RESUMO
Summary: DRESS (drug reaction with eosinophilia and systemic symptoms) is a rare but potentially life-threatening disorder characterized by fever, skin eruption, haematological abnormalities and multi-organ dysfunction after drug exposure. The pathophysiology is thought to be related to interactions between culprit drugs, viral reactivation and T-lymphocytes activation. We report 4 paediatric patients with DRESS who were treated at our centre over the past 12 years. Most cases improved after corticosteroids. Other immunosuppressive medications were attempted in refractory cases with varied outcomes. Patient 3 was the first reported case that involved the use of infliximab, a TNF-α inhibitor, for DRESS. Although clinical efficacy was not observed for this one patient, a previous study demonstrated that patients with DRESS, disease progression and HHV-6 reactivation had elevated pre-treatment TNF- α and IL-6 levels. Further research is needed to explore the role of these cytokines in DRESS.
Assuntos
Fármacos Dermatológicos/toxicidade , Síndrome de Hipersensibilidade a Medicamentos/tratamento farmacológico , Síndrome de Hipersensibilidade a Medicamentos/epidemiologia , Eosinofilia/induzido quimicamente , Infliximab/toxicidade , Adolescente , Corticosteroides/uso terapêutico , Pré-Escolar , Síndrome de Hipersensibilidade a Medicamentos/diagnóstico , Feminino , Hong Kong/epidemiologia , Humanos , Masculino , Testes Cutâneos , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
Proinflammatory cytokines such as TNF-α and type I interferons (IFN) are pathogenic signatures of systemic lupus erythematosus, and plasmacytoid dendritic cells (pDCs) play a major role by predominantly producing IFN-α. Given the rise of importance in identifying tumor necrosis stimulated gene 6 (TSG-6) as a key anti-inflammatory regulator, we investigate its function and its ability to counteract proinflammatory cytokine secretion by pDCs in vitro. CpG-A and R837 induced significant endogenous TSG-6 expression in the pDC cell-line GEN2.2. Following recombinant human TSG-6 treatment and CpG-A or R837 stimulation, significant reduction in IFN-α and TNF-α was observed in healthy donors' pDCs, and the same phenomenon was confirmed in GEN2.2. By CD44 blocking assay, we deduced that the suppressive effect of TSG-6 is mediated by CD44, by reducing IRF-7 phosphorylation. Our findings suggest that TSG-6 and its downstream signalling pathway could potentially be targeted to modulate proinflammatory cytokine expression in pDCs.
Assuntos
Moléculas de Adesão Celular/farmacologia , Células Dendríticas/metabolismo , Interferon-alfa/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Humanos , Fator Regulador 7 de Interferon , Fosforilação/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/efeitos dos fármacosRESUMO
A 7-year-old male with Stage 4 neuroblastoma was treated with chemotherapy and autologous hematopoietic stem cell transplantation (HSCT), resulting in partial response with residual bone and marrow disease. He proceeded to haploidentical-HSCT with his mother as donor and achieved remission. The patient developed marrow relapse 2 years after haploidentical-HSCT with cytopenia and dropping donor chimerism. Donor lymphocyte infusion (DLI) using mother's whole blood was given resulting in clearance of marrow disease, resolution of cytopenia, and full donor chimerism. This is the first report of successful treatment for neuroblastoma relapse after haploidentical-HSCT using DLI alone, supporting the role of adoptive cell therapy post-HSCT in neuroblastoma.
Assuntos
Antineoplásicos/uso terapêutico , Transplante de Células-Tronco Hematopoéticas/métodos , Imunoterapia Adotiva/métodos , Recidiva Local de Neoplasia/terapia , Neuroblastoma/terapia , Criança , Humanos , Masculino , Transplante HomólogoRESUMO
Familial multiple intestinal atresias is an autosomal recessive disease with or without combined immunodeficiency. In the last year, several reports have described mutations in the gene TTC7A as causal to the disease in different populations. However, exact correlation between different genotypes and various phenotypes are not clear. In this study, we report identification of novel compound heterozygous mutations in TTC7A gene in a Malay girl with familial multiple intestinal atresias and severe combined immunodeficiency (MIA-SCID) by whole exome sequencing. We found two mutations in TTC7A: one that destroyed a putative splicing acceptor at the junction of intron 17/exon 18 and one that introduced a stop codon that would truncate the last two amino acids of the encoded protein. Reviewing the recent reports on TTC7A mutations reveals correlation between the position and nature of the mutations with patient survival and clinical manifestations. Examination of public databases also suggests carrier status for healthy individuals, making a case for population screening on this gene, especially in populations with suspected frequent founder mutations.
