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1.
Transfus Med ; 25(5): 320-5, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26174724

RESUMEN

BACKGROUND AND OBJECTIVES: Irradiation of red cell concentrates RCCs) can lead to well-documented elevated extracellular potassium concentrations. Transfusion of these products has the potential, if given as a massive/rapid transfusion, to lead to transient hyperkalemia. A potassium absorption filter (PAF) has recently been developed and has been proven to effectively remove excess K(+) . However, data are lacking on the red cell quality parameters over storage after irradiation. METHODS: Thirty RCCs were pooled and split into 3 groups of 10. All RCCs were irradiated on day 14 and filtered on day 28 (group 1 control), day 15 (group 2) or day 17 (group 3). Pre-irradiation, pre- and post-filtration and day 28 samples were taken for each study. Standard red cell quality parameters were measured over storage at the above time points. RESULTS: Losses for haemoglobin, haematocrit and volume were minimal after filtration with all units containing >40 g Hgb unit(-1). Statistically, significant differences were observed for K(+) and Na(+) levels in groups filtered at either 24 or 72 h post-irradiation, and this was observed directly after filtration and remained by day 28. Filtration had no significant impact on any other parameters measured. CONCLUSIONS: PAF effectively removed supernatant potassium (93%) from all RCC units. Early removal of K(+) at either day 15 or 17 on RCCs subsequently stored to day 28 had no measurable effect on red cell quality, suggesting this may be a useful device to ensure further safety for at-risk immunocompromised patient groups requiring irradiated RCCs.


Asunto(s)
Conservación de la Sangre , Eritrocitos , Filtración/métodos , Rayos gamma , Potasio , Eritrocitos/química , Eritrocitos/citología , Eritrocitos/metabolismo , Femenino , Humanos , Masculino , Potasio/química , Potasio/metabolismo , Sodio/química , Sodio/metabolismo
2.
Clin Exp Immunol ; 167(2): 216-25, 2012 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-22235997

RESUMEN

Cancer is one of the most important pathological conditions facing mankind in the 21st century, and is likely to become the most important cause of death as improvements continue in health, diet and life expectancy. The immune response is responsible for controlling nascent cancer through immunosurveillance. If tumours escape this control, they can develop into clinical cancer. Although surgery and chemo- or radiotherapy have improved survival rates significantly, there is a drive to reharness immune responses to treat disease. As T cells are one of the key immune cells in controlling cancer, research is under way to enhance their function and improve tumour targeting. This can be achieved by transduction with tumour-specific T cell receptor (TCR) or chimaeric antigen receptors (CAR) to generate redirected T cells. Virus-specific cells can also be transduced with TCR or CAR to create bi-functional T cells with specificity for both virus and tumour. In this review we outline the development and optimization of redirected and bi-functional T cells, and outline the results from current clinical trials using these cells. From this we discuss the challenges involved in generating effective anti-tumour responses while avoiding concomitant damage to normal tissues and organs.


Asunto(s)
Inmunoterapia/métodos , Neoplasias/terapia , Subgrupos de Linfocitos T/inmunología , Animales , Antígenos de Neoplasias/inmunología , Ensayos Clínicos como Asunto , Vectores Genéticos , Humanos , Vigilancia Inmunológica , Inmunoterapia Adoptiva , Ratones , Terapia Molecular Dirigida , Neoplasias/inmunología , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/inmunología , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Especificidad del Receptor de Antígeno de Linfocitos T , Subgrupos de Linfocitos T/efectos de los fármacos , Subgrupos de Linfocitos T/trasplante , Transducción Genética , Acondicionamiento Pretrasplante/métodos , Escape del Tumor , Microambiente Tumoral/inmunología
3.
Clin Exp Immunol ; 163(1): 1-10, 2011 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-20964638

RESUMEN

T cell lines with defined cytokine profiles are an invaluable tool for assessing the control of immune responses both in vitro and in vivo. Production of such cell lines can be complex and time-consuming. Here we present a powerful technique to assay the cytokines produced by T cells activated polyclonally or with specific antigens. This paper presents a detailed methodology for the identification and isolation of cytokine-producing T cells activated with the artificial superantigen, CytoStim, or viral and fungal antigens. These cells can be analysed for different cytokines simultaneously, or cultured further to rapidly establish T cell lines making known cytokine types. We highlight the enumeration, isolation and phenotype of interleukin-17-producing T cells, and the rapid generation of virus-specific Th1 T cell lines.


