RESUMEN
Skill increase in motor performance can be defined as explicitly measuring task success but also via more implicit measures of movement kinematics. Even though these measures are often related, there is evidence that they represent distinct concepts of learning. In the present study, the effect of multiple tDCS-sessions on both explicit and implicit measures of learning are investigated in a pointing task in 30 young adults (YA) between 27.07 ± 3.8 years and 30 old adults (OA) between 67.97 years ± 5.3 years. We hypothesized, that OA would show slower explicit skill learning indicated by higher movement times/lower accuracy and slower implicit learning indicated by higher spatial variability but profit more from anodal tDCS compared with YA. We found age-related differences in movement time but not in accuracy or spatial variability. TDCS did not skill learning facilitate learning neither in explicit nor implicit parameters. However, contrary to our hypotheses, we found tDCS-associated higher accuracy only in YA but not in spatial variability. Taken together, our data shows limited overlapping of tDCS effects in explicit and implicit skill parameters. Furthermore, it supports the assumption that tDCS is capable of producing a performance-enhancing brain state at least for explicit skill acquisition.
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A diagnosis of idiopathic anaphylaxis following a detailed clinical assessment remains very challenging for patients and clinicians. Risk reduction strategies such as allergen avoidance are not possible. This study investigated whether the (ISAC) allergen array with 103 allergens would add diagnostic value in patients with idiopathic anaphylaxis. We extended the specific immunoglobulin (Ig)E testing in 110 patients with a diagnosis of idiopathic anaphylaxis from five UK specialist centres using ISAC arrays. These were divided into three groups: score I identified no new allergen sensitization beyond those known by previous assessment, score II identified new sensitizations which were not thought likely to explain the anaphylaxis and score III identified new sensitizations felt to have a high likelihood of being responsible for the anaphylaxis. A proportion (50%) of score III patients underwent clinical reassessment to substantiate the link to anaphylaxis in this group. The results show that 20% of the arrays were classified as score III with a high likelihood of identifying the cause of the anaphylaxis. A wide range of major allergens were identified, the most frequent being omega-5-gliadin and shrimp, together accounting for 45% of the previously unrecognized sensitizations. The ISAC array contributed to the diagnosis in 20% of patients with idiopathic anaphylaxis. It may offer additional information where a careful allergy history and follow-on testing have not revealed the cause of the anaphylaxis.
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Alérgenos , Anafilaxia/diagnóstico , Análisis por Micromatrices/métodos , Adulto , Anciano , Alérgenos/inmunología , Anafilaxia/inmunología , Femenino , Humanos , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
A cohort of injection drug users (IDU) have been identified who despite a long history of IDU and sharing of injecting equipment remain seronegative and aviraemic for hepatitis C virus (HCV). They have been termed HCV exposed uninfected (EU). The study of potential innate or adaptive immune mechanisms of resistance to HCV infection in this group is of interest. The aim of this study was to determine the levels of a broad range of cytokines in serum of exposed, uninfected individuals to ascertain whether there is a specific cytokine profile associated with apparent resistance to HCV. Sera from 22 EU individuals were analysed for a range of cytokines and chemokines, and compared to 16 treatment-naive chronic HCV cases (HCV Ab+ RNA+), 16 individuals with spontaneous resolution of HCV (HCV-Ab+ and HCV-RNA-) and 10 healthy unexposed controls. EU subjects had strikingly higher levels of both IL-6 (on average more than 100-fold, P = 0.001) and IL-8 (on average more than 10-fold, P < 0.001) than the comparison groups. Additionally higher levels of tumour necrosis factor-alpha (TNF-α; on average up to threefold, P = 0.02) were seen in EU individuals. The levels of interferon-alpha (IFN-α) were upregulated in all HCV exposed groups in comparison to healthy controls (P = 0.013). Adaptive immune cytokine levels were no different between the groups. Cytokine profiling demonstrated raised levels of pro-inflammatory innate immune cytokines and chemokines in EU IDU, in particular interleukin-6 and interleukin-8. These findings suggest innate immune activation may be the key to prevention of infection in this cohort.