Assuntos
Predisposição Genética para Doença/genética , Atresia Intestinal/genética , Mutação , Proteínas/genética , Imunodeficiência Combinada Severa/genética , Sequência de Aminoácidos , Sequência de Bases , Saúde da Família , Feminino , Heterozigoto , Humanos , Lactente , Masculino , Dados de Sequência Molecular , Linhagem , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
The vaginal microbiota is a determinant for the risk of preterm birth (PTB). Dominance of the vaginal niche by Lactobacillus crispatus associates with term delivery. This is the first observational clinical study of live vaginal biotherapeutics (Lactobacillus crispatus CTV-05 (LACTIN-V)) in pregnant women at high-risk of PTB. The primary aim was to explore safety, tolerability and acceptability of LACTIN-V in pregnancy. Women were offered a course of LACTIN-V at 14 weeks gestation for five consecutive days followed by weekly administration for six weeks. Participants were followed up at 15, 18-, 20-, 28- and 36-weeks' gestation and at delivery for assessment of adverse events, compliance and tolerability. Participants completed a questionnaire to gauge experience and acceptability. In total, 73 women were recruited, of whom eight withdrew, leaving a final cohort size of 61. Self-reported compliance to the course was high (56/60, 93%). Solicited adverse events were reported in 13 women (19%) including changes in vaginal discharge, odour, colour or consistency of urine, itching and vaginal bleeding. One unsolicited adverse event was reported as haematuria at 38 weeks gestation, but was judged to be unrelated to LACTIN-V. No serious adverse events occurred. One mild adverse event led to study withdrawal. Thirty-one women completed an experience and acceptability questionnaire. Women found LACTIN-V easy and comfortable to use and the majority (30/31, 97%) would use LACTIN-V in future pregnancies. Eight women (8/31, 26%) found the schedule of use difficult to remember. The rate of PTB <34 weeks in this cohort was 3.3% compared to 7% in a historical cohort of 2,190 women at similar background PTB risk. With satisfactory uptake and good compliance, we demonstrate that LACTIN-V is safe and accepted in pregnancy, with high tolerability. Further studies are needed to assess colonisation of Lactobacillus crispatus CTV-05 and clinical efficacy.
Assuntos
Lactobacillus crispatus , Nascimento Prematuro , Probióticos , Recém-Nascido , Feminino , Gravidez , Humanos , Gestantes , Probióticos/efeitos adversos , VaginaRESUMO
Inflammatory bowel disease is well recognized for a strong genetic involvement in its pathogenesis. Homozygous mutations in interleukin-10 receptor 1 (IL-10R1) identified by linkage analysis were shown to be involved in this disorder. However, the underlying molecular mechanism and the causal nature of the mutations in the disease process remain to be clarified. In this study, using whole exome sequencing, we identified novel compound heterozygous missense mutations in the extracellular domain of IL-10R1 in a Crohn's disease patient from a non-consanguineous family. These mutations did not affect IL-10R1 expression, nor IL-10 binding. However, they abrogated IL-10R1 phosphorylation induced by IL-10, therefore leading to impaired STAT3 activation and suppression of inflammatory responses. After reconstitution with wild-type IL-10R1, the patient cells showed fully restored IL-10R function including IL-10-induced STAT3 activation and expression of suppressor of cytokine signaling 3. Thus, our results demonstrated that the mutations in IL-10R1 extracellular domain impair IL-10R1 activation rather than IL-10 binding, indicating these residues are important in IL-10 signal transduction through IL-10R1. The reconstitution data also confirmed the causality of the IL-10R1 mutations.
