T-cell DNA hypomethylation can increase expression of genes that have potential relation to autoimmunity. CD70 overexpression may be associated with B-cell activation and immunoglobulin secretion, while perforin plays an important role in T-cell-mediated macrophage apoptosis. DNA hypomethylation can activate human endogenous retroviruses (HERV) sequences. Expression of HERV components may elicit autoantibodies production. Polyamines change structure of chromosome and interfere with DNA methylation process, which is involved in the pathogenesis of autoimmune diseases.
DNA Methylation ; Humans ; Lupus Erythematosus, Systemic ; etiology ; genetics ; immunology

The expression of inducible co-stimulator (ICOS) in peripheral blood T lymphocytes from the patients with systemic lupus erythematosus (SLE) and the role in the pathogenesis of SLE was investigated. By using two-color immunofluorescent staining and flow cytometric assay, the expression levels of ICOS in peripheal blood T lymphocytes from 33 patients with SLE and 16 healthy volunteers were detected. SLE diseases activity index (SLEDAI) of the patients with SLE was used to evaluate the disease activity. The correlation between the ICOS expression and SLEDAI was analyzed among the groups. The results showed that the expression levels of ICOS in T lymphocytes in active SLE group was markedly higher than those in the control and inactive SLE groups (both P< 0.01). There was no significant difference in the expression levels of ICOS between the inactive SLE and the control groups (P>0.05). In active SLE and inactive SLE groups, positive linear correlation was found between the levels of the ICOS expression in T lymphocytes and SLEDAI (r=0. 711, P=0.001; r=0.561, P=0.03). It was suggested that the expression of ICOS in peripheral blood T lymphocytes from the patients with active SLE was up-regulated and and ICOS might be related to the pathogenesis of SLE.
Antigens, Differentiation, T-Lymphocyte/*biosynthesis ; Antigens, Differentiation, T-Lymphocyte/genetics ; Lupus Erythematosus, Systemic/blood ; Lupus Erythematosus, Systemic/etiology ; Lupus Erythematosus, Systemic/*immunology ; T-Lymphocytes/*immunology

Antiphospholipid Syndrome ; complications ; Atherosclerosis ; etiology ; B-Lymphocytes ; immunology ; CD11a Antigen ; genetics ; CD27 Ligand ; genetics ; CD40 Ligand ; genetics ; DNA Methylation ; Epigenesis, Genetic ; Humans ; Lupus Erythematosus, Systemic ; etiology ; genetics ; immunology

BACKGROUNDPrevious studies indicate that CD43 plays a role in regulating the adhesion of lymphocytes, cell mutation and activation, however, little is known about its effect on systemic lupus erythematosus (SLE). This study was designed to explore the clinical significance of CD43 in SLE patients.

METHODSWe used microarray and real-time PCR to detect the mRNA and protein expression of magnetic bead sorted T cells and B cells from peripheral blood mononuclear cells (PBMCs) of SLE patients, and analyzed the relationship between CD43 and the clinical indexes.

RESULTSBoth microarray and real-time PCR results showed that CD43 mRNA was significantly decreased in PBMCs of SLE patients compared with healthy controls (P < 0.001). There were no significant differences between lupus nephritis and non-lupus nephritis patients, and neuropsychiatric and non-neuropsychiatric patients. CD43 mRNA expression was significantly reduced in T cells but not in B-cells in SLE patients compared to healthy controls (P < 0.01). Compared with healthy controls, the percentage of CD43(+) cells in the PBMCs of SLE was significantly decreased (P = 0.004), and the CD43 fluorescence intensity in CD3(+)/CD43(+) cells and CD19(+)/CD43(+) cells was also significantly weaker than in healthy controls (P = 0.039 and 0.003). There was no significant difference in the percentage of CD3(+)/CD43(+) cells, CD19(+)/CD43(+) cells between the two groups. The CD43 fluorescence intensity in CD3(+)/CD43(+) cells was inversely correlated with the levels of IgG and IgM (r = -0.8 and -0.6).

CONCLUSIONSCompared to healthy controls, both CD43 mRNA and protein expressions were reduced in T cells from patients with SLE, and were inversely correlated with IgG.

B-Lymphocytes ; immunology ; metabolism ; Humans ; Leukocytes, Mononuclear ; Leukosialin ; genetics ; metabolism ; Lupus Erythematosus, Systemic ; immunology ; metabolism ; Oligonucleotide Array Sequence Analysis ; Real-Time Polymerase Chain Reaction ; T-Lymphocytes ; immunology ; metabolism

OBJECTIVETo construct a personalized full-length fully human antibody mammalian display library for children with systemic lupus erythematosus (SLE).

