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Objective: To investigate the effect of pemphigus vulgaris (PV) serum on expression of Desmoglein (Dsg)1, Dsg3 and matrix metalloproteinase 9 (MMP) 9 in HaCaT cells, and its pathogenic role in acantholysis. Method: This study included four experimental groups, i.e., normal control group, healthy-serum group, PV-serum group, anti-Dsg3 monoclonal antibody group (positive group). Human HaCaT cells were cultured with high-glucose DMEM medium containing 5% PV serum, followed by extraction of RNA and protein for Q-PCR and Western Blot analyses, respectively. In addition, immunostaining was used to assess expression of Dsg1, Dsg3 and MMP-9 in keratinocytes. Results: In addition to elevations in expression levels of respective protein, culture of HaCaT cells with 5% PV-serum increased expression levels of mRNA for Dsg1, Dsg3 and MMP-9 by 432%, 402% and 402%, respectively. Likewise, incubation of HaCaT cells with anti-Dsg3 monoclonal antibody increased expression levels of mRNA for Dsg1, Dsg3 and MMP-9 by 495%, 488% and 604%, respectively (P