RESUMEN

OBJECTIVE: Chlamydia trachomatis and Chlamydophila (Chlamydia) pneumoniae are known triggers of reactive arthritis (ReA) and exist in a persistent metabolically active infection state in the synovium, suggesting that they may be susceptible to antimicrobial agents. The goal of this study was to investigate whether a 6-month course of combination antibiotics is an effective treatment for patients with chronic Chlamydia-induced ReA. METHODS: This study was a 9-month, prospective, double-blind, triple-placebo trial assessing a 6-month course of combination antibiotics as a treatment for Chlamydia-induced ReA. Eligible patients had to be positive for C trachomatis or C pneumoniae by polymerase chain reaction (PCR). Groups received 1) doxycycline and rifampin plus placebo instead of azithromycin; 2) azithromycin and rifampin plus placebo instead of doxycycline; or 3) placebos instead of azithromycin, doxycycline, and rifampin. The primary end point was the number of patients who improved by 20% or more in at least 4 of 6 variables without worsening in any 1 variable in both combination antibiotic groups combined and in the placebo group at month 6 compared with baseline. RESULTS: The primary end point was achieved in 17 of 27 patients (63%) receiving combination antibiotics and in 3 of 15 patients (20%) receiving placebo. Secondary efficacy end points showed similar results. Six of 27 patients (22%) randomized to combination antibiotics believed that their disease went into complete remission during the trial, whereas no patient in the placebo arm achieved remission. Significantly more patients in the active treatment group became negative for C trachomatis or C pneumoniae by PCR at month 6. Adverse events were mild, with no significant differences between the groups. CONCLUSION: These data suggest that a 6-month course of combination antibiotics is an effective treatment for chronic Chlamydia-induced ReA.

Asunto(s)

Artritis Reactiva/tratamiento farmacológico , Azitromicina/administración & dosificación , Infecciones por Chlamydia/complicaciones , Chlamydia trachomatis/aislamiento & purificación , Doxiciclina/administración & dosificación , Rifampin/administración & dosificación , Adulto , Anciano , Antibacterianos/administración & dosificación , Antibacterianos/efectos adversos , Antibióticos Antituberculosos/administración & dosificación , Antibióticos Antituberculosos/efectos adversos , Artritis Reactiva/microbiología , Artritis Reactiva/patología , Azitromicina/efectos adversos , Chlamydia trachomatis/genética , Chlamydophila pneumoniae/genética , Chlamydophila pneumoniae/aislamiento & purificación , Enfermedad Crónica , ADN Bacteriano/genética , Método Doble Ciego , Doxiciclina/efectos adversos , Quimioterapia Combinada , Femenino , Humanos , Articulaciones/patología , Masculino , Persona de Mediana Edad , Placebos , Prohibitinas , Estudios Prospectivos , Rifampin/efectos adversos , Resultado del Tratamiento

RESUMEN

Rarely, tumour necrosis factor (TNF)alpha antagonist therapy has been associated with de novo psoriasiform eruptions. This is unusual in that these same drugs are used to treat psoriasis. Most of these cases involve the palms and soles, yet palmoplantar pustular psoriasis represents only 1.7% of all cases of psoriasis. Keratoderma blenorrhagicum is a psoriasiform rash that occurs primarily on the palms and soles of some patients with reactive arthritis. It is grossly and histologically indistinguishable from pustular psoriasis. Chlamydia trachomatis is a common aetiological agent for reactive arthritis, and in vitro studies have shown that chlamydial replication is inversely proportional to TNFalpha levels. Three patients taking TNFalpha antagonists are presented who developed such lesions and who were found to be positive for C trachomatis DNA in the affected skin. It is proposed that these psoriasiform lesions may not be psoriasis, but rather keratoderma blenorrhagicum.

Asunto(s)

Antirreumáticos/efectos adversos , Artritis Reactiva/tratamiento farmacológico , Infecciones por Chlamydia/complicaciones , Queratosis/inducido químicamente , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Adalimumab , Adulto , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales Humanizados , Artritis Reactiva/patología , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/patología , Chlamydia trachomatis , Etanercept , Femenino , Humanos , Inmunoglobulina G/efectos adversos , Infliximab , Receptores del Factor de Necrosis Tumoral , Piel/patología

RESUMEN

Schistosomiasis mansoni is a major helminthic disease of the tropics characterised by chronic hepatic and intestinal granulomatous inflammation and fibrosis. The fibrotic response is regulated by the amount of collagen deposited in the tissues and the degradation of that collagen by matrix metalloproteinases (MMP). In the murine model of the disease, although hepatic granuloma formation and the ensuing fibrosis have been thoroughly examined, there is a dearth of information on the intestinal fibrotic process. The expression of fibrosis-related genes in the colons of chronically infected mice has therefore been investigated. Compared with that seen in uninfected mice, the expression of the genes coding for collagen of types I, III and IV was upregulated. Similarly, the messages for MMP-2, MMP-3 and MMP-8 were elevated, indicating the potential for collagen degradation. The genes for two tissue inhibitors of metalloproteinases (TIMP), TIMP-1 and TIMP-4, were, however, expressed at higher levels than those coding for the MMP. As a corollary, expression of the genes coding for three fibrogenic cytokines, transforming growth factor-beta, tumour necrosis factor and interleukin-4, was elevated. These data indicate that an imbalance in MMP:TIMP expression and enhanced levels of the messages for fibrogenic cytokines underlie the mechanism(s) of the colonic fibrosis seen in mice chronically infected with Schistosoma mansoni.

