RESUMO
OBJECTIVE: To test if interleukin-1beta (IL-1beta), IL-1 receptor antagonist (IL-1Ra), IL-4, or IL-10 gene polymorphisms could be used as markers of susceptibility or severity in rheumatoid arthritis (RA). METHODS: The study included 108 patients with early RA followed up for 2 years and 128 healthy controls. From genomic DNA, 6 polymorphisms in genes for IL-1beta, IL-1Ra, IL-10, and IL-4 were typed. Allelic frequencies and carriage rates were compared between RA patients and controls, between patients with erosive and nonerosive RA, and between patients with or without sustained remission. RESULTS: The RP1 allele of the IL-4 gene was found with a significantly higher frequency in RA patients compared with controls. The combination of an RA-related HLA-DR allele expressing shared epitope and the presence of allele E2 in IL-1beta exon 5 was found to expose patients to an increased risk of erosive disease, with an odds ratio of 8.20 (95% confidence interval 2.59-25.84, P < 0.0001). No significant association was observed between polymorphisms and the occurrence of sustained remission. CONCLUSION: This report, for the first time, indicates an association between RA and a polymorphic IL-4 gene sequence located in 5q31-33. In addition, the results show the prognostic value of a polymorphism in IL-1beta exon 5, which allowed prediction of erosive disease with a specificity of 91.8% in 42.1% of patients. Although these observations are very interesting, they have to be considered preliminary and will need to be confirmed.
Assuntos
Artrite Reumatoide/genética , Interleucina-10/genética , Interleucina-1/genética , Interleucina-4/genética , Polimorfismo Genético , Sialoglicoproteínas/genética , Artrite Reumatoide/fisiopatologia , DNA/análise , Primers do DNA/química , Suscetibilidade a Doenças , Frequência do Gene , Antígenos HLA-DR/genética , Cadeias HLA-DRB1 , Teste de Histocompatibilidade , Humanos , Proteína Antagonista do Receptor de Interleucina 1 , Reação em Cadeia da Polimerase , Índice de Gravidade de DoençaRESUMO
Rheumatoid arthritis (RA) is an autoimmune disease characterized by inflammation of the synovial membrane of multiple joints. This inflammatory microenvironment allows fibroblast-like synoviocytes (FLS) to express or enhance several adhesion or costimulatory molecules. This phenotypic shift, under proinflammatory cytokines, seems to be related to functional consequences for antigen presentation to T cells. The sensory neuropeptide substance P (SP), present at high levels, is able to act on FLS proliferation and enzyme secretion. These data led us to investigate whether SP could also provoke a phenotypic change of FLS. Using flow cytometry and a three-step cellular ELISA method, we determined whether SP has an influence on the expression of MHC class II, intercellular adhesion molecule-1 (ICAM-1), VCAM-1, LFA-3, CD40, B7.1 or B7.2 molecules on RA FLS incubated with interferon-gamma (IFN-gamma) or IL-1beta or tumour necrosis factor-alpha (TNF-alpha) with or without SP. Our results indicate that SP potentiates the effect of proinflammatory cytokines on the expression of VCAM-1 on RA FLS. We verified the presence of specific SP (NK1) receptor mRNA. Using reverse transcription-polymerase chain reaction, we showed that RA FLS of patients express NK1 receptor mRNA. These results suggest that SP increase of cytokine-induced VCAM-1 expression acts via this specific SP receptor. Thus, during chronic inflammation RA FLS are at the interface between the immune and the nervous systems.
Assuntos
Artrite Reumatoide/imunologia , Substância P/farmacologia , Membrana Sinovial/imunologia , Molécula 1 de Adesão de Célula Vascular/biossíntese , Antígeno B7-1/análise , Antígenos CD40/análise , Antígenos CD58/análise , Interações Medicamentosas , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/imunologia , Citometria de Fluxo , Antígenos de Histocompatibilidade Classe II/análise , Humanos , Molécula 1 de Adesão Intercelular/análise , Interferon gama/farmacologia , Interleucina-1/farmacologia , RNA Mensageiro/análise , Receptores de Taquicininas/análise , Receptores de Taquicininas/genética , Membrana Sinovial/citologia , Membrana Sinovial/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
Two closely-related molecules, DR(alpha,beta1*0101) and DR(alpha,beta1*0103), whose beta chains only differ by three amino acids at positions 67, 70, and 71, and six intermediate molecules obtained by site-directed mutagenesis were used to ascertain the respective roles of the three polymorphic residues. Substitutions at positions 70 (D-->Q), 71 (E-->R) and 67 (I or L-->F) strongly affected HA 306-318-specific T-cell recognition. The consequences of the substitution of residue 67 by a phenylalanine depended on the modified HLA-DR molecule. Although this substitution completely inhibited peptide-specific DR1-restricted T-cell recognition, its manifestations on the DR103-restricted T-cell response were variable (abolishing proliferation of some cell lines and not others), no matter what the peptide presented was (HA 306-319 or HIV P25 peptides). We also observed that inhibition of the proliferation of an alloreactive anti-DR103 T-cell clone, caused by a substitution at position 70, was completely cancelled by substitution of residue 67 by a phenylalanine. The observations based on functional experiments, thus, suggest that residue 67 plays an important role in determining conformation of the peptide presented to the T cells. Molecular modeling was used to predict changes induced by amino acid substitutions and highly supports functional data. Substitution of residue 67 by a phenylalanine could have repercussions on the structure of HLA-DR molecule/peptide complexes and affect T-cell recognition.
