RESUMEN

Fifty years have passed since anti-mitochondrial antibodies were found in patients with primary biliary cirrhosis (PBC). PBC is an autoimmune hepatic disease in which 85-90% of patient antibodies bind to mitochondrial antigens that include pyruvate dehydrogenase complex (PDC)-E2 and other members of the oxaloacid dehydrogenase family. In addition, indirect immunofluorescence (IIF) assays utilizing HEp-2 cell substrates have been used to identify anti-centromere antibodies in 20-30% of PBC sera. These antibodies are generally easily recognized, however, anti-nuclear envelope and anti-multiple nuclear dot antibodies are occasionally more difficult to recognize with certainty by IIF. The use of enzyme linked immunosorbent assays that utilize recombinant gp210 (an autoantigen of the nuclear envelope) and/or sp100 (a protein target represented by multiple nuclear dots) should be particularly considered in anti-mitochondrial antibody negative PBC sera. Although the clinical significance of these antibodies still remains to be determined, there is evidence that the existence of anti-gp210 antibodies are related to poorer prognosis and more aggressive disease progression.

Asunto(s)

Autoanticuerpos/sangre , Cirrosis Hepática Biliar/inmunología , Centrómero/inmunología , Humanos , Cirrosis Hepática Biliar/diagnóstico , Mitocondrias/inmunología

RESUMEN

One major pathogenic factor of Porphyromonas gingivalis is Arg-gingipain (Rgp), an arginine-specific cysteine proteinase. To clarify the effect of rgpA DNA vaccine, we immunized BALB/c mice via the abdomen with a Gene Gun or via the nasal cavity weekly for 6 weeks. After immunization, the mice were challenged orally with P. gingivalis. Immunization elicited IgG responses against P. gingivalis in both groups. Nasal immunization also induced sIgA against P. gingivalis, although Gene Gun immunization did not. Reduction of alveolar bone loss was observed in both groups at 42 days following initial infection. This effect was more pronounced in the intranasal immunization group than in the Gene Gun group. The results of this study suggest that immunization with rgpA DNA vaccine via the nasal cavity is an effective method for preventing alveolar bone loss incurred by infection with P. gingivalis.

Asunto(s)

Adhesinas Bacterianas/inmunología , Pérdida de Hueso Alveolar/microbiología , Pérdida de Hueso Alveolar/prevención & control , Vacunas Bacterianas/uso terapéutico , Cisteína Endopeptidasas/inmunología , Porphyromonas gingivalis/inmunología , Vacunas de ADN/uso terapéutico , Administración Intranasal , Análisis de Varianza , Animales , Anticuerpos Antibacterianos/biosíntesis , Vacunas Bacterianas/administración & dosificación , Vacunas Bacterianas/química , Biolística , Femenino , Cisteína-Endopeptidasas Gingipaínas , Ratones , Ratones Endogámicos BALB C , Porphyromonas gingivalis/enzimología , Proteínas Recombinantes , Estadísticas no Paramétricas , Vacunas de ADN/administración & dosificación

RESUMEN

We previously reported that 12.5% of primary biliary cirrhosis (PBC) sera reacted with a 95 kDa cytosol protein (p95c) that was subsequently identified as a p97/valosin-containing protein (VCP). The clinical features and course of the six anti-p97/VCP-positive PBC patients with Scheuer's stage 1 and 2 liver biopsies were monitored for an average of 15 years. This group was compared with 50 PBC patients that did not have detectable anti-VCP. Autoantibodies to a full-length recombinant p97/VCP were assayed by immunoprecipitation. All six PBC patients with anti-VCP had antibodies to the mitochondrial pyruvate dehydrogenase complex-E2 antigen as measured by an addressable laser bead immunoassay. The first was a male with no evidence of liver failure that died of cerebral infarction at the age of 85. The second was a 73-year-old female with Hashimoto's thyroiditis who has remained clinically stable without ursodeoxycolic acid (UDCA) treatment. Although the third had no HCV antibodies, he developed hepatocellular carcinoma at the age of 76 and died of renal failure at 78. The fourth was a 50-year-old female who remained clinically stable during follow-up and the fifth with Hashimoto's thyroiditis and stable liver function following UCDA treatment. The sixth was a male patient presenting a mild clinical course. The clinical course of these patients was in contrast to the 50 comparison group PBC patients who did not have anti-p97/VCP. As the six PBC patients with anti-p97/VCP antibodies had slowly progressive liver disease and no mortality related to autoimmune liver disease, our observations suggest that this autoantibody might be an indicator of a favourable prognosis.