Asunto(s)
Separación Celular/métodos , Citocinas/análisis , Subgrupos de Linfocitos T/citología , Células TH1/citología , Animales , Antígenos Fúngicos/inmunología , Antígenos Virales/inmunología , Técnicas de Cultivo de Célula , Humanos , Interferón gamma/inmunología , Interferón gamma/metabolismo , Interleucina-17/análisis , Interleucina-17/metabolismo , Activación de Linfocitos/inmunología , Ratones , Monocitos/inmunología , Superantígenos/inmunología , Subgrupos de Linfocitos T/inmunología , Células TH1/inmunología
4.
Clin Exp Allergy ; 40(7): 1025-35, 2010 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-20412135

RESUMEN

BACKGROUND: A sensitive measurement of low numbers of intracellular cytokine-expressing antigen-specific T cells from peripheral blood mononuclear cells (PBMC) is possible using CD154 as a marker of recently activated T cells. This technique may have potential for monitoring peripheral blood T cell responses to immunotherapy. OBJECTIVE: To evaluate the applicability of this method for measuring changes in cytokine production by allergen-specific T cells in a clinical trial setting. METHODS: Ex vivo ragweed-specific CD154 and intracellular cytokine expression were evaluated using a subset of subjects in an environmental chamber study of allergic rhinitis immunotherapy. PBMC were collected and cryopreserved from Amb a 1-immunostimulatory oligodeoxynucleotide conjugate (AIC)-treated (n=17) and placebo-treated (n=15) ragweed-allergic subjects both after pre- and post-treatment ragweed exposures. In vitro allergen-stimulated CD3(+)CD4(+)CD154(+) T cell intracellular IL-4, IL-5, IL-13, and IFN-gamma expression were evaluated by flow cytometry. RESULTS: Compared with the T helper type 2 (Th2) cytokine expression measured after pre-treatment ragweed exposures, placebo-treated subjects demonstrated a significantly elevated ragweed- and Amb a 1-specific T cell IL-4 and IL-13 co-expression (P=0.005 and P=0.022, respectively) and a significantly elevated ragweed-specific IL-5 expression (P<0.001) following post-treatment ragweed exposures. In contrast, AIC-treated subjects demonstrated no increases in allergen-specific Th2 cytokine expression following post-treatment ragweed exposures. IFN-gamma expression remained low and un-changed in both groups. Subject reported total nasal symptom scores demonstrated modest but significant correlations with Amb a 1- and ragweed-stimulated intracellular Th2 cytokine responses. CONCLUSION: Combined CD154 and intracellular cytokine staining in PBMC can be used to sensitively monitor changes in antigen-specific T cell subset frequencies in clinical studies. Antigen-specific cytokine expression moderately correlated with the reported levels of allergic symptoms.


Asunto(s)
Alérgenos , Ambrosia/inmunología , Ligando de CD40/sangre , Inmunoterapia , Células TH1/inmunología , Células Th2/inmunología , Alérgenos/inmunología , Citometría de Flujo , Humanos
5.
Allergy ; 65(9): 1141-8, 2010 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-20148804

RESUMEN

BACKGROUND: Omalizumab, an anti-immunoglobulin E antibody, reduces exacerbations and symptoms in uncontrolled allergic asthma. The study objective was to estimate the costs and consequences of omalizumab compared to usual care from a US payer perspective. METHODS: We estimated payer costs, quality-adjusted survival (QALYs), and the incremental cost-effectiveness ratio (ICER) of omalizumab compared to usual care using a state-transition simulation model that included sensitivity analyses. Every 2 weeks, patients could transition between chronic asthma and exacerbation health states. The best available evidence informed the clinical and cost input estimates. Five years of omalizumab treatment followed by usual care was assumed to estimate a lifetime horizon. Omalizumab responders (60.5% of treated) were modeled as a separate scenario where nonresponders reverted back to usual care after 16 weeks of active treatment. RESULTS: The mean lifetime discounted costs and QALYs were $83,400 and 13.87 for usual care and $174,500 and 14.19 for omalizumab plus usual care resulting in $287 200/QALY (95% interval: $219,300, $557, 900). The ICER was $172 300/QALY when comparing omalizumab to usual care in the responder scenario. One-way sensitivity analyses indicated that the results were sensitive to the difference in treatment-specific utilities for the chronic state, exacerbation-associated mortality, omalizumab price, exacerbation rates, and response definition. CONCLUSIONS: The results suggest that adding omalizumab to usual care improves QALYs at an increase in direct medical costs. The cost-effectiveness of omalizumab is similar to other chronic disease biologics. The value increases when omalizumab response is used to guide long-term treatment.


