RESUMEN
Studies of human mucosal lymphoid follicles are rare and have been limited to children's Peyer's patches, which are visible at endoscopy. We investigated lymphoid follicles in ileum biopsies of 87 patients and surgical colon specimens from 66 cancer patients, and examined phenotype and function of isolated follicular immune cells. Two (0-10) and 12 (0-117) follicles per patient were found in ileum and colon samples respectively (P < 0.001). The number of lymphoid follicles mononuclear cells (LFMC) that could be isolated per patient was higher from colon compared with ileum specimens [725 000 (0-23 Mio) versus 100 000 (0-1.3 Mio), P < 0.001]. T cells were predominant in both LFMC and lamina propria mononuclear cells (LPMC), but B cells were more and plasma cells less frequent in LFMC. T cells from mucosal follicles were more frequently CD4-positive and CD62L-positive, but less frequently CD8-positive, CD103-positive and CD69-positive than lamina propria T cells. LFMC from ileum compared with colon showed no differences in mononuclear cell composition. Anti-CD3/CD28 stimulation induced similar proliferation of LFMC and LPMC from ileum and colon, as well as secretion of high levels of interferon-gamma, tumour necrosis factor-alpha and interleukin (IL)-2, but lower levels of IL-4, IL-6 and IL-10. LFMC from colon secreted more IL-2 than those from ileum. Our study shows that mucosal lymphoid follicles can be identified clearly in adult human colon and yield viable immune cells sufficient for phenotypical and functional analysis. The cellular composition of LFMC from ileum and colon is similar, and both secrete predominantly T helper type 1 cytokines.
Asunto(s)
Colon/inmunología , Íleon/inmunología , Mucosa Intestinal/inmunología , Leucocitos Mononucleares/citología , Tejido Linfoide/citología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores/análisis , Proliferación Celular , Células Cultivadas , Citocinas/análisis , Femenino , Citometría de Flujo , Humanos , Masculino , Persona de Mediana Edad , Estadísticas no Paramétricas , Células TH1/inmunología , Adulto JovenRESUMEN
: In this study the TCR-Vß repertoire expressed in T cells of lamina propria mononuclear cells (LPMC) and peripheral blood mononuclear cells (PBMC) was examined using a reverse transcription-PCR (RT-PCR) technique for TCR-Vß mRNA. Using a qualitative RT-PCR method, LPMC of patients with IBD and control individuals were shown to contain mRNA for each of 20 TCR-Vß families, indicating that IBD is not associated with a major deletion or expansion of any TCR-Vß family. Subsequently, using a quantitative method for four frequently expressed TCR-Vß families, it was shown that the pattern of TCR-Vß expression was different in PBMC and LPMC of both IBD patients and control individuals. In addition, it was shown that the LPMC/PBMC ratio of mean mRNA values for TCR-Vß2, but not for TCR-Vß6, 7, and 14 was lower in IBD patients than control individuals. These results show that the TCR-Vß repertoire in PBMC and LPMC is different both in IBD patients and control individuals. In addition, they show that the TCR-Vß repertoire is altered in IBD, possibly due to an immune response to disease specific antigens, superantigens or neoantigens.
RESUMEN
It is increasingly recognized that the inability of the immune system to clear H. pylori infection is caused by an inadequate immune response and is associated with chronic gastric inflammation. To further investigate the cellular immune response to H. pylori, we studied PBMC from 31 H. pylori antibody-negative and 16 H. pylori antibody-positive individuals for H. pylori-induced DNA synthesis, secretion of the Th1-type cytokine IFN-gamma and secretion of IL-12, a cytokine produced by bacteria-stimulated monocyte/macrophages and a potent inducer of antibacterial immune responses and Th1-type T cells. All experiments were performed using Y. enterocolitica 03 as control. Our results demonstrate that DNA synthesis, IFN-gamma production and IL-12 production induced by H. pylori or Y. enterocolitica 03 did not differ significantly between H. pylori antibody-positive and H. pylori antibody-negative individuals. However, in the H. pylori-positive group there was a tendency, although not statistically significant, to produce less IFN-gamma in response to H. pylori but not Y. enterocolitica. These results demonstrate that monocyte/macrophages from H. pylori-positive individuals secrete normal amounts of IL-12 upon bacterial challenge and suggest that the decreased production of IFN-gamma in H. pylori-positive individuals observed in previous studies is selective for H. pylori and not caused by decreased IL-12 secretion.
