RESUMO
OBJECTIVE: To evaluate autoimmune hepatitis (AIH) in a multicenter cohort of childhood-onset systemic lupus erythematosus (cSLE) patients. METHODS: This retrospective multicenter study included 847 patients with cSLE, performed in 10 Pediatric Rheumatology services of São Paulo state, Brazil. AIH was defined according to the International Autoimmune Hepatitis Group criteria (IAHGC). The statistical analysis was performed using the Bonferroni's correction (p < 0.0033). RESULTS: AIH in cSLE patients confirmed by biopsy was observed in 7/847 (0.8%) and all were diagnosed during adolescence. The majority occurred before or at cSLE diagnosis [5/7 (71%)]. Antinuclear antibodies were a universal finding, 43% had concomitantly anti-smooth muscle antibodies and all were seronegative for anti-liver kidney microsomal antibodies. All patients with follow-up ≥18 months (4/7) had complete response to therapy according to IAHGC. None had severe hepatic manifestations such as hepatic failure, portal hypertension and cirrhosis at presentation or follow-up. Further comparison of 7 cSLE patients with AIH and 28 without this complication with same disease duration [0 (0-8.5) vs. 0.12 (0-8.5) years, p = 0.06] revealed that the frequency of hepatomegaly was significantly higher in cSLE patients in the former group (71% vs. 11%, p = 0.003) with a similar median SLEDAI-2 K score [6 (0-26) vs. 7 (0-41), p = 0.755]. No differences were evidenced regarding constitutional involvement, splenomegaly, serositis, musculoskeletal, neuropsychiatric and renal involvements, and treatments in cSLE patients with and without AIH (p > 0.0033). CONCLUSIONS: Overlap of AIH and cSLE was rarely observed in this large multicenter study and hepatomegaly was the distinctive clinical feature of these patients. AIH occurred during adolescence, mainly at the first years of lupus and it was associated with mild liver manifestations.
Assuntos
Hepatite Autoimune/epidemiologia , Hepatomegalia/epidemiologia , Lúpus Eritematoso Sistêmico/epidemiologia , Adolescente , Idade de Início , Anticorpos Antinucleares/análise , Autoantígenos/análise , Brasil/epidemiologia , Criança , Feminino , Hepatite Autoimune/imunologia , Humanos , Lúpus Eritematoso Sistêmico/imunologia , Masculino , Microssomos/imunologia , Músculo Liso/imunologia , Estudos RetrospectivosRESUMO
The aim of this study was to determine expression, not previously described, of PLUNC (palate, lung, and nasal epithelium clone) (BPI-fold containing) proteins in major and minor salivary glands from very early fetal tissue to the end of the second trimester and thus gain further insight into the function of these proteins. Early fetal heads, and major and minor salivary glands were collected retrospectively and glands were classified according to morphodifferentiation stage. Expression of BPI-fold containing proteins was localized through immunohistochemistry. BPIFA2, the major BPI-fold containing protein in adult salivary glands, was detected only in the laryngeal pharynx; the lack of staining in salivary glands suggested salivary expression is either very late in development or is only in adult tissues. Early expression of BPIFA1 was seen in the trachea and nasal cavity with salivary gland expression only seen in late morphodifferentiation stages. BPIFB1 was seen in early neural tissue and at later stages in submandibular and sublingual glands. BPIFA1 is significantly expressed in early fetal oral tissue but BPIFB1 has extremely limited expression and the major salivary BPIF protein (BPIFA2) is not produced in fetal development. Further studies, with more sensitive techniques, will confirm the expression pattern and enable a better understanding of embryonic BPIF protein function.