Asunto(s)

Colágeno/genética , Colon/química , Citocinas/genética , Metaloproteinasas de la Matriz/genética , Esquistosomiasis mansoni/genética , Inhibidores Tisulares de Metaloproteinasas/genética , Animales , Enfermedad Crónica , Colon/patología , Enfermedades del Colon/genética , Modelos Animales de Enfermedad , Femenino , Fibrosis/genética , Genes de Helminto/genética , Granuloma/genética , Íleon/química , Íleon/patología , Interleucina-4/genética , Ratones , Ratones Endogámicos CBA , Factor de Crecimiento Transformador beta/genética , Factor de Necrosis Tumoral alfa/genética , Regulación hacia Arriba/genética

RESUMEN

BACKGROUND: Synovial tissues in patients with Chlamydia associated arthritis are persistently infected by C trachomatis, an organism for which genetic manipulation is not possible. M tuberculosis also engages in persistent infection, and because this bacterium is genetically tractable many groups have been able to define transcriptional characteristics of mycobacterial growth and persistence. OBJECTIVE: To investigate whether the pattern of gene expression underlying chlamydial persistence is similar to that underlying mycobacterial persistence. METHODS: 194 genes in M tuberculosis that are transcriptionally up regulated to support in vivo growth and persistence of that organism have previously been identified. Each of those genes was compared with the C trachomatis genome to identify orthologues. Expression of selected chlamydial orthologues so identified was assessed by real time RT-PCR in an in vitro model of chlamydial persistence and synovial tissues from patients who were PCR positive for C trachomatis at that site. RESULTS: 67 C trachomatis genes were identified as being orthologous to mycobacterial persistence related genes, representing 35% of the genes tested. The chlamydial orthologues fell into similar metabolic and other categories as those in M tuberculosis. Expression of a majority of selected chlamydial orthologues was strongly up regulated in an in vitro model of chlamydial persistence and in synovial tissues of relevant patients, compared with their expression during active infection. CONCLUSIONS: These observations provide new insight into the molecular genetic basis underlying chlamydial persistence, and indicate that this information can be obtained, in some instances, by extrapolating observations made in other biological systems and/or organisms.

Asunto(s)

Artritis Reactiva/microbiología , Infecciones por Chlamydia/microbiología , Chlamydia trachomatis/genética , Genes Bacterianos , Membrana Sinovial/microbiología , Línea Celular Tumoral , Chlamydia trachomatis/crecimiento & desarrollo , Enfermedad Crónica , Regulación Bacteriana de la Expresión Génica , Humanos , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/crecimiento & desarrollo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Especificidad de la Especie , Regulación hacia Arriba

RESUMEN

Schistosomiasis mansoni disseminated worm eggs in mice and humans induce granulomatous inflammations and cumulative fibrosis causing morbidity and possibly mortality. In this study, intrahepatic and I.V. injections of a double-stranded oligodeoxynucleotide decoy containing the TGF-beta regulatory element found in the distal promoter of the COL1A1 gene into worm-infected mice suppressed TGF-beta1, COL1A1, tissue inhibitor of metalloproteinase-1, and decreased COL3A1 mRNAs to a lesser extent. Sequence comparisons within the mouse genome found homologous sequences within the COL3A1, TGF-beta1, and TIMP-1 5' flanking regions. Cold competition gel mobility shift assays using these homologous sequences with 5' and 3' flanking regions found in the natural COL1A1 gene showed competition. Competitive gel mobility assays in a separate experiment showed no competition using a 5-base mutated or scrambled sequence. Explanted liver granulomas from saline-injected mice incorporated 10.45 +/- 1.7% (3)H-proline into newly synthesized collagen, whereas decoy-treated mice showed no collagen synthesis. Compared with the saline control schistosomiasis mice phosphorothioate double-stranded oligodeoxynucleotide treatment decreased total liver collagen content (i.e. hydroxy-4-proline) by 34%. This novel molecular approach has the potential to be employed as a novel antifibrotic treatment modality.

