RESUMEN
Immunoglobulin (Ig) A deficiency has long been recognized in patients with chromosome 18 abnormalities. We present the case of a young girl in whom a chromosome 18p deletion syndrome (46,XX,del[18][p11.1]) was associated not only with IgA deficiency, but also with an inability to make antibody to the unconjugated pneumococcal polysaccharide vaccine, Pneumovax II, indicating a concomitant specific polysaccharide antibody deficiency. The patient suffered from recurrent upper respiratory tract and genitourinary infections, which were controlled by the use of prophylactic antibiotics. The association of specific polysaccharide antibody deficiency, IgA deficiency, and chromosome 18p deletion syndrome has not been described previously, and extends the immunological phenotype of antibody deficiencies associated with defects of chromosome 18. The presence of specific polysaccharide antibody deficiency should be investigated in patients with chromosome 18 abnormalities, as these patients may have a more severe spectrum of infections than patients with chromosome 18 abnormalities and selective IgA deficiency alone.
Asunto(s)
Aberraciones Cromosómicas , Deleción Cromosómica , Cromosomas Humanos Par 18/inmunología , Deficiencia de IgA/genética , Polisacáridos Bacterianos/inmunología , Anticuerpos Antibacterianos/biosíntesis , Niño , Femenino , Humanos , Deficiencia de IgA/inmunología , Vacunas Neumococicas/inmunología , Polisacáridos Bacterianos/genética , SíndromeRESUMEN
The interaction of chemokines and their receptors directs lymphocyte migration, and is involved in the distribution and organization of lymphocytes within lymphoid tissues. We reasoned that abnormal chemokine receptor expression might give rise to defects of lymphocyte migration into and within lymphoid tissues, and consequently be associated with defective antibody production in primary antibody deficiencies. In this study, we have investigated the expression of chemokine receptors CXCR4, CXCR5 and CCR7 on lymphocyte subpopulations (naive and memory B cells; CD4(+) and CD8(+) T cells) in a cohort of patients with primary antibody deficiency (n = 23), and compared these with a group of healthy controls (n = 19). We show that there were significant differences in both the proportions of lymphocytes expressing, and the levels of expression of, specific chemokine receptors on individual lymphocyte subpopulations between patients and controls. Furthermore, these changes appeared more pronounced in patients with more severe antibody deficiency. These data support the hypothesis that abnormal lymphocyte trafficking may be involved in the pathogenesis of primary antibody deficiencies.
Asunto(s)
Linfocitos B/química , Síndromes de Inmunodeficiencia/inmunología , Receptores de Quimiocina/análisis , Linfocitos T/química , Adulto , Linfocitos B/inmunología , Estudios de Casos y Controles , Inmunodeficiencia Variable Común/inmunología , Femenino , Citometría de Flujo/métodos , Humanos , Memoria Inmunológica , Masculino , Persona de Mediana Edad , Receptores CCR7/análisis , Receptores CXCR4/análisis , Receptores CXCR5/análisis , Estadísticas no Paramétricas , Linfocitos T/inmunologíaRESUMEN
The cell line KG-1 has been used as an in vitro model for human dendritic cell (DC) differentiation. We have investigated the response of KG-1 cells to stimulation with a number of factors known to induce differentiation and/or maturation of DCs in vitro. KG-1 cells showed no differentiation in response to LPS, CpG oligodeoxynucleotide or CD40 ligation. Culture in the presence of TNF-alpha induced some differentiation, but only treatment with PMA and ionomycin (with or without prior culture in GM-CSF and IL-4) induced morphological and phenotypic changes consistent with DC-like maturation, and even these maximally differentiated KG-1 cells showed lower levels of surface marker expression, macromolecular endocytosis, and ability to stimulate in allogeneic MLR compared with in vitro monocyte-derived DCs. Our data show that KG-1 cells differentiate in vitro into cells with DC-like functional characteristics under the influence of strong inducers of cellular activation, but lack the potency of mature DCs in key aspects of professional antigen presenting cells.