Asunto(s)

Autoanticuerpos/sangre , Proteínas de Ciclo Celular/inmunología , Cirrosis Hepática Biliar/diagnóstico , Adenosina Trifosfatasas , Anciano , Anciano de 80 o más Años , Biomarcadores/sangre , Femenino , Humanos , Inmunoprecipitación , Hígado/patología , Cirrosis Hepática Biliar/inmunología , Cirrosis Hepática Biliar/patología , Masculino , Persona de Mediana Edad , Pronóstico , Proteína que Contiene Valosina

RESUMEN

The nuclear envelope (NE) is one of many intracellular targets of the autoimmune response in patients with autoimmune liver disease, systemic lupus erythematosus, and related conditions. In eukaryotic organisms the NE consists of five interconnected regions: an outer nuclear membrane (ONM) that is continuous with the endoplasmic reticulum, an intermembrane or perinuclear space, an inner nuclear membrane (INM) with a unique set of integral membrane proteins, the underlying nuclear lamina, and the pore domains that are regions where the ONM and INM come together. The pore domains are sites of regulated continuity between the cytoplasm and nucleus that are occupied by supramolecular structures, termed nuclear pore complexes (NPCs). Human autoantibodies identified to date bind to specific components in three of the five NE compartments. Autoantigen targets include the lamins A, B, and C of the nuclear lamina, gp210, p62 complex proteins, Nup153, and Tpr within the NPC, and LBR, MAN1, LAP1, and LAP2 that are integral proteins of the INM. Autoantibodies to these NE targets have been shown to be correlated with various autoimmune diseases such as primary biliary cirrhosis, other autoimmune liver diseases and systemic rheumatic diseases. Now that the proteome of the NE is more clearly defined, other autoantibodies to components in this cell compartment are likely to be defined.

Asunto(s)

Autoantígenos/inmunología , Lámina Nuclear/metabolismo , Proteínas de Complejo Poro Nuclear/metabolismo , Poro Nuclear/metabolismo , Proteínas Nucleares/metabolismo , Animales , Anticuerpos Antinucleares/inmunología , Autoantígenos/metabolismo , Enfermedades Autoinmunes/inmunología , Humanos , Lámina Nuclear/inmunología , Poro Nuclear/inmunología , Poro Nuclear/ultraestructura , Proteínas de Complejo Poro Nuclear/inmunología , Proteínas Nucleares/inmunología

RESUMEN

We have reported previously that p95c, a novel 95-kDa cytosolic protein, was the target of autoantibodies in sera of patients with autoimmune hepatic diseases. We studied 30 sera that were shown previously to immunoprecipitate a 95 kDa protein from [(35)S]-methionine-labelled HeLa lysates and had a specific precipitin band in immunodiffusion. Thirteen sera were available to test the ability of p95c antibodies to inhibit nuclear envelope assembly in an in vitro assay in which confocal fluorescence microscopy was also used to identify the stages at which nuclear assembly was inhibited. The percentage inhibition of nuclear envelope assembly of the 13 sera ranged from 7% to 99% and nuclear envelope assembly and the swelling of nucleus was inhibited at several stages. The percentage inhibition of nuclear assembly was correlated with the titre of anti-p95c as determined by immunodiffusion. To confirm the identity of this autoantigen, we used a full-length cDNA of the p97/valosin-containing protein (VCP) to produce a radiolabelled recombinant protein that was then used in an immunoprecipitation (IP) assay. Our study demonstrated that 12 of the 13 (93%) human sera with antibodies to p95c immunoprecipitated recombinant p97/VCP. Because p95c and p97 have similar molecular masses and cell localization, and because the majority of sera bind recombinant p97/VCP and anti-p95c antibodies inhibit nuclear assembly, this is compelling evidence that p95c and p97/VCP are identical.

