RESUMO
OBJECTIVE: Dermatomyositis (DM) is an idiopathic inflammatory myopathy which often involves the lungs. DM is likely to be associated with aberrant T- and B-cell activation in the pathogenesis because of the proven effectiveness of T- and B-cell-targeted treatments. Assuming that the aberrant activation is reflected by biases in the lymphocyte subset repertoires, we aimed to elucidate these biases, especially in relation to clinical features of DM. METHOD: Based on the immunophenotyping standardized by the Human Immunology Project Consortium, untreated 13 DM patients, including seven patients with interstitial lung disease (ILD), and 18 age-matched healthy donors (HDs) were examined for proportions of peripheral blood lymphocyte subsets. Six DM patients were examined before and after successful induction of remission. RESULTS: Naïve CD4+ T cells and naïve B cells were more abundant, while there were fewer naïve CD8+ T cells, central memory CD8+ T cells, effector memory CD4+ T cells, Th1 cells, Tfh cells, and memory B cells in DM patients than in HDs. When the patients were subgrouped according to the presence of ILD, the lymphocyte subset repertoires in the patients with ILD contributed to the statistical differences in all the biased lymphocyte subset proportions. After treatment, transitional B cells vanished and there was an increase in memory B cells. CONCLUSION: The lymphocyte subset repertoires in the DM patients were biased, and were associated with the presence of ILD and disease activity of DM.
Assuntos
Dermatomiosite , Imunossupressores , Doenças Pulmonares Intersticiais , Subpopulações de Linfócitos/imunologia , Dermatomiosite/complicações , Dermatomiosite/diagnóstico , Dermatomiosite/tratamento farmacológico , Dermatomiosite/imunologia , Feminino , Humanos , Memória Imunológica/efeitos dos fármacos , Imunofenotipagem/métodos , Imunofenotipagem/estatística & dados numéricos , Imunossupressores/imunologia , Imunossupressores/uso terapêutico , Japão , Doenças Pulmonares Intersticiais/complicações , Doenças Pulmonares Intersticiais/diagnóstico , Doenças Pulmonares Intersticiais/imunologia , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde , Gravidade do PacienteAssuntos
Irmãos , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/imunologia , Análise Mutacional de DNA , Predisposição Genética para Doença , Humanos , Testes Imunológicos , Subunidade gama Comum de Receptores de Interleucina/genética , Masculino , Mutação , Fenótipo , Prognóstico , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/diagnóstico , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/genéticaAssuntos
Transplante de Células-Tronco Hematopoéticas , Herpesvirus Humano 4 , Células Matadoras Naturais/virologia , Transtornos Linfoproliferativos/terapia , Linfócitos T/virologia , Adulto , Proliferação de Células , Feminino , Humanos , Transtornos Linfoproliferativos/virologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Indução de Remissão , Fatores de Tempo , Condicionamento Pré-Transplante , Resultado do Tratamento , Adulto JovemRESUMO
Adoptive immunotherapies have been developed for antiviral agent-refractory cytomegalovirus (CMV) disease after stem cell transplantation (SCT). However, the application of such strategies is limited, particularly in terms of need for donor cooperation regarding blood sampling and inaccessibility in the setting of cord blood transplantation. Herein, we describe the first successful treatment of antiviral agent-refractory CMV enteritis after allogeneic SCT by the infusion of ex vivo-expanded donor-derived CD4(+) lymphocytes obtained from the recipient's peripheral blood.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Infecções por Citomegalovirus/imunologia , Infecções por Citomegalovirus/virologia , Citomegalovirus/isolamento & purificação , Enterite/tratamento farmacológico , Transplante de Células-Tronco de Sangue Periférico/efeitos adversos , Doadores de Sangue , Citomegalovirus/fisiologia , Infecções por Citomegalovirus/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Transplante Homólogo/efeitos adversos , Ativação ViralRESUMO
Hematopoietic cell transplantation (HCT) is used for treatment of hematopoietic diseases. Assessment of T- and B-cell reconstitution after HCT is crucial because poor immune recovery has a major effect on the clinical course. In this study, we retrospectively analyzed T-cell receptor excision circles (TRECs) as well as signal and coding joint kappa-deleting recombination excision circles (sjKRECs and cjKRECs, respectively) as markers of newly produced lymphocytes in 133 patients (56 primary immunodeficient and 77 malignant cases, median (range): 12 (0-62) years old). We analyzed the kinetics of TREC and KREC recovery and determined the factors that contributed to better immune recovery. KRECs became positive earlier than TRECs and increased thereafter. Younger recipient age had a favorable effect on recovery of sjKRECs and cjKRECs. Compared with BM and peripheral blood, our data suggested that cord blood (CB) provided rapid B-cell recovery. CB also provided better B-cell neogenesis in adult HCT recipients. Chronic GVHD was associated with low TRECs, but not increased sjKRECs/cjKRECs. Finally, positive sjKRECs 1 month after HCT were associated with fewer infectious episodes. Monitoring of TRECs and KRECs may serve as a useful tool for assessment of immune reconstitution post HCT.