Asunto(s)
Células Dendríticas/citología , Células Dendríticas/inmunología , Linfocitos T/metabolismo , Ligando de CD40/inmunología , Ligando de CD40/metabolismo , Adhesión Celular , Diferenciación Celular , Línea Celular , Proliferación Celular , Islas de CpG , Citocinas/inmunología , Citocinas/metabolismo , Células Dendríticas/metabolismo , Humanos , Prueba de Cultivo Mixto de Linfocitos , Oligodesoxirribonucleótidos/inmunología , Pinocitosis , Linfocitos T/inmunología , Acetato de Tetradecanoilforbol/farmacología , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
A panel of stable cell hybrids was generated by fusing a range of marrow-derived and solid tumour-derived human cell lines with the B-lymphoblastoid cell lines, HMy2 or KR4, and expression of immunologically relevant accessory and co-stimulatory molecules, and ability to stimulate allogeneic T-cell responses in vitro was investigated. Hybrid cell lines generated from three marrow-derived tumour cells consistently expressed both MHC class I and class II molecules, a range of accessory and T-cell co-stimulatory ligand molecules, including CD80 and CD86, and directly stimulated markedly enhanced T-cell proliferative responses in vitro, as compared with the parent tumour cell lines. The responses were blocked by addition of CTLA4-Ig fusion protein to the cultures, indicating a role of CD28/B7 interaction in induction of T-cell activation. By contrast, hybrid cells derived from three solid tumours only expressed MHC class II when the parent tumour cell line expressed MHC class II and consistently failed to express CD80 or CD86. These hybrid cells also stimulated greater T-cell proliferative responses in vitro than the parent tumour cell lines, although effective co-stimulation depended on the presence of responder non-T cells in the cultures. The expression of co-stimulatory ligand molecules and ability to directly stimulate strong allogeneic T-cell responses correlated with the EBV latency type of the hybrid cells. These data suggest that phenotypic and functional differences in fusion cells of professional antigen- presenting cells and tumour cells arise as a result of the parent tumour cell type.
Asunto(s)
Células Presentadoras de Antígenos/inmunología , Linfocitos B/inmunología , Fusión Celular , Células Híbridas/inmunología , Activación de Linfocitos , Linfocitos T/inmunología , Células Presentadoras de Antígenos/citología , Linfocitos B/citología , Linfocitos B/metabolismo , Linfocitos B/virología , Antígeno B7-1/inmunología , Antígeno B7-2/inmunología , Neoplasias de la Médula Ósea/inmunología , Línea Celular Tumoral , Proliferación Celular , Femenino , Herpesvirus Humano 4/fisiología , Antígenos de Histocompatibilidad Clase I/análisis , Antígenos de Histocompatibilidad Clase II/análisis , Humanos , Células Híbridas/citología , Melanoma/inmunología , Neoplasias del Cuello Uterino/inmunologíaRESUMEN
DiGeorge syndrome, which falls within a wider phenotypic spectrum associated with deletions of 22q11.2, is associated with a number of endocrine disorders. These include hypoparathyroidism, hypothyroidism and growth hormone deficiency. We report an unusual case of autoimmune hyperthyroidism (Graves' disease) presenting in a 3 year-old male with DiGeorge syndrome. The development of endocrine specific autoimmune disease in a syndrome associated with immune deficiency and the spectrum of endocrine autoimmunity associated with deletions of 22q11.2 are described. Paediatricians and patients with 22q11.2 deletions should be particularly aware of the risks of developing disorders of thyroid function.