Asunto(s)

Autoanticuerpos/inmunología , Proteínas de Ciclo Celular/inmunología , Cirrosis Hepática Biliar/inmunología , Adenosina Trifosfatasas , Reacciones Antígeno-Anticuerpo , Núcleo Celular/inmunología , Femenino , Humanos , Hígado/patología , Cirrosis Hepática Biliar/patología , Masculino , Microscopía Confocal , Pruebas de Precipitina , Proteína que Contiene Valosina

RESUMEN

We report on an 80-year-old man with rheumatoid arthritis (RA) who presented with chronic myelogenous leukemia (CML). Five years after the onset of RA, the CML diagnosis was made. The patient was treated for CML with 300 mg of imatinib mesylate (STI; signal transduction inhibitor 571) for 8 weeks. Laboratory tests showed that the C-reactive protein level, percentage of cells exhibiting the Philadelphia chromosome (Ph1), WBC count, and Lansbury index for RA all dropped respectively from 7.5 mg/dl to 1.0 mg/dl, 74.9% to 1%, 25, 100/microl to 9900/microl, and 51% to 14%. Administration of imatinib mesylate is felt to be effective in treating not only CML but also RA in the active stage.

Asunto(s)

Antineoplásicos/administración & dosificación , Artritis Reumatoide/complicaciones , Leucemia Mielógena Crónica BCR-ABL Positiva/complicaciones , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Piperazinas/administración & dosificación , Pirimidinas/administración & dosificación , Anciano , Anciano de 80 o más Años , Artritis Reumatoide/diagnóstico , Benzamidas , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Estudios de Seguimiento , Humanos , Mesilato de Imatinib , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Masculino , Dimensión del Dolor/efectos de los fármacos , Rango del Movimiento Articular/efectos de los fármacos , Medición de Riesgo , Índice de Severidad de la Enfermedad , Resultado del Tratamiento

RESUMEN

Telomeres of a specific length are essential for continuous cell proliferation. The length of telomeres must be maintained by telomerase action and the telomeric DNA-repeat binding protein must be protected. Therefore, there seems to be a relationship between cell immortality due to telomerase activity and telomeric DNA-repeat binding protein. We examined telomerase activity and the expression of telomeric-repeat binding factor 1 and 2 (TRF1 and TRF2) in gastric cancer. Telomerase activity was semi-quantified using the f-TRAP technique in 53 cancerous and non-cancerous gastric tissue specimens. TRF1 and TRF2 were also studied using an immunohistochemical method to determine the frequency of these factors in cell nuclei. Telomerase activity was observed in 79.2% of the cancerous tissue and in 39.6% of the non-cancerous tissue. The average semi-quantitative values for telomerase activity were 67.3 total product generated (TPG) unit/microg protein in cancerous tissue and 6.0 TPG unit/microg protein in non-cancerous tissue. Moreover, T0/1 tumor had the same incidence of telomerase activity as T2 or deeper tumors. These results indicated that the activation of telomerase begins at an early stage of carcinogenesis. TRF1 and TRF2 were detected in 45.1% and 42.9% of the cancerous tissue and in 70.6% and 65.6% of the non-cancerous tissue, respectively. In addition, low positive staining ratios were found for TRF1 and TRF2 when cancer had more deeply invaded. However, telomerase activity did not correlate with either TRF1 or TRF2. These findings suggest that optimal conditions for efficient telomerase are produced as cancer progresses, via suppression of TRFs.