Assuntos
Linfócitos B/citologia , Sangue Fetal/transplante , Doenças Hematológicas/terapia , Transplante de Células-Tronco Hematopoéticas/métodos , Adolescente , Adulto , Linfócitos B/imunologia , Criança , Pré-Escolar , Feminino , Doenças Hematológicas/imunologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Receptores de Antígenos de Linfócitos T/sangue , Receptores de Antígenos de Linfócitos T/imunologia , Estudos Retrospectivos , Condicionamento Pré-Transplante/métodos , Adulto JovemAssuntos
Ataxia Telangiectasia/diagnóstico , Proteínas de Ciclo Celular/metabolismo , Dano ao DNA , Proteínas de Ligação a DNA/metabolismo , Citometria de Fluxo/métodos , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Proteínas Mutadas de Ataxia Telangiectasia , Citometria de Fluxo/normas , Humanos , FosforilaçãoRESUMO
AIM: To measure the genomic DNA of human herpes viruses (HHV) in the ocular fluids and to analyse the clinical relevance of HHV in uveitis. METHODS: After informed consent was obtained, a total of 111 ocular fluid samples (68 aqueous humour and 43 vitreous fluid samples) were collected from 100 patients with uveitis. The samples were assayed for HHV-DNA (HHV1-8) by using two different polymerase chain reaction (PCR) assays, qualitative PCR (multiplex PCR) and quantitative PCR (real-time PCR). RESULTS: In all of the patients with acute retinal necrosis (n = 16) that were tested, either the HSV1 (n = 2), HSV2 (n = 3), or VZV (n = 11) genome was detected. In all patients, high copy numbers of the viral DNA were also noted, indicating the presence of viral replication. In another 10 patients with anterior uveitis with iris atrophy, the VZV genome was detected. When using multiplex PCR, EBV-DNA was detected in 19 of 111 samples (17%). However, real-time PCR analysis of EBV-DNA indicated that there were only six of the 19 samples that had significantly high copy numbers. The cytomegalovirus (CMV) genome was detected in three patients with anterior uveitis of immunocompetent patients and in one immunocompromised CMV retinitis patient. In addition, one patient with severe unilateral panuveitis had a high copy number of HHV6-DNA. There was no HHV7- or HHV8-DNA detected in any of the samples. CONCLUSIONS: A qualitative multiplex PCR is useful in the screening of viral infections. However, the clinical relevance of the virus infection needs to be evaluated by quantitative real-time PCR.
Assuntos
Alphaherpesvirinae/isolamento & purificação , DNA Viral/análise , Infecções Oculares Virais/diagnóstico , Infecções por Herpesviridae/diagnóstico , Uveíte/virologia , Alphaherpesvirinae/genética , Humor Aquoso/virologia , Infecções Oculares Virais/virologia , Genoma Viral , Infecções por Herpesviridae/virologia , Humanos , Reação em Cadeia da Polimerase/métodos , Corpo Vítreo/virologiaRESUMO
AIM: To investigative whether clinical manifestations of anterior uveitis are associated with the viral load of varicella zoster virus (VZV) in the aqueous humor in patients with herpes zoster ophthalmicus (HZO) and zoster sine herpete (ZSH). METHODS: After informed consent was given, an aliquot of aqueous humor was collected from patients with VZV anterior uveitis (n = 8). Genomic DNA of the human herpes viruses was measured in the aqueous humor by two PCR assays: a qualitative multiplex PCR and a quantitative real-time PCR. RESULTS: All patients had unilateral acute anterior uveitis with high intraocular pressure, mutton fat keratic precipitates with some pigmentation, and trabecular meshwork pigmentation. Multiplex PCR demonstrated VZV genomic DNA in all of the samples, but not in other human herpes virus samples (human simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus and human herpes virus types 6, 7 and 8). Real-time PCR revealed a high copy number of VZV DNA in the aqueous humor. After the initial onset of anterior uveitis, iris atrophy and distorted pupil with paralytic mydriasis developed. The intensity of iris atrophy and pupil distortion, but not ocular hypertension, correlated with the viral load of VZV in the aqueous humor. CONCLUSION: VZV viral load in the aqueous humor correlated significantly with damage to the iris (iris atrophy and pupil distortion) in patients with HZO and ZSH.