Asunto(s)
Anticuerpos Antibacterianos/inmunología , Infecciones por Helicobacter/inmunología , Helicobacter pylori/inmunología , Interleucina-12/biosíntesis , Humanos , Inmunidad Celular , Interferón gamma/biosíntesis , Persona de Mediana EdadRESUMEN
Quantitative determination of T-cell receptor (TCR) V beta expression is necessary to define the changes in TCR-V beta subfamily expression that occur during T-cell maturation and selection and to detect alterations of the TCR-V beta repertoire that may be associated with human diseases. Here we describe and validate a quantitative reverse transcription-polymerase chain reaction (RT-PCR) technique to determine human TCR-V beta subfamily mRNA levels (as well as other mRNA species), based on the use of synthetic poly(A) mRNA internal standards that are coprocessed with native (sample) mRNA transcripts. The technique allows simultaneous reverse transcription of sample and standard mRNA and thus obviates errors arising during reverse transcription. In addition, the technique allows coamplification of several concentrations of standard mRNA (cDNA) with sample mRNA (cDNA) under conditions in which these mRNAs amplify with equal efficiency; thus, it avoids errors resulting from saturation of competition effects. Finally, the technique is sensitive to lower than 1.5-fold differences in input mRNA. To apply the technique, we also describe methods for the generation of poly(A) mRNA internal standards that can be used to quantitate TCR-V beta 2/6/7 and TCR-C beta mRNA.
Asunto(s)
Toxinas Bacterianas , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Superantígenos , Antígenos Bacterianos/inmunología , Secuencia de Bases , Enterotoxinas/inmunología , Humanos , Cinética , Datos de Secuencia Molecular , Familia de Multigenes , Poli A/síntesis química , Reacción en Cadena de la Polimerasa , ARN Mensajero/análisis , ARN Mensajero/síntesis química , ADN Polimerasa Dirigida por ARN , Estándares de Referencia , Reproducibilidad de los Resultados , Linfocitos T/inmunologíaRESUMEN
Infection of humans with Helicobacter pylori results in the development of chronic gastritis and plays an important role in gastric ulcer pathogenesis. Despite the infiltration of the mucosa with specific immunocompetent cells and production of specific antibodies, the infection usually persists for life. This study was performed to investigate if immunologic mechanisms exist which could contribute to the inability of the host to terminate the infection. Therefore, we compared the in vitro immunoreactivity of peripheral blood mononuclear cells (PBMC) from H. pylori-infected patients after stimulation with sonicated H. pylori bacteria from the stomach of the patient (autologous bacterial strain) with stimulation by bacteria from other patients (heterologous bacteria). We measured cell proliferation, expression of T cell activation markers CD25, HLA-DR, and CD71, as well as production ofinterleukin-10 (IL-10), an inhibitory cytokine. We found that the proliferative response of PBMC was significantly lower after autologous than after heterologous stimulation. Furthermore, secretion of IL-10 in the culture supernatants was significantly higher when PBMC were incubated with autologous than with heterologous H. pylori antigens. No significant differences between autologous or heterologous stimulation were observed in the increased expression of T cell activation markers. These data indicate that systemic immunologic response to H. pylori are strain-dependent. For further studies of the immune responses towards H. pylori, the use of an autologous stimulatory system seems necessary.
