RESUMEN
Lyme disease (LD) is an increasing public health problem. Current laboratory testing is insensitive in early infection, the stage at which appropriate treatment is most effective in preventing disease sequelae. The Lyme Disease Biobank (LDB) collects samples from individuals with symptoms consistent with early LD presenting with or without erythema migrans (EM) or an annular, expanding skin lesion and uninfected individuals from areas of endemicity. Samples were collected from 550 participants (298 cases and 252 controls) according to institutional review board-approved protocols and shipped to a centralized biorepository. Testing was performed to confirm the presence of tick-borne pathogens by real-time PCR, and a subset of samples was tested for Borrelia burgdorferi by culture. Serology was performed on all samples using the CDC's standard two-tiered testing algorithm (STTTA) for LD. LD diagnosis was supported by laboratory testing in 82 cases, including positive results by use of the STTTA, PCR, or culture or positive results by two enzyme-linked immunosorbent assays for cases presenting with EM lesion sizes of >5 cm. The remaining 216 cases had negative laboratory testing results. For the controls, 43 were positive by at least one of the tiers and 6 were positive by use of the STTTA. The results obtained with this collection highlight and reinforce the known limitations of serologic testing in early LD, with only 29% of individuals presenting with EM lesion sizes of >5 cm yielding a positive result using the STTTA. Aliquots of whole blood, serum, and urine from clinically characterized patients with and without LD are available to investigators in academia and industry for evaluation or development of novel diagnostic assays for LD, to continue to improve upon currently available methods.
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Grupo Borrelia Burgdorferi , Borrelia burgdorferi , Enfermedad de Lyme , Bancos de Muestras Biológicas , Borrelia burgdorferi/genética , Ensayo de Inmunoadsorción Enzimática , Humanos , Enfermedad de Lyme/diagnóstico , Enfermedad de Lyme/epidemiología , Estados Unidos/epidemiologíaRESUMEN
Background Military personnel post-deployment to Iraq and Afghanistan have noted new-onset respiratory illness. This study's primary objective was to further develop an animal model of Iraq Afghanistan War Lung Injury (IAW-LI) and to test a novel class of anti-injury drug called RuX. Methods Particulate Matter (PM) samples were obtained in Iraq then characterized by spectromicroscopy. C57BL/6 mice underwent orotracheal instillation with PM, followed by drinkable treatment with RuX. Lung histology, inspiratory capacity (FlexiVent), thymic/splenic regulatory T cell (Treg) number, and whole-lung genomics were analyzed. Results Tracheal instillation of Iraq PM led to lung septate thickening and lymphocytic inflammation. PM-exposed mice had suppression of thymic/splenic regulatory T-cells (Tregs). Drinking RuX after PM exposure attenuated the histologic lung injury response, improved lung inspiratory capacity, and increased Tregs. Pooled whole lung genomics suggest differences among gene expression of IL-15 among control, PM, and PM + RuX groups. Conclusions RuX, a ruthenium and alpha-lipoic acid complex, attenuates lung injury by improving histology and inspiratory capacity via upregulation of Tregs in Iraq PM-exposed C57BL/6. Plausible genomic effects may involve IL-15 whole lung gene expression.
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Lesión Pulmonar/tratamiento farmacológico , Material Particulado/toxicidad , Linfocitos T Reguladores/citología , Campaña Afgana 2001- , Animales , Modelos Animales de Enfermedad , Interleucina-15/metabolismo , Irak , Ratones , Rutenio/uso terapéutico , Ácido Tióctico/uso terapéutico , Regulación hacia ArribaRESUMEN
GOALS: Contemporary management of epithelial ovarian cancer (EOC) uses biomarkers to monitor response to therapy. This study evaluates the role of invasive circulating tumor cells (iCTCs) in monitoring EOC treatment in comparison with serum cancer antigen 125 (CA125). METHODS: Molecular and microscopic analyses were used to identify seprase and CD44 as tumor progenitor (TP) markers. The iCTC flow cytometry assay was optimized using blood donated by 64 healthy donors, 49 patients with benign abdominal diseases and 123 EOC patients. Serial changes in iCTCs and CA125 were measured in 129 blood and 169 serum samples, respectively, from 31 EOC patients to assess their concordance during therapy and their relationship with risk of progressive disease (PD). RESULTS: The assay had 97% specificity and 83% sensitivity for detecting iCTCs in blood of EOC patients. iCTCs were detected in each monitoring patient (31/31, 100%) and in 110 of the 129 blood samples (85.3%). The concordance between changes in iCTCs/CA125 levels and changes in the intervals associated with no evidence of disease (NED) were markedly stronger (specificity: CA125 93.8%; iCTCs 90.6%), whereas increases in iCTCs (79.5%) were more sensitive than increases in CA125 (67.6%) to predict PD or relapse. Among the six patients who had greater than 6 measurements, iCTCs but not CA125 antedated changes in clinical status from PD to NED during and after chemotherapy and predated relapse. CONCLUSION: Serial measurements of iCTCs could predict therapeutic responsiveness in 31 EOC patients who underwent standard taxol/carboplatin therapy.