Asunto(s)

Neoplasias Gástricas/metabolismo , Telomerasa/metabolismo , Proteína 1 de Unión a Repeticiones Teloméricas/metabolismo , Proteína 2 de Unión a Repeticiones Teloméricas/metabolismo , Cartilla de ADN/química , Humanos , Técnicas para Inmunoenzimas , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Telomerasa/genética , Proteína 1 de Unión a Repeticiones Teloméricas/genética , Proteína 2 de Unión a Repeticiones Teloméricas/genética

RESUMEN

Abstract An 81-year-old man who had previously shown high levels of alkaline phosphatase (ALP), γ-glutamyltransferase (GTP), and total bilirubin presented with acute liver damage. He was positive for serum anti-gp210 and anti-p62 antibodies, but negative for serum antimitochondrial antibody. A liver biopsy revealed massive interstitial fibrosis and pseudolobulus, which were compatible with a diagnosis of primary biliary cirrhosis (PBC) at Scheuer's stage 4. He was given ursodeoxycolic acid at 600 mg/day. However, his condition deteriorated, and he eventually died of hepatic insufficiency in a state of malnutrition. We hypothesize that the presence of anti-gp210 and anti-p62 complex protein antibodies, rather than that of antimitochondrial antibodies, was correlated with the progression of PBC in this particular case.

RESUMEN

Abstract A 60-year-old woman visited the Keigu Clinic in January 1998 complaining of morning stiffness and flexion contracture of the distal interphalangeal joint. Blood tests showed the presence of antinuclear antibody at a 1 : 40 dilution with speckled staining. She was suspected of having Heberden's node. Nine months later, she developed Raynaud's phenomenon, sclerodactyly, and Gottron's sign, and was diagnosed with systemic sclerosis/dermatomyositis (SSc/DM) overlap. Blood tests revealed the presence of antinuclear antibody at a 1 : 5120 dilution, along with high titer of anti-Ku and anti-Ki antibodies. Subsequently, the patient developed interstitial pneumonia in January 2000. It is thought that the appearance of antinuclear antibody and development of other immunological events played an important role in determining this patient's limited SSc/DM overlap.

RESUMEN

OBJECTIVE: To identify antinuclear antibodies (ANA) specific for chronic fatigue syndrome (CFS), and in related conditions such as fibromyalgia (FM) or psychiatric disorders. METHODS: One hundred and fourteen CFS patients and 125 primary and secondary FM patients were selected based on criteria advocated by the Centers for Disease Control and Prevention and by the American College of Rheumatology, respectively. As controls, healthy subjects and patients with either various psychiatric disorders or diffuse connective tissue diseases were included. Autoantibodies were examined by immunoblot utilizing HeLa cell extracts as the antigen. RESULTS: Autoantibodies to a 68/48 kDa protein were present in 13.2 and 15.6% of patients with CFS and primary FM, respectively. In addition, autoantibodies to a 45 kDa protein were found in 37.1 and 21.6% of the patients with secondary FM and psychiatric disorders, respectively. Meanwhile, these two autoantibodies were not found at all in connective tissue disease patients without FM, nor in healthy subjects (P<0.05). As a group, the anti-68/48 kDa-positive CFS patients presented more frequently with hypersomnia (P<0.005), short-term amnesia (P<0.07) or difficulty in concentration (P<0.05) than those CFS patients without the antibodies. CONCLUSIONS: The presence of the anti-68/48 kDa protein antibodies in a portion of both CFS and primary FM patients suggests the existence of a common immunological background. These antibodies may find utility as possible markers for a clinicoserological subset of CFS/FM patients with hypersomnia and cognitive complaints.