Asunto(s)
Antígenos Bacterianos/inmunología , Helicobacter pylori/inmunología , Adulto , Anciano , Femenino , Antígenos HLA-DR/análisis , Humanos , Interleucina-10/metabolismo , Activación de Linfocitos , Masculino , Persona de Mediana EdadRESUMEN
OBJECTIVE: In reactive arthritis (ReA), synovial fluid-derived bacteria-specific CD4+ and CD8+ T cells have been studied intensively in recent years. We have addressed the question whether gamma delta-TCR+ lymphocytes could contribute to antibacterial or anti-self cytotoxicity in the affected joints of patients, with spondylarthropathies. METHODS: T cell clones were derived by random cloning from the synovial fluids of one patient with Yersinia-induced ReA, one patient with a Yersinia-induced flare up of pre-existing ankylosing spondylitis, and one patient with ankylosing spondylitis. Eight clones with a CD3+, alpha beta-TCR-, CD4-, CD8- and gamma delta-TCR+ phenotype (all expressing V gamma 9) were tested in a standard 52Cr-release assay using autologous or allogeneic B cell lines, CIR-B27, Daudi cells, and RJ.225 cells. RESULTS: Four gamma delta-TCR+ clones killed both autologous and allogeneic target cells when infected with live Yersinia or Salmonella and also uninfected Daudi cells expressing GroEL heatshock protein. One clone was specific for Yersinia-infected targets. Three gamma delta-TCR+ clones were cytotoxic when uninfected autologous or allogeneic targets were employed. Polymorphic "classical" MHC class I or class II molecules were not used as restriction elements. CONCLUSION: We conclude that, upon in vivo contact with bacteria such as Yersinia and Salmonella, synovial gamma delta-T lymphocytes are activated and contribute to antibacterial immunity via specific target cell lysis. Furthermore, anti-self cytolytic gamma delta-T cells could participate in the clearance of stressed and detrimental cells in the arthritic joint or, alternatively, could support the chronicity of autoimmune arthritis.
Asunto(s)
Artropatías/inmunología , Receptores de Antígenos de Linfocitos T gamma-delta/metabolismo , Enfermedades de la Columna Vertebral/inmunología , Líquido Sinovial/inmunología , Subgrupos de Linfocitos T/inmunología , Adulto , Artritis Reactiva/etiología , Artritis Reactiva/inmunología , Autoinmunidad , Bacterias/inmunología , Citotoxicidad Inmunológica , Humanos , Técnicas In Vitro , Masculino , Persona de Mediana Edad , Prohibitinas , Salmonella/inmunología , Espondilitis Anquilosante/inmunología , Líquido Sinovial/citología , Yersinia/inmunología , Yersiniosis/complicacionesRESUMEN
PURPOSE: To compare the value of hydro-MRI with follow-through examinations in the follow-up of Crohn's disease. METHOD: 22 patients known to be suffering from Crohn's disease were examined via 1.5 T-MR system; an oral contrast examination using 1000 ml of a 2.5% mannitol solution was performed in all patients. T2-weighted TSE sequences and T1-weighted SE sequences were performed before and after the intravenous injection of Gd-DTPA. To reduce movement artifacts caused by peristalsis of the gut, intravenous injection of 40 mg Buscopan was given. The findings of hydro-MRI were compared with the follow-through examinations. RESULTS: In the upper gastrointestinal tract, the follow-through examination showed clear advantages compared with hydro-MRI for the demonstration of inflammatory changes in the gut; Hydro-MRI was, however, somewhat more reliable in the ileum and colon. It was also more sensitive than the follow-through for the demonstration of enteric fistulae (four as compared with two cases), and in demonstration extraluminal changes (free fluid in six against zero, and inflammatory adherent loops (four against zero)). Amongst the 22 patients, hydro-MRI was equal (in 10) or better (in 8) than the follow-through examination for demonstrating the intestinal manifestations of Crohn's disease, and follow-through was better in only four. CONCLUSION: For follow-up of Crohn's disease, hydro-MRI is at least as good as follow-through examination, and is even preferable, because of the absence of radiation exposure of the usually young patients.
Asunto(s)
Enfermedad de Crohn/diagnóstico , Sistema Digestivo/diagnóstico por imagen , Imagen por Resonancia Magnética , Administración Oral , Adulto , Sulfato de Bario/administración & dosificación , Constricción Patológica , Medios de Contraste/administración & dosificación , Enfermedad de Crohn/diagnóstico por imagen , Enfermedad de Crohn/patología , Enema , Estudios de Evaluación como Asunto , Femenino , Estudios de Seguimiento , Tránsito Gastrointestinal , Humanos , Intestinos/patología , Imagen por Resonancia Magnética/métodos , Masculino , Radiografía , Factores de TiempoRESUMEN
Recent data indicate that cytotoxic T lymphocytes (CTL) are involved in the pathogenesis of HLA-B27-associated spondylarthropathies. In the absence of clearly defined "arthritogenic" bacterial or self peptides that are presented by HLA-B27 and recognized by such CD8+CTL, one approach has been to investigate the T cell repertoire of lesional cellular infiltrates by determining T cell receptor (TCR) variable (V) gene segment frequencies. Furthermore, the TCR V alpha and V beta chains of HLA-B27-restricted CTL clones, notably the putative peptide-contacting CDR3-regions of these TCRs, have been sequenced. This article will give a short review of the current literature on the topology of the TCR and its hypervariable CDR3 region, TCR repertoire diversity in rheumatic diseases and will concentrate on TCR V alpha and V beta gene usage in HLA-B27-restricted T cell responses.