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Citocinas/sangre , Resistencia a la Enfermedad , Hepatitis C/prevención & control , Abuso de Sustancias por Vía Intravenosa/complicaciones , Abuso de Sustancias por Vía Intravenosa/inmunología , Adulto , Estudios de Cohortes , Femenino , Hepatitis C/inmunología , Humanos , Masculino , Compartición de AgujasRESUMEN
Many researchers employed mammalian expression system to artificially express cannabinoid receptors,but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports.In present study,we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system.This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide.In addition,the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e.,≥95℃),forming a large molecular weight band when analyzed by immuno-blotting.Only denaturing temperatures ≤75℃ yielded a clear band at the predicted molecular weight.Collectively,we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence,and described the requirement for a low sample denaturing temperature in immuno-blot analysis.These findings provide very useful information for efficient mammalian expression and immuno-blotting of mcmbrane receptors.
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Cellular immunity with interferon gamma production could have a role in protection from hepatitis C virus (HCV). Interleukin (IL)-12 is a key cytokine in promoting such anti-viral T helper 1 (Th1) responses. We hypothesized that a genetic background able to promote cellular responses may be associated with apparent protection from infection and have investigated the distribution of the functional 1188A/C polymorphism of IL-12B in HCV exposed but uninfected cases. The frequency of the high IL-12-producing C allele was determined by restriction enzyme genotyping in 76 exposed-uninfected individuals and 105 healthy controls. Overall, the C allele was found in 27.6% of exposed-uninfected cases compared with 16.7% of healthy controls [chi(2) = 6.3, P = 0.02, odds ratio (OR) = 1.9, 95% confidence interval (CI) = 1.1-3.2]. CC genotype was found in 10.5% of exposed-uninfected cases compared with 0.9% controls (chi(2) = 9.3, P = 0.01, OR = 12, 95% CI = 1.5-100). Individuals at high risk of HCV infection yet who remain uninfected may be resistant in some way to infection. In our cohort of exposed-uninfected cases a genetic background of enhanced IL-12 production was associated with apparent resistance to HCV infection. This lends support to a central role for cellular immune responses in protecting from infection.
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Hepatitis C/genética , Hepatitis C/prevención & control , Subunidad p40 de la Interleucina-12/genética , Polimorfismo de Longitud del Fragmento de Restricción , Adulto , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Genotipo , Hepatitis C/inmunología , Hepatitis C/transmisión , Hepatitis C Crónica/genética , Hepatitis C Crónica/inmunología , Hepatitis C Crónica/transmisión , Humanos , Inmunidad Celular , Inmunidad Innata , Masculino , Abuso de Sustancias por Vía Intravenosa/complicacionesRESUMEN
Acute rejection remains a poor predictor of graft outcome. In this study, we measured serum levels of interferon (IFN)-gamma and neopterin by enzyme-linked immunosorbent assay and a single nucleotide polymorphism (SNP) within the 3' untranslated region of the interleukin (IL)-12 B gene (1188 A/C) to determine whether either of these factors could predict acute rejection in renal transplantation. Significantly higher early post-transplant neopterin levels (days 5-7; 35.7 versus 19.9 nmol/l) were observed in recipients who subsequently rejected their grafts. Post-transplant neopterin levels showed a strong positive correlation with 1-month creatinine levels (Spearman's correlation 0.62, P < 0.001), suggesting macrophage activation early after transplantation. Pretransplant neopterin and IFN-gamma levels and the IL-12B gene SNP did not predict acute rejection in this small retrospective study. The ability to predict acute rejection non-invasively early after transplantation could lead to individual tailoring of immunosuppressive regimens and perhaps lead eventually to longer graft survival.
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Rechazo de Injerto/diagnóstico , Interferón gamma/sangre , Subunidad p40 de la Interleucina-12/genética , Trasplante de Riñón/inmunología , Neopterin/sangre , Enfermedad Aguda , Adulto , Biomarcadores/sangre , Femenino , Rechazo de Injerto/genética , Rechazo de Injerto/inmunología , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Nucleótido Simple , Pronóstico , Estudios RetrospectivosRESUMEN
BACKGROUND: Chronic idiopathic urticaria (CIU) is a distressing skin condition involving recurrent itchy hives lasting 6 weeks or longer. The mechanism involves mast cell and basophil degranulation, which releases inflammatory mediators including histamine. In our clinical practice, we have observed that the onset of CIU is often preceded by a major life event. OBJECTIVE: To investigate the role of the hormones of the hypothalamic-pituitary-adrenal (HPA) axis in the link between psychological stress and CIU. METHODS: Thirty people with CIU and 30 normal controls were recruited. A flow cytometric CD63 expression assay was used to quantify basophil activation, and serum cortisol concentrations were measured as an indication of stress. RESULTS: Both corticotrophin releasing factor (CRF) and adrenocorticotrophic hormone (ACTH) were shown to activate basophils. There was no significant difference between numbers of CIU patients and normal controls responding to CFR, ACTH or cortisol. However, the responses in the CIU patients were stronger than those in normal controls. There was also a trend towards higher serum cortisol concentrations in CIU patients. The basophil response to CRF and ACTH correlated with the serum cortisol concentration in normal controls, but not in CIU patients. CONCLUSIONS: Although our data have not supported the hypothesis that stress makes a major contribution to CIU, the heightened basophil response to CFR and ACTH and higher levels of serum cortisol do suggest a derangement of the HPA axis in CIU.
