SummaryThe aim of this study was to quantify and evaluate the forkhead box P3 (FoxP3) expression regulatory T cells in new-onset systemic lupus erythematosus (SLE) patients before and after treatment. Forty-four newly diagnosed and untreated SLE patients, including 24 with active disease (SLEDAI Ն 10) and 20 with inactive disease (SLEDAI < 5), were enrolled in this study. Twenty-one age-and sex-matched healthy volunteers were also included as controls. Peripheral blood samples were collected and mononuclear cells isolated. The expression of CD25 and FoxP3 in CD4 + T cells were analysed with flow cytometry. CD4+ CD25 + (3·95-13·04%) and CD4 + CD25 high (0·04-1·34%) T cells in peripheral blood in untreated patients with new-onset active lupus were significantly lower than that in the patients with inactive lupus (7·27-24·48%, P < 0·05 and 0·14-3·07% P < 0·01 respectively) and that in healthy controls (5·84-14·84%, P < 0·05). Interestingly, the decrease in CD4 + CD25 high T cells was restored significantly in patients with active lupus after corticosteroid treatment. There was, however, a significantly higher percentage of CD4 + FoxP3 + T cells in patients with active (5·30-23·00%) and inactive (7·46-17·38%) new-onset lupus patients compared with healthy control subjects (2·51-12·94%) (P < 0·01). Intriguingly, CD25 expression in CD4 + FoxP3 + T cells in patients with active lupus (25·24-62·47%) was significantly lower than that in those patients with inactive lupus (30·35-75·25%, P < 0·05) and healthy controls (54·83-86·38%, P < 0·01). Most strikingly, the levels of FoxP3 expression determined by mean fluorescence intensity in CD4 + CD25 high cells in patients with active SLE were significantly downregulated compared with healthy subjects (130 Ϯ 22 versus 162 Ϯ 21, P = 0·012

BCSC-1 is dramatically upregulated in CNE-2L2 human nasopharyngeal carcinoma cells with reduced malignancy (AS cells) and isW e previously observed that α-mannosidase Man2c1 suppression causes profound inhibition of malignant activities of the human nasopharyngeal carcinoma-derived cell line CNE-2L2.(1) In order to explore possible mechanisms of malignancy inhibition, mRNA differential display analysis was carried out on CNE-2L2 cells with Man2c1 suppression (called AS cells) and CNE-2L2 cells transfected with mock (called M cells). Of the 1069 genes examined, 28 were upregulated and 31 were downregulated in AS cells. LOH11CR2A, also called BCSC-1 (GenBank no. NM_014622), was identified as the most dramatically upregulated gene. BCSC-1 cDNA was originally cloned and proposed to be a candidate tumor suppressor gene (TSG) by Martin et al.(2) Therefore, association of BCSC-1 with malignant behaviors of CNE-2L2 cells was examined in this study.It was observed that growth and tumorigenesis of wild-type CNE-2L2 cells (W cells) were inhibited by ectopic BCSC-1 whereas those of AS cells were promoted by BCSC-1 suppression, indicating that BCSC-1 is a TSG in CNE-2L2 cells. The tumor suppressor function was further confirmed by a study showing that intratumor BCSC-1 injection results in growth inhibition of tumors from W cells inoculated in nude mice. BCSC-1 expression was examined by means of immunohistochemistry in human nasopharyngeal carcinomas. Because only some carcinoma specimens showed marked reduction of BCSC-1 expression and BCSC-1 expression was poor in CNE-2L2 cells, the HNE-1 nasopharyngeal carcinoma cell line with rich BCSC-1 expression was used as a control in the present study. The data obtained suggest that BCSC-1 suppression might play roles in tumorigenesis of some nasopharyngeal carcinomas and BCSC-1 might be a potential target in gene therapy of those nasopharyngeal carcinomas with poor BCSC-1 expression.TSG have been reported to exhibit multiple inhibitive effects on tumorigenesis.(3) Tumorigenesis, including that of nasopharyngeal carcinoma, (4) is a very complicated process and is regulated by a number of factors.(5,6) Therefore, it could be postulated that the suppressive effect of BCSC-1 on malignant behaviors of CNE-2L2 cells would be at multiple points. Because reduced cell adhesion caused by decreased E-cadherin expression (7) and activation of the Wnt signaling pathway (8) were found in nasopharyngeal carcinoma, the effect of BCSC-1 expression on cell aggregation, E-cadherin expression, and Wnt signaling was examined. It was observed that ectopic BCSC-1 resulted in enhanced cell aggregation associated with increased expression of E-cadherin/α-catenin, and BCSC-1 suppression promoted Wnt signaling. Materials and MethodsCell lines, tissue sections, and animals. The CNE-2L2 human nasopharyngeal carcinoma cell line was developed by Yu and Gao.(9) The HNE-1 human nasopharyngeal carcinoma cell line was developed by Yao et al.(10) CNE-2L2 cells with Man2c1 suppression (AS cells) and CNE-2L2 cells wi...

