Combination of fixation using PLP fixative and embedding in paraffin by the AMeX method is useful for histochemical studies in assessment of immunotoxicity.

Suzuki, Masami; Katsuyama, Kiyoka; Adachi, Kenji; Ogawa, Yumie; Yorozu, Keigo; Fujii, Etsuko; Misawa, Yasuyuki; Sugimoto, Tetsuro

doi:10.2131/jts.27.165

Cited by 47 publications

(41 citation statements)

References 12 publications

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“…As for the GPC3 function relevant to macrophages, we have reported that macrophage infiltration is stimulated by GPC3 membranous expression in human clinical HCC as well removed and fixed and embedded in paraffin using the PLP-AMeX method. [26][27][28][29] Histopathological evaluation. Thin sections from the paraffin specimens were prepared and stained with HE to evaluate morphological changes.…”

Section: Methodsmentioning

confidence: 99%

Histopathological analyses of the antitumor activity of anti-glypican-3 antibody (GC33) in human liver cancer xenograft models: The essential role of macrophages

Takai¹,

Kato²,

Kinoshita³

et al. 2009

Cancer Biology & Therapy

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Previously, we demonstrated the antitumor efficacy of the antiglypican-3 (GPC3) antibody GC33 in several human liver cancer xenograft models and the important role of antibody-dependent cellular cytotoxicity (ADCC) in the antitumor mechanism of GC33. Involvement of other mechanisms such as modulation of the functions of GPC3 in antitumor activity remains to be elucidated. In this study, we investigated histopathologically time-course changes in xenografts in mice following a single administration of GC33 to clarify the morphological changes contributing to the tumor growth inhibition of GC33, including the changes in GPC3-related factors/components [proliferation, extracelluar matrices (ECMs) and macrophage]. Histopathological changes peaked 3-5 d after GC33 administration and included increased tumor cell death, tumor cells with round morphology, multinucleated tumor cells and small spindle/round-like cells (mostly F4/80-positive macrophages). No direct effects of GC33 on proliferation activity of tumor cells were observed. Meanwhile, alteration of ECM structures and a remarkable increase in macrophages was noted in the GC33-treated group. Increase in macrophages was observed mainly in the outer layer of tumor nodules; the area of the increase approximately included the area where the change in tumor cells and ECMs were observed. Interestingly, depletion of macrophages in the xenograft models resulted in a marked reduction of the antitumor activity of GC33. In the in vitro ADCC assay, ADCC was only slightly induced by mouse peritoneal macrophages. These data suggest that macrophages play an important role in the antitumor activity of GC33, which is not likely to be direct ADCC by macrophages themselves.

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Section: Methodsmentioning

confidence: 99%

Histopathological analyses of the antitumor activity of anti-glypican-3 antibody (GC33) in human liver cancer xenograft models: The essential role of macrophages

Takai¹,

Kato²,

Kinoshita³

et al. 2009

Cancer Biology & Therapy

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“…• C, processed and embedded in paraffin by the AMeX (PLPAMeX) method (38,39), sectioned at 3 microns, and stained with hematoxylin and eosin (HE), Azan or by an enzymehistochemical method.…”

Section: Tissue Preparationmentioning

confidence: 99%

Effects of Granulocyte Colony-Stimulating Factor (G-CSF) on Bleomycin-Induced Lung Injury of Varying Severity

Adachi¹,

Suzuki²,

Sugimoto³

et al. 2003

Toxicol Pathol

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We evaluated the effects of granulocyte colony-stimulating factor (G-CSF) on bleomycin (BLM)-induced lung injury that developed diffuse alveolar damage and subsequent pulmonary fibrosis (PF) of varying severity. G-CSF (100 µg/kg/day) was administered subcutaneously to BLM (0.2, 20, 2,000 µg)-treated or -untreated rats for 3 or 14 days. In the BLM 0.2 µg group, slight alveolar mononuclear cell infiltration was observed, although PF was not noted. In the BLM 20-µg and 2,000-µg groups, diffuse alveolar damage along with neutrophil infiltration and subsequent PF were observed. In the saline + G-CSF group and BLM 0.2 µg + G-CSF group, a marked increase in the number of alkaline phosphatase (ALP)-positive neutrophils was noted in the alveolar capillaries, although there was neither neutrophil infiltration in alveoli nor exacerbation of lung injury. In the BLM 20 µg + G-CSF and BLM 2,000 µg + G-CSF groups, an exacerbation of lung injury along with an increase in the number of ALP-positive neutrophils in the alveoli was observed. These results indicate that the administration of G-CSF to rats with slight lung injury bearing no PF does not exacerbate the lung injury. The exacerbating effects of G-CSF seem to be associated not only with the marked infiltration of activated neutrophils but also with the severity of underlying lung injury.

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“…Paraffin cross sections of the organotypic cocultures were prepared by the PLP-AMeX method. 19) Briefly, the gels were washed three times with phosphate-buffered saline (PBS) and immersed in a PLP fixative for 6 h at 4 C. They were then washed with PBS, dehydrated, and embedded in paraffin. The cross sections (4 mm thick) were deparaffinized with xylene and ethanol, and then stained with hematoxylin and eosin.…”

Section: Methodsmentioning

confidence: 99%

Mouse Epidermal Keratinocytes in Three-Dimensional Organotypic Coculture with Dermal Fibroblasts Form a Stratified Sheet Resembling Skin

Ikuta¹,

Sekino²,

Hara³

et al. 2006

Bioscience, Biotechnology, and Biochemistry

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We describe an organotypic model of mouse skin consisting of a stratified sheet of epidermal keratinocytes and dermal fibroblasts within a contracted collagen gel. The model was designed to maintain the polarity of stratified keratinocytes and permit their long-term culture at an air-liquid interface. After air exposure, the thickness of the keratinocyte sheet transiently increased and then decreased to two cell layers at 2 weeks. The two-cell-layer structure is similar to that of the adult mouse epidermis. Cytokeratin 5 was localized in the lowest cell layer in the epithelial sheet, but cytokeratin 1 and loricrin were localized in the outer cell layers, resembling mouse skin. The expressions of interleukin 1 and 1 in the keratinocytes and of keratinocyte growth factor 1 and 2 in the fibroblasts correlated with keratinocyte stratification. The mouse organotypic coculture is useful in studying epithelial cell-mesenchymal cell interactions in vitro.

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Combination of fixation using PLP fixative and embedding in paraffin by the AMeX method is useful for histochemical studies in assessment of immunotoxicity.

Cited by 47 publications

References 12 publications

Histopathological analyses of the antitumor activity of anti-glypican-3 antibody (GC33) in human liver cancer xenograft models: The essential role of macrophages

Histopathological analyses of the antitumor activity of anti-glypican-3 antibody (GC33) in human liver cancer xenograft models: The essential role of macrophages

Effects of Granulocyte Colony-Stimulating Factor (G-CSF) on Bleomycin-Induced Lung Injury of Varying Severity

Mouse Epidermal Keratinocytes in Three-Dimensional Organotypic Coculture with Dermal Fibroblasts Form a Stratified Sheet Resembling Skin

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