Assuntos
Humor Aquoso/virologia , Herpes Zoster Oftálmico/virologia , Herpesvirus Humano 3/isolamento & purificação , Uveíte Anterior/virologia , Zoster Sine Herpete/virologia , Doença Aguda , Adulto , Idoso , DNA Viral/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase/métodos , Carga ViralAssuntos
Transformação Celular Neoplásica , Dano ao DNA , Regulação Leucêmica da Expressão Gênica , Leucemia/genética , Leucemia/patologia , Síndromes Mielodisplásicas/genética , Síndromes Mielodisplásicas/patologia , Células da Medula Óssea/metabolismo , Histonas/metabolismo , Humanos , Imuno-Histoquímica/métodos , Perda de Heterozigosidade , MutaçãoAssuntos
Transplante de Células-Tronco de Sangue do Cordão Umbilical , Displasia Ectodérmica/terapia , Doenças Genéticas Ligadas ao Cromossomo X/terapia , Quinase I-kappa B/genética , Síndromes de Imunodeficiência/terapia , Condicionamento Pré-Transplante/métodos , Pré-Escolar , Displasia Ectodérmica/genética , Doenças Genéticas Ligadas ao Cromossomo X/genética , Humanos , Síndromes de Imunodeficiência/genética , Masculino , MutaçãoRESUMO
Haematopoietic stem cell transplantation (HSCT) is performed for treatment of a broad spectrum of illnesses. Reconstitution of an intact immune system is crucial after transplantation to avoid infectious complications, and above all, the establishment of T cell receptor (TCR) diversity is the most important goal in the procedure. Until recently, little has been known of the mechanism of T cell reconstitution in the very early period after HSCT. In this study, we analysed TCR repertoires sequentially in four patients with severe combined immunodeficiency (SCID) before and after HSCT. In all patients, the TCR repertoires were extremely abnormal before HSCT, whereas after transplantation there was progressive improvement in TCR diversity, based on analysis of the TCR Vbeta repertoire and CDR3 size distributions. Somewhat unexpectedly, there was a significant but transient expansion of TCR diversity 1 month after transplantation in all cases. Clonotypic analysis of TCRs performed in one case showed that many T cell clones shared identical CDR3 sequences at 1 month and that the shared fraction decreased progressively. These results indicate that early expansion of TCR diversity may reflect transient expansion of pre-existing mature T cells from the donor blood, independent of de novo T cell maturation through the thymus.
Assuntos
Transplante de Células-Tronco Hematopoéticas , Imunodeficiência Combinada Severa/imunologia , Imunodeficiência Combinada Severa/terapia , Subpopulações de Linfócitos T/imunologia , Sequência de Aminoácidos , Sequência de Bases , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Diferenciação Celular , Células Clonais/imunologia , Regiões Determinantes de Complementaridade/genética , Citometria de Fluxo , Humanos , Lactente , Masculino , Dados de Sequência Molecular , Receptores de Antígenos de Linfócitos T alfa-beta/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodosRESUMO
We retrospectively analyzed our results of 30 patients with three distinctive primary immunodeficiency diseases (PIDs)--severe combined immunodeficiency (SCID, n = 11), Wiskott-Aldrich syndrome (WAS, n = 11) and X-linked hyper-immunoglobulin M (IgM) syndrome (XHIM, n = 8)--who underwent hematopoietic SCT (HSCT) during the past 20 years. Until 1995, all donors were HLA-haploidentical relatives with T-cell depletion (TCD) (n = 8). Since 1996, the donors have been HLA-matched related donors (MRD) (n = 8), unrelated BM (UR-BM) (n = 7) and unrelated cord blood (UR-CB) (n = 7). Twenty-seven of 30 patients had various pre-existing infections with or without organ damages before HSCT. Conditioning regimen and GVHD prophylaxis were determined according to disease, donor and pretransplant status. Although one of eight patients transplanted with TCD is alive with full engraftment, the other seven died. On the other hand, 18 of 22 patients transplanted without TCD are alive and well, including six of eight transplanted from MRD, seven of seven from UR-BM and five of seven from UR-CB. All 19 survivors did not require Ig supplementation after HSCT. These results indicate that UR-CBT as well as UR-BMT provides good results for PID comparable to MRD-SCT, and that early diagnosis, HSCT at early stage, careful supportive therapy and monitoring for various pathogens are important for the successful HSCT.