Asunto(s)

Colágeno Tipo I/genética , Cirrosis Hepática/prevención & control , Cirrosis Hepática/parasitología , Oligodesoxirribonucleótidos/genética , Oligonucleótidos/genética , Oligonucleótidos/farmacología , Esquistosomiasis mansoni/complicaciones , Factor de Crecimiento Transformador beta/genética , Animales , Colágeno/antagonistas & inhibidores , Cadena alfa 1 del Colágeno Tipo I , Secuencia de Consenso , ADN , Femenino , Fibroblastos/metabolismo , Granuloma/metabolismo , Granuloma/patología , Hidroxiprolina/antagonistas & inhibidores , Hidroxiprolina/metabolismo , Hígado/metabolismo , Hepatopatías/metabolismo , Hepatopatías/patología , Ratones , Ratones Endogámicos C57BL , Mutación , Miocitos del Músculo Liso/metabolismo , Oligonucleótidos/síntesis química , Esquistosomiasis mansoni/metabolismo , Homología de Secuencia de Ácido Nucleico , Transducción de Señal/efectos de los fármacos , Inhibidor Tisular de Metaloproteinasa-1/genética , Transfección , Factor de Crecimiento Transformador beta/biosíntesis

RESUMEN

In schistosomiasis mansoni, granulomatous inflammation and fibrotic resolution are the major pathogenetic factors. The outcome of fibrosis is influenced by the deposition of collagen and degradation mediated by matrix metalloproteinases (MMP). There is a dearth of data on the expression of MMP and the tissue inhibitors of metalloproteinase (TIMP) during the fibrosis associated with schistosomiasis. In this study, the dynamics of collagen, MMP and TIMP gene expression were analysed during murine Schistosoma mansoni infection. Expression within the granulomatous liver tissue of the genes coding for collagen of types I, III and IV was up-regulated at the onset of granuloma development, and the dominant type-I expression peaked at the chronic, fibrotic stage. The amount of deposited hepatic collagen increased with the chronicity of the infection, indicating cumulative fibrosis. Collagenase, gelatinase, stromelysin, matrilysin-specific gene activities were similarly up-regulated, but only MMP-8 (collagenase-2) expression peaked at the height of fibrosis. TIMP-1 gene expression gradually increased during the course of the disease and, along with TIMP-2, peaked at the chronic, fibrotic stage. Granuloma myofibroblasts expressed both MMP and TIMP-1 genes. In ELISA of the splenic cytokines, high levels of fibrogenic interleukin-13 and moderate production of transforming growth factor-beta were found to be concurrent with fibrosis. These data indicate that an imbalance in MMP:TIMP expression and fibrogenic cytokine production are associated with cumulative fibrosis.

Asunto(s)

Colágeno/genética , Metaloproteinasas de la Matriz/genética , Esquistosomiasis mansoni/genética , Inhibidores Tisulares de Metaloproteinasas/genética , Animales , Secuencia de Bases , Células Cultivadas , Colágeno/metabolismo , Citocinas/biosíntesis , Femenino , Fibrosis/genética , Expresión Génica/genética , Granuloma/genética , Hígado/patología , Parasitosis Hepáticas/genética , Ratones , Ratones Endogámicos CBA , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Schistosoma mansoni/genética , Schistosoma mansoni/metabolismo , Esquistosomiasis mansoni/metabolismo , Bazo/patología , Inhibidores Tisulares de Metaloproteinasas/metabolismo

RESUMEN

During persistent infection, the intracellular bacterial pathogen Chlamydia trachomatis is viable but severely attenuates the production of new, infectious elementary bodies (EBs). To investigate the reasons for this lack of new EB output, we analysed the expression of chlamydial genes encoding products required for DNA replication and cell division, using in vitro models of active versus persistent infection and synovial tissue samples from patients with chronic Chlamydia-associated arthritis. Hep-2 cells were infected with K serovar C. trachomatis and harvested at t = 0-48 h post-infection (p.i; active). Human monocytes were infected similarly and harvested at t = 1-7 days p.i. (persistent). RNA preparations from infected/uninfected cells and patient samples were subjected to reverse transcription-polymerase chain reaction (RT-PCR) targeting polA, dnaA, mutS and parB mRNA, related to chlamydial DNA replication/segregation; these were expressed in infected Hep-2 cells from 11 to 48 h p.i; ftsK and ftsW, related to cell division, were expressed similarly. Real-time PCR analyses demonstrated that significant accumulation of chlamydial chromosome began at about 12 h p.i. in infected Hep-2 cells. In infected human monocytes, polA, dnaA, mutS and parB mRNA were produced from days 1-7 p.i. and were weakly expressed in patient samples. Real-time PCR indicated the continuing accumulation of chlamydial chromosome during the 7 day monocyte infection, although the rate of such accumulation was lower than that occurring during active growth. However, transcripts from ftsK and ftsW were detected only at 1 day p.i. in infected monocytes but not thereafter, and they were absent in all patient samples. Thus, genes whose products are required for chlamydial DNA replication are expressed during persistence, but transcription of genes whose products are required for cytokinesis is severely downregulated. These data explain, at least in part, the observed attenuation of new EB production during chlamydial persistence.