Assuntos
Apresentação de Antígeno , Antígeno HLA-DR1/imunologia , Peptídeos/imunologia , Linfócitos T/imunologia , Adulto , Animais , Sítios de Ligação , Divisão Celular , Linhagem Celular Transformada , Sobrevivência Celular , Células Cultivadas , Homólogo 5 da Proteína Cromobox , Células Clonais , Glutamina/genética , Glutamina/imunologia , Antígeno HLA-DR1/genética , Humanos , Isoleucina/genética , Isoleucina/imunologia , Leucina/genética , Leucina/imunologia , Camundongos , Modelos Moleculares , Fenilalanina/genética , Fenilalanina/imunologia , Conformação Proteica , Relação Estrutura-Atividade , Linfócitos T/citologiaRESUMO
Transformation of human T cells by herpesvirus saimiri allows the production of an unlimited number of T cells which express a functional T-cell receptor. In this study we transformed four T-cell lines derived from rheumatoid arthritis synovial membranes. The transformed T cells were mainly CD4+ and expressed the phenotype of activated T cells. They were grown for more than 1 year in the absence of mitogen or feeder cells, and three of them could be maintained without exogenous IL-2. The presence of viral DNA in the transformed cells was shown by in situ hybridization with a probe from the H-DNA region of the virus. No infectious virus could be recovered from the transformed cells. The relative proportion of the 24 different Vbeta families between the four transformed lines showed variations that increased with time. In the two T-cell lines transformed at an early stage of culture, the Vbeta2 family was maintained at about 10%. The dominant Vbeta2 clones that previously have been characterized in the patient were found in all transformed T-cell lines. We have thus shown the feasibility of obtaining transformed T cells from synovial membranes. They contain the dominant clones that are considered of potential importance for the disease, permitting further functional studies.
Assuntos
Artrite Reumatoide/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Membrana Sinovial/imunologia , Linfócitos T/patologia , Artrite Reumatoide/patologia , Antígenos CD4 , Linhagem Celular Transformada , Transformação Celular Viral , Células Clonais , Herpesvirus Saimiriíneo 2 , Humanos , Membrana Sinovial/patologia , Linfócitos T/imunologiaRESUMO
Human myelin oligodendrocyte glycoprotein (MOG), a myelin component of the central nervous system, is a candidate target antigen for autoimmune-mediated demyelination. We have isolated and sequenced part of a cosmid clone that contains the entire human MOG gene. The primary nuclear transcript, extending from the putative start of transcription to the site of poly(A) addition, is 15,561 nucleotides in length. The human MOG gene contains 8 exons, separated by 7 introns; canonical intron/exon boundary sites are observed at each junction. The introns vary in size from 242 to 6484 bp and contain numerous repetitive DNA elements, including 14 Alu sequences within 3 introns. Another Alu element is located in the 3'-untranslated region of the gene. Alu sequences were classified with respect to subfamily assignment. Seven hundred sixty-three nucleotides 5' of the transcription start and 1214 nucleotides 3' of the poly(A) addition sites were also sequenced. The 5'-flanking region revealed the presence of several consensus sequences that could be relevant in the transcription of the MOG gene, in particular binding sites in common with other myelin gene promoters. Two polymorphic intragenic dinucleotide (CA)n and tetranucleotide (TAAA)n repeats were identified and may provide genetic marker tools for association and linkage studies.
