Limits...
Preparation and characterization of anti-tissue factor single-chain variable fragment antibody for cancer diagnosis.

Sato R, Obonai T, Tsumura R, Tsumoto K, Koga Y, Yasunaga M, Matsumura Y - Cancer Sci. (2014)

Bottom Line: Tissue factor (TF), which serves as the initiator of the extrinsic blood coagulation cascade, has been found to be overexpressed in various solid tumors, especially brain tumors, pancreatic cancer, and gastric cancer.The data obtained showed that the affinity of the anti-TF scFv was 2.04 × 10(-8) (KD), and that the protein showed significant binding to the cancer cells.This study indicates anti-TF scFv may be suitable as an imaging probe for the diagnosis of solid tumors.

View Article: PubMed Central - PubMed

Affiliation: Division of Developmental Therapeutics, Research Center for Innovative Oncology, National Cancer Center Hospital East, Chiba, Japan; Laboratory of Cancer Biology, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Chiba, Japan.

Show MeSH

Related in: MedlinePlus

In vivo imaging of anti-mTF scFv and IgG. (a) Massive spontaneous cutaneous tumors were developed on mice back. These tumors showed partially ulcerative and hemorrhagic change. (b) HE staining for the tumor cell clusters and tumor stroma (left). These tumor had abundant tumor stroma. The immunohistochemistry with anti-mTF mAb showed remarkable mTF expression in the tumor region (right). Yellow arrow denotes tumor cells clusters. Black arrow denotes cancer stroma. Scale bar: 200 μm. (c) HE staining for the invasion site of the squamous cancer cell clusters. Arrows denote tumor cells invasion site into stroma (left). The immunohistochemistry with anti-mTF mAb showed remarkable mTF expression in the cancer invasion region (right). Arrows indicated tumor cells invasion site into stroma. Scale bar: 200 μm. (d) In vivo imaging for accumulation of control scFv, anti-mTF scFv and anti-mTF IgG. The tumors are indicated by yellow arrows, respectively. (e) Fluorescence intensities in the tumor for control scFv (blue), anti-mTF scFv (red) and anti-mTF IgG (green) were measured (n = 3 tumor areas per time-point). Selective accumulation of anti-mTF scFv was seen at 1 h after the injection, and eliminated from the body by 12 h after the injection. On the other hand, selective accumulation of anti-mTF IgG in the tumor was observed for over 72 h. (f) Comparison of the tumor to background ratio (TBR) between anti-mTF scFv and IgG (n = 3 tumor areas per time-point). The TBR of anti-mTF scFv was maximal at 3 h after the injection; on the other hand, that of anti-TF IgG was at 24 h after the injection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4317963&req=5

fig04: In vivo imaging of anti-mTF scFv and IgG. (a) Massive spontaneous cutaneous tumors were developed on mice back. These tumors showed partially ulcerative and hemorrhagic change. (b) HE staining for the tumor cell clusters and tumor stroma (left). These tumor had abundant tumor stroma. The immunohistochemistry with anti-mTF mAb showed remarkable mTF expression in the tumor region (right). Yellow arrow denotes tumor cells clusters. Black arrow denotes cancer stroma. Scale bar: 200 μm. (c) HE staining for the invasion site of the squamous cancer cell clusters. Arrows denote tumor cells invasion site into stroma (left). The immunohistochemistry with anti-mTF mAb showed remarkable mTF expression in the cancer invasion region (right). Arrows indicated tumor cells invasion site into stroma. Scale bar: 200 μm. (d) In vivo imaging for accumulation of control scFv, anti-mTF scFv and anti-mTF IgG. The tumors are indicated by yellow arrows, respectively. (e) Fluorescence intensities in the tumor for control scFv (blue), anti-mTF scFv (red) and anti-mTF IgG (green) were measured (n = 3 tumor areas per time-point). Selective accumulation of anti-mTF scFv was seen at 1 h after the injection, and eliminated from the body by 12 h after the injection. On the other hand, selective accumulation of anti-mTF IgG in the tumor was observed for over 72 h. (f) Comparison of the tumor to background ratio (TBR) between anti-mTF scFv and IgG (n = 3 tumor areas per time-point). The TBR of anti-mTF scFv was maximal at 3 h after the injection; on the other hand, that of anti-TF IgG was at 24 h after the injection.

Mentions: Initially we tried to use mouse pancreatic tumor cell LTPA and TF overexpressing LTPA to examine the distribution of the anti-TF scFv and IgG. These murine cells, however, could not grow in the body of any types of mice including nude, Scid, NOD-Scid, and NOG mice. These anti-mTF mAbs did not cross-react with the TF on human pancreatic cancer cells, BxPC3 (Fig. S1). Instead, mice bearing chemically induced spontaneous cutaneous tumors were selected for evaluation of the distribution of the anti-mTF IgG and scFv, because the spontaneous tumors contained abundant stroma, similar to the case of human cancer. These tumors showed ulcerative and hemorrhagic change in gross appearance (Fig. 4a). HE staining showed that these tumors had abundant tumor stroma (Fig. 4b, left). Moreover, the immunohistochemistry with anti-mTF mAb indicated remarkable mTF expression in the cancer region, especially at the invasion site (Fig. 4b,c, right). Systemic in vivo imaging revealed that while control scFv did not accumulate in the tumors, anti-mTF scFv showed selective accumulation in the tumor from 1 h after the injection, and was eliminated from the body by 12 h after the injection (Fig. 4d,e). On the other hand, anti-mTF IgG showed selective accumulation in the tumor for over 72 h (Fig. 4d,e). The TBR of anti-mTF scFv was maximal at 3 h after the injection. On the other hand, that of anti-TF IgG was maximal at 24 h after the injection (Fig. 4f).