Asunto(s)
Antígeno HLA-B27/inmunología , Receptores de Antígenos de Linfocitos T , Enfermedades Reumáticas , Citotoxicidad Inmunológica/genética , Citotoxicidad Inmunológica/inmunología , Receptores de Antígenos de Linfocitos T/análisis , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/inmunología , Enfermedades Reumáticas/inmunologíaRESUMEN
BACKGROUND: Enteropathic spondylarthropathies (SpA) are the most frequent extraintestinal manifestation of the chronic inflammatory bowel disease (IBD), Crohn's disease (CD) and Ulcerative Colitis (UC). It was the aim of the present study, to analyze a large number of IBD patients for the prevalence and pattern of joint manifestation and the association of SpA with the extend of bowel involvement and HLA-haplotype. PATIENTS AND METHODS: 521 patients (409 CD and 112 UC) were prospectively analyzed over a period of one year. SpA was diagnosed on the basis of an appropriate patient history as well as clinical, radiological and immunoserological parameters. RESULTS: SpA was diagnosed in 10.7% of all CD and 14.4% of all UC patients. In 26.8% of all patients symptoms of SpA occurred prior to and in 14.4% simultaneously with IBD. 28.1% of all patients presented with isolated peripheral arthritis, 26.8% of all patients showed an isolated involvement of the spine or sacroiliic joints and 45.1% of all patients presented with combined involvement. 2/12 UC patients with SpA suffered from rectosigmoiditis, 5/12 from partial colitis and 5/12 had pancolitis. In CD patients with SpA, 8/59 had isolated colitis, 8/59 ileocolitis and 31/59 isolated small bowel involvement. There was a positive correlation between SpA and HLA-B27 (p < 0.01). CONCLUSION: Enteropathic spondylarthropathies are an important extraintestinal manifestation of IBD. Spondylarthropathies occur irrespective of the extend of IBD and frequently become symptomatic prior to IBD. These and recent data describing inflammatory bowel disease in patients with SpA of unknown etiology suggest that both diseases have a common pathogenetic background.
Asunto(s)
Enfermedad de Crohn/diagnóstico , Antígenos de Histocompatibilidad Clase II/análisis , Antígenos de Histocompatibilidad Clase I/análisis , Osteoartritis/diagnóstico , Espondilitis Anquilosante/diagnóstico , Adulto , Enfermedad de Crohn/inmunología , Femenino , Antígeno HLA-B27/análisis , Humanos , Masculino , Persona de Mediana Edad , Osteoartritis/inmunología , Articulación Sacroiliaca/inmunología , Columna Vertebral/inmunología , Espondilitis Anquilosante/inmunologíaRESUMEN
Crohn's disease (CD) and ulcerative colitis (UC) are clinical entities characterized by spontaneous relapses and are thought to be caused in large part by a dysregulated immune response to inflammatory stimuli. Specific infectious agents or antigens inducing or perpetuating inflammation, however, are not known. Recent results in contrast support the hypothesis, that the normal intestinal flora plays a central role in the pathogenesis of both diseases. Studies performed with E. coli Nissle 1917 demonstrated that this bacterium can positively affect the course of disease in UC and CD patients. The clinical efficacy of probiotics can yield valuable information about disease pathogenesis and, as a modification of current standard therapy, opens new and interesting therapeutic alternatives.