Assuntos
Autoantígenos/análise , Feto/química , Glicoproteínas/análise , Fosfoproteínas/análise , Proteínas/análise , Glândulas Salivares/química , Proteínas e Peptídeos Salivares/análise , Epitélio/química , Proteínas de Ligação a Ácido Graxo , Desenvolvimento Fetal , Idade Gestacional , Cabeça/embriologia , Humanos , Imuno-Histoquímica , Pescoço/embriologia , Palato/química , Palato/embriologia , Estudos Retrospectivos , Glândulas Salivares/embriologia , Fatores de Tempo , Língua/química , Língua/embriologiaRESUMO
Abstract The aim of this study was to determine expression, not previously described, of PLUNC (palate, lung, and nasal epithelium clone) (BPI-fold containing) proteins in major and minor salivary glands from very early fetal tissue to the end of the second trimester and thus gain further insight into the function of these proteins. Early fetal heads, and major and minor salivary glands were collected retrospectively and glands were classified according to morphodifferentiation stage. Expression of BPI-fold containing proteins was localized through immunohistochemistry. BPIFA2, the major BPI-fold containing protein in adult salivary glands, was detected only in the laryngeal pharynx; the lack of staining in salivary glands suggested salivary expression is either very late in development or is only in adult tissues. Early expression of BPIFA1 was seen in the trachea and nasal cavity with salivary gland expression only seen in late morphodifferentiation stages. BPIFB1 was seen in early neural tissue and at later stages in submandibular and sublingual glands. BPIFA1 is significantly expressed in early fetal oral tissue but BPIFB1 has extremely limited expression and the major salivary BPIF protein (BPIFA2) is not produced in fetal development. Further studies, with more sensitive techniques, will confirm the expression pattern and enable a better understanding of embryonic BPIF protein function.
Assuntos
Humanos , Fosfoproteínas/análise , Glândulas Salivares/química , Proteínas e Peptídeos Salivares/análise , Autoantígenos/análise , Glicoproteínas/análise , Proteínas/análise , Feto/química , Palato/embriologia , Palato/química , Glândulas Salivares/embriologia , Fatores de Tempo , Língua/embriologia , Língua/química , Imuno-Histoquímica , Estudos Retrospectivos , Idade Gestacional , Desenvolvimento Fetal , Epitélio/química , Cabeça/embriologia , Pescoço/embriologiaRESUMO
BACKGROUND: Bladder cancer is the second most common urological malignancy worldwide. CIP2A is a newly identified inhibitor of PP2A. Recent studies have highlighted a potential role for CIP2A in promoting the proliferation of several cancer cells. However, the role of CIP2A in bladder cancer still remains unclear. METHODS: The expression of CIP2A was detected by quantitative real-time polymerase chain reaction and IHC in bladder cancer tissues and bladder cancer cell lines. In addition, silencing of CIP2A with siRNA was performed in T24 cells, and the impact on proliferation, and apoptosis of T24 cells was analyzed. RESULTS: Our results found that CIP2A expression levels were higher in bladder cancer tissues and cell lines. Furthermore, CIP2A siRNA significantly reduced the proliferation rate of T24 cells, induced a significant population of early and late apoptosis, and could reverse EMT in T24 cells, indicates that CIP2A expression is increased in bladder cancer and implies a role of the protein in bladder cancer progression. CONCLUSIONS: These results suggest that CIP2A is involved in tumor progression, and thus CIP2A could represent selective targets for the targeted treatments of bladder cancer.