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Basófilos/inmunología , Urticaria/inmunología , Urticaria/patología , Adolescente , Adulto , Anciano , Antígenos CD/metabolismo , Basófilos/efectos de los fármacos , Basófilos/metabolismo , Antagonistas de los Receptores Histamínicos H1/farmacología , Hormonas/farmacología , Humanos , Hidrocortisona/sangre , Persona de Mediana Edad , Modelos Biológicos , N-Formilmetionina Leucil-Fenilalanina/farmacología , Hipófisis/efectos de los fármacos , Hipófisis/inmunología , Glicoproteínas de Membrana Plaquetaria/metabolismo , Tetraspanina 30 , Urticaria/metabolismoRESUMEN
Loss of pancreatic beta-cells in type I diabetes is associated with an increase in T helper 1 (Th1) proinflammatory cytokines in the islet milieu, with a concomitant reduction in Th2 anti-inflammatory cytokines. In animal models, manoeuvres designed to polarize Th1 responses towards Th2, particularly involving interleukin (IL)-4, have been shown to protect against insulitis and diabetes. The aim of this study was to determine whether IL-4 can exert a direct effect on beta-cell viability. The rat pancreatic beta-cell line, BRIN-BD11, was used. IL-4R mRNA expression was assayed by reverse transcription-polymerase chain reaction and DNA sequencing and protein expression measured using anti-IL-4R antibodies and confocal microscopy. Cells were pretreated in vitro with IL-4, incubated with IL-1beta and interferon (IFN)-gamma and DNA fragmentation and nitrite production analysed by flow cytometry and Griess assay, respectively. Expression of type I (IL-4R alpha and common gamma-chain) and type II (IL-4R alpha, IL-13R alpha-1) IL-4R mRNA transcripts, together with cell surface expression of IL-4R, was demonstrated. Pre-incubation with IL-4 reduced significantly cell death induced by IL-1beta alone or by a combination of IL-1beta and IFN-gamma, although this was not accompanied by a reduced production of nitrite. The protective effect of IL-4 was not seen when all three cytokines were added simultaneously. These results demonstrate, for the first time, expression of IL-4 receptor components on rat pancreatic beta-cells and reveal a direct protective effect on the loss of viability mediated by proinflammatory cytokines when beta-cells are pre-incubated with IL-4.
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Mediadores de Inflamación/inmunología , Células Secretoras de Insulina/inmunología , Interleucina-4/inmunología , Animales , Apoptosis/inmunología , Células Cultivadas , Citocinas/inmunología , Relación Dosis-Respuesta Inmunológica , Microscopía Confocal , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosRESUMEN
Cannabinoids have been suggested as possessing immunomodulatory properties, and cannabinoid receptors are present on leucocytes. Clinically, there is some evidence that cannabinoids may be therapeutically useful in treating multiple sclerosis, which is generally believed to be an autoimmune condition. This paper reports data derived from the Cannabinoids in MS (CAMS) study, which was the largest randomized controlled trial yet conducted to evaluate the therapeutic efficacy of cannabinoids. We found no evidence for cannabinoid influence on serum levels of interferon (IFN)-gamma, interleukin (IL)-10, IL-12 or C-reactive protein as measured using enzyme-linked immunosorbent assay (ELISA), in comparison to control values. Mitogenic stimulation experiments also failed to demonstrate any significant reduction in percentage of CD3+, IFN-gamma producing cells after exposure to cannabinoids in vivo, although numbers were small. Further work is needed to establish the functional significance of cannabinoid receptors on immune cells.
