ASSOCIATION BETWEEN PEMPHIGUS VULGARIS AND HUMAN LEUKOCYTE ANTIGEN IN HAN NATION OF NORTHEAST CHINA

Objective To investigate the relationship between pemphigus vulgaris （PV） and human leukocyte antigen （HLA） in Han nation of northeast China. Mothods Standard microcytotoxicity test and polymerase chain reaction-sequence specific primers method were used to detect the HLA class Ⅰ antigens and HLA-DRBI and DQBI alleles in 27 patients with PV and results were compared with control group. Gene and phenotype frequencies of HLA-A3, A26（10）, B60（40）, and B13 （27.99%, 48%; 16.11%, 30%; 23.02%, 41%; 16.11%, 30%, respectively） increased significantly in PV group compared with control （1.01%, 2%; 0.5%, 1%; 4.61%, 9%; 5,13%, 10%, respectively）. After P value correction, the difference of A3, A26 （10）, and B60 （40） between the two groups was still significant. The gene frequencies of HLA-DRB 1^＊ 140x （1401, 1404, 1405, 1407, 1408）, DRBI^＊I20x, and DQBI＊0503 alleles in PV group （42.26%, 25.46%, and 23.02%） were significantly higher than control group （5.09%, 7.74%, and 1.89%）. After P value correction, the difference was still significant between the two groups. Collusion PV significantly relates with HLA in PV patients of Han nation of northeast China.

Effects of chimeric molecule of recombinant Dsg3EC_(1-2) with toxin PE40 on T and B lymphocytes in Pemphigus Vulgaris

Objective:To observe the effects of the recombinant chimeric toxin Dsg3EC 1-2PE40 on T and B lymphocytes isolated from Pemphigus Vulgaris (PV) patients to further study its biological therapeutic function for PV. Methods:Recombinant chimeric toxin Dsg3EC 1-2PE40 was first identified, expressed and purified, and then its effects on T and B lymphocytes of PV patients in vitro were detected and quantified by ELISPOT assay and MTT assay.Results:The purity of the expressed protein Dsg3EC 1-2PE40 was up to 80%. In ELISPOT assay, with Dsg3EC 1-2PE40, the overall number of B cells that produce anti-Dsg3 antibodies among PV patients was only about 60% of the comparable number with Dsg3EC 1-2. The proliferation of T cells of PV patients was inhibited markedly by Dsg3EC 1-2PE40. There was significant difference between the different groups with Dsg3EC 1-2PE40 and Dsg3EC 1-2.Conclusion:The recombinant chimeric toxin Dsg3EC 1-2PE40 decrease the number of B cells that produce anti-Dsg3 antibodies in PV patients and can inhibit or kill T cells of PV patients in vitro.

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.