Assuntos
Autoantígenos/biossíntese , Transição Epitelial-Mesenquimal/fisiologia , Proteínas de Membrana/biossíntese , Neoplasias da Bexiga Urinária/patologia , Apoptose/fisiologia , Autoantígenos/análise , Western Blotting , Linhagem Celular Tumoral , Proliferação de Células/fisiologia , Progressão da Doença , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas de Membrana/análise , RNA Interferente Pequeno , Reação em Cadeia da Polimerase em Tempo Real , Transfecção , Regulação para CimaRESUMO
INTRODUCTION: The identification of molecules expressed selectively on the surface of retinoblastoma cells would allow applying targeted therapies. The Ganglioside, N-Glycolyl-GM3 (NeuGc-GM3), is an attractive candidate, as it has been detected in other paediatric neuroectodermic tumours, and it is not expressed in human normal tissues. The 14F7 antibody recognizes specifically the ganglioside NeuGc-GM3. PURPOSE: To characterize the expression of NeuGc-GM3 in retinoblastoma cell lines and in retinoblastoma tumours using the 14F7 monoclonal antibody. METHODS: We studied WERI-Rb1 and Y79 cell lines, 24 retinoblastoma primary tumours from unilateral and bilateral cases and two bone marrow biopsies from metastatic retinoblastoma. Tumours were classified into three groups: non-invasive (n = 13), invasive (n = 9) and metastatic (n = 2). Three eyes enucleated because of non-tumoural conditions were used as controls. Cell lines and tumour sections were studied by immunohistochemistry using the 14F7 antibody. NeuGc-GM3 expression was evaluated by analysing the percentage of positive tumoural cells and the staining intensity. These parameters were analysed comparatively among the three groups. RESULTS: Both retinoblastoma cell lines showed immunoreactivity to NeuGc-GM3 but WERI-Rb1 presented higher intensity than Y79. All the tumours studied showed strong immunoreactivity to NeuGc-GM3 with no significant differences among groups. In both bone marrow specimens, NeuGc-GM3 immunoreactivity was observed in retinoblastoma cells. In bilaterally enucleated cases, NeuGc-GM3 immunoreactivity was not altered before and after chemotherapy. Non-tumoural retinas were negative. CONCLUSIONS: NeuGc-GM3 is highly expressed in retinoblastoma cell lines, tumours and metastatic cells to the bone marrow, and it is not detectable in control eyes. There were no significant differences in the immunoreactivity to 14F7 among tumours from different disease stages. Its immunoreactivity did not change after chemotherapy.
Assuntos
Autoantígenos/análise , Gangliosídeo G(M3)/análogos & derivados , Neoplasias da Retina/química , Retinoblastoma/química , Anticorpos Monoclonais/imunologia , Linhagem Celular Tumoral , Gangliosídeo G(M3)/análise , Gangliosídeo G(M3)/imunologia , Humanos , Técnicas ImunoenzimáticasRESUMO
Systemic sclerosis (SSc) is a multisystem disease of unknown etiology. Esophageal involvement affects 50-90% of patients and is characterized by abnormal motility and hypotonic lower esophageal sphincter. Data on the association of esophageal abnormalities and age, gender, SSc subset or duration, autoantibody profile, esophageal symptoms, and medication are lacking or conflicting. The aim of this study was the evaluation of these associations in Brazilian sclerodermic patients from the Rheumatology Division, Clinics Hospital, Federal University, Minas Gerais. They underwent medical records review, clinical interview, and esophageal manometry. The normal cutoff level for lower esophageal sphincter pressure was 14 mmHg. Abnormal peristalsis occurred when less than 80% of peristaltic waves were propagated. P-values less than 0.05 were considered significant. Twenty-eight patients were included: 71% were women. The population presented medium age and disease duration of 46 years and 12 years, respectively. Cutaneous diffuse SSc occurred in 39% and its limited form in 61%. Dysphagia, pyrosis, and regurgitation occurred, respectively, in 71%, 43%, and 61% of patients. Lower esophageal sphincter pressure and number of peristaltic waves-propagated medias were, respectively, 17.2 mmHg and 2.3. SSc-related manometric abnormalities were present in 86% of patients. Manometry revealed distal esophageal body hypomotility, hypotonic lower esophageal sphincter, or both, respectively, in 82%, 39%, and 36% of patients. One patient presented the manometric pattern of esophageal achalasia. Male patients more frequently presented hypotonic inferior esophageal sphincter. Manometric findings have had no relationship with the other variables. Nifedipine use did not influence manometric findings.