Assuntos
Transplante de Células-Tronco Hematopoéticas/métodos , Síndromes de Imunodeficiência/terapia , Adolescente , Adulto , Criança , Pré-Escolar , Intervalo Livre de Doença , Transplante de Células-Tronco Hematopoéticas/mortalidade , Humanos , Síndromes de Imunodeficiência/complicações , Síndromes de Imunodeficiência/mortalidade , Lactente , Infecções , Depleção Linfocítica , Masculino , Estudos Retrospectivos , Taxa de Sobrevida , Doadores de Tecidos , Condicionamento Pré-Transplante/métodosRESUMO
The social amoebae are exceptional in their ability to alternate between unicellular and multicellular forms. Here we describe the genome of the best-studied member of this group, Dictyostelium discoideum. The gene-dense chromosomes of this organism encode approximately 12,500 predicted proteins, a high proportion of which have long, repetitive amino acid tracts. There are many genes for polyketide synthases and ABC transporters, suggesting an extensive secondary metabolism for producing and exporting small molecules. The genome is rich in complex repeats, one class of which is clustered and may serve as centromeres. Partial copies of the extrachromosomal ribosomal DNA (rDNA) element are found at the ends of each chromosome, suggesting a novel telomere structure and the use of a common mechanism to maintain both the rDNA and chromosomal termini. A proteome-based phylogeny shows that the amoebozoa diverged from the animal-fungal lineage after the plant-animal split, but Dictyostelium seems to have retained more of the diversity of the ancestral genome than have plants, animals or fungi.
Assuntos
Dictyostelium/genética , Genoma , Genômica , Comportamento Social , Transportadores de Cassetes de Ligação de ATP/genética , Animais , Composição de Bases , Adesão Celular/genética , Movimento Celular/genética , Centrômero/genética , Sequência Conservada/genética , Elementos de DNA Transponíveis/genética , DNA Ribossômico/genética , Dictyostelium/citologia , Dictyostelium/enzimologia , Dictyostelium/metabolismo , Células Eucarióticas/metabolismo , Duplicação Gênica , Transferência Genética Horizontal/genética , Humanos , Dados de Sequência Molecular , Filogenia , Proteoma , Proteínas de Protozoários/química , Proteínas de Protozoários/genética , RNA de Transferência/genética , Sequências Repetitivas de Ácido Nucleico/genética , Análise de Sequência de DNA , Transdução de Sinais/genética , Telômero/genéticaRESUMO
We have identified a gene encoding a eukaryotic initiation factor 4E-binding protein (4E-BP) in the EST database of the Dictyostelium cDNA project. The Dictyostelium 4E-BP, designated febA (four e-binding), showed significant similarity to mammalian 4E-BPs. Northern blot analysis revealed that febA was expressed at a high level in the vegetative growth phase but the level of expression decreased during late development. The gene was shown to be non-essential since disruption of the gene had no severe effect; the null mutant proliferated normally and formed normal fruiting bodies. However, strains overexpressing the gene could not be established, suggesting that an excess of FebA protein may have a lethal effect on the cells.
Assuntos
Proteínas de Transporte/genética , Dictyostelium/genética , Fosfoproteínas , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Complementar/química , DNA Complementar/isolamento & purificação , Vetores Genéticos , Dados de Sequência Molecular , Alinhamento de SequênciaRESUMO
The spatial expression patterns of genes involved in cyclic adenosine monophosphate (cAMP) responses during morphogenesis in Dictyostelium discoideum were analyzed by in situ hybridization. Genes encoding adenylyl cyclase A (ACA), cAMP receptor 1, G-protein alpha2 and beta subunits, cytosolic activator of ACA (CRAC and Aimless), catalytic subunit of protein kinase A (PKA-C) and cAMP phosphodiesterases (PDE and REG-A) were preferentially expressed in the anterior prestalk (tip) region of slugs, which acts as an organizing center. MAP kinase ERK2 (extracellular signal-regulated kinase-2) mRNA, however, was enriched in the posterior prespore region. At the culmination stage, the expression of ACA, CRAC and PKA-C mRNA increased in prespore cells in contrast with the previous stage. However, no alteration in the site of expression was observed for the other mRNA analyzed. Based on these findings, two and four classes of expression patterns were catalogued for these genes during the slug and culmination stages, respectively. Promoter analyses of genes in particular classes should enhance understanding of the regulation of dynamic and coordinated gene expression during morphogenesis.
