<b>Background:</b> Among matrix metalloproteinases (MMPs), MMP-2 and MMP-9 are considered to play important roles in the tissue metabolism. <b>Objective:</b> To study the production of MMP-2 and MMP-9 in cultured keratinocytes. <b>Methods:</b> We examined the effect of cell passage number and cell differentiation on these enzymes by zymography and immunocytochemistry. <b>Results:</b> Both MMP-2 and MMP-9 gelatinases were detected in keratinocyte-conditioned media using zymography. The level of MMP-2 in the medium decreased after sequential passage, whereas MMP-9 was detected at a constant level. Treatment of keratinocytes with Ca<sup>2+</sup>, a potent stimulator for cell differentiation, induced secretion of MMP-9, which was confirmed immunocytochemically. Inversely, treatment with retinoic acid, which inhibits cell differentiation, increased the level of MMP-2. <b>Conclusion:</b> These results suggest that keratinocytes regulate their secretion of MMP-2 and MMP-9 gelatinase in distinct and independent patterns during differentiation.
beta 1H globulin is a plasma protein which regulates the biologic activities of the major fragment of the 3rd complement component, C3b. The role of beta 1H globulin in pemphigus was investigated using immunofluorescence in the present study. Lesional skin biopsies from patients with confirmed pemphigus demonstrated in-vivo deposition of beta 1H in addition to C3 in all of four biopsies. Eight serum samples containing C3 fixing intercellular antibodies were then tested for the capacity to fix beta 1H and other complement components. All eight pemphigus sera showed fixation of beta 1H to the intercellular areas of normal human skin. C1q and C4 fixation by pemphigus sera was also demonstrated in 7 of 8 sera, respectively. The experiment using C2-deficient serum indicated that the fixation of beta 1H by intercellular antibodies requires the activation of the classical complement pathway. These data suggested that beta 1H, a co-factor of C3b inactivator, plays a role in the in-vivo regulation of complement activity and supplies additional evidence for the participation of complement system in the pathogenesis of pemphigus.
The fungal elements observed by direct microscopic examination sometimes fail to grow on subsequent culture. To elucidate the cause of this discrepancy, the viability of dermatophytes and Candida in skin scales was evaluated by neutral red staining. Autoradiographic study using 3H-thymidine confirmed that grain-positive cells (viable cells) were stained with neutral red, whereas negative cells (non-viable cells) were not stained. Taking this as a baseline, the correlation between neutral red-positive fungal elements in scales taken from 211 patients with tinea and 27 patients with cutaneous candidosis and cultures on Sabouraud glucose agar was studied. Strong positive correlations were found in both mycoses. These findings suggest that neutral red staining provides a useful method for evaluating the viability of dermatophytes and Candida in human skin scales.
To determine the cell surface autoimmune target of herpetiform pemphigus (HP).
Design
Serum samples of HP were examined by immunoblot studies with human epidermal extracts, enzyme-linked immunosorbent assay with baculovirus-expressed recombinant desmoglein (rDsg) 1 and rDsg3, and immunoadsorption assay with rDsg.
Patients
Twenty serum samples were obtained from patients with HP who have typical clinical and histological features. All serum samples showed positive staining against keratinocyte cell surfaces by indirect immunofluorescence studies with healthy human skin.
Results
Immunoblot results showed that of 17 HP serum samples, only 5 reacted with a 160-kd band and 1 reacted with a 130-kd band. Results of enzyme-linked immunosorbent assays with rDsg1 and rDsg3 demonstrated that of 20 HP serum samples, 16 were positive against Dsg1 and 4 were positive against Dsg3. No serum samples reacted with both. Furthermore, in 19 of 20 HP serum samples, immunoreactivity against keratinocyte cell surfaces was completely removed by preincubation with rDsg1 and rDsg3 as shown by indirect immunofluorescence, excluding a possibility that these HP sera contain autoantibodies against other cell surface molecules.
Conclusions
Dsg1 and Dsg3 are the major cell surface target molecules of HP, suggesting that most cases of HP are clinical variants of pemphigus foliaceus and that the rest might be variants of pemphigus vulgaris.
Polymorphisms of the 5′‐flanking promoter/enhancer region of the TNFA gene were determined in 80 Japanese patients with pulmoplantar pustulosis (PPP). The 5′‐flanking region of the TNFA gene from –1107 to –66 was amplified by polymerase chain reaction (PCR) method. Nucleotide sequencing data from the PCR products revealed that 5 single nucleotide polymorphisms at position –1031, –863, –857, –307 and –237. None of the nucleotide substitutions were significantly increased in PPP patients when compared with those in controls. To clarify the linkage among the neighboring genetic marker, we analyzed the association between the polymorphisms in the TNFA promoter region and the Nco I polymorphism in the first intron of the TNFB gene as well as HLA‐DR9 . The genotype at –1031C is strongly associated with TNFB1 and negatively associated with TNFB2 which is reported to be associated with PPP. These data indicate that TNFA gene centromeric to TNFB is not associated with PPP and the susceptible gene of PPP is located between TNFB and HLA‐B .
Journal Article Comparative study of autoantigens for various bullous skin diseases by immunoblotting using different dermo‐epidermal separation techniques Get access Y. OHATA, Y. OHATA Department of Dermatology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, 160 Tokyo, Japan Search for other works by this author on: Oxford Academic Google Scholar T. HASHIMOTO, T. HASHIMOTO Department of Dermatology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, 160 Tokyo, Japan Search for other works by this author on: Oxford Academic Google Scholar T. NISHIKAWA T. NISHIKAWA Department of Dermatology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, 160 Tokyo, Japan Search for other works by this author on: Oxford Academic Google Scholar Clinical and Experimental Dermatology, Volume 20, Issue 6, 1 November 1995, Pages 454–458, https://doi.org/10.1111/j.1365-2230.1995.tb01376.x Published: 01 November 1995 Article history Accepted: 21 August 1995 Published: 01 November 1995
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