Asunto(s)
Anticuerpos Antiidiotipos/economía , Anticuerpos Antiidiotipos/uso terapéutico , Anticuerpos Monoclonales/economía , Anticuerpos Monoclonales/uso terapéutico , Asma/tratamiento farmacológico , Asma/economía , Corticoesteroides/economía , Corticoesteroides/uso terapéutico , Anticuerpos Monoclonales Humanizados , Asma/mortalidad , Asma/fisiopatología , Enfermedad Crónica , Análisis Costo-Beneficio , Hospitalización/estadística & datos numéricos , Humanos , Cadenas de Markov , Modelos Económicos , Omalizumab , Años de Vida Ajustados por Calidad de Vida , Ensayos Clínicos Controlados Aleatorios como Asunto , Resultado del Tratamiento , Estados Unidos
6.
Knee Surg Sports Traumatol Arthrosc ; 18(10): 1379-84, 2010 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-20020101

RESUMEN

Rotational stability of the knee has been traditionally difficult to quantify, limiting the ability of the orthopedic community to determine the potential role of rotational laxity in the etiology of anterior cruciate ligament (ACL) injuries. The purposes of this multicenter cohort study were to evaluate the reliability of a robotic axial rotation measurement system, determine whether the uninjured knees of patients that had previous contralateral ACL reconstruction demonstrated different rotational biomechanical characteristics than a group of healthy volunteers, and determine whether knee rotational biomechanical characteristics differ between male and female non-injured limbs in groups of both healthy volunteers and patients with a previous contralateral ACL injury. Fourteen healthy volunteers and 79 patients with previous unilateral ACL injury participated in this study. Patients were tested using a computerized tibial axial rotation system. Only the normal (non-operated) knee data were used for analysis. In order to assess the reliability of the robotic measurement system, 10 healthy volunteers were tested daily over four consecutive days by four different examiners. Rotational laxity and compliance measures demonstrated excellent reliability (ICC = 0.97). Patients with a contralateral ACL injury demonstrated significantly increased tibial internal rotation (20.6° vs. 11.4°, P < 0.001) and reduced external rotation (16.7° vs. 26.6°, P < 0.001) compared to healthy volunteers. Females demonstrated significantly increased internal and external rotation, as well as significantly increased rotational compliance compared with males (P < 0.05). Computer-assisted measurement techniques may offer clinicians an accurate, reliable, non-invasive method to select the most appropriate preventative or surgical interventions for patients with increased knee rotational laxity.


Asunto(s)
Ligamento Cruzado Anterior/cirugía , Inestabilidad de la Articulación/diagnóstico , Rango del Movimiento Articular/fisiología , Robótica/métodos , Adulto , Lesiones del Ligamento Cruzado Anterior , Fenómenos Biomecánicos , Estudios de Casos y Controles , Estudios de Cohortes , Fenómenos Electromagnéticos , Diseño de Equipo , Femenino , Humanos , Inestabilidad de la Articulación/epidemiología , Traumatismos de la Rodilla/cirugía , Masculino , Persona de Mediana Edad , Procedimientos de Cirugía Plástica/métodos , Valores de Referencia , Factores de Riesgo , Sensibilidad y Especificidad , Factores Sexuales
7.
Eur Respir J ; 32(5): 1237-42, 2008 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-18684845

RESUMEN

The aim of the present study was to assess the response of high-dose salmeterol/fluticasone combination (SFC) and low-dose SFC compared with regimens without inhaled corticosteroid (ICS) plus long-acting beta-agonist (LABA) in a large cohort with severe or difficult-to-treat asthma. Subjects were administered low-dose SFC (100/50 or 250/50 microg) or high-dose SFC (500/50 microg), and a control group received medications that could include ICS or LABA but not both. The present authors calculated unadjusted and propensity score-adjusted differences in outcomes consistent with components of asthma control, comparing high-dose and low-dose SFC cohorts with controls. The low-dose SFC cohort had higher asthma-related quality of life and fewer asthma control problems compared with controls. The high-dose SFC cohort had higher forced expiratory volume in one second but higher odds of having severe asthma compared with controls. The present results support the evidence that some asthmatics achieve better outcomes while receiving a low-dose salmeterol/fluticasone combination, but also suggest that those on a high-dose salmeterol/fluticasone combination fail to achieve significant improvement in many control-related health outcomes as compared with similar patients not receiving salmeterol/fluticasone combination. These findings suggest a limited value of high-dose salmeterol/fluticasone combination compared with the alternatives. While additional studies are needed, the present findings call for alternative therapeutic approaches in severe/difficult-to-treat asthma for those unable to attain asthma control with or without salmeterol/fluticasone combination.