Assuntos
Doença de Crohn/genética , Exoma , Heterozigoto , Subunidade alfa de Receptor de Interleucina-10/genética , Mutação , Substituição de Aminoácidos , Sequência de Bases , Doença de Crohn/diagnóstico , Doença de Crohn/tratamento farmacológico , Humanos , Recém-Nascido , Interleucina-10/metabolismo , Subunidade alfa de Receptor de Interleucina-10/química , Subunidade alfa de Receptor de Interleucina-10/metabolismo , Masculino , Modelos Moleculares , Dados de Sequência Molecular , Ligação Proteica , Transdução de SinaisRESUMO
OBJECTIVE: Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease with complex genetic inheritance. CD247 (CD3Z, TCRZ) plays a vital role in antigen recognition and signal transduction in antigen-specific immune responses, and is known to be involved in SLE pathogenesis. Weak disease association was reported for genetic variants in this gene in Caucasian studies for SLE, Crohn's disease and systemic sclerosis, but its role as a genetic risk factor was never firmly established. METHODS: In this study, using a collection of 612 SLE patients and 2193 controls of Chinese ethnicity living in Hong Kong in a genome-wide study, single nucleotide polymorphisms (SNPs) in and around CD247 were identified as being associated with SLE. The two most significant SNPs in this locus were selected for further replication using TaqMan genotyping assay in 3339 Asian patients from Hong Kong, Mainland China, and Thailand, as well as 4737 ethnically and geographically matched controls. RESULTS: The association of CD247 with SLE in Asian populations was confirmed (rs704853: odds ratio [OR] = 0. 81, p = 2.47 × 10(-7); rs858543: OR = 1.10, p = 0.0048). Patient-only analysis suggested that rs704853 is also linked to oral ulcers, hematologic disorders and anti-double-stranded DNA (dsDNA) antibody production. CONCLUSION: A significant association between variants in CD247 and SLE was demonstrated in Asian populations. Understanding the involvement of CD247 in SLE may shed new light on disease mechanisms and development of new treatment paradigms.
Assuntos
Povo Asiático/genética , Complexo CD3/genética , Lúpus Eritematoso Sistêmico/genética , Lúpus Eritematoso Sistêmico/imunologia , Adulto , China , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Genótipo , Hong Kong , Humanos , Desequilíbrio de Ligação , Razão de Chances , Polimorfismo de Nucleotídeo Único , TailândiaRESUMO
UHRF1BP1 encodes a highly conserved protein with unknown function. Previously, a coding variant in this gene was found to be associated with systemic lupus erythematosus (SLE) in populations of European ancestry (rs11755393, R454Q, P=2.22 x 10â»8, odds ratio=1.17). In this study, by a combination of genome-wide study and replication involving a total of 1230 patients and 3144 controls, we confirmed the association of this coding variant to SLE in Hong Kong Chinese. We also identified another coding variant in this gene that independently contributes to SLE susceptibility (rs13205210, M1098T, P=4.44 x 10â»9, odds ratio=1.49). Cross-population confirmation establishes the involvement of this locus in SLE and indicates that distinct alleles are contributing to disease susceptibility.
Assuntos
Povo Asiático/genética , Proteínas Estimuladoras de Ligação a CCAAT/genética , Lúpus Eritematoso Sistêmico/genética , Mutação de Sentido Incorreto/genética , Alelos , Sequência de Aminoácidos , Frequência do Gene , Ordem dos Genes , Predisposição Genética para Doença/genética , Estudo de Associação Genômica Ampla , Hong Kong , Humanos , Desequilíbrio de Ligação , Polimorfismo de Nucleotídeo Único/genética , Ubiquitina-Proteína LigasesRESUMO
Although the immune system has long been implicated in the control of cancer, evidence for specific and efficacious immune responses in human cancer has been lacking. In the case of chronic myelogenous leukemia (CML), either allogeneic bone marrow transplant (BMT) or interferon-alpha2b (IFN-alpha2b) therapy can result in complete remission, but the mechanism for prolonged disease control is unknown and may involve immune anti-leukemic responses. We previously demonstrated that PR1, a peptide derived from proteinase 3, is a potential target for CML-specific T cells. Here we studied 38 CML patients treated with allogeneic BMT, IFN- alpha2b or chemotherapy to look for PR1-specific T cells using PR1/HLA-A*0201 tetrameric complexes. There was a strong correlation between the presence of PR1-specific T cells and clinical responses after IFN-alpha and allogeneic BMT. This provides for the first time direct evidence of a role for T-cell immunity in clearing malignant cells.