Asunto(s)

Proteínas Bacterianas/metabolismo , Chlamydia trachomatis/genética , Replicación del ADN/genética , Genes Bacterianos/genética , Monocitos/microbiología , Proteínas Bacterianas/genética , División Celular/genética , Chlamydia trachomatis/citología , Chlamydia trachomatis/crecimiento & desarrollo , Chlamydia trachomatis/metabolismo , Cromosomas Bacterianos/genética , Cartilla de ADN , Regulación Bacteriana de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Humanos , Modelos Biológicos , ARN Bacteriano/análisis , ARN Bacteriano/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Líquido Sinovial/microbiología , Transcripción Genética , Células Tumorales Cultivadas

RESUMEN

OBJECTIVE: We and others have reported the presence of Chlamydia and other bacterial species in joint specimens from patients with reactive arthritis (ReA). The present study was conducted to investigate whether bacteria other than those specified by diagnostic criteria for ReA could be identified in synovial fluid (SF) or tissue from patients with various arthritides, and whether the presence of such organisms corresponds to particular clinical characteristics in any patient set or subset. METHODS: DNA in synovial biopsy samples and SF obtained from 237 patients with various arthritides, including ReA, rheumatoid arthritis, and undifferentiated oligoarthritis, was assayed by polymerase chain reaction (PCR) using "panbacterial" primers; we chose only samples known to be PCR negative for Chlamydia, Borrelia, and Mycoplasma species. PCR products were cloned, and cloned amplicons from each sample were sequenced; DNA sequences were compared against all others in GenBank for identification of bacterial species involved. RESULTS: Ten percent of patient samples were PCR positive in panbacterial screening assays. Bacterial species identified belonged to the genera Neisseria, Acinetobacter, Moraxella, Salmonella, Pseudomonas, and others. Thirty-five percent of PCR-positive patients showed the presence of DNA from more than a single bacterial species in synovium; overall, however, we could identify no clear relationship between specific single or multiple bacterial species in the synovium and any general clinical characteristics of any individual or group of patients. CONCLUSION: This analysis provides the first systematic attempt to relate bacterial nucleic acids in the synovium to clinical characteristics, joint findings, and outcomes. Many patients with arthritis have bacterial DNA in the joint, and, in some cases, DNA from more than a single species is present. However, except for 1 case of a control patient with staphylococcal septic arthritis, it is not clear from the present study whether the synovial presence of such organisms is related to disease pathogenesis or evolution in any or all cases.

Asunto(s)

Artritis Reumatoide/microbiología , ADN Bacteriano/aislamiento & purificación , Bacilos y Cocos Aerobios Gramnegativos/aislamiento & purificación , Membrana Sinovial/microbiología , Acinetobacter/genética , Acinetobacter/aislamiento & purificación , Adulto , Anciano , Artritis Psoriásica/microbiología , Artritis Reactiva/microbiología , Biopsia , Niño , Clonación Molecular , Femenino , Bacilos y Cocos Aerobios Gramnegativos/genética , Humanos , Masculino , Persona de Mediana Edad , Moraxella/genética , Moraxella/aislamiento & purificación , Neisseria/genética , Neisseria/aislamiento & purificación , Reacción en Cadena de la Polimerasa , Prohibitinas , Pseudomonas/genética , Pseudomonas/aislamiento & purificación , Salmonella/genética , Salmonella/aislamiento & purificación , Membrana Sinovial/patología

RESUMEN

It has been reported recently that the bacterial respiratory pathogen Chlamydia pneumoniae is present in the cerebrospinal fluid of a subset of multiple sclerosis (MS) patients. However, it is not known whether this organism is a causative agent of MS, or merely an opportunistic pathogen that takes advantage of a disease process initiated by some other means. We report identification of a 20-mer peptide from a protein specific to C. pneumoniae which shares a 7-aa motif with a critical epitope of myelin basic protein, a major CNS Ag targeted by the autoimmune response in MS. This bacterial peptide induces a Th1 response accompanied by severe clinical and histological experimental autoimmune encephalomyelitis in Lewis rats, a condition closely reflective of many aspects of MS. Studies with peptide analogues suggest that different populations of encephalitogenic T cells are activated by the C. pneumoniae and myelin basic protein Ags. Mild experimental autoimmune encephalomyelitis was also observed when rats were immunized with sonicated C. pneumoniae in CFA.

Asunto(s)

Proteínas Bacterianas/administración & dosificación , Proteínas Bacterianas/inmunología , Chlamydophila pneumoniae/inmunología , Encefalomielitis Autoinmune Experimental/inmunología , Secuencia de Aminoácidos , Sustitución de Aminoácidos/genética , Sustitución de Aminoácidos/inmunología , Animales , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Células Cultivadas , Chlamydophila pneumoniae/genética , Encefalomielitis Autoinmune Experimental/microbiología , Femenino , Regulación Bacteriana de la Expresión Génica , Cobayas , Inyecciones Subcutáneas , Datos de Secuencia Molecular , Proteína Básica de Mielina/química , Proteína Básica de Mielina/inmunología , Fragmentos de Péptidos/química , Fragmentos de Péptidos/inmunología , Isoformas de Proteínas/administración & dosificación , Isoformas de Proteínas/genética , Isoformas de Proteínas/inmunología , Ratas , Ratas Endogámicas Lew , Homología de Secuencia de Aminoácido , Linfocitos T/inmunología , Linfocitos T/microbiología , Células Tumorales Cultivadas/inmunología , Células Tumorales Cultivadas/microbiología , Células Tumorales Cultivadas/trasplante

