In fluorescence diagnosis with 5-aminolevulinic acid (ALA)-induced porphyrins (FDAP), protoporphyrin IX (PpIX)-accumulation can be macroscopically visualized. Interpretation of these data is still problematic because ...In fluorescence diagnosis with 5-aminolevulinic acid (ALA)-induced porphyrins (FDAP), protoporphyrin IX (PpIX)-accumulation can be macroscopically visualized. Interpretation of these data is still problematic because of the low reproducibility of the procedure and poor understanding of the mechanisms involved in PpIX tumor selectivity. In this study, PpIX accumulation is investigated in patients with psoriasis and actinic keratosis (AK) following FDAP. For this purpose, desquamated lesional and non-lesional skin were incubated with 20% ALA ointment for 3 h, FDAP was performed, and highly fluorescing lesional skin and non-lesional skin were biopsied. In extracts from these biopsies, PpIX, protein, and dsDNA were quantified by spectrofluorometry. Digital images acquired with FDAP were analyzed using image analysis software. PpIX per biopsy in lesional skin in both psoriasis and AK was significantly higher than in non-lesional skin (p < 0.05). When corrected for epidermal involvement, only lesional psoriatic skin showed significantly higher PpIX levels than non-lesional skin. The PpIX-ratio lesional:non-lesional skin (mean(pmol per mL) ± SEM) was 4.12 ± 0.91 in psoriasis and 1.96 ± 0.24 in AK. In FDAP, the ratio of lesional: non-lesional skin was 1.77 ± 0.06 in psoriasis and 1.37 ± 0.07 in AK. Macroscopic fluorescence and PpIX content appeared to be well correlated (r = 0.73), thus making FDAP a good predictor of PpIX content.展开更多
Background: Depletion of CD4+CD25+Foxp3+naturally occurring regulatory T cells (Treg) induces autoimmune phenomena. These cells have not yet been fully characterized in the skin of psoriatic patients. Objectives: To p...Background: Depletion of CD4+CD25+Foxp3+naturally occurring regulatory T cells (Treg) induces autoimmune phenomena. These cells have not yet been fully characterized in the skin of psoriatic patients. Objectives: To prove that the Zenon immunofluorescent labeling technique is suitable for the demonstration of co-localization of T-cell markers and in particular to show the distribution of Treg in psoriatic skin. Methods: In biopsies derived from normal and psoriatic skin, CD4+CD25+, CD4+CD45RO+, CD8+CD25+, CD8+CD45RO+and CD4+CD25+Foxp3+cells in the dermis and in the epidermis were immunophenotyped, using a quantitative immunofluorescent labeling technique (Zenon), analyzed and compared using image analysis. Results: The immuno-fluorescent labeling technique was shown to be an easy and reliable tool to demonstrate co-localization of T-cell markers.In psoriasis, all pathogenic T-cell subsets (CD4+CD25+,CD4+CD45RO+, CD8+CD25+and CD8+CD45RO+cells) were significantly increased in the dermis and in the epidermis, as compared to normal skin (all p < 0.05). Using this labeling technique we were able to reveal CD4+CD25+Foxp3+Treg in psoriatic dermis, but not in the dermis of normal skin (p < 0.0001). Conclusions: The Zenon immunofluorescence technique in combination with image analysis is suitable for the demonstration of co-localization of T-cell markers in tissue. Increased numbers of pathogenic T cells (CD4+CD25+,CD4+CD45RO+, CD8+CD25+and CD8+CD45RO+) were shown in the dermis and epidermis, whereas CD4+CD25+Foxp3+Treg were identified in psoriatic skin with a predilection for the upper dermis.展开更多
文摘In fluorescence diagnosis with 5-aminolevulinic acid (ALA)-induced porphyrins (FDAP), protoporphyrin IX (PpIX)-accumulation can be macroscopically visualized. Interpretation of these data is still problematic because of the low reproducibility of the procedure and poor understanding of the mechanisms involved in PpIX tumor selectivity. In this study, PpIX accumulation is investigated in patients with psoriasis and actinic keratosis (AK) following FDAP. For this purpose, desquamated lesional and non-lesional skin were incubated with 20% ALA ointment for 3 h, FDAP was performed, and highly fluorescing lesional skin and non-lesional skin were biopsied. In extracts from these biopsies, PpIX, protein, and dsDNA were quantified by spectrofluorometry. Digital images acquired with FDAP were analyzed using image analysis software. PpIX per biopsy in lesional skin in both psoriasis and AK was significantly higher than in non-lesional skin (p < 0.05). When corrected for epidermal involvement, only lesional psoriatic skin showed significantly higher PpIX levels than non-lesional skin. The PpIX-ratio lesional:non-lesional skin (mean(pmol per mL) ± SEM) was 4.12 ± 0.91 in psoriasis and 1.96 ± 0.24 in AK. In FDAP, the ratio of lesional: non-lesional skin was 1.77 ± 0.06 in psoriasis and 1.37 ± 0.07 in AK. Macroscopic fluorescence and PpIX content appeared to be well correlated (r = 0.73), thus making FDAP a good predictor of PpIX content.
文摘Background: Depletion of CD4+CD25+Foxp3+naturally occurring regulatory T cells (Treg) induces autoimmune phenomena. These cells have not yet been fully characterized in the skin of psoriatic patients. Objectives: To prove that the Zenon immunofluorescent labeling technique is suitable for the demonstration of co-localization of T-cell markers and in particular to show the distribution of Treg in psoriatic skin. Methods: In biopsies derived from normal and psoriatic skin, CD4+CD25+, CD4+CD45RO+, CD8+CD25+, CD8+CD45RO+and CD4+CD25+Foxp3+cells in the dermis and in the epidermis were immunophenotyped, using a quantitative immunofluorescent labeling technique (Zenon), analyzed and compared using image analysis. Results: The immuno-fluorescent labeling technique was shown to be an easy and reliable tool to demonstrate co-localization of T-cell markers.In psoriasis, all pathogenic T-cell subsets (CD4+CD25+,CD4+CD45RO+, CD8+CD25+and CD8+CD45RO+cells) were significantly increased in the dermis and in the epidermis, as compared to normal skin (all p < 0.05). Using this labeling technique we were able to reveal CD4+CD25+Foxp3+Treg in psoriatic dermis, but not in the dermis of normal skin (p < 0.0001). Conclusions: The Zenon immunofluorescence technique in combination with image analysis is suitable for the demonstration of co-localization of T-cell markers in tissue. Increased numbers of pathogenic T cells (CD4+CD25+,CD4+CD45RO+, CD8+CD25+and CD8+CD45RO+) were shown in the dermis and epidermis, whereas CD4+CD25+Foxp3+Treg were identified in psoriatic skin with a predilection for the upper dermis.