Assuntos
Leucemia Mielogênica Crônica BCR-ABL Positiva/imunologia , Serina Endopeptidases/imunologia , Linfócitos T Citotóxicos/imunologia , Circulação Sanguínea , Transplante de Medula Óssea , Citotoxicidade Imunológica , Efeito Enxerto vs Leucemia , Humanos , Interferon-alfa/uso terapêutico , Leucemia Mielogênica Crônica BCR-ABL Positiva/terapia , Mieloblastina , Fragmentos de Peptídeos/imunologia , Indução de RemissãoRESUMO
We identified circulating CD8+ T-cell populations specific for the tumor-associated antigens (TAAs) MART-1 (27-35) or tyrosinase (368-376) in six of eleven patients with metastatic melanoma using peptide/HLA-A*0201 tetramers. These TAA-specific populations were of two phenotypically distinct types: one, typical for memory/effector T cells; the other, a previously undescribed phenotype expressing both naive and effector cell markers. This latter type represented more than 2% of the total CD8+ T cells in one patient, permitting detailed phenotypic and functional analysis. Although these cells have many of the hallmarks of effector T cells, they were functionally unresponsive, unable to directly lyse melanoma target cells or produce cytokines in response to mitogens. In contrast, CD8+ T cells from the same patient were able to lyse EBV-pulsed target cells and showed robust allogeneic responses. Thus, the clonally expanded TAA-specific population seems to have been selectively rendered anergic in vivo. Peptide stimulation of the TAA-specific T-cell populations in other patients failed to induce substantial upregulation of CD69 expression, indicating that these cells may also have functional defects, leading to blunted activation responses. These data demonstrate that systemic TAA-specific T-cell responses can develop de novo in cancer patients, but that antigen-specific unresponsiveness may explain why such cells are unable to control tumor growth.
Assuntos
Antígenos de Neoplasias/metabolismo , Melanoma/imunologia , Melanoma/patologia , Linfócitos T/imunologia , Antígenos de Neoplasias/imunologia , Antineoplásicos/uso terapêutico , Antígenos CD28/imunologia , Antígenos CD28/metabolismo , Antígenos CD57/imunologia , Antígenos CD57/metabolismo , Linfócitos T CD8-Positivos/imunologia , Feminino , Citometria de Fluxo/métodos , Antígeno HLA-A2/genética , Antígeno HLA-A2/imunologia , Humanos , Receptores de Hialuronatos/imunologia , Receptores de Hialuronatos/metabolismo , Antígenos Comuns de Leucócito/imunologia , Antígenos Comuns de Leucócito/metabolismo , Metástase Linfática/imunologia , Metástase Linfática/patologia , Antígeno MART-1 , Melanoma/tratamento farmacológico , Melanoma/secundário , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Monofenol Mono-Oxigenase/genética , Monofenol Mono-Oxigenase/imunologia , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/imunologia , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/imunologia , Receptores de IgG/imunologia , Receptores de IgG/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Antígeno gp100 de MelanomaRESUMO
Current strategies for the immunotherapy of melanoma include augmentation of the immune response to tumor antigens represented by melanosomal proteins such as tyrosinase, gp100, and MART-1. The possibility that intentional targeting of tumor antigens representing normal proteins can result in autoimmune toxicity has been postulated but never demonstrated previously in humans. In this study, we describe a patient with metastatic melanoma who developed inflammatory lesions circumscribing pigmented areas of skin after an infusion of MART-1-specific CD8(+) T cell clones. Analysis of the infiltrating lymphocytes in skin and tumor biopsies using T cell-specific peptide-major histocompatibility complex tetramers demonstrated a localized predominance of MART-1-specific CD8(+) T cells (>28% of all CD8 T cells) that was identical to the infused clones (as confirmed by sequencing of the complementarity-determining region 3). In contrast to skin biopsies obtained from the patient before T cell infusion, postinfusion biopsies demonstrated loss of MART-1 expression, evidence of melanocyte damage, and the complete absence of melanocytes in affected regions of the skin. This study provides, for the first time, direct evidence in humans that antigen-specific immunotherapy can target not only antigen-positive tumor cells in vivo but also normal tissues expressing the shared tumor antigen.