METHODSThe total RNA was isolated from the PBMCs of SLE children. The heavy chain variable region and kappa light chain (VH and LCκ) of the antibody genes were amplified by RT-PCR and inserted into the pDGB-HC-TM vector separately to construct the heavy chain and light chain libraries. The library DNAs were transfected into 293T cells and the expression of full-length fully human antibody on the surface of 293T cells was analyzed by flow cytometry.

RESULTSUsing 0.8 µg total RNA as the template, the VH and LCκ were amplified and the full-length fully human antibody mammalian display library was constructed. The VH and LCκ gene libraries had a size of 9.4×10(4) and 8.4×10(4), respectively. Sequence analysis of 10 clones randomly selected from the VH and LCκ gene libraries each showed that 8 heavy chain clones and 7 light chain clones contained correct open reading frames, and flow cytometry demonstrated that all the 15 clones express full-length antibodies on 293T cell surfaces. 293T cells co-transfected with the VH and LCκ gene libraries expressed the full-length antibodies on the cell surface.

CONCLUSIONThe personalized full-length fully human antibody library for SLE children constructed allows display of the full-length antibodies on mammalian cell surfaces, thus providing a valuable platform for analyzing the autoantibodies, their etiological role, and their clinical implications in SLE.

Amino Acid Sequence ; Child ; Gene Library ; Genetic Vectors ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Lupus Erythematosus, Systemic ; genetics ; immunology ; Membrane Proteins ; genetics

OBJECTIVETo investigate the expressions of chemokine receptors and interleukin (IL) receptors on the peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients and their correlations with clinical features as well as SLE disease activity index (SLEDAI).

METHODSThe mRNA expressions of chemokine receptors and IL receptors on PBMCs of 93 SLE patients and 30 healthy controls were detected by reverse transcription-polymerase chain reaction, including CCR2, CCR3, CCR4, CCR5, CCR6, CCR8, CXCR3, CXCRS, CX3CR1, XCR1, IL-4R, and IL-10R. The clinical features of SLE patients were recorded. The correlations of chemokine receptors and IL receptors mRNA expressions with clinical features as well as SLEDAI were assayed using linear regression analysis.

RESULTSThe level of CCR5 mRNA in SLE patients (including active and inactive SLE) was significantly higher than that in healthy controls (P < 0.05), and there was no significant difference between active and inactive patients in this respect (P > 0.05). CX3CR1 mRNA expression significantly increased from healthy control to inactive SLE to active SLE in sequence. The others (except for CCR8, CXCR3, and IL-10R) in active SLE patients were significantly higher than those in both inactive SLE patients and healthy controls (all P < 0.05). There were positive correlations between SLEDAI and CCR2 (r = 0.424, t = 4.313, P < 0.001), CCR3 (r = 0.518, t = 5.410, P < 0.001), CCR4 (r = 0.376, t = 3.851, P < 0.001), CCR6 (r = 0.457, t = 4.513, P < 0.001), CXCR5 (r = 0.455, t = 4.629, P < 0.001), CX3CR1 (r = 0.445, t = 4.523, P < 0.001), as well as XCR1 (r = 0.540, t = 5.445, P < 0.001). And CCR5 mRNA expression level was positively correlated with IL-4R mRNA (r = 0.313, t = 2.353, P < 0.05). The patients with myositis and cutaneous vasculitis simultaneously showed lower levels of CCR5 and CX3CR1, and CCR5 expression was negatively correlated with the scores of SLEDAI in SLE cases accompanied by photosensitivity (r = 0.426, t = -2.155, P < 0.05).

CONCLUSIONIncreased expressions of CCR5 and CX3CR1 on PBMCs may be indicators in clinical survey for SLE.

Adolescent ; Adult ; CX3C Chemokine Receptor 1 ; Child ; Female ; Humans ; Leukocytes, Mononuclear ; immunology ; Lupus Erythematosus, Systemic ; etiology ; immunology ; Male ; Middle Aged ; RNA, Messenger ; blood ; Receptors, CCR5 ; genetics ; Receptors, Chemokine ; genetics ; Receptors, Interleukin-10 ; genetics ; Receptors, Interleukin-4 ; genetics

OBJECTIVE: To investigate the effect of total glucosides of peony (TGP) on expression and DNA methylation status of ITGAL gene (CD11a) in CD4(+) T cells from patients with systemic lupus erythematosus (SLE). METHODS: CD4(+) T cells were isolated by positive selection using CD4 beads. CD4(+) T cells were treated by TGP at 0, 62.5, 312.5 and 1562.5 mg/L for 48 h. The MTT method was used to assess cell viability; mRNA expression level was measured by realtime-PCR; protein level of CD11a was measured by flow cytometric analysis; DNA methylation status was assayed by bisulfite sequencing. RESULTS: No significant change in cell viability was found in CD4(+) T cells among the different concentration groups (P>0.05). Compared with control, the mRNA and protein levels of ITGAL were down-regulated significantly in SLE CD4(+) T cells treated with TGP (1562.5 mg/L) (P< 0.01). Furthermore, the extent of DNA methylation of ITGAL promoter was increased in TGP (1562.5 mg/L) treated CD4(+) T cells compared with control group (P<0.01). CONCLUSION TGP can repress CD11a gene expression through enhancing DNA methylation of ITGAL promoter in CD4(+) T cells from patients with SLE. This observation represents a preliminary step in understanding the mechanism of TGP in SLE therapy.
CD11a Antigen ; genetics ; metabolism ; CD4-Positive T-Lymphocytes ; immunology ; metabolism ; DNA Methylation ; drug effects ; Down-Regulation ; drug effects ; Glucosides ; pharmacology ; Humans ; Lupus Erythematosus, Systemic ; genetics ; immunology ; Paeonia ; chemistry ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; genetics ; metabolism

Tissue and organ differentiation is tightly controlled to ensure proper development and function of the growing embryo as well as cells such as lymphocytes that differentiate throughout the adult stage. Therefore it is vital that the genes and the protein they encode that are involved in these processes function accurately. Hence, any mutation or error that occurs along the way can result in extensive damage, which is expressed in various ways in the embryo and can result in immune pathogenesis, including immunodeficiency and autoimmune diseases, when lymphocyte development is altered. A number of studies have been carried out to look at the genes regulating transcription in tissue differentiation, including the transcription factors Pbx1. This gene is of particular interest to us as we have identified that it is associated with systemic lupus erythematosus susceptibility (Cuda et al., in press). This perspective summarizes the known roles of Pbx1 in tissue differentiation as well as our recent findings associating genetic variations in Pbx1 to lupus susceptibility, and we will speculate on how this gene controls the maintenance of immune tolerance in T cells.
Animals ; Cell Differentiation ; Chromatin Immunoprecipitation ; DNA-Binding Proteins ; genetics ; immunology ; Genetic Loci ; immunology ; Genetic Predisposition to Disease ; Homeodomain Proteins ; genetics ; immunology ; Humans ; Immune Tolerance ; Lupus Erythematosus, Systemic ; genetics ; immunology ; Lymphocyte Activation ; Mice ; Mice, Transgenic ; Pre-B-Cell Leukemia Transcription Factor 1 ; Protein Structure, Tertiary ; Proto-Oncogene Proteins ; genetics ; immunology ; Signal Transduction ; T-Lymphocytes, Regulatory ; cytology ; immunology ; Transcription Factors ; genetics ; immunology ; Tretinoin ; metabolism

Animals ; DNA-Binding Proteins ; genetics ; immunology ; Genetic Loci ; immunology ; Genetic Predisposition to Disease ; Homeodomain Proteins ; genetics ; immunology ; Humans ; Lupus Erythematosus, Systemic ; genetics ; immunology ; Lymphocyte Activation ; Mice ; Mice, Transgenic ; Pre-B-Cell Leukemia Transcription Factor 1 ; Proto-Oncogene Proteins ; genetics ; immunology ; T-Lymphocytes, Regulatory ; cytology ; immunology ; Transcription Factors ; genetics ; immunology

OBJECTIVETo analyze the drift of the complementarity-determining region 3 (CDR3) of T cell receptor (TCR) beta chain variable region (TCR BV) in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus.

METHODSImmunoscope spectratyping techniques was used to analyze the distribution of TCRbeta chain CDR3 in 5 normal blood donors and the dominant CDR3 in the PBMCs in 5 SLE patients. Sequence analysis of the CDR3 region in monoclonal or oligoclonal T cells was performed.

RESULTSThe spectratypes of TCR BV gene CDR3 region showed Gaussian distribution in the 5 normal blood donors. The 5 SLE patients, however, displayed anomalous proliferation and oligoclonal expansion of the T cells was observed in different TCR BV families with different CDR3 sequences.

CONCLUSIONNoticeable drift of TCRbeta chain CDR3 can be seen in active SLE, indicating possible association of selective expression of TCR with immune pathogenesis in SLE. Determination of specific TCR CDR3 sequence provides a new means for studying the pathogenesis and personalized treatment of SLE.

Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Complementarity Determining Regions ; genetics ; Female ; Genes, T-Cell Receptor beta ; genetics ; Humans ; Leukocytes, Mononuclear ; cytology ; metabolism ; Lupus Erythematosus, Systemic ; genetics ; immunology ; Molecular Sequence Data ; Sequence Analysis, DNA