Assuntos
Transtornos da Motilidade Esofágica/etiologia , Escleroderma Sistêmico/complicações , Adulto , Anticorpos Antinucleares/análise , Autoantígenos/análise , Brasil , Bloqueadores dos Canais de Cálcio/farmacologia , DNA Topoisomerases Tipo I , Transtornos da Motilidade Esofágica/imunologia , Feminino , Humanos , Masculino , Manometria , Pessoa de Meia-Idade , Nifedipino/farmacologia , Proteínas Nucleares/análise , Estudos Retrospectivos , Esclerodermia Difusa/complicações , Esclerodermia Limitada/complicações , Escleroderma Sistêmico/imunologiaRESUMO
Trypanosoma cruzi expresses several proteins containing antigenic amino acid repeats. Here we characterized TcRpL7a and TcRBP28, which carry similar repeat motifs and share homology to the eukaryotic L7a ribosomal protein and to a Trypanosoma brucei RNA binding protein, respectively. Analyses of the full length and truncated recombinant TcRpL7a showed that the humoral response of patients with Chagas disease is directed towards its repetitive domain. Sequence analyses of distinct copies of TcRpL7a genes present in the genome of six T. cruzi strains indicate that the number of repeats is higher in proteins from T. cruzi II than T. cruzi I strains. A serum panel of 59 T. cruzi infected patients showed that 73% reacted with TcRpL7a, 71% reacted with TcRBP28 and 80% reacted with 1:1 mixture of both antigens. Synthetic peptides harboring the TcRpL7a repeat motif reacted with 46% of the serum samples. Antibodies raised against both antigens identified equivalent amounts of the native proteins in all three stages of the parasite life cycle. Analyses of subcellular fractions indicated that TcRBP28 is present in the cytoplasm whereas TcRpL7a co-fractionates with polysomes. Confirming their predicted cellular localization, GFP fusions showed that, whereas GFP::TcRBP28 localizes in the cytoplasm, GFP::TcRpL7a accumulates in the nucleus, where ribosome biogenesis occurs.
Assuntos
Anticorpos Antiprotozoários/imunologia , Antígenos de Protozoários/imunologia , Autoantígenos/imunologia , Sequências Repetitivas de Aminoácidos , Trypanosoma cruzi/imunologia , Sequência de Aminoácidos , Animais , Antígenos de Protozoários/análise , Antígenos de Protozoários/genética , Autoantígenos/análise , Autoantígenos/genética , Núcleo Celular/química , Doença de Chagas/imunologia , Citoplasma/química , Mapeamento de Epitopos , Humanos , Dados de Sequência Molecular , Peptídeos/imunologia , Alinhamento de Sequência , Trypanosoma cruzi/genética , Proteínas Centrais de snRNPRESUMO
INTRODUCTION: The frequency and severity of salivary and lacrymal gland human T-cell lymphotropic virus type 1 (HTLV-1) infection were assessed in HTLV-1 plus patients, presenting with neurological deficit (tropical spastic paraparesis/HTLV-1 associated myelopathy [TSP/HAM]) or not. The mechanism of this deficit was investigated. MATERIAL AND METHODS: A case-control study was made from April 2002 to December 2005, in an area strongly endemic for HTLV-1. The patients were classified in three groups: group 1 with 16 patients presenting with TSP/HAM; group 2 with 67 HTLV-1 carriers and group 3 with 29 healthy volunteers. The dry syndrome was investigated by history taking and by oral and ophthalmological clinical examination. Immunological and biological screening for rhumatoid factors, antinuclear antibodies, and antibodies against soluble nuclear antigens (SSA, SSB). Peripheral blood was separated by density gradient and mononuclear cells were recovered to dose interferon-gamma and tumor necrosis factor-alpha. Patients in the three groups were assessed for salivary flow by stimulated weighing using Saxon's test. A Chi-2 test, a variance analysis (Anova), and the Spearman rank correlation test were used for the statistical analysis. RESULTS: The dry syndrome was mild and more common in group 1 patients (75%). In group 2, 22% of the patients presented with functional signs of buccal mucosa dryness comparable to those observed in group 1. No correlation was found between salivary flow and screened pro-inflammatory cytokines. DISCUSSION: Our results show that hyposialia is an important part of the disease induced by HTLV-1, even in virus carriers without neurological deficit. Its mechanism seems different than that of the Gougerot-Sjögren syndrome.
