Molecular cloning and protein expression of EC1-2 and EC3-4 epitopes of pemphigus vulgaris antigen

To clone and express EC1 2 and EC3 4 epitopes of PVA (pemphigus vulgaris antigen, desmoglein 3) in order to diagnose pemphigus and study the relationship between epitopes of PVA and anti PVA antibody Methods RNA was extracted from keratinocytes and the cDNA of epitopes EC1 2 and EC3 4 was synthesized by reverse transcription Amplified genes of EC1 2 and EC3 4 were inserted into the expression plasmid, PGEX 4T 1, and transformed into E coli BL21 by electric transduction Recombinant fusion proteins of EC1 2 and EC3 4 epitopes were expressed by IPTG induction These proteins were separated on SDS PAGE gels and electroblotted to nitrocellulose to detect the anti PVA antibody Results The sequences of cloned EC1 2 and EC3 4 genes were identical to the sequence registered in PC/GENE Expressed recombinant proteins reacted only to sera from patients with pemphigus vulgaris, not to sera from patients with bullous pemphigoid, systemic lupus erythematosus or normal persons Conclusions These recombinant proteins are very specific in antigenicity This may provide a new method for the diagnosis of pemphigus vulgaris (PV) or the differential diagnosis of other bullous cutaneous diseases via patient sera It is also helpful in understanding the relationship between adhesion molecules and the pathogenic mechanism of pemphigus vulgaris

Dapsone-induced infectious mononucleosis-like syndrome in a patient with pemphigus vulgaris

Dapsone syndrome is characterized by high fever, skin rash, methemoglobinemia, liver toxicity, and generalized lymphadenopathy.The incidence of me dapsone syndrome is increasing with the extensive application of dapsone. However it is rarely reported in pemphigus. We present a case of ＂infectious mononucleosis-like＂ dapsone syndrome in a patient with pemphigus vulgaris. The syndrome appeared when he was being treated methylprednisolone, which was commonly used as a treatment for dapsone syndrome.

Effects of Pemphigus Vulgaris Serum on the Expression of ATP2C1 and PKP3 in HaCaT Cells

Objective:To investigate the effects of serum from patients with pemphigus vulgaris(PV)on the transcription and protein expression level of calcium-transporting ATPase type 2C member 1(ATP2C1)and plakophilin 3(PKP3)in HaCaT cells.Methods:The HaCaT cells were divided into four groups:PV sera group,anti-Dsg3 monoclonal antibody group(AK23,positive control group),normal healthy serum group,and blank cell group.The groups were treated with corresponding different conditions for 24 hours.Quantitative polymerase chain reaction and Western blot were used to detect mRNA and protein levels of ATP2C1 and PKP3.Results:Compared with the blank group,the mRNA level of theATP2C1 and PKP3 genes in PV sera group was significantly increased by 384%and 404%,respectively(bothP<0.001).The treatment of PV sera and anti-Dsg3 antibody increased PKP3 protein expression(P=0.03 andP=0.004)but decreased protein expression of ATP2C1 in HaCaT cells(bothP<0.001).Conclusions:Our study indicates that serum from patients with PV promotes bothATP2C1 andPKP3 transcription in HaCaT cells,implying that the two genes may be involved in the pathological process of PV.