Related Experiment Video
Updated: Sep 20, 2026
Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment
Published on: November 23, 2017
Urokinase plasminogen activator mRNA is induced by IL-1alpha and TNF-alpha in in vitro acantholysis
Claudio Feliciani1, Paola Toto, Binghe Wang
1Department of Dermatology, University 'G. d'Annunzio', Chieti, Italy.
Abstract:
The role of urokinase type plasminogen activator (uPA) has been well documented in the pathogenesis of pemphigus vulgaris (PV). Activation of plasminogen into active serine protease plasmin initiates extracellular proteolysis leading to acantholysis but the mechanisms underlying this process are not clearly understood. We have previously shown that keratinocyte derived cytokines IL-1alpha and TNF-alpha are involved in PV-induced acantholysis. In the present study we sought to examine whether keratinocyte-derived IL-1alpha and TNF-alpha are correlated with uPA induction in keratinocytes during acantholysis. Normal human keratinocytes were incubated with diluted PV serum. mRNAs for IL-1alpha, TNF-alpha and uPA were examined with RT-PCR at various time points and acantholysis was measured. IL-1alpha, TNF-alpha and uPA mRNAs were all induced in keratinocytes following PV serum stimulation; IL-1alpha/TNF-alpha mRNAs' expression was earlier than the expression of uPA mRNA. To further examine the role of IL-1alpha, TNF-alpha and uPA in acantholysis, we performed antibody blocking studies. Anti-IL-1alpha, anti-TNF-alpha and anti-uPA antibodies suppressed acantholysis by 76%, 80% and 90%, respectively. In addition, anti-IL-1alpha and anti-TNF-alpha antibodies inhibited uPA mRNA induction, whereas anti-uPA antibodies did not alter IL-1alpha/TNF-alpha mRNAs' expression. Our results confirm the role of uPA in acantholysis and suggest an involvement of IL-1alpha/TNF-alpha in uPA induction.
Insights
Pemphigus vulgaris (PV) involves urokinase plasminogen activator (uPA) in acantholysis. Keratinocyte cytokines, interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha), are linked to uPA induction, suggesting a pathway for PV pathogenesis.
Area of Science:
- Dermatology
- Molecular Biology
- Immunology
Background:
- Pemphigus vulgaris (PV) pathogenesis involves urokinase type plasminogen activator (uPA) and acantholysis.
- Keratinocyte-derived cytokines, IL-1alpha and TNF-alpha, have been implicated in PV-induced acantholysis.
Purpose of the Study:
- To investigate the correlation between keratinocyte-derived IL-1alpha and TNF-alpha and uPA induction during PV-induced acantholysis.
- To elucidate the mechanistic relationship between these cytokines and uPA in PV.
Main Methods:
- Normal human keratinocytes were stimulated with diluted PV serum.
- Messenger RNA (mRNA) expression of IL-1alpha, TNF-alpha, and uPA was analyzed using RT-PCR.
- Antibody blocking studies were performed to assess the functional roles of IL-1alpha, TNF-alpha, and uPA in acantholysis.
Main Results:
- PV serum stimulation induced IL-1alpha, TNF-alpha, and uPA mRNA expression in keratinocytes, with cytokine expression preceding uPA.
- Blocking antibodies against IL-1alpha, TNF-alpha, and uPA significantly suppressed PV-induced acantholysis (76%, 80%, and 90%, respectively).
- Anti-IL-1alpha and anti-TNF-alpha antibodies inhibited uPA mRNA induction, while anti-uPA antibodies did not affect cytokine mRNA levels.
Conclusions:
- uPA plays a confirmed role in PV-induced acantholysis.
- Keratinocyte-derived IL-1alpha and TNF-alpha are involved in the induction of uPA during PV pathogenesis.
- This study suggests a sequential pathway where IL-1alpha and TNF-alpha drive uPA expression, contributing to acantholysis in pemphigus vulgaris.