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Cannabinoides/inmunología , Citocinas/sangre , Factores Inmunológicos/inmunología , Esclerosis Múltiple/inmunología , Proteína C-Reactiva/análisis , Proteína C-Reactiva/inmunología , Cannabinoides/uso terapéutico , Cannabis/inmunología , Células Cultivadas , Citocinas/inmunología , Método Doble Ciego , Dronabinol/inmunología , Dronabinol/uso terapéutico , Femenino , Humanos , Factores Inmunológicos/uso terapéutico , Interferón gamma/sangre , Interferón gamma/inmunología , Interleucina-10/sangre , Interleucina-10/inmunología , Interleucina-12/sangre , Interleucina-12/inmunología , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/sangre , Esclerosis Múltiple/tratamiento farmacológico , Fitoterapia/métodos , Aceites de Plantas/uso terapéuticoRESUMEN
The benefits of providing accurate and culturally sensitive information for patients are now undisputable and information strategies are generally being adopted nationally. Measuring their benefits and adapting strategies to local needs is now a new challenge. Although questionnaires to measure global satisfaction are available, no concise tool has previously existed to specifically measure the information aspects of care. The aims of this study are: to establish which aspects of information were most important to the patients during their cancer pathway; to develop a robust single-page questionnaire that is sensitive to patient priorities to measure and compare satisfaction between patient groups; and to audit information provision within routine clinical practice. Seventy-seven consecutive patients were asked to complete a priority list of which aspects of information were most important to them during their recent cancer care. A total of 303 answers were categorized into five final categories of equal weighting. The data were combined with demographic issues to produce a robust one-page questionnaire which has been adopted formally by the West Anglia Cancer Network. One hundred consecutive patients were then audited to highlight the information shortcomings of a busy community oncology department and the findings are reforming information services. We recommend the use of this practical audit tool in routine clinical practice, which can be downloaded free from cancernet.co.uk in seven languages.
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Neoplasias , Educación del Paciente como Asunto/normas , Satisfacción del Paciente , Encuestas y Cuestionarios/normas , Adulto , Anciano , Servicios de Salud Comunitaria/normas , Femenino , Humanos , Masculino , Persona de Mediana Edad , Sensibilidad y EspecificidadRESUMEN
The generation of an effective immune response is dependent on the efficient capture and presentation of antigen by antigen-presenting cells. The most potent antigen-presenting cells are dendritic cells (DC). These cells have the capability of activating naive helper and cytotoxic T cells. In recent years it has been demonstrated that in vivo responses to a number of solid tumours can be generated by DC pulsed with either purified tumour antigen or whole tumour cell lysate. In addition, a number of in vivo studies using DC have also been attempted in solid tumours, with some encouraging results. In haematological malignancies, there is now strong evidence that previous T cell anergy can be reversed and significant anti-tumour immune responses generated, in vitro, against the majority of leukaemias. As far as in vivo studies in haematological malignancies are concerned, although T cell responses have been demonstrated in the majority of cases and some dramatic early clinical responses reported, overall results appear disappointing. However, considering the fact that many of these studies were performed in patients with advanced disease and that such therapeutic strategies are still in their infancy, the overall results are actually quite encouraging. Although there is a real potential for DC immunotherapy in the future, it is important to be realistic about the limitations and obstacles to its development. It is highly unlikely that any form of immunotherapy is going to be effective in advanced disease due to the physical bulk of tumour, the immunosuppressive effects of tumours themselves and to any secondary immunosuppression following standard cancer therapy. The potential for immunotherapy is likely to lie either in adjunctive therapy or for treating minimal residual disease. Even in those situations, one of the major obstacles to be overcome is the state of immunological anergy or tolerance that many tumours seem able to induce. Indeed, there is evidence that, under certain circumstances, DC themselves can present antigen in such a way as to produce this state of anergy. Although, in vitro manipulation of DC and T cells can generate tumour-specific T cells from previously "anergic" cells, once reintroduced in vivo, these cells will be re-exposed to the tumour environment with the risk of being rendered anergic again.