Asunto(s)
Colitis Ulcerosa/microbiología , Enfermedad de Crohn/microbiología , Escherichia coli , Mucosa Intestinal/microbiología , Colitis Ulcerosa/inmunología , Colitis Ulcerosa/terapia , Enfermedad de Crohn/inmunología , Enfermedad de Crohn/terapia , Humanos , Mucosa Intestinal/inmunología , Probióticos/uso terapéutico , RecurrenciaRESUMEN
The normal intestinal flora is required for the development of intestinal inflammation in animal models of inflammatory bowel disease (IBD). In humans, several studies indicated a potential association of Escherichia coli (E. coli) with IBD. In addition, we have shown that T-cell clones of IBD patients cross react toward different enteric bacterial species and thus likely respond to conserved bacterial antigens. Therefore, we hypothesized that highly conserved E. coli proteins might be a reasonable candidate to screen for abnormal T-cell responses in IBD. We used high-throughput techniques for cloning, expression, and purification under native conditions of a set of 271 conserved proteins of E. coli, of which 196 were used for whole blood stimulations to assess peripheral T helper (T(H))-cell responses. In addition, because of the association of an adherent-invasive E. coli with Crohn's disease (CD), we included 13 pathogenicity factors of E. coli in the study. We observed that pools of these conserved E. coli proteins less frequently induced interferon-gamma (IFNgamma) production in peripheral T(H) cells in patients with CD and ankylosing spondylitis (AS) compared with healthy controls. In addition, lower percentage of patients with CD and AS responded toward single proteins. The reason for the decreased frequency of an in vitro T(H)-cell IFNgamma response toward E. coli proteins in peripheral blood of CD and AS patients, e.g., increased suppression needs to be clarified.
Asunto(s)
Antígenos/análisis , Epítopos/análisis , Enfermedades Inflamatorias del Intestino/inmunología , Enfermedades Inflamatorias del Intestino/microbiología , Intestinos/inmunología , Intestinos/microbiología , Adolescente , Adulto , Anciano , Animales , Antígenos/inmunología , Antígenos/aislamiento & purificación , Epítopos/inmunología , Epítopos/aislamiento & purificación , Escherichia coli/genética , Escherichia coli/inmunología , Escherichia coli/metabolismo , Escherichia coli/patogenicidad , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/inmunología , Proteínas de Escherichia coli/aislamiento & purificación , Proteínas de Escherichia coli/metabolismo , Femenino , Humanos , Enfermedades Inflamatorias del Intestino/sangre , Masculino , Persona de Mediana Edad , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/aislamiento & purificación , Linfocitos T Colaboradores-Inductores/inmunología , Adulto JovenRESUMEN
Hepatitis E virus (HEV) is a common cause of acute hepatitis in endemic areas. Yet reports on autochthonous cases in other areas such as middle Europe are increasing. Here we report on a patient, who obviously acquired his HEV infection in Germany. Sequence analysis of the virus gained from his serum revealed homologies to other European isolates and swine isolates.
Asunto(s)
Virus de la Hepatitis E/fisiología , Hepatitis E/fisiopatología , Enfermedades Transmisibles , Europa (Continente)/epidemiología , Alemania , Hepatitis E/epidemiología , Virus de la Hepatitis E/clasificación , Virus de la Hepatitis E/genética , Virus de la Hepatitis E/aislamiento & purificación , Humanos , Hígado/metabolismo , Masculino , Persona de Mediana EdadRESUMEN
Ankylosing spondylitis (AS) is a complex genetic disease in which both MHC and non-MHC genes determine disease susceptibility. To determine whether the T cell repertoires of individuals with AS show signs of increased stimulation by exogenous antigens, CD4+ and CD8+ T cell subsets of five monozygotic HLA-B27+ twins (two concordant and three discordant for AS) and CD8+ T cell repertoires of three healthy HLA-B27+ individuals were characterized by TCR beta-chain (TCRB) CDR3 size spectratyping. Selected TCRB-CDR3 spectra were further analysed by BJ-segment analysis and TCRB-CDR3 from expanded T cell clones were sequenced. In an analysis of all data (519/598 possible TCRB-CDR3 spectra), AS was associated with increased T cell oligoclonality in both CD8+ (P = 0.0001) and CD4+ (P = 0.033) T cell subsets. This was also evident when data were compared between individual twins. Nucleotide sequence analysis of expanded CD8+ or CD4+ T cell clones did not show selection for particular TCRB-CDR3 amino acid sequence motifs but displayed sequence homologies with published sequences from intra-epithelial lymphocytes or synovial T cells from rheumatoid arthritis patients. Together, these results provide support for the hypothesis that responses to T cell-stimulating exogenous or endogenous antigens are involved in the induction and/or maintenance of AS.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Receptores de Antígenos de Linfocitos T/inmunología , Espondilitis Anquilosante/inmunología , Adulto , Anciano , Linfocitos T CD4-Positivos/patología , Linfocitos T CD8-positivos/patología , Diferenciación Celular , Antígeno HLA-B27/genética , Humanos , Persona de Mediana Edad , Espondilitis Anquilosante/genética , Espondilitis Anquilosante/patología , GemelosRESUMEN