Assuntos
Antígenos de Neoplasias/imunologia , Imunoterapia Adotiva/efeitos adversos , Melanócitos/imunologia , Melanoma/terapia , Proteínas de Neoplasias/imunologia , Neoplasias Cutâneas/terapia , Vitiligo/imunologia , Antígenos de Neoplasias/biossíntese , Feminino , Humanos , Imunoterapia Adotiva/métodos , Antígeno MART-1 , Melanócitos/citologia , Melanoma/complicações , Melanoma/imunologia , Melanoma/patologia , Pessoa de Meia-Idade , Proteínas de Neoplasias/biossíntese , Pele/citologia , Pele/imunologia , Pele/patologia , Neoplasias Cutâneas/complicações , Neoplasias Cutâneas/imunologia , Neoplasias Cutâneas/patologia , Linfócitos T Citotóxicos/classificação , Linfócitos T Citotóxicos/imunologia , Vitiligo/etiologia , Vitiligo/patologiaRESUMO
The method of oncotripsy, first proposed in Heyden & Ortiz (Heyden & Ortiz 2016 J. Mech. Phys. Solids 92, 164-175 (doi:10.1016/j.jmps.2016.04.016)), exploits aberrations in the material properties and morphology of cancerous cells in order to ablate them selectively by means of tuned low-intensity pulsed ultrasound. We propose the dynamical model of oncotripsy that follows as an application of cell dynamics, statistical mechanical theory of network elasticity and 'birth-death' kinetics to describe the processes of damage and repair of the cytoskeleton. We also develop a reduced dynamical model that approximates the three-dimensional dynamics of the cell and facilitates parametric studies, including sensitivity analysis and process optimization. We show that the dynamical model predicts-and provides a conceptual basis for understanding-the oncotripsy effect and other trends in the data of Mittelstein et al. (Mittelstein et al. 2019 Appl. Phys. Lett. 116, 013701 (doi:10.1063/1.5128627)), for cells in suspension, including the dependence of cell-death curves on cell and process parameters.
RESUMO
In this study, we compared the association of several newly discovered susceptibility genes for systemic lupus erythematosus (SLE) between populations of European origin and two Asian populations. Using 910 SLE patients and 1440 healthy controls from Chinese living in Hong Kong, and 278 SLE patients and 383 controls in Thailand, we studied association of STAT4, BLK and PXK with the disease. Our data confirmed association of STAT4 (rs7574865, odds ratio (OR) =1.71, P=3.55 x 10(-23)) and BLK (rs13277113, OR=0.77, P=1.34 x 10(-5)) with SLE. It was showed that rs7574865 of STAT4 is also linked to hematologic disorders and potentially some other subphenotypes of the disease. More than one genetic variant in STAT4 were found to be associated with the disease independently in our populations (rs7601754, OR=0.59, P=1.39 x 10(-9), and P=0.00034 when controlling the effect of rs7574865). With the same set of samples, however, our study did not detect any significant disease association for PXK, a risk factor for populations of European origin (rs6445975, joint P=0.36, OR=1.06, 95% confidence interval: 0.93-1.21). Our study indicates that some of the susceptibility genes for this disease may be population specific.