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Vacunas contra el Cáncer/uso terapéutico , Células Dendríticas/fisiología , Neoplasias Hematológicas/terapia , Inmunoterapia/métodos , Linfocitos T/inmunología , Ensayos Clínicos como Asunto , Neoplasias Hematológicas/inmunología , HumanosRESUMEN
There is increasing evidence of T cell dysfunction in B cell chronic lymphocytic leukaemia (B-CLL) which may contribute to the aetiology and progress of the disease. An absolute CD8+ lymphocytosis correlates with disease progression and low expression of CD4 and CD8 (as found in autoimmune disease) is seen with abnormal expression of other surface molecules. Although the expression of T cell surface activation markers, CD25 and CD152, may be increased on culture in B-CLL serum, response to the common mitogens, PHA and PWM, is reduced. This and the excess of CD8 cells may explain partly the variable cooperation of T cells with B cell production of immunoglobulin in B-CLL. In the context of T cell cross-talk with antigen presenting cells, B-CLL B cells are poor antigen presenters. But the T cells themselves have significant abnormalities of expression of the many antigens and ligands necessary for this process. In particular, they exhibit variable expression of the low affinity and non-specific adhesion molecules LFA-1 and ICAM-1, variable, clonally restricted and skewed expression of the TCR repertoire (implying repeated antigenic stimulation possibly by CLL antigens), reduced CD28 and CD152 expression (implying impairment of ability to start or stop an immune response) and reduced IL2 and CD25 (IL2 R) expression (critical for positive feed-back in maintenance and expansion of the T cell response to antigen presentation). Although the production of IL2 and other cytokines by the T cell in B-CLL may be impaired, production of the anti-apoptotic cytokine IL4 is not and there may be a unique and expanded subset of CD8/CD30 cells capable of releasing IL4. The relationship of this T cell subset to the malignant B cell in vivo is unknown. However, T cells which are CD4+/CD152+/CCR4+ migrate selectively in vitro in response to the chemokine CCL22 (specific for the receptor CCR4) produced by the malignant B cells and are always seen amongst the malignant cells in bone marrow and lymph nodes from B-CLL patients. Other abnormalities of cytokine secretion are described. These findings suggest that the T cell in B-CLL may be unable to start, maintain and complete an immune response to the malignant B cell and other antigens and may be involved directly in sustaining the tumour. However, autologous tumour specific cytotoxicity has been shown in vitro and T cells which recognise tumour-derived heavy chain fragments circulate in vivo. If adoptive immunotherapy of any nature is to succeed in B-CLL, manipulation to optimise these CTL responses is needed to overcome the profound and variable T cell dysfunction in this disease.
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Leucemia Linfocítica Crónica de Células B/inmunología , Subgrupos de Linfocitos T/inmunología , Formación de Anticuerpos , Antígenos CD/fisiología , Antígenos de Neoplasias/inmunología , Antígenos de Superficie/fisiología , Moléculas de Adhesión Celular/fisiología , Ensayo de Unidades Formadoras de Colonias , Citocinas/metabolismo , Citotoxicidad Inmunológica , Progresión de la Enfermedad , Humanos , Síndromes de Inmunodeficiencia/etiología , Síndromes de Inmunodeficiencia/inmunología , Leucemia Linfocítica Crónica de Células B/complicaciones , Activación de Linfocitos/efectos de los fármacos , Cooperación Linfocítica , Recuento de Linfocitos , Mitógenos/farmacología , Proteínas de Neoplasias/inmunología , Proteínas de Neoplasias/fisiología , Complejo Receptor-CD3 del Antígeno de Linfocito T/inmunología , Subgrupos de Linfocitos T/metabolismoRESUMEN
HLA class II-restricted proliferative and cytotoxic T cell (CTL) responses to B cell chronic lymphocytic leukaemia (B-CLL) can be generated using autologous dendritic cells (DCs) pulsed with tumour cell lysate. In this study a number of different approaches were used to optimize further the in vitro system. First, the effects of a variety of maturation agents were studied. The addition of TNF-alpha, polyriboinosinic polyribocytidylic acid (Poly(I:C)) and LPS to autologous DCs resulted in the emergence of only a small percentage of CD83+ DCs, IFN-alpha having no demonstrable effect. Only the addition of Poly(I:C) to DCs resulted in modestly increased specific cytotoxicity to B-CLL targets, IFN-alpha and LPS having no effect. Secondly, T cells were pretreated with IL-15, prior to culturing with lysate-pulsed autologous DCs. A significant increase in T cell activation (P = 0.038), IFN-gamma secretion (P = 0.030) and specific cytotoxicity to B-CLL targets (P = 0.006) was demonstrated compared to untreated T cells. Thirdly, monocyte derived DCs electrofused with B-CLL B cells were compared with lysate-pulsed DCs. T cells stimulated by fused DCs generated higher levels of specific cytotoxicity to autologous B-CLL B cell targets than those stimulated by lysate pulsed DCs (P = 0.013). Blocking studies demonstrated inhibition of this cytotoxicity by both anti-CD4 (P = 0.062) and anti-CD8 monoclonal antibodies (P = 0.018), suggesting the generation of both HLA class I- and HLA class II-restricted CTL responses. In summary, in vitro B-CLL-specific T cell responses can be enhanced further by preincubating T cells with IL-15 and using autologous fused DC-B-CLL hybrids instead of autologous lysate-pulsed DCs. These preliminary data require confirmation with larger numbers of patients. Such an approach, however, may eventually provide effective immunotherapy for treatment of B-CLL.