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Neoplasias Glandulares y Epiteliales/sangre , Neoplasias Glandulares y Epiteliales/tratamiento farmacológico , Células Neoplásicas Circulantes/patología , Neoplasias Ováricas/sangre , Neoplasias Ováricas/tratamiento farmacológico , Biomarcadores de Tumor/sangre , Antígeno Ca-125/sangre , Carcinoma Epitelial de Ovario , Estudios de Casos y Controles , Estudios de Cohortes , Monitoreo del Ambiente/métodos , Femenino , Citometría de Flujo , Humanos , Receptores de Hialuranos/sangre , Proteínas de la Membrana/sangre , Recurrencia Local de Neoplasia/sangre , Recurrencia Local de Neoplasia/patología , Estadificación de Neoplasias , Neoplasias Glandulares y Epiteliales/patología , Células Neoplásicas Circulantes/metabolismo , Neoplasias Ováricas/patología , Serina Endopeptidasas/sangreRESUMEN
Radiation (RT), temozolomide (TMZ), and dexamethasone in newly diagnosed high grade gliomas (HGG) produces severe treatment-related lymphopenia (TRL) that is associated with early cancer-related deaths. This TRL may result from inadvertent radiation to circulating lymphocytes. This study reinfused lymphocytes, harvested before chemo-radiation, and assessed safety, feasibility, and trends in lymphocyte counts. Patients with newly diagnosed HGG and total lymphocyte counts (TLC) ≥ 1000 cells/mm(3) underwent apheresis. Cryopreserved autologous lymphocytes were reinfused once radiation was completed. Safety, feasibility, and trends in TLC, T cell subsets and cytokines were studied. Serial TLC were also compared with an unreinfused matched control group. Ten patients were harvested (median values: age 56 years, dexamethasone 3 mg/day, TLC/CD4 1980/772 cells/mm(3)). After 6 weeks of RT/TMZ, TLC fell 69 % (p < 0.0001) with similar reductions in CD4, CD8 and NK cells but not Tregs. Eight patients received lymphocyte reinfusions (median = 7.0 × 10(7) lymphocytes/kg) without adverse events. A post-reinfusion TLC rise of ≥300 cells/mm(3) was noted in 3/8 patients at 4 weeks and 7/8 at 14 weeks which was similar to 23 matched controls. The reduced CD4/CD8 ratio was not restored by lymphocyte reinfusion. Severe lymphopenia was not accompanied by elevated serum interleukin-7 (IL-7) levels. This study confirms that severe TRL is common in HGG and is not associated with high plasma IL-7 levels. Although lymphocyte harvesting/reinfusion is feasible and safe, serial lymphocyte counts are similar to unreinfused matched controls. Studies administering higher lymphocyte doses and/or IL-7 should be considered to restore severe treatment-related lymphopenia in HGG.
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Astrocitoma/terapia , Glioblastoma/terapia , Transfusión de Linfocitos/métodos , Adulto , Astrocitoma/sangre , Transfusión de Sangre Autóloga/efectos adversos , Transfusión de Sangre Autóloga/métodos , Quimioradioterapia/efectos adversos , Quimioradioterapia/métodos , Estudios de Factibilidad , Femenino , Glioblastoma/sangre , Humanos , Interleucina-7/sangre , Recuento de Linfocitos , Transfusión de Linfocitos/efectos adversos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Factor de Crecimiento Transformador beta/sangre , Resultado del TratamientoRESUMEN
GOALS: Circulating tumor cells (CTCs) have been introduced as a biomarker in detecting advanced epithelial ovarian cancer (EOC). The goals are to examine the prevalence of the invasive subpopulation of CTCs (iCTCs) in patients at high risk of EOC and to compare this biomarker to serum CA125. METHODS: We used a unique cell adhesion matrix (CAM)-based, functional cell enrichment and identification platform to isolate iCTCs from 129 preoperative patients. We confirmed the identity of iCTCs using positive epithelial (Epi+) markers and negative hematopoietic lineage (HL-) markers. Sensitivity and specificity of the assays were examined and iCTCs/CA125 were correlated with overall survival (OS), progression-free survival (PFS) and clinical parameters. RESULTS: We found a 41.2% sensitivity, 95.1% specificity and 77.8% positive predictive value (PPV) of the iCTC assay in detecting patients with stage I and II EOC malignancy, and a 83% sensitivity and 97.3% PPV in detecting all stages of EOC malignancy. However, a positive CA125 test provided weak evidence to detect stage I and II malignancy (61.6% PPV) and all EOC (92.1% PPV), because of its 76.2% specificity. A significantly stronger concordance in OS and PFS of clinical factors (tumor stage, debulking and platinum sensitivity) was noted for elevated iCTCs than for serum CA125. CONCLUSION: The CAM-initiated CTC enrichment/identification method enabled the detection of early stage EOC. iCTCs were better correlated with worse OS and PFS, more specific and better PPV than CA125 in detecting EOC malignancy in patients at high risk of EOC.