HISTORY AND ADMISSION FINDINGS: A 79-year-old local resident, presenting with abdominal pain, sweating and weight loss and suspected of having cancer of the pancreas was referred for diagnosis and treatment. Physical examination was negative except for pain on pressure over the right upper abdomen and the epigastrium. INVESTIGATIONS: Erythrocyte sedimentation rate was increased; as were the transaminases and cholestasis parameters. Ultrasonography and computed tomography of the abdomen revealed an echo-poor mass with cystic areas in the region of the head of the pancreas, as well as extra- and intrahepatic dilatation of the biliary tract. Endoscopic retrograde cholangiopancreatography failed to demonstrate a ductal pancreatic carcinoma. Biopsies of a macroscopically peculiar-looking duodenal ulcer demonstrated a noncaseous epithelioid granuloma. A fine-needle biopsy was performed for further diagnosis. DIAGNOSIS, TREATMENT AND COURSE: Histological examination of the needle biopsy revealed a caseous granuloma and acid-fast bacteria. The tuberculin test (GTI) was strongly positive (14-15 mm), indicating tuberculosis of the pancreas and duodenum. Multiple tuberculostatics rapidly improved the patient's symptoms, and the further course was without complications. CONCLUSION: Tuberculosis should be included in the differential diagnosis of consumptive disease with an atypical presentation, especially because treatment could well be curative.
Asunto(s)
Enfermedades Duodenales/diagnóstico , Enfermedades Pancreáticas/diagnóstico , Neoplasias Pancreáticas/diagnóstico , Tuberculosis Gastrointestinal/diagnóstico , Tuberculosis/diagnóstico , Anciano , Antituberculosos/uso terapéutico , Biopsia con Aguja , Colangiopancreatografia Retrógrada Endoscópica , Diagnóstico Diferencial , Enfermedades Duodenales/tratamiento farmacológico , Femenino , Estudios de Seguimiento , Humanos , Páncreas/patología , Enfermedades Pancreáticas/tratamiento farmacológico , Radiografía Abdominal , Radiografía Torácica , Factores de Tiempo , Tomografía Computarizada por Rayos X , Prueba de Tuberculina , Tuberculosis/tratamiento farmacológico , Tuberculosis Gastrointestinal/tratamiento farmacológicoRESUMEN
The intestinal mucosa constitutes an important barrier as it separates each individual from a large array of antigens within the bowel lumen. These luminal antigens may either be derived from pathogens or may be derived from harmless constituents such as ingested food or the normal intestinal flora. The dichotomy of potentially harmful and potentially harmless antigens encountered by the mucosal immune system poses the important task that, with regard to bacteria-derived antigens, the gut associated immune system is required to mount an efficient host defense against pathogenic bacteria but to maintain at the same time the regulatory control mechanisms which protect the human organism from hyperresponsiveness, and thus chronic inflammation, towards antigens from the normal intestinal flora. In the present review, we discuss variable host and bacterial factors which are likely to determine whether the immune response to pathogenic or normal intestinal bacteria will have beneficial or detrimental consequences for the human organism. Using infections with the prototype enteropathogens V. cholerae and enteropathogenic E. coli (ETEC), Y. enterocolitica induced reactive arthritis (ReA) and in more detail, inflammatory bowel diseases (IBD) as exemplary clinical situations, we review current hypotheses of how bacteria or their products are encountered by cellular components of the specific immune system and how this may relate to disease pathogenesis and the development of new treatment strategies.
Asunto(s)
Antígenos Bacterianos/inmunología , Enterobacteriaceae/inmunología , Mucosa Intestinal/inmunología , Artritis Reactiva/inmunología , Artritis Reactiva/microbiología , Cólera/inmunología , Cólera/microbiología , Escherichia coli/inmunología , Infecciones por Escherichia coli/inmunología , Infecciones por Escherichia coli/microbiología , Humanos , Inmunidad Celular/inmunología , Enfermedades Inflamatorias del Intestino/inmunología , Enfermedades Inflamatorias del Intestino/microbiología , Mucosa Intestinal/microbiología , Subgrupos Linfocitarios/inmunología , Ganglios Linfáticos Agregados/inmunología , Ganglios Linfáticos Agregados/microbiología , Prohibitinas , Vibrio cholerae/inmunologíaRESUMEN
We present the case of a female patient with a diagnosis of systemic lupus erythematosus (SLE) at the age of 54 years. At the age of 63 years, she suffered from malignant thymoma and 3 years after removal of the thymoma a diagnosis of pure red cell aplasia (PRCA) was established. This is, to our knowledge, the first report of the occurrence of SLE, thymoma and PRCA in the same patient. The case is discussed with regard to the already known associations between these diseases.