Assuntos
Predisposição Genética para Doença , Peptídeos e Proteínas de Sinalização Intracelular/genética , Lúpus Eritematoso Sistêmico/genética , Proteínas do Tecido Nervoso/genética , Proteínas Serina-Treonina Quinases/genética , Fator de Transcrição STAT4/genética , Adulto , Feminino , Genótipo , Hong Kong , Humanos , Desequilíbrio de Ligação , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genéticaRESUMO
Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease with complex genetic inheritance. Recently, single nucleotide polymorphisms (SNPs) in BANK1 and TNFSF4 have been shown to be associated with SLE in Caucasian populations, but it is not known whether they are also involved in the disease in other ethnic groups. Recent data from our genome-wide association study (GWAS) for 314 SLE cases and 920 controls collected in Hong Kong identified SNPs in and around BANK1 and TNFSF4 to be associated with SLE risk. On the basis of the results of the reported studies and our GWAS, SNPs were selected for further genotyping in 949 SLE patients (overlapping with the 314 cases in our GWAS) and non-overlapping 1042 healthy controls. We confirmed the associations of BANK1 and TNFSF4 with SLE in Chinese (BANK1, rs3733197, odds ratio (OR)=0.84, P=0.021; BANK1, rs17266594, OR=0.61, P=4.67 x 10(-9); TNFSF4, rs844648, OR=1.22, P=2.47 x 10(-3); TNFSF4, rs2205960, OR=1.30, P=2.41 x 10(-4)). Another SNP located in intron 1 of BANK1, rs4522865, was separately replicated by Sequenom in 360 cases and 360 controls and was also confirmed to be associated with SLE (OR=0.725, P=2.93 x 10(-3)). Logistic regression analysis showed that rs3733197 (A383T in ankyrin domain) and rs17266594 (a branch point-site SNP) from BANK1 had independent contributions towards the disease association (P=0.037 and 6.63 x 10(-8), respectively). In TNFSF4, rs2205960 was associated with SLE independently from the effect of rs844648 (P=6.26 x 10(-3)), but not vice versa (P=0.55). These findings suggest that multiple independent genetic variants may be present within the gene locus, which exert their effects on SLE pathogenesis through different mechanisms.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Povo Asiático/genética , Lúpus Eritematoso Sistêmico/etnologia , Lúpus Eritematoso Sistêmico/genética , Proteínas de Membrana/genética , Ligante OX40/genética , Epistasia Genética , Estudo de Associação Genômica Ampla , Hong Kong/epidemiologia , Humanos , Polimorfismo de Nucleotídeo Único , População Branca/genéticaRESUMO
OBJECTIVE: To estimate and compare the rate of pedestrian injuries in primary school-attending children of urban Uganda using different data sources. DESIGN: Data collection from a hospital-based trauma registry, police data, teacher reports, and a cross-sectional community-based survey. SETTING: Kawempe, the largest urban district in the capital Kampala, Uganda. Patients or SUBJECTS: Primary school-attending children aged 4-12 from 39 randomly selected schools were included in the trauma registry, police data, and teacher reports. 1828 households randomly selected from the 39 schools were interviewed for the community survey. MAIN OUTCOME MEASURE: A pedestrian injury. For the trauma registry-defined as a pedestrian injury resulting in a visit to the hospital. For the police data-defined as a pedestrian injury reported to the police. For the teacher reports and survey-defined as a pedestrian injury resulting in at least a day off school. RESULTS: The estimated pedestrian injury rates per 100 000 person-years were 54.0 (95% CI 25.3 to 117.4), <53.97 (95% CI 23.8 to 125.9), 1878.8 (95% CI 1513.1 to 2322.4), and 764.0 (95% CI 523.3 to 1117.2) from the trauma registry, police data, teacher reports, and community survey, respectively. CONCLUSIONS: Pedestrian injury rates differed significantly between different data sources. Users must be aware of the different target populations, definitions, and limitations of the data sources before direct comparisons are made. Injury reports by volunteer teachers may be a feasible source of injury data in other low/middle-income countries.