Asunto(s)
Enfermedades Autoinmunes/etiología , Deleción Cromosómica , Cromosomas Humanos Par 22/genética , Síndrome de DiGeorge/complicaciones , Síndrome de DiGeorge/genética , Enfermedades del Sistema Endocrino/etiología , Enfermedad de Graves/etiología , Enfermedad de Graves/genética , Antitiroideos/uso terapéutico , Aorta Torácica/anomalías , Autoanticuerpos/análisis , Enfermedades Autoinmunes/genética , Carbimazol/uso terapéutico , Enfermedades del Sistema Endocrino/genética , Enfermedad de Graves/tratamiento farmacológico , Defectos de los Tabiques Cardíacos/complicaciones , Defectos de los Tabiques Cardíacos/cirugía , Humanos , Recién Nacido , Recuento de Linfocitos , Masculino , RecurrenciaRESUMEN
Recent studies have demonstrated that angiogenesis and suppressed cell-mediated immunity (CMI) play a central role in the pathogenesis of malignant disease facilitating tumour growth, invasion and metastasis. In the majority of tumours, the malignant process is preceded by a pathological condition or exposure to an irritant which itself is associated with the induction of angiogenesis and/or suppressed CMI. These include: cigarette smoking, chronic bronchitis and lung cancer; chronic oesophagitis and oesophageal cancer; chronic viral infections such as human papilloma virus and ano-genital cancers, chronic hepatitis B and C and hepatocellular carcinoma, and Epstein-Barr virus (EBV) and lymphomas; chronic inflammatory conditions such as Crohn's disease and ulcerative colitis and colorectal cancer; asbestos exposure and mesothelioma and excessive sunlight exposure/sunburn and malignant melanoma. Chronic exposure to growth factors (insulin-like growth factor-I in acromegaly), mutations in tumour suppressor genes (TP53 in Li Fraumeni syndrome) and long-term exposure to immunosuppressive agents (cyclosporin A) may also give rise to similar environments and are associated with the development of a range of solid tumours. The increased blood supply would facilitate the development and proliferation of an abnormal clone or clones of cells arising as the result of: (a) an inherited genetic abnormality; and/or (b) acquired somatic mutations, the latter due to local production and/or enhanced delivery of carcinogens and mutagenic growth factors. With progressive detrimental mutations and growth-induced tumour hypoxia, the transformed cell, to a lesser or greater extent, may amplify the angiogenic process and CMI suppression, thereby facilitating further tumour growth and metastasis. There is accumulating evidence that long-term treatment with cyclo-oxygenase inhibitors (aspirin and indomethacin), cytokines such as interferon-alpha, anti-oestrogens (tamoxifen and raloxifene) and captopril significantly reduces the incidence of solid tumours such as breast and colorectal cancer. These agents are anti-angiogenic and, in the case of aspirin, indomethacin and interferon-alpha have proven immunomodulatory effects. Collectively these observations indicate that angiogenesis and suppressed CMI play a central role in the development and progression of malignant disease.
Asunto(s)
Neoplasias/irrigación sanguínea , Neovascularización Patológica/etiología , Progresión de la Enfermedad , Silenciador del Gen , Genes p53/inmunología , Humanos , Higiene , Neoplasias/tratamiento farmacológico , Neoplasias/inmunología , Neovascularización Patológica/inmunología , Prostaglandina-Endoperóxido Sintasas/inmunología , Receptores de Interferón/inmunologíaAsunto(s)
Inmunodeficiencia Variable Común/diagnóstico , Tos/etiología , Enfermedades Profesionales/diagnóstico , Policia , Sarcoidosis Pulmonar/diagnóstico , Adulto , Biopsia , Inmunodeficiencia Variable Común/patología , Tos/patología , Diagnóstico Diferencial , Humanos , Pulmón/patología , Ganglios Linfáticos/patología , Masculino , Enfermedades Profesionales/patología , Sarcoidosis Pulmonar/patologíaRESUMEN
BACKGROUND AND METHODS: We have used the erosion of telomeric DNA as a measure of cellular division to study the replicative history of isolated T-lymphocyte subpopulations from a group of HIV-infected long-term survivors and age-matched healthy controls. RESULTS: In keeping with previous studies, we found that CD45RO+ (memory) T-cells showed greater telomere erosion than CD45RA+ (naive) T-cells. We did not, however, find any significant differences in the telomere lengths of isolated CD4+, CD8+, CD45RA+ or CD45RO+ T-cells between HIV-infected long-term survivors and age-matched controls. Further, we found no evidence of telomerase activation in T-cells from the HIV-infected groups to account for the lack of telomere erosion. CONCLUSIONS: Our data show no evidence, through telomere shortening, of clonal exhaustion or replicative senescence due to an increased rate of immune cell turnover in HIV-infected long-term survivors.