Asunto(s)
Lupus Eritematoso Sistémico/complicaciones , Aplasia Pura de Células Rojas/complicaciones , Timoma/complicaciones , Neoplasias del Timo/complicaciones , Adulto , Médula Ósea/patología , Femenino , Humanos , Timoma/patología , Neoplasias del Timo/patologíaRESUMEN
Analysis of formal amino acid sequence identity between different TCRB chain (TCRB) hypervariable regions (CDR3) is commonly used to localize relevant sites of TCR antigen interaction or to yield indirect information on unknown corresponding antigens. However, this analysis sometimes fails to demonstrate expected concordances, e.g. between CDR3 from T cell clones of identical reactivity. Since this may be due to ignorance of physico-chemical parameters, we have now use hydropathy profile analysis as an additional method to examine TCRB-CDR3 and putative peptide antigens. Superimposed hydropathy plots (SHOP) of 20 TCRB-CDR3 from HLA-B27-restricted autoreactive and Yersinia enterocolitica-specific synovial cytotoxic T lymphocytes (CTL) isolated from patients with reactive arthritis (ReA) revealed restricted distribution of polar amino acids resulting in characteristically different SHOP profiles between the two CTL groups. Similarly, Yersinia-derived and self nonapeptides known to bind HLA-B27 differed in SHOP profiles. To validate the method we have extended SHOP analysis to published TCRB sequence data from additional TCRB-CDR3 from peptide-specific CTL but not in TCRB from HLA-B27-alloreactive CTL or non-HLA-B27-restricted control CTL. We here demonstrate that SHOP may improve TCR-CDR3 sequence analysis by detection of structural constraints which remain cryptic by conventional sequence analysis. Our data suggest that electrostatic properties rather than rigid sequence motifs determine T cell specificities.
Asunto(s)
Secuencia Conservada/inmunología , Antígeno HLA-B27/inmunología , Antígeno HLA-B27/metabolismo , Péptidos/metabolismo , Receptores de Antígenos de Linfocitos T alfa-beta/análisis , Linfocitos T Citotóxicos/inmunología , Secuencia de Aminoácidos , Evolución Molecular , Antígeno HLA-B27/genética , Humanos , Mapeo Peptídico , Prohibitinas , Unión Proteica/inmunología , Conformación Proteica , Complejo Receptor-CD3 del Antígeno de Linfocito T/inmunología , Homología de Secuencia de AminoácidoRESUMEN
BACKGROUND AND AIMS: One major problem in the management of steroid refractory attacks of patients with inflammatory bowel disease (IBD) is the establishment of a rapidly acting immunosuppressive regimen. Based on its well known efficacy in systemic vasculitis, intravenous cyclophosphamide pulse therapy was used in refractory IBD patients to evaluate both its efficacy and safety. METHODS: Between December 1998 and May 2001, seven patients (Crohn's disease, n=5; indeterminate colitis, n=1) with severe steroid refractory IBD (Crohn's disease activity index (CDAI) 264-479 points) received 4-6 cycles of monthly treatment with intravenous cyclophosphamide (750 mg) in a prospective uncontrolled pilot study. RESULTS: All patients improved after two intravenous pulses of cyclophosphamide and six of seven patients achieved complete remission (CDAI <150 points). One patient with Crohn's disease of the small and large bowel showed an impressive clinical response but did not enter into remission. Tapering to low dose steroids was possible in all responders. Remission was maintained in all patients for 18 months (median) but required a second course of intravenous pulse cyclophosphamide in one patient. The drug was well tolerated except for two episodes of candida oesophagitis. CONCLUSIONS: Intravenous pulse cyclophosphamide may be a safe and effective treatment in patients with severe IBD unresponsive to steroid treatment and merits evaluation in a controlled trial.