Assuntos
Acidentes de Trânsito/estatística & dados numéricos , Coleta de Dados/métodos , Estudantes/estatística & dados numéricos , Ferimentos e Lesões/epidemiologia , Criança , Pré-Escolar , Estudos Transversais , Feminino , Humanos , Masculino , Polícia/estatística & dados numéricos , Pobreza , Fatores de Risco , Instituições Acadêmicas/estatística & dados numéricos , Uganda/epidemiologia , Saúde da População UrbanaRESUMO
1. SARS coronavirus has low transmissibility at the community level. 2. Subclinical SARS coronavirus infection is rare in children.
Assuntos
Surtos de Doenças/estatística & dados numéricos , Transmissão de Doença Infecciosa/estatística & dados numéricos , Síndrome Respiratória Aguda Grave/epidemiologia , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/isolamento & purificação , Adolescente , Distribuição por Idade , Criança , Pré-Escolar , Intervalos de Confiança , Doenças Endêmicas/estatística & dados numéricos , Feminino , Hong Kong/epidemiologia , Humanos , Masculino , Prevalência , Probabilidade , Prognóstico , Valores de Referência , Fatores de Risco , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/imunologia , Estudos Soroepidemiológicos , Síndrome Respiratória Aguda Grave/sangue , Índice de Gravidade de Doença , Distribuição por Sexo , Taxa de SobrevidaRESUMO
Local IgA levels in 36 normal, 92 nonmalignant atypical, and 47 malignant human breast tissue specimens were analyzed by three methods: direct immunofluorescence, modified radioimmunoassay for IgA in cryostat sections, and the Immuno-Fluor (Bio-Rad Laboratories, Richmond, Calif.) assay for the cell surface IgA in tissue eluates. The proportion of epithelium in each sample was estimated microscopically. For malignant specimens the proportion of epithelial cells was significantly higher than that for nonmalignant specimens. The immunofluorescence test showed no qualitative difference in epithelial IgA content between normal and nonmalignant atypical tissues, but the frequency of positive specimens was markedly lower in malignant tissues. The modified radioimmunoassay and the Immuno-Fluor assay indicated little or no difference in IgA concentration per tissue weight among normal, nonmalignant atypical, and malignant breast tissues. However, when the data were corrected for proportion of glandular epithelium, nonmalignant atypical tissues had less IgA per cell than normal tissues, and malignant mammary carcinomas had significantly less IgA per cell than both normal and nonmalignant atypical tissues.
Assuntos
Neoplasias da Mama/imunologia , Mama/imunologia , Imunoglobulina A/metabolismo , Adenocarcinoma/imunologia , Carcinoma/imunologia , Carcinoma Intraductal não Infiltrante/imunologia , Epitélio/imunologia , Feminino , Imunofluorescência , Secções Congeladas , Humanos , Radioimunoensaio/métodosRESUMO
We previously showed (E. Clave et al., J. Immunother., 22: 1-6, 1999; J. Molldrem et al., Blood, 88: 2450-2457, 1996) that PR1, a human-lymphocyte-antigen (HLA)-A2.1-restricted peptide from proteinase 3, could be used to elicit CTLs from normal individuals. These CTLs showed HLA-restricted cytotoxicity and colony inhibition of myeloid leukemia cells that overexpress proteinase 3. In this study, we constructed a phycoerythrin-labeled PR1-HLA-A2 tetramer to identify PR1-specific CTLs by flow cytometry. No peripheral blood lymphocytes from three HLA-2.1+ donors stained with the tetramer, but, after 20 days in culture with weekly PR1 stimulation, 2-8% became tetramer+. Tetramer staining identified up to 40-fold more PR1-specific CTLs than were identified by limiting dilution analysis and correlated better with lysis of PR1-coated T2 cells (R2 = 0.95 versus R2 = 0.76). Tetramer+ CTLs were memory phenotype (91% CD45RO+), and most (58% CD95+) were activated. Tetramer-sorted allogeneic CTLs produced 83% lysis of HLA-A2.1+ chronic myelogenous leukemia (CML) blasts at an E:T ratio of 2.5:1, compared with 23% lysis by nonsorted CTLs, with no background lysis of HLA-A2.1+ normal cells. Cytoplasmic proteinase-3 expression was one log greater in CML blasts than in normal granulocytes. These results show that a PR1-HLA-A2 tetramer can be used to identify and select CTLs from normal donors that preferentially lyse CML cells, which could be used for leukemia-specific adoptive immunotherapy.
