The Immunogold-Silver Staining Procedure in the Study of Freshly Suspended Langerhans Cells at the Transmission Electron Microscopic Level

Manara, G. C.; Ferrari, C.; Torresani, Claudio; Panfilis, Giuseppe De

doi:10.1159/000247799

Cited by 4 publications

(4 citation statements)

References 7 publications

(8 reference statements)

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“…4b). The above data indicate that these cells could be of DC origin, as they had many characteristic features often described for DC [21]. Furthermore, our results indicate that MSG biopsies with a lymphocytic focus score > 1·5…”

Section: Morphological Evaluation Of DC By Transmission Electron Micrsupporting

confidence: 73%

“…The expression of the DRC molecule by DC indicated that these cells might belong to the follicular DC lineage and comprise an FDC network in the residing lymphoid follicles. Moreover, the formation of desmosome junctions between DC and lymphocytes is a characteristic feature of follicular dendritic cells (FDC) and emphasizes the FDC origin of these cells [21]. This FDC network could also participate in the ectopic formation of GC.…”

Section: Discussionmentioning

confidence: 99%

“…A pre-embedding immunogold labelling technique was applied as described by Manara et al [21]. Briefly, MSG biopsies were cut in small pieces (< 0·5 mm 3 ) and prefixed in 2% paraformaldehyde in PBS pH 7·4 for 20 min.…”

Section: Methodmentioning

confidence: 99%

“…The expression of the CD11c þ marker that is present on DC, mainly those of the mature phenotype, was examined [21]. Using serial sections it was observed that the CD11c þ molecule was mainly expressed by CD4 þ lymphocytes in the infiltrates of the salivary gland tissue of SS patients ( Fig.…”

Section: Existence and Localization Of Cells With Dendritic Morphologmentioning

confidence: 99%

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CD4 cytotoxic and dendritic cells in the immunopathologic lesion of Sjögren’s syndrome

Xanthou¹,

Tapinos²,

Polihronis³

et al. 1999

Clinical and Experimental Immunology

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The existence of CD4+ T lymphocytes with cytotoxic activity in minor salivary gland (MSG) biopsies from Sjögren's syndrome (SS) patients was investigated using in situ double immunohistochemistry technique. The presence of dendritic cells (DC) in SS lesions was examined by using single and double immunohistochemistry methods and a panel of different MoAbs to specific cell surface markers (i.e. CD3, CD11c, DRC). Furthermore, the ultrastructural morphology of DC was characterized by electron microscopy (EM). Immunogold labelling technique using the DRC surface marker was also applied. Finally, we investigated the existence of germinal centres (GC) in the salivary gland lesions of SS patients. Seven patients with primary SS and five patients with non-specific sialadenitis were the subjects of this study. Our results indicate the existence of a CD4+ cytotoxic cell population that utilizes perforin-mediated cell destructions as they expressed perforin mRNA. Quantitative analysis of these cells revealed that they comprised approximately 20% of the existing T lymphocytes. We also identified a population of CD4+ T cells that expressed the CD11c activation marker. Furthermore, we observed a distinct cell subtype which expressed the DRC cell surface marker. These cells had the characteristic ultrastructural morphology of DC and were DRC+ when examined by immunoelectron microscopy. Finally, the formation of GC structures in the histopathologic lesions of the salivary glands was observed. The above findings indicate that both CD4+ cytotoxic T lymphocytes (CTL) and DC may be involved in the initiation and perpetuation of SS pathogenesis. Moreover, the formation of GC in the lesions reveals a possible mechanism for in situ differentiation and proliferation of activated B lymphocytes.

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Section: Morphological Evaluation Of DC By Transmission Electron Micrsupporting

confidence: 73%

Section: Discussionmentioning

confidence: 99%

Section: Methodmentioning

confidence: 99%

Section: Existence and Localization Of Cells With Dendritic Morphologmentioning

confidence: 99%

See 2 more Smart Citations

CD4 cytotoxic and dendritic cells in the immunopathologic lesion of Sjögren’s syndrome

Xanthou¹,

Tapinos²,

Polihronis³

et al. 1999

Clinical and Experimental Immunology

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Human epidermal Langerhans cells express the ICAM-3 molecule. Immunohistochemical and immunoelectron microscopical demonstration

Manara¹,

Pasquinelli

Giorgi

et al. 1996

British Journal of Dermatology

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Three ligands have been described for the leucocyte integrin LFA-1, namely intercellular adhesion molecule (ICAM)-1, ICAM-2, and ICAM-3. ICAMs show differences in tissue distribution and inducibility. The recently described ICAM-3 is highly expressed on resting lymphocytes, monocytes and neutrophils. Here, we demonstrate that the whole human epidermal Langerhans cell (LC) population expresses this molecule. Immunohistochemical staining of skin sections with an anti-ICAM-3 monoclonal antibody displayed reactivity with dendritic epidermal cells regularly distributed along the epidermis. Highly-sensitive immunoelectron microscopy procedures, performed on freshly suspended epidermal cells both at transmission and scanning electron microscopic levels, enabled demonstration that the whole LC population expresses cell surface ICAM-3. In contrast, keratinocytes and melanocytes were consistently negative. The prominent expression of ICAM-3 by resident LC could imply a crucial part for this molecule in leucocyte-intercellular interactions in the skin.

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CD95-ligand (Fas-L)-expressing epidermal lymphocytes

Panfilis

Marcelli

Pasolini

et al. 2000

British Journal of Dermatology

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The Immunogold-Silver Staining Procedure in the Study of Freshly Suspended Langerhans Cells at the Transmission Electron Microscopic Level

Cited by 4 publications

References 7 publications

CD4 cytotoxic and dendritic cells in the immunopathologic lesion of Sjögren’s syndrome

CD4 cytotoxic and dendritic cells in the immunopathologic lesion of Sjögren’s syndrome

Human epidermal Langerhans cells express the ICAM-3 molecule. Immunohistochemical and immunoelectron microscopical demonstration

CD95-ligand (Fas-L)-expressing epidermal lymphocytes

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