Assuntos
Infecções por HTLV-I/complicações , Doenças do Aparelho Lacrimal/complicações , Doenças das Glândulas Salivares/complicações , Adulto , Idoso , Anticorpos Antinucleares/análise , Autoantígenos/análise , Brasil , Portador Sadio , Estudos de Casos e Controles , Síndromes do Olho Seco/complicações , Doenças Endêmicas , Feminino , Humanos , Interferon gama/análise , Ceratoconjuntivite/complicações , Ceratoconjuntivite/fisiopatologia , Doenças do Aparelho Lacrimal/fisiopatologia , Masculino , Pessoa de Meia-Idade , Paraparesia Espástica Tropical/complicações , Fator Reumatoide/análise , Ribonucleoproteínas/análise , Saliva/metabolismo , Doenças das Glândulas Salivares/fisiopatologia , Taxa Secretória/fisiologia , Fator de Necrose Tumoral alfa/análise , Xerostomia/complicações , Xerostomia/fisiopatologia , Antígeno SS-BRESUMO
OBJECTIVE: To determine whether ultraviolet B (UVB) irradiation induces novel modifications in autoantigens targeted during experimental photoinduced epidermal damage. METHODS: To search for novel UVB-induced autoantigen modifications, lysates made from UVB-irradiated human keratinocytes or HeLa cells were immunoblotted using human autoantibodies that recognize ribonucleoprotein autoantigens. Novel autoantigen structures identified were further characterized using nucleases and RNA hybridization. RESULTS: Human sera that recognize U1-70 kd (U1-70K) and La by immunoblotting also recognized multiple novel species when they were used to immunoblot lysates of UVB-irradiated keratinocytes or HeLa cells. These species were not present in control cells and were not observed when apoptosis was induced by Fas ligation or cytotoxic lymphocyte granule contents. Biochemical analysis using multiple assays revealed that these novel UVB-induced molecular species result from the covalent crosslinking between the U1 RNA and the hYRNA molecules with their associated proteins, including U1-70K, La, and likely components of the Sm particle. CONCLUSION: These data demonstrate that UVB irradiation of live cells can directly induce covalent RNA-protein complexes, which are recognized by human autoantibodies. As previously described for other autoantigens, these covalent complexes of RNA and proteins may have important consequences in terms of antigen capture and processing.
Assuntos
Autoantígenos/análise , Queratinócitos/efeitos da radiação , RNA Nuclear Pequeno/efeitos da radiação , Ribonucleoproteína Nuclear Pequena U1/efeitos da radiação , Ribonucleoproteínas/efeitos da radiação , Apoptose/efeitos da radiação , Autoanticorpos/imunologia , Células HeLa/imunologia , Células HeLa/patologia , Células HeLa/efeitos da radiação , Humanos , Queratinócitos/imunologia , Queratinócitos/patologia , RNA Nuclear Pequeno/imunologia , Ribonucleoproteína Nuclear Pequena U1/imunologia , Ribonucleoproteínas/imunologia , Raios Ultravioleta , Antígeno SS-BRESUMO
BACKGROUND: Besides Brazilian endemic pemphigus foliaceus (EPF), we have described another focus of EPF in Colombia. Our previous study suggested that Colombian EPF seemed to react various plakin family proteins, such as envoplakin, periplakin and BP230. OBJECTIVE: To further characterize the Colombian EPF and study the difference from Brazilian EPF, we examined the antigen profile of the two types of EPF. METHODS AND RESULTS: Immunoblotting using normal human epidermal extracts revealed that 38% Colombian EPF sera and 25% Brazilian EPF sera showed IgG antibodies reactive with desmoglein (Dsg) 1, pemphigus foliaceus antigen. The sera of both types of EPF showed protein bands co-migrating with plakin family proteins, particularly periplakin. Immunoblotting analyses using recombinant proteins of various domains of envoplakin, periplakin and BP230 revealed that a considerable number of Colombian EPF sera reacted with recombinant proteins of periplakin, while only few Brazilian sera reacted with some of the recombinant proteins of any plakins. Enzyme-linked immunosorbent assay (ELISA) for Dsg1 and Dsg3 showed that Dsg1 was reacted by almost all sera of both types of EPF. However, unexpectedly, while none of Colombian EPF sera reacted with Dsg3, about half of Brazilian EPF sera reacted with Dsg3. CONCLUSION: These results suggested that the Colombian EPF is basically similar to Brazilian EPF in terms that major antigen is Dsg1, but there were some different antigen profiles between the two types of EPF.