Assuntos
Antígeno HLA-A2/imunologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/imunologia , Fragmentos de Peptídeos/imunologia , Linfócitos T Citotóxicos/imunologia , Citometria de Fluxo , Humanos , Mieloblastina , Fenótipo , Serina Endopeptidases/imunologiaRESUMO
DNA methyltransferase I (Dnmt1), the maintenance enzyme for DNA cytosine methylation, is expressed at high levels in the CNS during embryogenesis and after birth. Because embryos deficient for Dnmt1 die at gastrulation, the role of Dnmt1 in the development and function of the nervous system could not be studied by using this mutation. We therefore used the cre/loxP system to produce conditional mutants that lack Dnmt1 in neuroblasts of embryonic day 12 embryos or in postmitotic neurons of the postnatal animal. Conditional deletion of the Dnmt1 gene resulted in rapid depletion of Dnmt1 proteins, indicating that the enzyme in postmitotic neurons turns over quickly. Dnmt1 deficiency in postmitotic neurons neither affected levels of global DNA methylation nor influenced cell survival during postnatal life. In contrast, Dnmt1 deficiency in mitotic CNS precursor cells resulted in DNA hypomethylation in daughter cells. Whereas mutant embryos carrying 95% hypomethylated cells in the brain died immediately after birth because of respiratory distress, mosaic animals with 30% hypomethylated CNS cells were viable into adulthood. However, these mutant cells were eliminated quickly from the brain within 3 weeks of postnatal life. Thus, hypomethylated CNS neurons were impaired functionally and were selected against at postnatal stages.
Assuntos
Sistema Nervoso Central/metabolismo , DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA , Doenças Neurodegenerativas/metabolismo , Neurônios/metabolismo , Animais , Animais Recém-Nascidos , Contagem de Células , Morte Celular , Sobrevivência Celular/fisiologia , Células Cultivadas , Sistema Nervoso Central/crescimento & desenvolvimento , Sistema Nervoso Central/patologia , Cruzamentos Genéticos , DNA (Citosina-5-)-Metiltransferase 1 , DNA (Citosina-5-)-Metiltransferases/deficiência , DNA (Citosina-5-)-Metiltransferases/genética , Eletrofisiologia , Marcação de Genes , Camundongos , Camundongos Transgênicos , Mosaicismo/genética , Doenças Neurodegenerativas/genética , Doenças Neurodegenerativas/patologia , Neurônios/citologia , Insuficiência Respiratória/genética , Células-Tronco/citologia , Células-Tronco/metabolismoRESUMO
Phospho-N-acetylmuramyl-pentapeptide translocase, the initial membrane enzyme in the biosynthesis of peptidoglycan, requires a lipid microenvironment for function. n-Butanol was reversibly intercalated into membranes to perturb the hydrophobic interactions in this microenvironment in order to define further the role of lipid. In the concentration range for maximal stimulation of enzymic activity (0.12-0.18 M), n-butanol causes a 40% decrease in the fluorescence emission of the dansylated product, undecaprenyl diphosphate-(N epsilon-dansyl)pentapeptide. Since no change in emission maximum occurs below 22 degrees C in the presence of 0.12 M n-butanol, it is concluded that intercalation of this alkanol causes an increase in fluidity. Above 22 degrees C this concentration of n-butanol causes both a decrease in the fluorescence emission and a red shift in the emission maximum. It is concluded that a polarity change as well as fluidity change occurs above 22 degrees C. n-Butanol also causes a significant change in the phase transition experienced by the dansylated lipid product. Thus, it is possible with n-alkanols, e.g. n-butanol, to perturb lipid-translocase interactions resulting in an increase in fluidity in the microenvironment of the enzyme. This change in fluidity correlates with a stimulation of enzymic activity.