Pemphigus erythematosus, initially described as a combination of pemphigus with lupus erythematosus, and pemphigus foliaceus are now frequently considered localized and generalized variants of superficial pemphigus. Yet diagnostic criteria for pemphigus erythematosus remain controversial. Distinct from pemphigus foliaceus, pemphigus erythematosus displays immune depositions at the dermal-epidermal junction, which suggests additional immunopathological mechanisms. We present three patients with clinical and histopathologic signs of superficial pemphigus, who all exhibited an immunomorphology characteristic of pemphigus erythematosus. Complement depositions in a granular-linear fashion were consistently found at the dermal-epidermal junction besides in vivo bound and circulating antikeratinocyte cell-surface autoantibodies. Histopathology showed subcorneal acantholysis, and all sera contained antidesmoglein 1 but not antidesmoglein 3 autoantibodies detected by enzyme-linked immunosorbent assays (ELISA). Additional autoantibodies against a 230-kDa protein and against a 190-kDa protein comigrating with bullous pemphigoid antigen 1 (BP230) and periplakin, respectively, were present in all the patients' sera. As two sera specifically reacted with BP230 by ELISA, the presence of BP230-specific autoantibodies could be associated with dermal-epidermal immune staining in these patients. In pemphigus erythematosus, dermal-epidermal immune staining is generally attributed to the deposition of immune complexes, while the presence of BP230-specific autoantibodies has not been reported in this disease previously. Perhaps, the unique autoantibody profile of the patients in the study permits discrimination between patients with superficial pemphigus that display additional dermal-epidermal immune staining from those with conventional pemphigus foliaceus on a molecular basis. Further studies will be required to substantiate the frequency of this occurrence and to unravel its pathogenic significance.

Delivering a gene into the Epstein±Barr virus (EBV)-transformed B cells is useful in studying effects of the gene on B-cell functions. However, although people have been able to efficiently transfer genes into and get them expressed in B-lympho blastoidcells for a time probably long enough to kill the cells using vectors harbouring oriP, the expression time of the delivered gene is not long enough in order to study the gene function in B cells. To solve this problem, we constructed an adeno-associated virus (AAV) plasmid pAGX(1) based on plasmids pSub201 and pRc/CMV. We developed and packaged recombinant AAV (rAAV) expression vectors containing an antisense or a sense DNA fragment of 6A8 cDNA encoding a human a-mannosidase, or an antisense fragment of 5D4 cDNA encoding a human cell membrane protein, or EYFP DNA. EBV-transformed B cell SKW6 and 3D5 were transduced with those rAAV or the mock. Transduction with the rAAV-EYFP showed an infection frequency of 64^3.5% and 58^6.2% for SKW6 and 3D5 cell, respectively. Genomic polymerase chain reaction (PCR) for neo R gene indicated an integration of the transferred gene into the host DNA. After being cultured and propagated for over 12 months, the cells were detected for the expression of the transferred gene. The RT±PCR, enzymatic assay and Con A binding test demonstrated an inhibition of 6A8 a-mannosidase in both SKW6 and 3D5 cells transduced with the antisense 6A8 DNA. Immunofluorescence staining with monoclonal antibodies (MoAb) 5D4 showed a reduction of the 5D4 protein expression on both the cells transduced with the antisense 5D4 DNA. The DNA fragmentation assay showed a resistance of the cells with 6A8 a-mannosidase inhibition to apoptosis induction by anti-Fas antibody. The data indicate that the AAV vector pAGX(1) can efficiently introduce genes into EBV-transformed B cells and the delivered gene can be expressed in the cells for more than 12 months which may be long enough for the study of gene functions in B cells.

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