Asunto(s)
Albuterol/análogos & derivados , Androstadienos/administración & dosificación , Asma/tratamiento farmacológico , Quimioterapia Combinada , Corticoesteroides/metabolismo , Anciano , Albuterol/administración & dosificación , Estudios de Cohortes , Femenino , Fluticasona , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Calidad de Vida , Xinafoato de Salmeterol , Encuestas y Cuestionarios , Resultado del Tratamiento
8.
Allergy ; 63(12): 1581-92, 2008 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19032230

RESUMEN

The aim of this systematic review was to summarize and assess the quality of asthma intervention health economic studies from 2002 to 2007, compare the study findings with clinical management guidelines, and suggest avenues for future improvement of asthma health economic studies. Forty of the 177 studies met our inclusion criteria. We assessed the quality of studies using The Quality of Health Economic Studies validated instrument (total score range: 0-100). Six studies (15%) had quality category 2, 26 studies (65%) achieved quality category 3, and the remaining eight (20%) studies were scored as the highest quality level, category 4. Overall, the findings from this review are in line with the Global Initiative for Asthma clinical guidelines. Many asthma health economic studies lacked appropriate long term time horizons to match the chronic nature of the disease and suffered from using effectiveness measures that did not capture all disease related risks and benefits. We recommend that new asthma simulation models: be flexible to allow for long term time horizons, focus on using levels of asthma control in their structure, and estimate both long term asthma specific outcomes like well-controlled time as well as generic outcomes such as quality adjusted survival.


Asunto(s)
Antiasmáticos/economía , Antiasmáticos/farmacología , Asma/tratamiento farmacológico , Asma/economía , Guías de Práctica Clínica como Asunto , Animales , Análisis Costo-Beneficio , Humanos , Guías de Práctica Clínica como Asunto/normas
9.
J Leukoc Biol ; 80(4): 862-9, 2006 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-16895973

RESUMEN

Production of the anti-inflammatory cytokine IL-10 by monocytes has been implicated as a probable negative regulator of graft-versus-host disease (GvHD) in patients undergoing allogeneic stem cell transplants (SCT). Monocytes from G-CSF-mobilized peripheral blood stem cell (gmPBSC) collections have been reported to produce more IL-10 than unmobilized monocytes in response to proinflammatory factors such as LPS. Why this should occur is unclear. In this study, monocyte phenotype and IL-10 localization and release were investigated in PB mononuclear cells (MNC) from 27 healthy donors mobilized for allogeneic SCT and from 13 patients with hematological malignancies mobilized for autologous SCT. All isolates contained elevated total percentages of monocytes in comparison with unmobilized PB, a high proportion of which displayed an immature phenotype. Stimulation of gmPB MNC with an inflammatory stimulus [fixed Staphylococcus aureus cells (SAC)] induced rapid up-regulation of CD14, indicating conversion to mature status. Localization studies indicated that IL-10 was predominantly present, bound on the surface of CD64(+)/CD14(low/neg) immature monocytes. Inflammatory stimuli (LPS, polyinosinic:polycytidylic acid, or SAC) induced release of variable quantities of IL-10 from the cell surface. MNC, separated into surface IL-10-positive or -negative fractions, differed in their ability to stimulate alloreactivity in MLR, and IL-10(+) MNC induced significantly lower levels of proliferation than IL-10(-) MNC. Thus, the subset of immature monocytes carrying surface-bound IL-10 in gmPB has the potential to modulate alloreactivity and GvHD after allogeneic SCT through cell-to-cell contact and released IL-10.


Asunto(s)
Factor Estimulante de Colonias de Granulocitos/farmacología , Movilización de Célula Madre Hematopoyética/métodos , Células Madre Hematopoyéticas/inmunología , Interleucina-10/biosíntesis , Monocitos/inmunología , Selección de Donante , Enfermedad Injerto contra Huésped/inmunología , Trasplante de Células Madre Hematopoyéticas , Células Madre Hematopoyéticas/efectos de los fármacos , Humanos , Monocitos/efectos de los fármacos , Fenotipo , Trasplante Homólogo
10.
J Leukoc Biol ; 66(6): 981-8, 1999 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-10614781