RESUMEN

OBJECTIVE: Molecular biology techniques such as polymerase chain reaction (PCR) and ligase chain reaction (LCR) are routinely used in research for detection of C trachomatis DNA in synovial samples, and these methods are now in use in some clinical laboratories. This study aimed at determining the method best suited to molecular diagnosis of C trachomatis by examining four standard DNA preparation methods using chlamydia spiked synovial tissue and chlamydia infected monocytes. METHODS: Synovial tissue from a chlamydia negative patient with rheumatoid arthritis was spiked with defined numbers of C trachomatis elementary bodies (EB). Purified human peripheral monocytes from normal donors were infected with the organism at a multiplicity of infection 1:1 in vitro and harvested after four days. DNA was prepared from all samples by four methods: (1) QIAmp tissue kit; (2) homogenisation in 65 degrees C phenol; (3) incubation at 97 degrees C; (4) proteinase K digestion at 97 degrees C. DNA from methods 1 and 2 was subjected to PCR using two different primer sets, each targeting the C trachomatis omp1 gene. LCR was done on DNA prepared by each method. RESULTS: In synovial tissue samples spiked with EB, and in monocytes persistently infected with the organism, preparation of template using the QIAmp tissue kit (method 1) and the hot phenol extraction technique (method 2) allowed sensitive detection of C trachomatis DNA. These methods also produced template from both sample types for LCR. DNA prepared by heat denaturation (method 3) allowed only low sensitivity chlamydia detection in LCR and did not work at all for PCR. Proteinase K digestion plus heat denaturation (method 4) gave template that did not allow amplification in either PCR or LCR assays. CONCLUSIONS: The sensitivity of detection for C trachomatis DNA in synovial tissue by PCR and LCR depends strongly on the method used for preparation of the amplification template. LCR targeting the multicopy chlamydial plasmid and two nested PCR assay systems targeting the single copy omp1 gene showed roughly equivalent sensitivity. Importantly, template preparation method and the specific PCR primer system used for screening must be optimised in relation to one another for highest sensitivity.

Asunto(s)

Infecciones por Chlamydia/diagnóstico , Chlamydia trachomatis/aislamiento & purificación , ADN Bacteriano/análisis , Técnicas de Amplificación de Ácido Nucleico , Membrana Sinovial/microbiología , Chlamydia trachomatis/genética , Humanos , Leucocitos Mononucleares/microbiología , Reacción en Cadena de la Ligasa , Reacción en Cadena de la Polimerasa , Sensibilidad y Especificidad

RESUMEN

PURPOSE: The presence of Chlamydia trachomatis has been previously shown in the temporomandibular joint (TMJ). This study investigated whether the presence of other bacteria associated with reactive arthritis (ReA) can be identified in the TMJ. MATERIALS AND METHODS: Posterior bilaminar tissue removed during TMJ surgery from 26 patients (24 F, 2 M) was evaluated for the presence of C. trachomatis, Mycoplasma fermentans, Mycoplasma genitalium, Campylobacter jejuni, Yersinia enterocolitica, Salmonella spp, and Shigella spp by highly specific polymerase chain reaction (PCR) assays. RESULTS: Bacterial DNA was identified in the TMJ as follows: C. trachomatis, 11 of 26 (42%); M. fermentans/orale, 6 of 26 (23%); M. genitalium, 9 of 26 (35%). Nine of 26 TMJs (35%) had the presence of a single bacterial species. Eight of 26 TMJs (31%) had more than 1 species, as follows: C. trachomatis with either M. genitalium or M. fermentans/orale in 5 of 26 (19%), M. fermentans/orale with M. genitalium 2 of 26 (8%), and C. trachomatis/M. fermentans/orale/M. genitalium, 1 of 26 (4%). A total of 17 of 26 (65%) of TMJs had the presence of bacteria identified in the TMJ. Campylobacter jejuni, Y. enterocolitica, Salmonella spp, and Shigella spp were not identified in any samples. CONCLUSIONS: The presence of M. genitalium in the human TMJ has not been previously reported. The presence of bacteria in the TMJ, either singly or concurrently with other bacteria, may serve as the pathogenetic mechanism of TMJ inflammation. The presence of 2 bacteria from the urogenital tract in the TMJ suggests that internal derangement of the TMJ may occur as a result of a sexually acquired infection.

Asunto(s)

Artritis Reactiva/microbiología , Trastornos de la Articulación Temporomandibular/microbiología , Adolescente , Adulto , Técnicas de Tipificación Bacteriana , Campylobacter jejuni/aislamiento & purificación , Chlamydia trachomatis/aislamiento & purificación , ADN Bacteriano/análisis , Enterobacteriaceae/aislamiento & purificación , Femenino , Humanos , Masculino , Persona de Mediana Edad , Mycobacterium/aislamiento & purificación , Reacción en Cadena de la Polimerasa , Prohibitinas