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Biomarcadores de Tumor/sangre , Antígeno Ca-125/sangre , Neoplasias Glandulares y Epiteliales/sangre , Neoplasias Glandulares y Epiteliales/patología , Células Neoplásicas Circulantes/patología , Neoplasias Ováricas/sangre , Neoplasias Ováricas/patología , Carcinoma Epitelial de Ovario , Recuento de Células , Supervivencia sin Enfermedad , Femenino , Humanos , Persona de Mediana Edad , Neoplasias Glandulares y Epiteliales/mortalidad , Neoplasias Ováricas/mortalidad , Valor Predictivo de las Pruebas , Pronóstico , Sensibilidad y Especificidad , Tasa de Supervivencia , Células Tumorales CultivadasRESUMEN
A 6-year-old male presented with a testicular mass, hepatosplenomegaly, and a pleural effusion while undergoing maintenance chemotherapy for treatment of T-cell acute lymphoblastic leukemia (T-ALL). He was subsequently diagnosed with a lymphoproliferative disorder that resembled hepatosplenic lymphoma (HSL). While the extranodal presentation and the protracted yet aggressive clinical course are consistent with HSL, the findings of monosomy 8 and polymorphic cell populations are unique and have not been previously described in this type of lymphoma.
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Trastornos Linfoproliferativos/patología , Neoplasias Primarias Secundarias/patología , Leucemia-Linfoma Linfoblástico de Células T Precursoras/patología , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Niño , Preescolar , Citometría de Flujo , Humanos , Inmunofenotipificación , Neoplasias Hepáticas/patología , Linfoma/patología , Quimioterapia de Mantención , Masculino , Leucemia-Linfoma Linfoblástico de Células T Precursoras/tratamiento farmacológico , Neoplasias del Bazo/patologíaRESUMEN
Little is known about in vivo cytogenetic effects of protons delivered at the dose and dose rates encountered in space. We determined the effects of 100MeV protons, one of the most abundant type of protons produced during solar particle events (SPE), on the induction of chromosome aberrations (CAs) in bone marrow (BM) cells collected at early (3 and 24h) and late (6 months) time-points from groups of BALB/cJ mice (a known radiosensitive strain) exposed whole-body to 0 (sham-controls), 0.5, or 1.0Gy of 100MeV protons, delivered at 0.5 or 1.0cGy/min. These doses and dose-rates are comparable to those produced during SPE events. Additionally, groups of mice were exposed to 0 or 1Gy of (137)Cs γ rays (delivered at 1cGy/min) as a reference radiation. The kinetics of formation/reduction of gamma-histone 2-AX (γH2AX) were determined in BM cells collected at 1.5, 3, and 24h post-irradiation to assess the early-response. There were five mice per treatment-group per harvest-time. Our data indicated that the kinetics of γH2AX formation/reduction differed, depending on the dose and dose rate of protons. Highly significant numbers of abnormal cells and chromatid breaks (p<0.01), related to those in sham-control groups, were detected in BM cells collected at each time-point, regardless of dose or dose-rate. The finding of significant increases in the frequencies of delayed non-clonal and clonal CAs in BM cells collected at a late time-point from exposed mice suggested that 0.5 or 1Gy of 100MeV protons is capable of inducing genomic instability in BM cells. However, the extent of effects induced by these two low dose rates was comparable. Further, the results showed that the in vivo cytogenetic effects induced by 1Gy of 100MeV protons or (137)Cs γ rays (delivered at 1cGy/min) were similar.