RESUMEN

Multiple myeloma (MM) is a cancer of plasma cells, characterized by profound suppression of host immune responses. Here we show that MM cell lines significantly suppress the proliferation, blasting, response to interleukin-2 (IL-2), and expression of CD25 by concanavalin A (Con A)-activated or allostimulated peripheral blood T lymphocytes. T cells arrest in the G1 stage of the cell cycle, and do not enter the IL-2 autocrine growth pathway. T cell inhibition was mediated by a soluble factor. MM cell lines did not produce IL-10 but did produce large amounts of transforming growth factor beta1 (TGF-beta1). T cells were assessed for their ability to respond to IL-2 when co-cultured with MM cells in the presence or absence of the TGF-beta inhibitor, TGF-beta latency-associated peptide (LAP). MM cells suppressed IL-2 responses but this inhibition was completely reversed by TGF-beta LAP. A CD25-, IL-2-dependent blast cell line was not inhibited by MM cells or rhTGF-beta, confirming the specificity of the inhibition mechanism for the IL-2 autocrine growth pathway. We conclude that MM cells suppress T cells in their entry into the autocrine IL-2/CD25 pathway and in response to IL-2, and that TGF-beta has a significant role to play.


Asunto(s)
Adyuvantes Inmunológicos/antagonistas & inhibidores , Interleucina-2/antagonistas & inhibidores , Activación de Linfocitos/inmunología , Mieloma Múltiple/inmunología , Fragmentos de Péptidos , Precursores de Proteínas , Linfocitos T/inmunología , Factor de Crecimiento Transformador beta/farmacología , Adyuvantes Inmunológicos/fisiología , Apoptosis/inmunología , Comunicación Celular/inmunología , Ciclo Celular/inmunología , Técnicas de Cocultivo , Concanavalina A/antagonistas & inhibidores , Humanos , Interleucina-2/fisiología , Activación de Linfocitos/efectos de los fármacos , Prueba de Cultivo Mixto de Linfocitos , Mitógenos/antagonistas & inhibidores , Mieloma Múltiple/metabolismo , Mieloma Múltiple/patología , Proteínas/farmacología , Receptores de Interleucina-2/biosíntesis , Proteínas Recombinantes/farmacología , Linfocitos T/citología , Linfocitos T/metabolismo , Factor de Crecimiento Transformador beta/biosíntesis , Factor de Crecimiento Transformador beta1 , Células Tumorales Cultivadas
11.
J Leukoc Biol ; 63(5): 567-74, 1998 May.
Artículo en Inglés | MEDLINE | ID: mdl-9581800

RESUMEN

In this study we present data on a novel cell surface antigen recognized by monoclonal antibody (mAb) VPM30, originally thought to recognize only bovine and ovine sIg+ B cells from peripheral blood. Here we show that the antigen, molecular mass 28 kDa, is not only found in B cell follicles in frozen sections, but when used on paraffin sections VPM30 specifically stains B cells in the light zone of germinal centers but not in the mantle or dark zones. In addition we show that the antigen is also expressed by 90% of T cells after activation, with kinetics of antigen expression mirroring those of proliferation. By both size and distribution, the antigen appears to be novel, corresponding to no known cluster of differentiation, and will be of great use in the study of ruminant cellular immune responses.


Asunto(s)
Antígenos de Diferenciación de Linfocitos B/química , Antígenos de Diferenciación de Linfocitos T/química , Linfocitos B/inmunología , Linfocitos T/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Linfocitos B/química , Linfocitos B/citología , Bovinos , Citometría de Flujo , Centro Germinal/citología , Ganglios Linfáticos/citología , Activación de Linfocitos , Peso Molecular , Linfocitos T/química , Linfocitos T/citología
12.
Blood Rev ; 14(4): 228-39, 2000 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-11124110

RESUMEN

The finding that T cell immune responses could be divided into those promoting cell mediated immunity (Th1) and humoral responses (Th2) has had a profound effect on the understanding of immune response generation over the last 15 years. With ever increasing knowledge of the immune system, the model has come under criticism, as not all responses easily fit the classification. Nonetheless, the model still provides a valuable framework on which to base immunological research. In this review we update the model with current thinking regarding the generation and maintenance of immune responses. We then examine how the Th1-Th2 paradigm may be applied in developing new understanding of several topical issues in haematological malignancy-control of graft-versus-host disease; cytokine control of proliferating clones in B and T cell diseases; and suppression of T cell responses in multiple myeloma.