Assuntos
Genes , Glicoproteína Associada a Mielina/genética , Sequência de Bases , Encéfalo/metabolismo , Cromossomos Humanos Par 6 , Regulação da Expressão Gênica , Humanos , Repetições de Microssatélites , Dados de Sequência Molecular , Proteínas da Mielina , Glicoproteína Mielina-Oligodendrócito , Sequências Reguladoras de Ácido Nucleico , Sequências Repetitivas de Ácido Nucleico , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Transcrição GênicaRESUMO
Gallium nitrate (GaN) reduces cancer-related hypercalcemia and inhibits bone resorption in vitro. This study investigated the effects of chronic GaN administration on bone, kidney, and parathyroid gland activity of growing rats. Experimental animals received GaN (1.75 mg elemental gallium i.p. QOD X 8, Ga+), and controls received the solvent (Ga-). In the bone of Ga+ rats the number of osteoclasts was increased (Ga+: 70.4 +/- 2.31 osteoclasts/mm2; Ga-: 46.5 +/- 1.61 osteoclasts/mm2, P less than 0.001), and apposition rate and osteoid width were unchanged. Ga was concentrated in bone (2.4 mumol/g cortical bone) and detected by electron microprobe on the surface of a few trabeculae. Alkaline (Alp) and acid (Acp) phosphatase activities were higher in Ga+ than in Ga- calvaria (Ga+: Alp 223 +/- 23.4 U/mg prot, Ga-: Alp 145 +/- 13.3 U/mg prot, P less than 0.02; Ga+: Acp 69.5 +/- 4.7 U/mg prot, Ga-: 57.5 +/- 2.8 U/mg prot, P less than 0.05). Serum iPTH was increased (Ga+: 112.9 +/- 17.6 pg/ml, Ga-: 41.4 +/- 7.4 pg/ml, P less than 0.01), serum calcium was reduced (Ga+: 2.4 +/- 0.02 mmol/l, Ga-: 2.6 +/- 0.03 mmol/l, P less than 0.001); calciuria remained comparable to controls. Relative to the hypocalcemia this suggests renal loss of Ca. The calcemic response to hPTH 1-34 (i.v. 50 micrograms/kg) was decreased 2 hours after injection of the hormone (delta Ca: TPTX Ga+: 0.11 +/- 0.04 mmol/l, Ga-: 0.33 +/- 0.03 mmol/l P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Reabsorção Óssea/efeitos dos fármacos , Osso e Ossos/efeitos dos fármacos , Gálio/farmacologia , Fosfatase Ácida/metabolismo , Fosfatase Alcalina/metabolismo , Animais , Cálcio/sangue , Contagem de Células , Rim/efeitos dos fármacos , Rim/metabolismo , Masculino , Osteoclastos/citologia , Hormônio Paratireóideo/sangue , Hormônio Paratireóideo/farmacologia , RatosRESUMO
The initial mineralization of the tibial bone collar of 17-day-old rat fetuses has been investigated. Images obtained after glutaraldehyde-paraformaldehyde-OsO4 fixation were compared to those obtained after K-pyroantimonate (PAO) fixation. Ca, P and Sb were identified and Ca/P intensity ratios evaluated by wavelength dispersive X-ray microanalysis. Alkaline phosphatase was detected on decalcified sections. Some osteoblasts showed degenerative changes and free mitochondria could be seen within the osteoid on the prolongation of their cytoplasmic processes. The mitochondria contained mineralized granules and clusters. Similar granules, numerous clusters and few matrix vesicles were observed within osteoid. The Ca/P intensity ratios (PAO fixed sections) of mitochondrial mineral (11.5 +/- 2.54) were different from the ratio of crystalline mineral in matrix vesicles (1.52 +/- 0.07). Alkaline phosphatase was present along plasmalemma of osteoblasts and around mineral deposits. These results show that in the rat fetus osteoblast mitochondria may be extruded from the cells, and that mitochondrial granules may represent the first mineral deposits in osteoid.
Assuntos
Osso e Ossos/metabolismo , Cálcio/metabolismo , Animais , Osso e Ossos/ultraestrutura , Feto/metabolismo , Minerais/metabolismo , Mitocôndrias/metabolismo , Osteoblastos/metabolismo , Ratos , Ratos EndogâmicosRESUMO
Over a period of 10 months (august 1975 to may 1976), the authors observed 6 successive accidents during the Shirodkar manoeuvre during operations for sterility of tubal origin: 1) 4 cases involved disturbances in haemostasis only, clinical manifestations being seen in only one case. In the laboratory, these quite atypical manifestations suggested either fibrinolysis or disseminated intravascular coagulation. 2) In 2 cases there was cardiovascular collapse with cardiac arrest. In the first patient, cardiac arrest was followed by a period of coagulopathy quite similar to those seen previously. In the second case resuscitation failed, leading the authors to abandon use of the Shirodkar manoeuvre, which brought the study to an end. We are unable to provide any satisfactory pathological interpretation of these complications, thought two factors are worthy of consideration:--The hydrocortisone suspension used;--The uterine trauma related to peroperative manipulations.
Assuntos
Transtornos da Coagulação Sanguínea/etiologia , Testes de Obstrução das Tubas Uterinas/efeitos adversos , Parada Cardíaca/etiologia , Infertilidade Feminina/terapia , Testes de Obstrução das Tubas Uterinas/métodos , Feminino , Hemostasia , Humanos , Hidrocortisona , Infertilidade Feminina/diagnóstico , Infertilidade Feminina/etiologia , MétodosRESUMO
We have found a series of 14 cases of chronic suppuration of the breast in 10 years in the Surgical and Gynaecological Clinic of the Salpêtrière Hospital. The symptomatology at the outset is usually innocuous, showing itself as a tumour that appears either gradually or suddenly. Diagnosis is not difficult and is usually solved by tapping the tumour. The progress of these lesions to become chronic with the formation of fistulae is the main risk, and the incident is not negligible. Thus the true problem is to find the correct therapy so as to avoid this possible unfavourable outcome.