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Vacunas contra el Cáncer , Células Dendríticas/inmunología , Interleucina-15/inmunología , Leucemia Linfocítica Crónica de Células B/inmunología , Linfocitos T Citotóxicos , Linfocitos B/inmunología , Linfocitos T CD8-positivos/inmunología , Fusión Celular , Antígenos de Histocompatibilidad Clase I/inmunología , Humanos , Hibridomas , Interferón-alfa/farmacología , Lipopolisacáridos/farmacología , Polinucleótidos/farmacología , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
Bronchiectasis is a common complication of primary antibody deficiency but the incidence of antibody deficiency as an underlying cause of bronchiectasis is largely undefined. In this study the humoral immune status of a cohort of bronchiectatic patients was investigated to detect the frequency of significant antibody deficiency and to determine the extent of immunological investigation which is appropriate for routine assessment of bronchiectasis patients. Fifty-six out-patients (with a mean age of 59.6 years) had serum immunoglobulins, IgG subclasses and specific antibodies to capsular polysaccharides of Haemophilus influenzae and Streptococcus pneumoniae measured. Where specific antibody -levels were low, where possible, appropriate immunization with pneumococcal or conjugated Haemophilus polysaccharide vaccines was offered and the responses quantified. Three of 56 patients had low total serum IgG levels. Thirteen of 56 had deficiencies of either a single IgG subclass or combinations of two or more subclasses, with IgG4 being most frequently implicated (9/56). Twenty-nine of 56 had low basal specific polysaccharide antibody levels. Test immunization, where performed, produced satisfactory responses in all cases except one, where a specific defect of responsiveness to pneumococcal polysaccharide was identified. This study indicates that antibody deficiency is an uncommon aetiological/underlying factor in the causation of bronchiectasis beyond the fourth decade and that detailed investigation of humoral immune status as a routine in bronchiectasis patients, at least at this age, is not generally justified.
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Anticuerpos Antibacterianos/sangre , Bronquiectasia/inmunología , Síndromes de Inmunodeficiencia/complicaciones , Adulto , Anciano , Vacunas Bacterianas/uso terapéutico , Femenino , Haemophilus influenzae tipo b/inmunología , Humanos , Inmunoglobulina G/sangre , Inmunoglobulinas/sangre , Síndromes de Inmunodeficiencia/diagnóstico , Síndromes de Inmunodeficiencia/terapia , Masculino , Persona de Mediana Edad , Polisacáridos Bacterianos/inmunología , Streptococcus pneumoniae/inmunologíaRESUMEN
AIMS: alpha1 Antitrypsin was undetectable in several patient samples treated with 0.5% beta propiolactone, which was used as a virucidal agent. This study was designed to confirm beta propiolactone as the cause and determine why it might have such an effect. METHODS: Volumes of 0, 5, 10, and 20 micro l of beta propiolactone were added to 2 ml aliquots of serum to make final concentrations of 0%, 0.25%, 0.5%, and 1% of beta propiolactone. alpha1 Antitrypsin concentrations and the pH were measured at different time intervals. The effects of adding buffer before the addition of beta propiolactone, NaOH after beta propiolactone, and 6M HCl instead of beta propiolactone were also measured. RESULTS: The addition of beta propiolactone to a volunteer's serum showed a fall in both alpha1 antitrypsin values and pH with increasing time and concentration of beta propiolactone. This effect was also seen when adding HCl, but was partially prevented by buffering the serum or adding NaOH. CONCLUSIONS: These results suggest that it is the acidity of the degradation products of beta propiolactone that is responsible for the fall in alpha1 antitrypsin values. This fall in alpha1 antitrypsin values was dependent on the concentration of beta propiolactone used and the length of time before the test was performed. The effect of beta propiolactone on laboratory tests should be re-evaluated, with attention being paid to sample pH, storage time, and storage temperature.