Asunto(s)

Anticuerpos Antinucleares/sangre , Trastornos del Conocimiento/sangre , Trastornos de Somnolencia Excesiva/sangre , Síndrome de Fatiga Crónica/inmunología , Fibromialgia/inmunología , Adulto , Biomarcadores/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Immunoblotting , Masculino , Persona de Mediana Edad , Membrana Nuclear/inmunología , Proteínas Nucleares/análisis

RESUMEN

Detection of anti-nuclear antibodies (ANA) is essential for diagnosing autoimmune diseases including autoimmune liver diseases. An indirect immunofluorescence (IIF) method with a cell line (HEp-2) derived from human laryngeal carcinoma has been used as a standard substrate. Recently, an enzyme-linked immunosorbent assay (ELISA) using multiple solid-phase antigens has been developed. We assayed sera from 272 cases of chronic liver diseases, 91 cases of healthy subjects and studied clinical significance of ANA. The sensitivity of IIF method in detection of ANA (fluorescence-ANA: FANA) and that of ELISA (ELISA-ANA: EANA) were 19.2% and 17.3% in chronic hepatitis B (CH-B), 16.7% and 17.3% in chronic hepatitis C (CH-C), 84.2% and 50.9% in primary biliary cirrhosis (PBC), 100% and 85.7% in autoimmune hepatitis (AIH) and 15.4% and 18.7% in healthy subjects. The sensitivity of EANA was considerably lower than that of FANA in PBC and AIH, but the sensitivity was the same in CH-C, CH-B, and healthy subjects. Because the solid-phase target antigens do not include nuclear antigen components recognized only by patients with PBC or AIH, ELISA can not detect all the species of ANA. This accounts for the low sensitivity of EANA in PBC and AIH. In conclusion, the current EANA is useful for screening of ANA, but FANA should be performed when PBC or AIH is suspected.

Asunto(s)

Anticuerpos Antinucleares/sangre , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente Indirecta , Hepatopatías/inmunología , Enfermedad Crónica , Femenino , Humanos , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad

RESUMEN

Sera from patients with primary biliary cirrhosis recognize various cellular components, such as mitochondria, centromere, nuclear envelope, and multiple nuclear dot antigens. There also appears to be a novel antibody reacting with a particular protein in these sera. The presence of this antibody was investigated by double immunodiffusion using rat liver cytoplasmic antigens, by immunoprecipitation of [35S]-methionine labelled HeLa cell extracts, and by immunoblot using disrupted HeLa cell extracts. Test sera were obtained from 491 patients with various liver diseases. Nine of the 491 sera were found to react with a 95-kDa protein as determined by immunoprecipitation of [35S]-methionine labelled HeLa cell extracts and by double immunodiffusion using a rat liver microsomal preparation. However, these same nine sera showed no reaction in the immunoblot assay. On the basis of its molecular mass and its presence in the cytoplasmic fraction, this antigen was named p95 C. This anti-p95 C antibody was detected in six of 50 (12%) sera from patients with primary biliary cirrhosis, and in three of 31 (9.7%) sera from patients with autoimmune hepatitis, but not in any of the remaining 410 sera obtained from patients with other hepatic diseases. It is concluded that anti-p95 C antibody reacts primarily with the native form of the 95-kDa protein, and represents another possible analyte for diagnosing autoimmune liver diseases.

Asunto(s)

Anticuerpos/inmunología , Enfermedades Autoinmunes/inmunología , Hepatopatías/inmunología , Anciano , Anciano de 80 o más Años , Secuencia de Aminoácidos , Animales , Anticuerpos/sangre , Reacciones Antígeno-Anticuerpo , Enfermedades Autoinmunes/sangre , Femenino , Células HeLa , Humanos , Immunoblotting , Inmunodifusión , Hepatopatías/sangre , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Pruebas de Precipitina , Conformación Proteica , Ratas

Asunto(s)

Enfermedades del Colágeno/diagnóstico , Patología Clínica , Antiinflamatorios/administración & dosificación , Anticuerpos Antinucleares/análisis , Biomarcadores/análisis , Enfermedades del Colágeno/fisiopatología , Enfermedades del Colágeno/terapia , Humanos , Inmunosupresores/administración & dosificación , Factor Reumatoide/análisis , Transducción de Señal , Esteroides , Linfocitos T/inmunología

RESUMEN

OBJECTIVE: To identify a Golgi complex autoantigen bound by Sjögren's syndrome (SS) autoantibodies. METHODS: Serum from a patient with secondary SS and anti-Golgi antibodies was used as a probe to isolate a complementary DNA (cDNA) insert from a HeLa cDNA library. RESULTS: A 3.7-kb cDNA encoding a 56-kd recombinant protein was immunoprecipitated by the human anti-Golgi serum and immune rabbit serum. Western blot analysis showed that the immune rabbit sera recognized a protein of 97 kd (golgin-97), suggesting that the isolated clone contained a partial cDNA. The 5' upstream sequence was obtained by rapid amplification of the cDNA ends. The complete cDNA contained 4,860 basepairs, encoding a protein with a calculated Mr of 88 kd. Antibodies to golgin-97 were found in 12 (20%) of 60 sera known to have anti-Golgi autoantibodies, and the majority of these sera (8 of 12, or 75%) were from patients who had secondary SS. CONCLUSION: Golgin-97 is a unique Golgi complex antigen that appears to be a target of SS autoantibodies.

Asunto(s)

Autoantígenos/genética , Aparato de Golgi/inmunología , Síndrome de Sjögren/inmunología , Adulto , Secuencia de Aminoácidos , Autoanticuerpos/análisis , Autoantígenos/inmunología , Secuencia de Bases , Línea Celular , Clonación Molecular , Cartilla de ADN/química , ADN Complementario/aislamiento & purificación , Femenino , Proteínas de la Matriz de Golgi , Humanos , Datos de Secuencia Molecular , Peso Molecular

RESUMEN

Two cases of systemic lupus erythematosus (SLE) with autoimmune hepatitis are reported. Both patients had a mild form of SLE without central nervous system or renal involvement and showed a rapid response to corticosteroid therapy. Neither of them had antibodies to mitochondria, smooth muscle, and liver/kidney microsome-1 related to autoimmune hepatitis. Instead, novel autoantibodies which react with transfer RNA-related antigen were detected. These autoantibodies could be a useful marker for classification of SLE associated with autoimmune hepatitis.

Asunto(s)

Autoanticuerpos/análisis , Enfermedades Autoinmunes/complicaciones , Hepatitis Crónica/complicaciones , Lupus Eritematoso Sistémico/complicaciones , ARN de Transferencia/inmunología , Adulto , Enfermedades Autoinmunes/diagnóstico , Enfermedades Autoinmunes/inmunología , Biomarcadores/sangre , Diagnóstico Diferencial , Femenino , Hepatitis Crónica/diagnóstico , Hepatitis Crónica/inmunología , Humanos , Hígado/inmunología , Hígado/patología , Lupus Eritematoso Sistémico/diagnóstico , Lupus Eritematoso Sistémico/inmunología

Asunto(s)

Anemia Hemolítica/etiología , Prueba de Coombs , Deficiencia del Factor V/etiología , Hemorragia/etiología , Anciano , Anemia Hemolítica/sangre , Anemia Hemolítica/diagnóstico , Antibacterianos/efectos adversos , Autoinmunidad , Hemorragia/sangre , Humanos , Inmunoglobulina G/sangre , Infecciones/complicaciones , Masculino

RESUMEN

A 41-year-old woman was admitted to Tokyo Women's Medical College Aoyama Hospital in January 1994. She presented cough and dyspnea in September 1991. The diagnosis of interstitial pneumonia was made based on TBLB. Interstitial pneumonia was responsive to initial prednisolone of 40 mg daily. When dose of prednisolone was reduced to 15 mg daily, she complained of cough and dyspnea again. She was referred to Institute of Rheumatology, Tokyo Women's Maedical College in November 1993. She was diagnosed as systemic sclerosis associated with interstitial pneumonia based on proximal scleroderma and digital pitting scar. Double immunodiffusion and immunoprecipitation assay revealed the presence of anti-Ki, anti-Wa, and anti-RNA polymerases antibodies in the serum. On admission in March 1994, she was treated with intravenous cyclophosphamide therapy at 500 mg/day once a mouth. After the third infusion, respiratory symptoms, pulmonary function test values, findings of chest X-ray and CT scan were improved without adverse drug effects. Intravenous cyclophosphamide therapy seems to be useful in this case. The efficacy of Intravenous cyclophosphamide therapy in the treatment of the interstitial pneumonia associated with systemic sclerosis was discussed.

Asunto(s)

Autoanticuerpos/análisis , Ciclofosfamida/administración & dosificación , Inmunosupresores/administración & dosificación , Enfermedades Pulmonares Intersticiales/complicaciones , Esclerodermia Sistémica/complicaciones , Adulto , Autoantígenos/inmunología , ARN Polimerasas Dirigidas por ADN/inmunología , Femenino , Humanos , Infusiones Intravenosas , Enfermedades Pulmonares Intersticiales/tratamiento farmacológico , Enfermedades Pulmonares Intersticiales/inmunología , Proteínas Nucleares/inmunología , Complejo de la Endopetidasa Proteasomal , Esclerodermia Sistémica/tratamiento farmacológico , Esclerodermia Sistémica/inmunología

RESUMEN

Anti-centromere antibody (ACA) have been recognized in sera of patients with primary biliary cirrhosis (PBC) and CREST syndrome. The major reactive antigen of ACA have been identified as CENP-B (80kDa). Using an indirect immunofluorescence (IIF) method and ELISA method, we detected ACA and anti-CENP-B antibody in patients with PBC and various liver diseases and collagen diseases. We tested sera of 44 patients with PBC, 8 patients with autoimmune hepatitis (AIH), 51 patients with chronic hepatitis B (CH-B), 312 patients with chronic hepatitis C(CH-C), 12 patients with progressive systemic sclerosis (PSS), 10 patients with systemic lupus erythematosus (SLE), 10 patients with rheumatoid arthritis (RA), and 30 with healthy subjects (HS). ACA was detected by IIF technique, using HEp-2 cell and fluoro-CENTRO slides (MBL) as substrates. Anti-CENP-B antibody was detected by ELISA method using recombinant CENP-B (MBL) as the antigen. ACA was detected in sera of 12 (27%) patients with PBC, two (25%) patients with AIH, five (2%) patients with CH-C, nine (75%) patients with PSS, and one (10%) patients with RA. ACA was not detected in sera of patients with CH-B and SLE and in HS. The results of IIF test for ACA, using HEp -2 cells and fluoro-CENTRO slides, were completely agreed. Anti-CENP-B antibody was detected in 28(97%) out of 29 patients sera positive for ACA. The titers of ACA and anti-CENP-B antibody did not show a correlation (r = 0.24). Out of 12 sera, in which, the titers of anti-CENP-B antibody was over 400. Among them, eight were patients with PBC and four were PSS. Later, out of four patients with PSS, three (75%) were found to be positive for anti-mitochondrial antibody. Out of five patients, in which the titer of anti-CENP-B antibody showed over 800, all were patients with PBC. The titers of ACA have no relationship with PBC. However, the titers of anti-CENP-B antibody have closed relationship with PBC. The reason why the titers of ACA and anti-CENP-B antibody were not correlated is unknown. We consider anti-CENP-B antibody is a new marker of a subset of PBC, because almost all the patients were PBC when this antibody showed over 400.

Asunto(s)

Autoanticuerpos/sangre , Síndrome CREST/diagnóstico , Centrómero/inmunología , Proteínas de Unión al ADN , Cirrosis Hepática Biliar/diagnóstico , Artritis Reumatoide/diagnóstico , Autoantígenos/inmunología , Enfermedades Autoinmunes/diagnóstico , Proteína B del Centrómero , Proteínas Cromosómicas no Histona/inmunología , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Hepatitis/diagnóstico , Hepatitis C/diagnóstico , Humanos

RESUMEN

OBJECTIVE: To identify and characterize a novel autoantibody, anti-WS, that binds total transfer RNA (tRNA). METHODS: Serum from patient WS, who had polyarthritis, Sjögren's syndrome, Raynaud's phenomenon, and interstitial pulmonary fibrosis, was used in this study. Characteristics of anti-WS and antibody-reactive determinants of tRNA were investigated by 32P immunoprecipitation using HeLa cell RNA and deletion mutants of tRNA transcribed in vitro. RESULTS: WS serum produced nucleolar and cytoplasmic staining on indirect immunofluorescence. 32P immunoprecipitation assays demonstrated that this serum immunoprecipitated total tRNAs and 5.8S and 5S ribosomal RNAs from 32P-labeled HeLa cell extract. When deproteinized RNA was used as antigen source, total tRNAs were still precipitated by WS serum. An immunoprecipitation study, using various deletion mutants of Escherichia coli tRNA, demonstrated that both D and T psi C loops were needed for antibody binding. Substitution of nucleotide 18G with 18A of E coli tRNA(Trp), which is essential in the formation of the tertiary "L" shape of tRNA, inhibited binding by anti-WS antibodies. CONCLUSION: Anti-WS antibodies are novel autoantibodies directed against tRNAs. The antibody binding site is the common L-shaped tertiary structure conformed by the D loop and T psi C loop of tRNA, suggesting that the antibodies are induced by a conserved sequence among all species. Furthermore, these antibodies could be a marker for a newly recognized subset of connective tissue disease.

Asunto(s)

Autoanticuerpos/análisis , Enfermedades Pulmonares Intersticiales/inmunología , ARN de Transferencia/inmunología , Escherichia coli/química , Femenino , Técnica del Anticuerpo Fluorescente Indirecta , Eliminación de Gen , Humanos , Persona de Mediana Edad , Mutagénesis/inmunología , Radioisótopos de Fósforo , Pruebas de Precipitina , Conformación Proteica , Estructura Terciaria de Proteína , ARN de Transferencia/química , ARN de Transferencia/genética , ARN de Transferencia de Aspártico/química , ARN de Transferencia de Aspártico/genética , ARN de Transferencia de Aspártico/inmunología , ARN de Transferencia de Histidina/química , ARN de Transferencia de Histidina/genética , ARN de Transferencia de Histidina/inmunología , ARN de Transferencia de Serina/química , ARN de Transferencia de Serina/genética , ARN de Transferencia de Serina/inmunología , ARN de Transferencia de Triptófano/química , ARN de Transferencia de Triptófano/genética , ARN de Transferencia de Triptófano/inmunología , ARN de Transferencia de Valina/química , ARN de Transferencia de Valina/genética , ARN de Transferencia de Valina/inmunología

RESUMEN

We have recently observed reactivity of primary biliary cirrhosis (PBC) sera with several proteins bearing N-acetylglucosamine residues from rat liver nuclear envelopes. The aim of this study was to characterize the reactive antigens. Sera from 31 patients with PBC, 30 with rheumatoid arthritis (RA) and 30 with Sjögren's syndrome (SS) were examined. Rim-like immunofluorescence staining was observed in 15 of 31 (48%) sera from patients with PBC, in 1 of 30 with RA and in 1 of 30 with SS. Upon immunoblotting using preparations of whole rat liver nuclear envelopes and their Triton X 100-KCl extract as antigen sources, a 200 kDa protein band was observed in 9 of sera with PBC. Furthermore, upon immunoblotting using the wheat germ aggulutinin-bound fraction of rat liver envelope as antigen, 62, 60 and 54 kDa protein bands corresponding to components of the p62 complex in the nuclear pore complex (Kita et al. Biochem. 113, 377-382) were observed in 7, 5 and 6 samples respectively, of the 31 PBC sera. Our data suggest that PBC sera recognize not only the 210 kDa protein but also the p62 complex proteins.

Asunto(s)

Cirrosis Hepática Biliar/inmunología , Hígado/inmunología , Glicoproteínas de Membrana/inmunología , Proteínas Nucleares/inmunología , Acetilglucosamina/inmunología , Animales , Anticuerpos Antinucleares/inmunología , Humanos , Cirrosis Hepática Biliar/sangre , Proteínas de Complejo Poro Nuclear , Ratas