Asunto(s)
Infecciones por VIH/genética , Infecciones por VIH/inmunología , Sobrevivientes de VIH a Largo Plazo , Linfocitos T , Telómero/genética , Adulto , Biomarcadores , Estudios de Casos y Controles , Femenino , Infecciones por VIH/patología , Humanos , Masculino , Persona de Mediana Edad , PronósticoRESUMEN
Most tumours do not stimulate effective antitumour immune responses in vivo. In order to enhance the immunogenicity of human tumour cells, we fused a variety of tumour cell lines with an Epstein-Barr virus transformed B-lymphoblastoid cell line (EBV B-LCL) in vitro, to produce stable hybrid cells. Hybrid cell lines showed a marked increase in their ability to stimulate primary allogeneic T-cell responses in vitro, as compared with the parent tumour cells. The hybrid cells induced proliferation of naive (CD45RA+) as well as memory (CD45RO+) T lymphocytes, and both CD4+ and CD8+ subpopulations of T cells were directly stimulated. The stimulatory hybrids expressed human leucocyte antigen (HLA) class I and II, and a wide range of surface accessory molecules, including the T-cell co-stimulatory ligand molecules CD40, CD80 (B7.1) and CD86 (B7.2), the expression of which was required for optimal stimulation of T-cell responses. Fusion of the EBVB-LCL with a melanoma cell line (518.A2) yielded hybrid cells that expressed the melanoma-associated antigens MAGE-1 and MAGE-3, and presented these antigens to antigen-specific, HLA class I-restricted cytotoxic T-lymphocyte clones with greater efficiency than the parent melanoma cell line. These findings suggest that the generation of human antigen-presenting cell/tumour cell hybrids offers promise as an approach to cancer immunotherapy.
Asunto(s)
Células Presentadoras de Antígenos/inmunología , Antígenos HLA/inmunología , Hibridomas/inmunología , Inmunoterapia/métodos , Células Tumorales Cultivadas/inmunología , Proteínas Adaptadoras Transductoras de Señales , Presentación de Antígeno , Antígenos CD/inmunología , Antígenos de Neoplasias/inmunología , Antígeno B7-1/inmunología , Antígeno B7-2 , Antígenos CD40/inmunología , Proteínas Portadoras/análisis , Proteínas Portadoras/genética , Proteínas del Citoesqueleto , Pruebas Inmunológicas de Citotoxicidad , Citometría de Flujo , Herpesvirus Humano 4/inmunología , Humanos , Hibridomas/virología , Péptidos y Proteínas de Señalización Intracelular , Proteínas con Dominio LIM , Activación de Linfocitos , Melanoma/inmunología , Glicoproteínas de Membrana/inmunología , Neoplasias Cutáneas/inmunología , Subgrupos de Linfocitos TRESUMEN
The purpose of this study was to determine the mechanism by which adenosine, inosine, and guanosine delay cell death in glial cells (ROC-1) that are subjected to glucose deprivation and mitochondrial respiratory chain inhibition with amobarbital (GDMI). ROC-1 cells are hybrid cells formed by fusion of a rat oligodendrocyte and a rat C6 glioma cell. Under GDMI, ATP was depleted rapidly from ROC-1 cells, followed on a much larger time scale by a loss of cell viability. Restoration of ATP synthesis during this interlude between ATP depletion and cell death prevented further loss of viability. Moreover, the addition of adenosine, inosine, or guanosine immediately before the amobarbital retarded the decline in ATP and preserved cell viability. The protective effects on ATP and viability were dependent on nucleoside concentration between 50 and 1,500 microM. Furthermore, protection required nucleoside transport into the cell and the continued presence of nucleoside during GDMI. A significant positive correlation between ATP content at 16 min and cell viability at 350 min after the onset of GDMI was established (r = 0.98). Modest increases in cellular lactate levels were observed during GDMI (1.2 nmol/mg/min lactate produced); however, incubation with 1,500 microM inosine or guanosine increased lactate accumulation sixfold. The protective effects of inosine and guanosine on cell viability and ATP were >90% blocked after treatment with 50 microM BCX-34, a nucleoside phosphorylase inhibitor. Accordingly, lactate levels also were lower in BCX-34-treated cells incubated with inosine or guanosine. We conclude that under GDMI, the ribose moiety of inosine and guanosine is converted to phosphorylated glycolytic intermediates via the pentose phosphate pathway, and its subsequent catabolism in glycolysis provides the ATP necessary for maintaining plasmalemmal integrity.