Assuntos
Autoantígenos/análise , Doenças Endêmicas , Pênfigo/epidemiologia , Pênfigo/imunologia , Autoanticorpos/análise , Bioquímica/métodos , Brasil , Caderinas/análise , Colômbia , Desmocolinas , Desmogleína 1 , Desmogleína 3 , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Humanos , Immunoblotting , Imunoglobulina A/análise , Glicoproteínas de Membrana/análise , Biologia Molecular/métodosRESUMO
GPIIb/IIIa, the human platelet glycoprotein complex, is the autoantigen most commonly recognized by autoantibodies in autoimmune thrombocytopenic purpura (AITP). Two murine monoclonal antibodies (mAbs), namely Y2/51 and 5B12, directed against gpIIIa and gpIIb/IIIa, respectively, and rabbit anti-human platelet polyclonal antibodies have been used to select AITP-related epitopes from a phage display peptide library expressing random dodecapeptides in the pIII coat protein of M13 phage. The selected phage clones were tested by ELISA for binding to rabbit anti-human platelet antibodies as well as to sera from AITP patients. Seven clones reacted strongly with rabbit anti-human platelet antibodies, and four clones reacted with sera from AITP patients. Some homology between peptide inserts sequences of selected clones and human platelet gpIIIa and gpIb were found.
Assuntos
Autoantígenos/análise , Plaquetas/imunologia , Epitopos/análise , Mimetismo Molecular , Púrpura Trombocitopênica Idiopática/imunologia , Animais , Bacteriófago M13/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Camundongos , Oligopeptídeos/análise , Oligopeptídeos/imunologia , Biblioteca de Peptídeos , Mapeamento de Peptídeos , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/imunologia , Coelhos , Homologia de Sequência de AminoácidosAssuntos
Autoanticorpos/sangue , Caderinas/imunologia , Doenças Endêmicas , Pênfigo/imunologia , Autoantígenos/análise , Autoantígenos/imunologia , Brasil , Caderinas/análise , Desmogleína 1 , Desmogleína 3 , Humanos , Indígenas Sul-Americanos , Recém-Nascido , Queratinócitos/química , Pênfigo/sangueRESUMO
Pemphigus describes a group of autoimmune diseases characterized by blisters and erosions of the skin and mucous membranes, acantholysis by histology, and autoantibodies directed against epidermal cell surface components. Since the early 1970s, the following new clinical variants of pemphigus have been reported: pemphigus herpetiformis, IgA pemphigus, and paraneoplastic pemphigus. In recent years, significant data have been obtained from laboratory investigation on these rare and atypical variants, especially regarding their specific target antigens. We review these variants, their clinical presentations, histologic findings, immunopathology, target antigens, theories of pathogenesis, treatment modalities, and clinical courses.