Preparation and characterization of anti-tissue factor single-chain variable fragment antibody for cancer diagnosis.

Sato R, Obonai T, Tsumura R, Tsumoto K, Koga Y, Yasunaga M, Matsumura Y - Cancer Sci. (2014)

In vivo imaging of anti-mTF scFv and IgG. (a) Massive spontaneous cutaneous tumors were developed on mice back. These tumors showed partially ulcerative and hemorrhagic change. (b) HE staining for the tumor cell clusters and tumor stroma (left). These tumor had abundant tumor stroma. The immunohistochemistry with anti-mTF mAb showed remarkable mTF expression in the tumor region (right). Yellow arrow denotes tumor cells clusters. Black arrow denotes cancer stroma. Scale bar: 200 μm. (c) HE staining for the invasion site of the squamous cancer cell clusters. Arrows denote tumor cells invasion site into stroma (left). The immunohistochemistry with anti-mTF mAb showed remarkable mTF expression in the cancer invasion region (right). Arrows indicated tumor cells invasion site into stroma. Scale bar: 200 μm. (d) In vivo imaging for accumulation of control scFv, anti-mTF scFv and anti-mTF IgG. The tumors are indicated by yellow arrows, respectively. (e) Fluorescence intensities in the tumor for control scFv (blue), anti-mTF scFv (red) and anti-mTF IgG (green) were measured (n = 3 tumor areas per time-point). Selective accumulation of anti-mTF scFv was seen at 1 h after the injection, and eliminated from the body by 12 h after the injection. On the other hand, selective accumulation of anti-mTF IgG in the tumor was observed for over 72 h. (f) Comparison of the tumor to background ratio (TBR) between anti-mTF scFv and IgG (n = 3 tumor areas per time-point). The TBR of anti-mTF scFv was maximal at 3 h after the injection; on the other hand, that of anti-TF IgG was at 24 h after the injection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317963&req=5

fig04: In vivo imaging of anti-mTF scFv and IgG. (a) Massive spontaneous cutaneous tumors were developed on mice back. These tumors showed partially ulcerative and hemorrhagic change. (b) HE staining for the tumor cell clusters and tumor stroma (left). These tumor had abundant tumor stroma. The immunohistochemistry with anti-mTF mAb showed remarkable mTF expression in the tumor region (right). Yellow arrow denotes tumor cells clusters. Black arrow denotes cancer stroma. Scale bar: 200 μm. (c) HE staining for the invasion site of the squamous cancer cell clusters. Arrows denote tumor cells invasion site into stroma (left). The immunohistochemistry with anti-mTF mAb showed remarkable mTF expression in the cancer invasion region (right). Arrows indicated tumor cells invasion site into stroma. Scale bar: 200 μm. (d) In vivo imaging for accumulation of control scFv, anti-mTF scFv and anti-mTF IgG. The tumors are indicated by yellow arrows, respectively. (e) Fluorescence intensities in the tumor for control scFv (blue), anti-mTF scFv (red) and anti-mTF IgG (green) were measured (n = 3 tumor areas per time-point). Selective accumulation of anti-mTF scFv was seen at 1 h after the injection, and eliminated from the body by 12 h after the injection. On the other hand, selective accumulation of anti-mTF IgG in the tumor was observed for over 72 h. (f) Comparison of the tumor to background ratio (TBR) between anti-mTF scFv and IgG (n = 3 tumor areas per time-point). The TBR of anti-mTF scFv was maximal at 3 h after the injection; on the other hand, that of anti-TF IgG was at 24 h after the injection.
Mentions: Initially we tried to use mouse pancreatic tumor cell LTPA and TF overexpressing LTPA to examine the distribution of the anti-TF scFv and IgG. These murine cells, however, could not grow in the body of any types of mice including nude, Scid, NOD-Scid, and NOG mice. These anti-mTF mAbs did not cross-react with the TF on human pancreatic cancer cells, BxPC3 (Fig. S1). Instead, mice bearing chemically induced spontaneous cutaneous tumors were selected for evaluation of the distribution of the anti-mTF IgG and scFv, because the spontaneous tumors contained abundant stroma, similar to the case of human cancer. These tumors showed ulcerative and hemorrhagic change in gross appearance (Fig. 4a). HE staining showed that these tumors had abundant tumor stroma (Fig. 4b, left). Moreover, the immunohistochemistry with anti-mTF mAb indicated remarkable mTF expression in the cancer region, especially at the invasion site (Fig. 4b,c, right). Systemic in vivo imaging revealed that while control scFv did not accumulate in the tumors, anti-mTF scFv showed selective accumulation in the tumor from 1 h after the injection, and was eliminated from the body by 12 h after the injection (Fig. 4d,e). On the other hand, anti-mTF IgG showed selective accumulation in the tumor for over 72 h (Fig. 4d,e). The TBR of anti-mTF scFv was maximal at 3 h after the injection. On the other hand, that of anti-TF IgG was maximal at 24 h after the injection (Fig. 4f).

Bottom Line: Tissue factor (TF), which serves as the initiator of the extrinsic blood coagulation cascade, has been found to be overexpressed in various solid tumors, especially brain tumors, pancreatic cancer, and gastric cancer.The data obtained showed that the affinity of the anti-TF scFv was 2.04 × 10(-8) (KD), and that the protein showed significant binding to the cancer cells.This study indicates anti-TF scFv may be suitable as an imaging probe for the diagnosis of solid tumors.

View Article: PubMed Central - PubMed

Affiliation: Division of Developmental Therapeutics, Research Center for Innovative Oncology, National Cancer Center Hospital East, Chiba, Japan; Laboratory of Cancer Biology, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Chiba, Japan.

Show MeSH
Related in: MedlinePlus