RESUMEN

OBJECTIVE: Recent studies suggest that giant cell arteritis (GCA) may be an antigen-driven disease. Since Chlamydia pneumoniae has been identified in arterial vessel walls, it was hypothesized that this organism might be associated with GCA. METHODS: Fourteen paraffin-embedded temporal artery biopsy specimens from 9 patients with GCA were examined by immunohistochemistry and by polymerase chain reaction (PCR) for the presence of C pneumoniae; for 5 patients, specimens were available from both the left and right arteries. Four temporal artery specimens from 3 patients with polymyalgia rheumatica (PMR) and 9 temporal artery specimens from 5 patients without GCA or PMR served as controls. RESULTS: C pneumoniae was detected by both immunohistochemistry and PCR in 6 GCA patient samples. One GCA patient sample was immunopositive only; another was PCR positive only. Thus, C pneumoniae was found in 8 of 9 GCA patients. One of 4 control samples from the PMR patients was immunopositive, but PCR negative, for C pneumoniae. The C pneumoniae-positive PMR patient also had respiratory symptoms. The remaining 9 control samples were negative for C pneumoniae by both immunohistochemistry and PCR. Immunohistochemistry showed that bacteria predominate in the adventitial layer of temporal arteries, in granulomatous infiltrates. Dendritic cells were examined by immunohistochemistry for their presence and localization in consecutive temporal artery specimens, and showed a strong topographic relationship with C pneumoniae. Like the bacterium, dendritic cells predominate in the adventitial layer of the arteries. CONCLUSION: C pneumoniae was found in temporal artery specimens from most GCA patients, in 1 specimen from a PMR patient, and in no other control specimens; thus, it may play a role in the pathogenesis of the disease. Dendritic cells may represent the antigen-presenting cells in this situation.

Asunto(s)

Infecciones por Chlamydia/microbiología , Chlamydophila pneumoniae/aislamiento & purificación , Células Dendríticas/microbiología , Arteritis de Células Gigantes/microbiología , Anciano , Infecciones por Chlamydia/patología , Chlamydophila pneumoniae/genética , ADN Bacteriano/análisis , Células Dendríticas/patología , Femenino , Técnica del Anticuerpo Fluorescente Indirecta , Arteritis de Células Gigantes/patología , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polimialgia Reumática/microbiología , Polimialgia Reumática/patología , Arterias Temporales/microbiología , Arterias Temporales/patología

RESUMEN

We demonstrated that chromosomal DNA from Chlamydia pneumoniae is present in synovial tissue in at least some patients with reactive arthritis/Reiter's syndrome and other arthritides. Here, we provide initial molecular evidence that the bacterium is viable and metabolically active when present in the synovium. We used reverse transcription-polymerase chain reaction (RT-PCR) assays targeting primary transcripts from the chlamydial rRNA operons, and mRNA from several C. pneumoniae genes (hsp60, ompA, KDO transferase, Mr=76000 protein), to analyse RNA preparations from synovial tissue of 10 patients with various forms of arthritis; each patient was known to be PCR-positive for C. pneumoniae DNA in synovium prior to RT-PCR assays. Two PCR-negative patients served as controls for RT-PCR assays. In the 10 patients PCR-positive for C. pneumoniae DNA, RT-PCR assays targeting primary transcripts from the rRNA operons of the organism showed that these molecules were present in each sample, as were transcripts from the bacterial hsp60 gene. Assays targeting mRNAs from the Mr=76000 protein and the KDO transferase genes of C. pneumoniae gave positive results for 6/10 preparations. We were unable to identify mRNA from the chlamydial major outer membrane protein gene (ompA) in any preparation. RNA preparations from the two control patients were negative in all RT-PCR assays targeting C. pneumoniae transcripts. These results indicate that in patients infected with the organism, synovial C. pneumoniae are viable and metabolically active, as are C. trachomatis cells in the same context. Such viability is consistent with a role in long-term contribution to pathogenesis in joint disease.

Asunto(s)

Artritis/microbiología , Chlamydophila pneumoniae , Membrana Sinovial/microbiología , Adulto , Anciano , Artritis Reactiva/microbiología , Artritis Reumatoide/microbiología , Proteínas de la Membrana Bacteriana Externa/metabolismo , Chaperonina 60/metabolismo , Chlamydophila pneumoniae/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Operón , Osteoartritis/microbiología , ARN Bacteriano/análisis , ARN Ribosómico/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa

RESUMEN

OBJECTIVE: To assess the presence of Chlamydia pneumoniae DNA in the joints of patients with reactive arthritis (ReA) and other arthritides. METHODS: DNA was prepared from synovial tissue (ST) and several synovial fluid (SF) samples from 188 patients with either ReA, undifferentiated oligoarthritis, or other forms of arthritis, and from 24 normal (non-arthritis) individuals. Preparations were screened using polymerase chain reaction (PCR) assays that independently targeted the C. pneumoniae 16S ribosomal RNA and major outer membrane protein genes. RESULTS: Twenty-seven of 212 ST samples (12.7%) were PCR positive for C. pneumoniae DNA; 10 SF samples from these 27 patients were similarly positive. Among the PCR-positive patients, 3 had ReA, 2 had Reiter's syndrome, 7 had undifferentiated oligoarthritis, 4 had undifferentiated monarthritis, 6 had rheumatoid arthritis, and 5 had other forms of arthritis. No samples from normal control individuals were PCR positive. CONCLUSION: DNA of C pneumoniae is present in synovial specimens from some arthritis patients. The prevalence of this organism in the joints was lower than that of C trachomatis, and synovial presence of the organism was not associated with any distinct clinical syndrome. Widely disseminated nucleic acids such as those of C. pneumoniae might have some role in the pathogenesis of several arthritides, since the organism was not found in the ST from normal control individuals.