Asunto(s)
Citotoxicidad Inmunológica , Neoplasias Hematológicas/inmunología , Células TH1/inmunología , Células Th2/inmunología , Humanos , Células TH1/clasificación , Células Th2/clasificación
13.
Blood Rev ; 13(3): 151-62, 1999 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-10527267

RESUMEN

Multiple myeloma (MM) remains essentially incurable by conventional anti-tumour therapy. This has led to increased interest in the possibility that forms of immune therapy might be effective. The successful use of donor lymphocyte infusions (DLI) in a few cases of MM relapse following allogeneic bone marrow transplantation have added weight to this view. MM is characterized by several defects in the host's immune system. The influence of the malignant clone on the function of the immune effector cells results from both passive and active suppression. Despite an array of functional adhesion molecules and HLA class I and II molecules on their surface and the secretion of a tumour-specific peptide, they fail to express adequate levels of co-stimulatory molecules thus inducing anergy in potentially tumour-specific T cells. In addition to this passive evasion of immune regulation, MM tumour cells are capable of producing a number of immunologically active agents which can induce immunosuppression such as transforming growth factor-beta, Fas ligand (FasL), vascular endothelial growth factor and Muc-1. It is postulated that these agents may be produced by the tumour cell to influence the microenvironment to support growth and differentiation of the clone but may have the additional benefit of altering the function of the host immune effector cells and thus preventing tumour rejection. This duality of function is important if clinicians are to design immunotherapy strategies which will achieve the true potential and result in improved survival in MM.


Asunto(s)
Sistema Inmunológico/fisiología , Mieloma Múltiple/inmunología , Antígenos de Neoplasias/inmunología , Humanos , Huésped Inmunocomprometido/inmunología , Mieloma Múltiple/fisiopatología
14.
J Immunol Methods ; 247(1-2): 111-8, 2001 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-11150542

RESUMEN

Many available ELISAs lack the sensitivity required to reliably quantify levels of cytokines released in response to antigenic stimulation. In an effort to increase sensitivity of these assays, we compare the sensitivity of standard colorimetric ELISAs and corresponding chemiluminescent assays for three cytokines which are usually produced in very low quantities: mouse IL-12 p70, human IL-4 and mouse IL-4. Use of a chemiluminescent substrate enhanced the sensitivity of these assays 12-29-fold as compared to current colorimetric ELISAs. Accompanying this increase in sensitivity was an increase in dynamic range, a decrease in the time required to obtain maximum sensitivity and a decrease in the concentration of reagents required. These findings are of particular interest to those wanting to quantitate levels of any cytokine which is typically produced in low levels.


Asunto(s)
Interleucina-2/análisis , Interleucina-4/análisis , Animales , Colorimetría/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Interleucina-2/inmunología , Interleucina-4/inmunología , Mediciones Luminiscentes , Ratones , Sensibilidad y Especificidad , Factores de Tiempo
15.
Hum Immunol ; 62(7): 668-78, 2001 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-11423172

RESUMEN

Cross-sectional analyses of human PBMC, plasma, and tissue have reported altered chemokine and/or chemokine receptor expression in several inflammatory diseases. Interpretation of such studies is difficult without data on the in vivo stability of such parameters. Using four color flow cytometry, we longitudinally followed CXCR3, CCR5 (Th1-associated), and CCR3 (Th2-associated) expression within CD4+/CD45RO+ and CD8+/CD45RO+ T cell populations in peripheral blood of healthy individuals over a 21 day period. In parallel, we quantified plasma levels of IP-10, Mig, eotaxin and TARC. Chemokine and receptor expression differed markedly between subjects but was highly stable, varying by <5% within individuals. Differences in chemokine receptor expression between subjects were markedly altered when quantified as absolute cell numbers rather than frequencies. Finally, CCR3 expression by CD4+/CD45RO+ T cells was positively correlated with plasma levels of its ligand, eotaxin, whereas strong negative correlations were evident between CXCR3 expression and IP-10 or Mig. These data demonstrate longitudinal stability of chemokine receptor and ligand expression among healthy individuals; reveal that both frequency and absolute cell count analysis is essential for accurate assessment of chemokine receptor expression; and identify inverse relationships between type 1 and type 2 immunity-associated receptors and their ligands in vivo.