Assuntos
Humanos , Acantólise/patologia , Autoanticorpos/imunologia , Autoantígenos/análise , Dermatite Herpetiforme/classificação , Doenças Autoimunes/classificação , Epiderme/imunologia , Imunoglobulina A/imunologia , Prognóstico , Pênfigo/classificação , Pênfigo/imunologia , Pênfigo/patologia , Síndromes Paraneoplásicas/classificação , Síndromes Paraneoplásicas/imunologiaRESUMO
Background. Guttate psoriasis is associated with infections by Streptococcus pyogenes and cross-reaction between skin and streptococcal antigens have been reported, suggesting an autoimmune component in the disease. Methods. In this work, the authors looked for antibodies against S. pyogenes M-5 antigens by immunoblot in 52 sera of psoriasis patients and in 52 sera of normal individuals. Histological and immunohistochemical analysis in skin biopsies from lesions of another group of 16 clinically diagnosed guttate psoriasis patients and four healthy controls were also carried out. Results. All guttate psoariasis patients studied (11) had IgG antibodies that intensively recognized three different proteins of 70,60 and 14 kDa, as compared to sera from patients with other forms of psoriasis or from healthy controls. The diagnosis of psoariasis was confirmed in 14 of the patients by hematoxylin-eosin staining. Of the other two patients, one was diagnosed as parapsoriasis and the other as liquen. By indirect immunofluorescence (IFI), all 14 psoriatic patients had autoantibodies against their own lesional skin that did not recognized normal skin from control subjects or from the two non-psoriatic patients. The parapsoriatic and the liquen patients did not have autoantibodies. A rabbit immune serum against S. pyogenes antigens reacted with lesional skin from the 14 guttate psoriatic patients, but not with normal skin from controls or with lesional skin from the 2 non-psoriatic patients. Conclusions. The recognition by immunoblot of streptococcal antigens by serum of guttate psoriasis patients, the presence of autoantibodies against their own skin, and recognition of the same skin antigens by anti-streptococcal rabbit antibodies confirm the participation of the immune system and of streptococcal infection in guttate psoriasis
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Autoantígenos/análise , Estudos de Casos e Controles , Infecções Estreptocócicas/imunologia , Psoríase/imunologia , Psoríase/microbiologia , Pele/imunologiaRESUMO
PURPOSE AND METHODS: We investigated the expression and localization of topoisomerase I by Western blot and indirect fluorescent antibody assay, respectively, using anti-Scl-70/topo I from patients with diffuse scleroderma. The contribution of topoisomerase I to DNA replication was assessed using cells treated with the topoisomerase I inhibitor camptothecin. RESULTS: Scl-topo I was detected at all cell cycle phases as a single immunoreactive band of 100 kDa. Extracts from cells in the S phase contained the largest amount of immunoreactive Scl-70/topo I. Variations in the subcellular distribution of Scl-70/topo I were seen throughout the cell cycle, with a speckled nucleoplasmic distribution during G1 contrasting with concentration within the nucleolus during S. Camptothecin exposure blocked topoisomerase I expression and caused a significant decrease in DNA production. CONCLUSION: These data suggest (1) that topomerase I is active mainly during the S phase and contributes to DNA replication, and (2) that topoisomerase I may be involved in ribosomal gene transcription.
Assuntos
DNA Topoisomerases Tipo I/metabolismo , Proteínas Nucleares/metabolismo , Antineoplásicos Fitogênicos/farmacologia , Autoantígenos/análise , Autoantígenos/genética , Autoantígenos/metabolismo , Western Blotting , Camptotecina/farmacologia , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/imunologia , Divisão Celular/efeitos dos fármacos , Replicação do DNA/efeitos dos fármacos , Replicação do DNA/imunologia , DNA Topoisomerases Tipo I/análise , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Hidroxiureia/farmacologia , Índice Mitótico , Proteínas Nucleares/antagonistas & inibidores , Proteínas Nucleares/genética , Inibidores da Síntese de Ácido Nucleico/farmacologia , Fase S , Escleroderma Sistêmico/enzimologia , Escleroderma Sistêmico/metabolismo , Inibidores da Topoisomerase I , Células Tumorais Cultivadas/química , Células Tumorais Cultivadas/citologia , Células Tumorais Cultivadas/enzimologiaRESUMO
We describe a myofibroblastic proliferation in the neck and lower part of the face involving skin and muscle of a 68-year-old female patient with an IgG kappa myeloma. Biopsies showed a fusocellular proliferation with scarce pseudoganglion cells involving the superficial fascia and the cutaneous muscle of the neck. The proliferative cells showed immunohistochemical and ultrastructural features characteristic of myofibrobasts with a proliferating cell nuclear antigen index of 48%; 42% of the cells displayed HLADR-positive membrane staining. Cellular proliferation subsided following the use of immunosuppressive drugs. Eight months after initial consultation, the patient developed polymyositis without a proliferative component and died of aplastic anemia.