Asunto(s)
Adenosina Trifosfato/metabolismo , Glucosa/farmacología , Mitocondrias/efectos de los fármacos , Oligodendroglía/citología , Nucleósidos de Purina/farmacología , Nucleótidos de Adenina/metabolismo , Nucleótidos de Adenina/farmacología , Adenosina/análogos & derivados , Adenosina/farmacología , Amobarbital/farmacología , Anaerobiosis , Animales , Astrocitos/citología , Astrocitos/efectos de los fármacos , Astrocitos/enzimología , Hipoxia de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Coformicina/farmacología , Relación Dosis-Respuesta a Droga , Transporte de Electrón/fisiología , Inhibidores Enzimáticos/farmacología , Moduladores del GABA/farmacología , Glioma , Glucólisis/fisiología , Nucleótidos de Guanina/metabolismo , Guanosina/farmacología , Células Híbridas/citología , Células Híbridas/efectos de los fármacos , Células Híbridas/metabolismo , Inosina/farmacología , Inosina Monofosfato/metabolismo , Isquemia/metabolismo , Ácido Láctico/biosíntesis , Ácido Láctico/metabolismo , Fármacos Neuroprotectores/farmacología , Oligodendroglía/efectos de los fármacos , Oligodendroglía/enzimología , Pentosiltransferasa/metabolismo , RatasRESUMEN
We have synthesized a recombinant gene encoding a single-chain HLA-A2/beta 2-microglobulin (beta 2m) molecule by linking beta 2m through its carboxyl terminus via a short peptide spacer to HLA-A2 (A*0201). This gene has been expressed in the beta 2m-deficient colorectal tumor cell line DLD-1. Transfection of this cell with the single-chain construct was associated with conformationally correct cell surface expression of a class I molecule of appropriate molecular mass. The single-chain HLA class I molecule presented either exogenously added peptide or (after interferon-gamma treatment) endogenously processed antigen to an influenza A matrix-specific, HLA-A2-restricted cytotoxic T-lymphocyte line. The need for interferon gamma for the processing and presentation of endogenous antigen suggests that DLD-1 has an antigen-processing defect that can be up-regulated, a feature that may be found in other carcinomas. Our data indicate that single-chain HLA class I constructs can form functional class I molecules capable of presenting endogenously processed antigens. Such molecules should be of use for functional studies, as well as providing potential anticancer immunotherapeutic agents or vaccines.
Asunto(s)
Células Presentadoras de Antígenos/fisiología , Antígeno HLA-A2/química , Linfocitos T Citotóxicos/inmunología , Microglobulina beta-2/química , Secuencia de Aminoácidos , Secuencia de Bases , Línea Celular , Cartilla de ADN/química , Humanos , Cooperación Linfocítica , Datos de Secuencia Molecular , Oligopéptidos/química , Oligopéptidos/inmunología , Proteínas Recombinantes de Fusión , Transducción de Señal , TransfecciónRESUMEN
We have developed a single DNA typing method which uses 144 sequence-specific primer (SSP) reactions to simultaneously detect all known HLA-A, B, C, DRB1, DRB3, DRB4, DRB5 and DQB1 specificities in an allele specific or group specific manner using the same method, reagents, PCR parameters and protocols for all loci. The results from this integrated class I & II method can be visualized on a single photographic or electronic image and hence is described as "Phototyping". Phototyping has an overall resolution greater than or equivalent to good serology and results can be obtained in under 3 hours making the method suitable for genotyping potential cadaver donor peripheral blood without serological backup. This in turn produces the potential for reducing cold ischaemia times in renal transplantation as well as the application of prospective matching to cardiac and liver transplantation. The method has capacity to detect new alleles, for example, novel amplification patterns suggestive of 4 new HLA-B alleles have been detected. The Phototyping set has been used as the sole method of HLA typing for over 1010 individuals. Phototyping is not problem-free; deviations from the standard protocol, poor quality DNA and unsuitable PCR machines can result in individual PCR failures or in incorrect assignment of antigens. Approximately 5% of genotypes were repeated (either partially or fully) because of incomplete or equivocal results.