Asunto(s)

Artritis Reumatoide/genética , Infecciones por Chlamydia/genética , Chlamydia trachomatis/genética , Chlamydophila pneumoniae/genética , Líquido Sinovial/microbiología , Membrana Sinovial/microbiología , Artritis Reactiva/etiología , ADN Bacteriano/análisis , Humanos , Articulaciones/química , Reacción en Cadena de la Polimerasa , Prohibitinas , Líquido Sinovial/química , Membrana Sinovial/química

RESUMEN

PURPOSE: Reactive arthritis (ReA) as a consequence of triggering Chlamydia trachomatis infections has been extensively studied to better understand inflammatory arthritis. This study investigated whether the presence of C trachomatis can be shown in the TMJ of patients with internal derangement. PATIENTS AND METHODS: Posterior bilaminar tissue removed from 31 patients (29 F, 2 M) during TMJ articular disc repositioning and posterior ligament repair was tested for the presence of C trachomatis. Cryosections were stained using a monoclonal antibody that identifies all chlamydial serovars. Highly specific polymerase chain reaction (PCR) assays independently targeting two genes of C trachomatis also were performed; these assays also identify all serovars of this organism. RESULTS: TMJ tissue from 6 of 30 patients (20%) showed the presence of C trachomatis in the posterior bilaminar tissue on immunostaining. PCR screening identified 12 of 31 patients (39%) as having C trachomatis DNA in tissue, including four of six positive by immunostaining. All chlamydia-positive patients were female, with an average age of 36.7 years (15 to 48 years). CONCLUSIONS: The presence of C trachomatis in the human TMJ has not been previously shown. The presence of this organism may serve as the pathogenetic mechanism for TMJ dysfunction, as demonstrated in other joints. Nonapparent chlamydial infection in females may also explain the marked prevalence of TMJ symptoms in women.

Asunto(s)

Artritis Infecciosa/microbiología , Chlamydia trachomatis/patogenicidad , Trastornos de la Articulación Temporomandibular/microbiología , Articulación Temporomandibular/microbiología , Adolescente , Adulto , Técnicas de Tipificación Bacteriana , Distribución Binomial , Chlamydia trachomatis/aislamiento & purificación , ADN Bacteriano/análisis , Femenino , Humanos , Técnicas para Inmunoenzimas , Luxaciones Articulares/microbiología , Masculino , Persona de Mediana Edad , Prohibitinas , Disco de la Articulación Temporomandibular/patología

RESUMEN

Genetically determined differences in interleukin-10 (IL-10) and gamma interferon (IFN-gamma) responses in mice correlate with clearance of Chlamydia pneumonitis infection. We measured the synovial expression of IL-10 and IFN-gamma and additional cytokine genes in patients who had recent-onset Chlamydia-associated arthritis (Chl-AA). IL-10 and IFN-gamma mRNA were relatively abundant in recent-onset Chl-AA.

Asunto(s)

Artritis Infecciosa/genética , Artritis Infecciosa/inmunología , Infecciones por Chlamydia/genética , Infecciones por Chlamydia/inmunología , Chlamydia/patogenicidad , Interferón gamma/genética , Interleucina-10/genética , Adulto , Animales , Artritis/genética , Artritis/inmunología , Artritis Infecciosa/etiología , Estudios de Casos y Controles , Chlamydia/genética , Chlamydia/aislamiento & purificación , Infecciones por Chlamydia/etiología , Citocinas/genética , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , Femenino , Expresión Génica , Humanos , Masculino , Ratones , Persona de Mediana Edad , ARN Mensajero/genética , ARN Mensajero/metabolismo , Membrana Sinovial/inmunología

Asunto(s)

Artritis Reactiva/microbiología , Infecciones por Chlamydia/complicaciones , Chlamydia/patogenicidad , Animales , Artritis Reactiva/patología , Chlamydia/metabolismo , Infecciones por Chlamydia/patología , Conjuntivitis Bacteriana/complicaciones , Conjuntivitis Bacteriana/microbiología , ADN Bacteriano/análisis , Modelos Animales de Enfermedad , Femenino , Articulación de la Rodilla/microbiología , Articulación de la Rodilla/patología , Ratones , Ratones Endogámicos BALB C , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Membrana Sinovial/microbiología , Membrana Sinovial/patología , Sinovitis/microbiología , Sinovitis/patología , Vaginosis Bacteriana/complicaciones , Vaginosis Bacteriana/microbiología