Asunto(s)
Quimiocinas/biosíntesis , Receptores de Quimiocina/biosíntesis , Adulto , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Quimiocinas/sangre , Quimiocinas/metabolismo , Quimiocinas CXC/biosíntesis , Quimiocinas CXC/sangre , Quimiocinas CXC/metabolismo , Humanos , Inflamación/sangre , Inflamación/inmunología , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Recuento de Linfocitos , Persona de Mediana Edad , Receptores CCR3 , Receptores CCR5/biosíntesis , Receptores CXCR3 , Receptores de Quimiocina/metabolismo
16.
Ann N Y Acad Sci ; 849: 152-4, 1998 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-9668460

RESUMEN

The tick-borne protozoan parasite Theileria annulata causes tropical theileriosis, a severe leukoproliferative disease of cattle, which naive susceptible animals fail to control. The parasite infects and transforms macrophages, developing in the local draining lymph node. IFN gamma has been shown to block parasite development in newly infected cells, and inhibits the growth of fully differentiated macroschizont stage-infected cells in vitro. However, the parasite has been found to specifically induce IFN gamma production by T cells and appears to flourish in the face of this T cell-derived response in vivo. Here we show that the production of IFN gamma in vivo is tightly controlled by the parasite. Induction of cytokine production by T cells is not initiated until the parasite has developed beyond the IFN gamma sensitive trophozoite stage. Cytokine production is kept high as infected macrophages develop, and IFN gamma appears to play an active role in maintaining the growth of these cells. Once the infection is fully established, IFN gamma is down regulated, avoiding potential inhibitory effects. Thus by controlling T cell IFN gamma production, the parasite induces a "window" of cytokine expression which promotes its own growth, but avoids potential inhibitory effects of the cytokine.


Asunto(s)
Interferón gamma/biosíntesis , Theileria annulata , Theileriosis/inmunología , Animales , Bovinos , Células Cultivadas , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/parasitología , Macrófagos/inmunología , Macrófagos/parasitología , Linfocitos T/inmunología , Theileria annulata/crecimiento & desarrollo , Theileria annulata/inmunología , Garrapatas/inmunología
17.
Metabolism ; 47(10): 1199-204, 1998 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-9781621

RESUMEN

The spontaneously hypertensive/NIH-corpulent (SHR/N-cp) rat is a genetic model that exhibits both non-insulin-dependent diabetes mellitus (NIDDM) and hypertension. To determine the impact of long-term treatment with the long-acting angiotensin-converting enzyme (ACE) inhibitor perindopril (PE) on the glucose metabolism, lipid levels, and heart in this model, studies were performed in three groups of SHR/N-cp rats maintained on a diet containing 54% carbohydrate with 18% sucrose and 36% starch. One group of obese rats received PE (0.5 to 1.0 mg/kg body weight/d) for 3 to 4 months, a second group of obese rats received no treatment, and a third group of lean rats were used as controls. The mean systolic blood pressure (SBP) increased gradually in both untreated obese and lean rats, with lean animals showing slightly higher levels compared with untreated obese rats. By contrast, SBP was reduced to normal levels in PE-treated obese rats throughout the treatment period. Compared with lean rats, obese rats showed significantly higher body weight and fasting serum levels of glucose, insulin, total cholesterol (TC), and triglyceride (TG). However, no significant differences were observed in these metabolic parameters between PE-treated and untreated obese rats. Plasma renin activity measured at the end of the treatment period was significantly higher in PE-treated rats compared with untreated obese and untreated lean rats. The mean heart weight and left ventricular weight, expressed in absolute terms or indexed to body weight, were significantly lower in PE-treated versus untreated obese and untreated lean rats. To further determine whether glucose metabolism is directly affected by PE treatment, in vitro glycogen synthesis was evaluated in isolated soleus muscles obtained from three additional groups of animals. The basal rate of muscle glycogen synthesis was significantly lower in obese compared with lean rats (P < .05), but did not differ between PE-treated and untreated obese rats. Maximal insulin-stimulated glycogen synthesis increased threefold in PE-treated obese rats, but this increase did not differ from the increases observed in untreated obese and lean rats. In conclusion, the present study shows that long-term PE treatment in obese SHR/N-cp rats with NIDDM and hypertension effectively controlled systemic arterial pressure and resulted in a significant reduction in left ventricular weight. However, these favorable effects of PE were not associated with significant improvement in glucose tolerance, hyperinsulinemia, and hyperlipidemia in this model. PE also had no direct stimulatory effects on either basal or insulin-mediated glycogen synthesis in the isolated soleus muscle of obese rats, perhaps because of the severe insulin-resistant state of the animals. Our results support the clinical observations that antihypertensive therapy with ACE inhibitors has neutral effects on glucose metabolism and insulin sensitivity in patients with combined hypertension and NIDDM.


Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Antihipertensivos/farmacología , Diabetes Mellitus Tipo 2/metabolismo , Corazón/efectos de los fármacos , Hipertensión/metabolismo , Indoles/farmacología , Animales , Glucógeno/biosíntesis , Masculino , Perindopril , Ratas , Ratas Endogámicas SHR
18.
Cladistics ; 13(3): 207-224, 1997 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34911231

RESUMEN

Previous morphologically based studies by several taxonomists disagree on whether the Tettigometridae represent a sister group to all other fulgoromorphans or whether they are a relatively derived family within the Fulgoromorpha. In this study, several parsimony-based analyses using data sets composed of nucleotide sequences of 18S rDNAs (genes encoding 18S rRNAs) support a monophyletic Fulgoromorpha. All analyses depict Tettigometridae as a relatively derived lineage in a monophyletic relationship with Tropiduchidae. Unscored and unweighted data sets position the tettigometrid + tropiduchid clade as sister to Flatidae. The tettigometrid + tropiduchid clade is supported by four synapomorphic sites, two deletions and two transversions. Constraining tettigometrids to a basal fulgoromorphan lineage significantly reduces parsimony, with several hundreds to thousand of trees being more parsimonious. An analysis employing the Barriel method for scoring potential phylogenetic information in regions containing deletions also supports a non-basal tettigometrid + tropiduchid clade. However, this method results in three equally most parsimonious trees and a sister relationship of the tettigometrid + tropiduchid clade to Flatidae becomes ambiguous. The molecular-based results showing a derived Tettigometridae agree with previous morphological interpretations of Bourgoin. The current biogeographical distribution of tettigometrids and morphological features supporting the 18S rDNA-based phylogeny are discussed.

19.
Acta Trop ; 43(3): 263-70, 1986 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-2430433

RESUMEN

We determined the prevalence of Babesia microti and Borrelia burgdorferi in four populations of Ixodes dammini in eastern Massachusetts. The Feulgen's reaction was more sensitive than the Giemsa method for detecting salivarian Babesia. A combination of darkfield and direct-fluorescent-antibody examination proved more sensitive than either method alone for detecting spirochetal infection. The prevalence of spirochetes was greater than the prevalence of Babesia in each of the tick populations studied. Overall, 24% of nymphs and 47% of adults examined were infected with spirochetes; in contrast, 11% of nymphs and 14% of adults were infected with Babesia. The difference between the spirochetal and babesial prevalence was greatest in the most recently infested site. The rising incidence of Lyme disease, as compared to the stable incidence of human babesiosis, may result from the relatively greater abundance of Bo. burgdorferi infected I. dammini ticks in newly infested locations.


Asunto(s)
Vectores Arácnidos/parasitología , Babesia/aislamiento & purificación , Borrelia/aislamiento & purificación , Colorantes de Rosanilina , Garrapatas/parasitología , Animales , Vectores Arácnidos/microbiología , Colorantes Azulados , Babesiosis/epidemiología , Colorantes , Técnica del Anticuerpo Fluorescente , Humanos , Enfermedad de Lyme/epidemiología , Massachusetts , Coloración y Etiquetado , Garrapatas/microbiología
20.
J Pers Soc Psychol ; 50(2): 281-94, 1986 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-3701578

RESUMEN

On the basis of a self-validation perspective, it was predicted that distortions in consensus estimates would vary as a function of attribute type (opinions versus abilities), relevance of the attribute, and individual differences in self-esteem and depression. Students rated themselves on seven opinion and ability dimensions using 5-point Likert scales. Then they estimated the percentage of the other students who held each opinion/ability position, and rank ordered the opinions/abilities for personal relevance. Absolute and directional accuracy scores were computed (comparing estimated percentages with actual percentages in the sample), as well as false consensus (FCE) scores (comparing estimates of subjects holding and not holding a particular position). Subjects overestimated consensus for their opinions and low abilities, but underestimated consensus for their high abilities. Although subjects exhibited a larger FCE on opinions than abilities, there was a reliable FCE for both attributes. Relevance affected the magnitude of these biases. Higher opinion relevance was associated with increased accuracy, lower FCE scores, and smaller overestimates. Higher ability relevance was associated with decreased accuracy, greater overestimation on low abilities, and greater underestimation on high abilities. Finally, low self-esteem and depressed subjects overestimated consensus on opinions and underestimated consensus on abilities less than high self-esteem and nondepressed subjects.


Asunto(s)
Depresión/psicología , Individualidad , Autoimagen , Disposición en Psicología , Adulto , Aptitud , Actitud , Femenino , Humanos , Masculino , Pruebas Psicológicas , Conducta Social , Percepción Social
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