstrated the superiority of the NIR probes over the tdTomato reporter protein at enhancing tissue penetration. In another xenograft model, patient-derived xenografts (PDX) of LC11-JCK (human non-small cell lung cancer) were successfully visualized using the NIR-Conjugated Macromolecule probe without any genetic modification. These results suggested that NIR-Conjugated Macromolecule, preferably, anti-HLA antibody probe is a valuable tool for the detection of human tumors in experimental metastasis models using whole-body imaging.

  • Validation of in vivo imaging of human tumors with the NIR-Conjugated Macromolecule probes.
    2013
    Co-Authors: Hiroshi Suemizu, Kenji Kawai, Yuichiro Higuchi, Haruo Hashimoto, Tomoyuki Ogura, Toshio Itoh, Erika Sasaki, Masato Nakamura
    Abstract:

    (A) Bright-field images and fluorescence images of the T-HCT 116 cells (which express tdTomato) and HCT 116 cells in vitro. Fluorescent signal from the orange-red fluorescent protein tdTomato and the NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively. The absence or presence of the NIR-αHLA antibody is indicated as NIR-αHLA (–) or (+), respectively. (B) In vivo fluorescence images of T-HCT 116 and HCT 116 tumor-bearing BRG nude mice. The NIR fluorescence intensity 2 days after iv injection of the NIR-αHLA probe can be observed. Fluorescent signal from tdTomato and NIR-αHLA probe were specifically detected at wavelengths of 535/600 nm and 720/790 nm, respectively, using the Kodak In-Vivo Imaging System FX. The absence or presence of the NIR-αHLA probe is indicated as NIR-αHLA (–) or (+), respectively. The red and yellow arrows indicate engraftment sites of T-HCT 116 cells and HCT 116 cells, respectively. (C) Fluorescent signal of the NIR-Conjugated Macromolecule probes co-localized with tdTomato in T-HCT 116 cells in tumor-bearing BRG mice. The fluorescent signals at 535/600 nm and 745/800 nm were overlaid (composite) using Living Image software 4.1.3. Li; liver, Sp; spleen. (D) Immunohistochemical staining of dissected tumors; anti-RFP (RFP; left), anti-mouse IgG2a (MIgG2a; center), and anti-CD31 (CD31; right); Enlarged view of boxed area shown below. Arrowheads indicate same position the on serial section. Scale bar, 200 µm.