Assuntos
AMP Cíclico/metabolismo , Dictyostelium/genética , Expressão Gênica , Animais , AMP Cíclico/genética , Dictyostelium/crescimento & desenvolvimento , Dictyostelium/metabolismo , Perfilação da Expressão Gênica , Genes de Protozoários , Hibridização In Situ , Modelos Biológicos , MorfogêneseRESUMO
h-Caldesmon is considered a novel specific marker for tumors with smooth muscle differentiation. To reassess its diagnostic use, the authors evaluated the immunohistochemical expression of h-caldesmon and other myogenic markers (calponin, alpha-smooth muscle actin, HHF35, and desmin) in 30 leiomyosarcomas (external soft tissues [15], retroperitoneum [8], uterus [5], other sites [2]), 26 myofibroblastic lesions, and 26 fibrohistiocytic tumors of varying biologic potential and histology. In contrast with previous data, h-caldesmon was expressed only in 11 (36%) of the 30 leiomyosarcomas analyzed, whereas they consistently expressed actins and frequently expressed calponin (86%) and desmin (76%). Leiomyosarcomas with the expression of h-caldesmon were well or moderately differentiated and primarily confined to the retroperitoneum or uterus. All but one leiomyosarcomas in the external soft tissues examined were negative for h-caldesmon, and the h-caldesmon-negative tumors showed moderately to poorly differentiated morphology. All myofibroblastic lesions examined were negative for h-caldesmon despite their constant expressions of at least one of the other markers. h-Caldesmon was not expressed in fibrohistiocytic tumors either, although focal positivity for the other markers was seen in subsets of the tumors. Thus, h-caldesmon can be regarded as a specific myogenic marker. However, one should be aware that the expression of h-caldesmon in leiomyosarcomas can be more variable according to their locations and/or extent of smooth muscle differentiation than considered previously.
Assuntos
Proteínas de Ligação a Calmodulina/metabolismo , Leiomiossarcoma/diagnóstico , Proteínas de Neoplasias/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/metabolismo , Diagnóstico Diferencial , Feminino , Humanos , Imuno-Histoquímica , Leiomiossarcoma/metabolismo , Leiomiossarcoma/patologia , Masculino , Pessoa de Meia-Idade , Músculo Liso/química , Neoplasias de Tecido Muscular/diagnóstico , Neoplasias de Tecido Muscular/metabolismo , Neoplasias de Tecido Muscular/patologiaRESUMO
We report here that soluble CD40 ligand (sCD40L) is released from human platelets when activated with collagen or thrombin. The sCD40L was detectable in the culture supernatants of platelets within 30 min after stimulation in vitro, and reached maximal levels in 3 h. The release was blocked by the metalloproteinase inhibitor, KB8301, indicating that the soluble CD40L is made by cleaving the membrane bound CD40L expressed on activated platelets. The sCD40L was undetectable in the supernatant of the activated platelets obtained from patients with X-linked hyper IgM syndrome (XHIM), who have defects in CD40L gene. Since sCD40L has been shown to have biologic function on the activation of vascular endothelial cells and B cells, these findings suggest that platelets play some roles in both inflammation and humoral immune response by releasing soluble CD40L.
Assuntos
Plaquetas/metabolismo , Ligante de CD40/análise , Ativação Plaquetária/fisiologia , Formação de Anticorpos/imunologia , Plaquetas/efeitos dos fármacos , Plaquetas/imunologia , Ligante de CD40/genética , Células Cultivadas , Colágeno/farmacologia , Ligação Genética/genética , Humanos , Ácidos Hidroxâmicos/farmacologia , Hipergamaglobulinemia/sangue , Hipergamaglobulinemia/genética , Imunidade Celular/imunologia , Imunoglobulina M/sangue , Células Jurkat , Inibidores de Metaloproteinases de Matriz , Mutação/genética , Ativação Plaquetária/efeitos dos fármacos , Solubilidade , Estatísticas não Paramétricas , Síndrome , Trombina/farmacologia , Fatores de Tempo , Cromossomo X/genéticaRESUMO
X-linked severe combined immunodeficiency (X-SCID) is a rare fatal disease that is caused by mutations in the gene encoding the gammac chain. In this study, 27 unrelated Japanese patients with X-SCID were examined in terms of their genetic mutations and surface expression of the gammac chain. Among 25 patients examined, excluding two patients with large deletions, 23 different mutations were identified in the IL2RG gene, including 10 novel mutations. One patient bearing an extracellular mutation and all three of the patients bearing intracellular mutations after exon 7 expressed the gammac chain on the cell surface. Overall, 84% of patients lacked surface expression of the gammac chain leading to a diagnosis of X-SCID.