Asunto(s)
Cartilla de ADN , ADN/genética , Genes MHC Clase II/genética , Genes MHC Clase I/genética , Prueba de Histocompatibilidad/métodos , Análisis de Secuencia de ADN , Secuencia de Bases , ADN/aislamiento & purificación , Sondas de ADN de HLA , Electroforesis en Gel de Agar , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la PolimerasaRESUMEN
HLA-A30 is present in the Sardinian population at a frequency of 23%. We have designed a system using nested ARMS-PCR to determine the relative frequencies of the HLA-A*30 allelic variants (A*3001, A*3002, and A*3003) within this population. The use of a nested PCR approach, in which the first-round reaction provides HLA-A*30 specificity and template DNA for the subsequent nested reactions, is a powerful means of discriminating between alleles of very similar sequence. Using this method, we performed subtyping of 35 serologically defined HLA-A30 Sardinian individuals, and taking into account homozygotes, identified 38 A*30 alleles. Of these, 33 typed as A*3002, four typed as A*3001, and one sample did not conform to the patterns of reactivity of any of the published A*30 alleles. Haplotype information showed strong linkage disequilibrium between A*3002 and B18. This study underlines the potential of DNA-based methods for typing HLA class I in terms of adding further levels of definition to studies of population structure and also as a means of identifying new alleles.
Asunto(s)
Alelos , Antígenos HLA-A/genética , Prueba de Histocompatibilidad/métodos , Reacción en Cadena de la Polimerasa/métodos , Secuencia de Aminoácidos , Secuencia de Bases , Secuencia de Consenso , Análisis Mutacional de ADN , Frecuencia de los Genes , Variación Genética , Genotipo , Humanos , Focalización Isoeléctrica , Italia/epidemiología , Desequilibrio de Ligamiento , Datos de Secuencia Molecular , Alineación de Secuencia , Homología de Secuencia de AminoácidoAsunto(s)
Alelos , Mapeo Cromosómico , Genes MHC Clase I , Antígenos HLA-A/genética , Reacción en Cadena de la Polimerasa/métodos , Antígenos de Neoplasias/genética , Secuencia de Bases , Linfoma de Burkitt/genética , Linfoma de Burkitt/patología , Exones , Humanos , Datos de Secuencia Molecular , Células Tumorales CultivadasRESUMEN
Daudi, a lymphoblastoid B cell line derived from an African Burkitt lymphoma does not express HLA-A,B,C antigens at the cell surface. Although HLA-A,B,C heavy chains are made normally they do not assemble into functional molecules because beta 2-microglobulin is absent. Previous serological analysis of somatic cell hybrids indicated that the HLA haplotypes of Daudi encoded HLA-A1, A10(A26), B17, and B16(38) antigens. Here we describe the application of molecular methods: ARMS-PCR, cDNA cloning and sequencing, immunoprecipitation and gel electrophoresis, to define the class I genotype of the Daudi cell line which is HLA-A*0102, A*6601, B*5801, B*5802, Cw*0302 and Cw*0602. With the exception of the B38 antigen, which is not a product of the alleles defined, the genotype is consistent with the serological description. Two previously undiscovered alleles emerged from this analysis: A*0102 and B*5802. The A*0102 allele differs from A*0101 by 5 nucleotide substitutions within exon 2 where it has a motif shared with A*30 alleles; the B*5802 allele differs from B*5801 by 3 substitutions in exon 3 where it has a motif shared with B*14 alleles. Subtyping HLA-A1 alleles showed A*0102 was well represented amongst individuals typed serologically as A1 in an African population but was absent from caucasoids. B*5802 has been found in a second individual. Thus the novel A and B alleles are not specific to the Daudi tumor. Overall, this analysis of a single East African cell illustrates the power of molecular methods to define new class I HLA alleles in non-caucasoid populations.