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Desinfectantes/farmacología , Propiolactona/farmacología , alfa 1-Antitripsina/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Concentración de Iones de Hidrógeno/efectos de los fármacos , Factores de Tiempo , alfa 1-Antitripsina/metabolismoRESUMEN
Immunotherapy using dendritic cells has shown encouraging results in both haematological and non-haematological malignancies. In this study, monocyte-derived dendritic cells from patients with B-CLL were cultured for 6 days in the presence of IL-4 and GM-CSF. Autologous B-CLL T-cells were cultured alone or with B-CLL lysate-pulsed and unpulsed autologous dendritic cells. IFN-gamma secretion was assessed using ELISA. Cytotoxicity was assessed, after 21 days in culture and re-stimulation, using flow cytometry with and without blockade by anti-HLA class I, anti-HLA class II, anti-CD4, anti-CD8 and anti-TCRalphabeta monoclonal antibodies. B-CLL T cells stimulated with B-CLL lysate-pulsed autologous dendritic cells showed a significant (P = 0.0004) increase in IFN-gamma secretion and a significant (P = 0.0008) increase in specific cytotoxicity to autologous B-cell targets, but none to autologous T cell or B cell targets from healthy individuals. B-CLL T cells cultured with (non-B-CLL) B-cell lysate-pulsed B-CLL dendritic cells showed no significant response. Pulsing dendritic cells from healthy volunteers with an autologous (non-B-CLL) B-cell lysate did not stimulate proliferation, cytokine production or cytotoxicity by autologous T cells. Pulsing B-CLL dendritic cells with allogeneic B-CLL lysates and culturing with autologous T-cells elicited cytotoxicity against autologous B-CLL targets in some cases, but not in others. Cytotoxicity was significantly reduced by blocking with anti-HLA class II (P = 0.001), anti-TCRalphabeta (P = 0.03) and anti-CD4 (P = 0.046) antibodies. Phenotyping of the responding T-cell population demonstrated the majority to be CD4 positive. Our data demonstrate that HLA class II-restricted proliferative and cytotoxic T-cell responses to B-CLL can be generated using autologous dendritic cells pulsed with tumour cell lysate.
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Antígenos de Neoplasias/inmunología , Células Dendríticas/inmunología , Antígenos de Histocompatibilidad Clase II/fisiología , Leucemia Linfocítica Crónica de Células B/inmunología , Activación de Linfocitos , Linfocitos T Citotóxicos/inmunología , Anticuerpos Monoclonales/farmacología , Extractos Celulares/inmunología , Células Cultivadas , Citocinas/biosíntesis , Pruebas Inmunológicas de Citotoxicidad , Células Dendríticas/clasificación , Antígenos de Histocompatibilidad Clase II/inmunología , Humanos , Inmunofenotipificación , Inmunoterapia Adoptiva , Cinética , Leucemia Linfocítica Crónica de Células B/terapia , Subgrupos de Linfocitos T/clasificaciónRESUMEN
Although there is evidence that cytokine gene polymorphisms are associated with varying quantities of cytokine protein production, the exact role of these polymorphisms in allograft rejection remains unclear. In a previous study, we demonstrated a significant association between high IL-10 secretion in mixed lymphocyte culture (MLC), together with HLA mismatching for at least 4-6 antigens, with the occurrence of acute rejection following renal transplantation. We, therefore, wished to ascertain whether cytokine gene polymorphisms are associated with varying levels of protein secretion and/or allograft rejection in the same group of patients. Cytokine protein secretion in MLC for IL-4, IL-6, IL-10 and IFN-gamma was measured by ELISA in 49 patient-donor pairs. Protein secretion for the above cytokines was also measured in phytohaemagglutinin (PHA) stimulated cultures in 30 normal controls. In both patient and control groups, single nucleotide polymorphism analysis for IL-4 G(-590)T, IL-6 G(-174)C, IL-10 G(-1082)A, IL-10 C(-819)T, IL-10 C(-592)A, TNF-alpha G(-308)A and microsatellite analysis for IFNG (CA repeat) was performed. No correlation was found between cytokine gene polymorphisms and cytokine protein secretion in either mitogen stimulated cultures (control group) or MLC (patient group). In addition, no correlation was demonstrated between cytokine gene polymorphisms and renal allograft rejection.