Asunto(s)
Ciclofosfamida/uso terapéutico , Inmunosupresores/uso terapéutico , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Enfermedad Aguda , Adulto , Antiinflamatorios/uso terapéutico , Enfermedad de Crohn/tratamiento farmacológico , Resistencia a Medicamentos , Femenino , Humanos , Infusiones Intravenosas , Masculino , Proyectos Piloto , Prednisolona/uso terapéutico , Estudios ProspectivosRESUMEN
There is now increasing evidence that hyperresponsiveness towards intestinal flora is a crucial event in the pathogenesis of inflammatory bowel disease (IBD). In support of this hypothesis, we recently described in humans that tolerance exists towards indigenous intestinal flora but is broken in active IBD lesions. In the present study, we have attempted to transfer this model into mice from different genetic backgrounds (BALB/c, SJL/J, C3H/HeJ). We found that mononuclear cells from spleen, small bowel and large bowel of mice do not proliferate, i.e. are tolerant when exposed to bacterial sonicates derived from autologous intestine (BsA) but do proliferate, i.e. are immune when exposed to bacterial sonicates derived from the heterologous intestine of syngenic littermates (BsH). Furthermore, we demonstrate that both local and systemic tolerance to BsA is broken in a murine model of chronic intestinal inflammation induced by the hapten reagent 2, 4, 6-trinitrobenzene sulfonic acid (TNBS), which mimics several important characteristics of Crohn's disease. Tolerance to BsA was restored and TNBS-induced colitis was abrogated in mice systemically treated with interleukin (IL)-10 or antibodies to IL-12. Treatment specifically restored tolerance to BsA, but did not suppress proliferation to BsH. In summary, we here report a new model for the study of immunity and tolerance towards bacterial products. Our data suggest that tolerance to BsA is an important protective mechanism and that restoration of tolerance intestinal flora by IL-10 and antibodies to IL-12 may be of potential therapeutic utility in patients with inflammatory bowel disease.
Asunto(s)
Bacterias/inmunología , Colitis/inmunología , Tolerancia Inmunológica , Factores Inmunológicos/uso terapéutico , Interleucina-10/uso terapéutico , Interleucina-12/antagonistas & inhibidores , Intestinos/microbiología , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/uso terapéutico , Bacterias/aislamiento & purificación , Colitis/inducido químicamente , Colitis/microbiología , Colitis/terapia , Colon/citología , Colon/inmunología , Enfermedad de Crohn/inmunología , Modelos Animales de Enfermedad , Humanos , Tolerancia Inmunológica/genética , Interleucina-12/inmunología , Intestino Delgado/citología , Intestino Delgado/inmunología , Leucocitos Mononucleares/inmunología , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ganglios Linfáticos Agregados/citología , Ganglios Linfáticos Agregados/inmunología , Ratas , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/uso terapéutico , Especificidad de la Especie , Organismos Libres de Patógenos Específicos , Bazo/citología , Bazo/inmunología , Ácido Trinitrobencenosulfónico/toxicidadRESUMEN
In this study, we report that differences between T-cell receptor (TCR) V beta gene family usage in CD4+ and CD8+ T cells are significantly greater in a subgroup of patients with common variable immunodeficiency (CVI) and high levels of activated CD8+ T cells (CD8hi CVI) than in controls (P < 0.001). In CD8hi CVI patients, such differences were also significantly greater for V beta 12 than for other V beta families. As the causes of the differential usage of V beta gene families by CD4+ and CD8+ T cells are under investigation, it was interesting that the combined differences between V beta gene family usage in the CD4+ and CD8+ T-cell subpopulations as a whole were significantly lower than the combined differences between individual V beta gene family usage in either CD4+ or CD8+ T-cell subpopulations (P < 0.001 in both control and CD8hi CVI patients). Further, the pattern of V beta gene family usage in CD4+ T cells was remarkably similar to that in CD8+ T cells in both groups. These data strongly suggest that differences in V beta gene family usage arising from coselection by major histocompatibility complex (MHC) class I versus MHC class II restriction elements do not fundamentally distort 'basic' V beta gene family usage patterns. They also support the concept that differences in CD4+ and CD8+ T-cell V beta gene family usage, which were increased in CD8hi CVI, can arise from high-affinity interactions between disease-associated antigens or superantigens and T cells in the post-thymic T-cell compartment.