RESUMEN

Genetic background is important in determining whether certain infecting bacteria disseminate to the joint and cause arthritis. We assessed whether APOE genotype is associated with the presence of DNA from Chlamydia or other bacteria in synovial tissues of patients with various arthritides. Nucleic acids from synovial tissues of 135 patients were screened by PCR for DNA from Chlamydia trachomatis, C. pneumoniae and other bacteria (pan-bacteria). APOE genotype was determined by a PCR-based method for all patients in each of four resulting groups comprised of about 35 individuals each, positive for C. trachomatis only, C. pneumoniae only, other bacteria, or no bacteria. RT-PCR was used to assess synovial APOE expression. The latter assays confirmed that APOE mRNA is present in synovial tissue. Determination of APOE genotype showed that patients PCR-negative in all assays, and those positive in the C. trachomatis - and pan-bacteria- (excluding Chlamydia) directed assays, had distributions of the APOE epsilon2, epsilon3 and epsilon4 alleles mirroring those of the general population (i.e. about 8%, 79% and 13%, respectively). In contrast, 68% of patients with C. pneumoniae DNA in synovium possessed a copy of the epsilon4 allele. These results indicate that no association exists between APOE genotype and synovial presence of C. trachomatis or other bacteria. However, individuals bearing at least one copy of the APOE epsilon4 allele may be at increased risk for synovial infection by C. pneumoniae.

Asunto(s)

Apolipoproteínas E/genética , Artritis Infecciosa/genética , Artritis/genética , Infecciones por Chlamydia/genética , Chlamydia/aislamiento & purificación , Líquido Sinovial/microbiología , Alelos , Artritis/microbiología , Artritis Infecciosa/microbiología , Bacterias/genética , Bacterias/aislamiento & purificación , Chlamydia/genética , Infecciones por Chlamydia/microbiología , Chlamydia trachomatis/genética , Chlamydia trachomatis/aislamiento & purificación , Chlamydophila pneumoniae/genética , Chlamydophila pneumoniae/aislamiento & purificación , Humanos , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa

RESUMEN

The principal host cell for persistently infecting synovial Chlamydia trachomatis is the macrophage. During infection of human monocytes/macrophages in culture this bacterium displays aberrant morphology and produces no new elementary bodies, reflecting the situation in synovium. Here we investigate the metabolic status of C. trachomatis (serovar K) during an extended infection of human peripheral monocytes in vitro. Using reverse transcription-polymerase chain reaction assays, we have shown that primary transcripts from the chlamydial rRNA operons are present throughout a 10-day course of infection. Other assays targeting mRNAs from chlamydial genes encoding r-proteins S5 and L5, the glycyl-tRNA synthetase, the 60-kDa cysteine-rich outer membrane protein, and the KDO transferase indicate that these messengers are also present throughout the entire 10-day period. The gene encoding the 57-kDa heat-shock protein (hsp60) is expressed by the bacterium throughout the 10-day infection of cultured monocytes, but transcript levels from the gene encoding the major outer membrane protein (omp1) appear to be attenuated. Western analyses targeting these latter proteins confirm the presence of the hsp60 gene product, and the virtual absence of major outer membrane protein, in chlamydia-infected cultured human monocytes. Thus, during extended infection of human monocytes in vitro, chlamydia are non-productive but transcriptionally active; the pattern of transcriptional activity reflects that known for persistent C. trachomatis infection in vivo in synovial tissue.

Asunto(s)

Proteínas Bacterianas/biosíntesis , Chlamydia trachomatis/metabolismo , Monocitos/microbiología , Anticuerpos Monoclonales , Especificidad de Anticuerpos , Proteínas de la Membrana Bacteriana Externa/biosíntesis , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Bacterianas/genética , Western Blotting , Chaperonina 60/biosíntesis , Chaperonina 60/genética , Chlamydia trachomatis/genética , Electroforesis en Gel de Agar , Humanos , Reacción en Cadena de la Polimerasa , ARN Bacteriano/genética , ARN Bacteriano/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transcripción Genética

RESUMEN

OBJECTIVE: To use standard molecular methods to define the prevalence and metabolic characteristics of Chlamydia trachomatis during infection of fallopian tubes in women with ectopic pregnancies. DESIGN: Polymerase chain reaction (PCR)- and reverse transcription-PCR (RT-PCR)-based assessment of presence of chlamydial DNA and various RNA species in fallopian tube biopsy samples. SETTING: Hospital and molecular genetics laboratory. PATIENTS: Ten women of varying ages, each presenting with ectopic pregnancy. MAIN OUTCOME MEASURE(S): Positive signal in specific chlamydia-directed PCR and RT-PCR assays. RESULT(S): Nucleic acid preparations from 7 of the 10 fallopian tube patient samples were PCR-positive for C. trachomatis DNA. Each of the 7 PCR-positive samples also showed the presence of several transcripts from the bacterium, including primary transcripts from the ribosomal RNA operons. CONCLUSION(S): A higher proportion of ectopic pregnancies than was believed previously may be attributable to infection of the fallopian tubes by C. trachomatis. The presence of various chlamydial RNA molecules suggests that viable, metabolically active bacteria were present in fallopian tubes of the patients studied.

Asunto(s)

Chlamydia trachomatis/crecimiento & desarrollo , Trompas Uterinas/microbiología , Embarazo Ectópico/microbiología , Adulto , Femenino , Células HeLa , Humanos , Reacción en Cadena de la Polimerasa/métodos , Embarazo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa