ABSTRACT
Down-regulation of soluble or membrane-bound co-stimulatory molecules by RNAi in dendritic cells can prevent the activation of immune responses. Therefore, this study was designed to evaluate the therapeutic efficacy of bone marrow-derived DCs (BMDCs) transduced with lentiviral vectors to permanently expressed shRNA specific for CD40 (CD40LV-DCs) and/or p19 subunit of interleukin (IL)-23 (p19LV-DCs) mRNAs in experimental autoimmune encephalomyelitis (EAE). In-vitro studies showed that double-transduced BMDCs (CD40(+) p19LV-DCs) resemble tolerogenic DCs due to profound down-regulation of CD40, lower expression of proinflammatory cytokines (IL-6 and IL-12), increased IL-10 production and stronger stimulation of myelin oligodendrocyte glycoprotein (MOG)35-55 -specific T cells for production of IL-10 compared with CD40LV-DCs, p19LV-DCs and BMDCs transduced with control lentiviral vector (CoLV-DCs). Moreover, injection of transduced CD40(+) p19LV- BMDCs in EAE mice resulted in more reduction in clinical score, significant reduction in IL-17 or increased production of IL-10 by mononuclear cells derived from the lymph nodes or spinal cord compared with CoLV-DCs-treated EAE mice. In conclusion, simultaneous knock-down of CD40 and IL-23 production by BMDCs may represent a promising therapeutic tool for the treatment of IL-17-dependent autoimmune diseases, including multiple sclerosis.
Subject(s)
CD40 Antigens/immunology , Dendritic Cells/immunology , Encephalomyelitis, Autoimmune, Experimental/immunology , Interleukin-23 Subunit p19/immunology , Th17 Cells/immunology , Animals , CD4-Positive T-Lymphocytes/immunology , CD4-Positive T-Lymphocytes/metabolism , CD40 Antigens/genetics , CD40 Antigens/metabolism , Cell Proliferation , Coculture Techniques , Cytokines/immunology , Cytokines/metabolism , Dendritic Cells/metabolism , Dendritic Cells/transplantation , Disease Models, Animal , Encephalomyelitis, Autoimmune, Experimental/metabolism , Encephalomyelitis, Autoimmune, Experimental/therapy , Flow Cytometry , Humans , Interleukin-10/immunology , Interleukin-10/metabolism , Interleukin-17/immunology , Interleukin-17/metabolism , Interleukin-23 Subunit p19/genetics , Interleukin-23 Subunit p19/metabolism , Lentivirus/genetics , Mice , Mice, Inbred C57BL , Multiple Sclerosis/immunology , Multiple Sclerosis/metabolism , Multiple Sclerosis/therapy , Myelin-Oligodendrocyte Glycoprotein/immunology , Peptide Fragments/immunology , RNA Interference/immunology , RNA, Small Interfering/genetics , Th17 Cells/metabolism , Treatment OutcomeABSTRACT
Although, the cytotoxic T lymphocyte antigen-4 gene polymorphism at position 49 of exon-1 has been strongly elucidated in different autoimmune diseases, but its role in predisposition to systemic sclerosis (SSc) is yet controversial. This study intends to analyze the genetic correlation of the ctla-4 gene locus with diffuse systemic sclerosis (dSSc), as well as to understand the influence of these genotypes in disease expression. Seventy known cases of SSc, and 151 age-matched healthy controls, were participated in this investigation. The frequencies of AA, GG and AG genotypes were found to be 26 (37.1%), 5 (7.2%) and 39 (55.7%) in patients, and 60 (39.7%), 19 (12.6%) and 72 (47.7%) in controls, respectively. As indicated, the differences in genotype and allele frequencies between patients and controls were insignificant (P>0.05). Moreover, the distribution of CTLA-4 polymorphism between patients did not differ significantly according to clinical and serologic features. In Iranian patients, susceptibility to SSc is not influenced by a bi-allelic ctla-4 gene (A49G) polymorphism.
Subject(s)
Antigens, CD/genetics , Antigens, Differentiation/genetics , Exons , Polymorphism, Genetic , Scleroderma, Systemic/genetics , Adult , Age of Onset , Alleles , CTLA-4 Antigen , Case-Control Studies , Female , Gene Frequency , Humans , Iran/epidemiology , Male , Middle Aged , Scleroderma, Systemic/pathologyABSTRACT
OBJECTIVE: One of the major complications of pregnancy, preeclampsia makes pregnancy termination inevitable in most cases. Similarities exist between the mechanisms that maintain normal pregnancy, allograft transplants, and, it is postulated, peripheral self-tolerance. In addition, the critical role of the cytotoxic T-lymphocyte antigen-4 (CTLA-4) molecule in maintaining self-tolerance has been established. Therefore, the frequency of CTLA-4 A49G polymorphism was investigated in severe preeclampsia. PATIENTS AND METHODS: Genomic DNA extracted from mononuclear cells of the peripheral blood of 36 pregnant women with severe preeclampsia and 151 healthy women was analyzed. A49G polymorphism in position 49 of exon-1 of the CTLA-4 gene was studied by the polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) method. RESULTS: The frequency of the GG genotype was 2 (5.6%) in patients and 19 (12.6%) in controls, while the frequency of the AA genotype was 4 (11.1%) and 60 (39.7%). Interestingly, the frequency of the AG genotype was significantly higher in preeclamptic than in healthy women from the general population (83.3% vs. 47.7%; P=0.0005). CONCLUSION: These data suggest that heterozygosity in the CTLA-4 A49G allele might be a predisposing factor for severe preeclampsia. Whether the observed association results from linkage imbalance with other loci on chromosome 2 or other polymorphisms of the CTLA-4 gene or even from a preferential transfer and/or expression of one allele from a heterozygous mother to the fetus will be the subject of future investigations.
Subject(s)
Antigens, Differentiation/genetics , Genetic Predisposition to Disease , Polymorphism, Genetic/genetics , Pre-Eclampsia/genetics , Adult , Antigens, CD , CTLA-4 Antigen , Female , Genetic Linkage , Heterozygote , Humans , Polymorphism, Single-Stranded Conformational , Pregnancy , White People/geneticsABSTRACT
Polymorphisms in ctla-4 gene have been shown to be associated with the Graves' disease (GD) susceptibility in different populations in the world. This study was undertaken to disclose the probable association of exon-1 polymorphism of ctla-4 with GD in Iranian patients. A49G polymorphism was investigated in 90 patients and 90 age/sex matched normal healthy controls, using PCR-SSCP and PCR-RFLP methods. Frequencies of AA, AG and GG genotypes among patients were found to be 21 (23.3%), 49 (54.5%) and 20 (22.2%) while these frequencies among healthy controls were 30 (33.3%), 53 (58.9%) and 7(7.8%), respectively. A significant increase of GG genotype and G allele was observed in patients (p = 0.012 and p = 0.025). In conclusion, consistent with the results of most other studies, the presence of a G allele in position 49 of ctla-4 exon-1 is associated with susceptibility to GD in Iranian population.