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Aberraciones Cromosómicas/efectos de la radiación , Rayos gamma , Inestabilidad Genómica/efectos de la radiación , Protones , Animales , Células de la Médula Ósea/efectos de la radiación , Ciclo Celular/efectos de la radiación , Células Cultivadas , Radioisótopos de Cesio , Citometría de Flujo , Histonas/metabolismo , Hibridación Fluorescente in Situ , Ratones , Ratones Endogámicos BALB CRESUMEN
BACKGROUND: Mounting evidence supports a key role for VIP as an anti-inflammatory agent and promoter of immune tolerance. It suppresses TNF-α and other inflammatory cytokines and chemokines, upregulates anti-inflammatory IL-10, and promotes immune tolerant cells called T regulatory (Treg) cells. VIP KO mice have recently been demonstrated to have spontaneous airway and pulmonary perivascular inflammatory responses, as part of asthma-like and pulmonary hypertension phenotypes, respectively. Both inflammatory responses are correctable with VIP. Focusing on this model, we have now investigated the influence of VIP not only on inflammatory cells but also on Treg cells. METHODS: Using flow cytometric analysis, we examined the relative preponderance of CD25+CD4+ cells and anti-inflammatory Treg cells, in extracts of thymus and spleen from VIP KO mice (5 VIP KO; 5 VIP KO+ VIP; 10 wild-type). This method allowed antibody-based flow cytometric identification of Treg cells using surface markers CD25 and CD4, along with the: 1) intracellular activation marker FoxP3; and 2) Helios, which distinguishes cells of thymic versus splenic derivation. CONCLUSIONS: Deletion of the VIP gene results in: 1) CD25+CD4- cell accumulation in the thymus, which is corrected by VIP treatment; 2) more Treg in thymus lacking Foxp3 expression, suggesting VIP is necessary for immune tolerance; and, 3) a tendency towards deficiency of Treg cells in the spleen, which is normalized by VIP treatment. Treg lacking Helios are induced by VIP intrasplenically rather than by migration from the thymus. These results confirm the dual role of VIP as an anti-inflammatory and immune tolerance-promoting agent.
RESUMEN
Recent research advances show that tumor cell intravasation (entry into the circulation) and metastasis occur very early in breast cancer progression. Clinical studies also illustrate the potential importance of detection of circulating tumor cells (CTCs) in outcomes of patients with metastatic breast cancer. Whether these cells exhibit the invasiveness and express tumor stem or progenitor markers, hallmark of the metastatic phenotype, is less well characterized. To detect CTCs with the invasive phenotype and to explore their molecular features, we applied a functional cell separation method, called collagen adhesion matrix (CAM) assay, as enrichment and identification steps. The CAM-coated device successfully recovered tumor cells spiked in 1 ml of blood with a 54% +/- 9% (n = 18) recovery rate and 0.5-35% purity, and detected invasive tumor cells in 10/10 blood samples (100% yield) from patients with metastatic breast cancer with a range of 18-256 CTCs/ml and average of 126 +/- 25 (mean +/- SD) CTCs/ml. CTCs were detected in blood samples of 28/54 (52%) Stage I-III breast cancer patients with a mean count of 61 CTCs/ml. Furthermore, the relative frequency of these cells correlated to the staging, lymph node-status and survival of patients with early stage breast cancer. CAM-captured cells were capable of propagation in culture. Gene expression and multiplex flow cytometric analyses on CAM-captured cells demonstrated the existence of distinct populations of CTCs including these of epithelial lineage and stem or progenitor cells. Thus, CAM-initiated CTC detection provides advantages for examining invasiveness and tumor progenitor phenotypes.
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Neoplasias de la Mama/patología , Carcinoma Ductal de Mama/patología , Carcinoma Intraductal no Infiltrante/patología , Separación Celular/métodos , Células Epiteliales/química , Citometría de Flujo/métodos , Células Neoplásicas Circulantes/química , Células Madre Neoplásicas/química , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Biomarcadores de Tumor/análisis , Neoplasias de la Mama/sangre , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/mortalidad , Carcinoma Ductal de Mama/sangre , Carcinoma Ductal de Mama/tratamiento farmacológico , Carcinoma Ductal de Mama/mortalidad , Carcinoma Intraductal no Infiltrante/sangre , Carcinoma Intraductal no Infiltrante/tratamiento farmacológico , Adhesión Celular , Línea Celular Tumoral/química , Línea Celular Tumoral/patología , Colágeno , Progresión de la Enfermedad , Células Epiteliales/efectos de los fármacos , Femenino , Humanos , Metástasis Linfática , Persona de Mediana Edad , Invasividad Neoplásica , Proteínas de Neoplasias/análisis , Estadificación de Neoplasias , Células Neoplásicas Circulantes/clasificación , Células Neoplásicas Circulantes/efectos de los fármacos , Células Madre Neoplásicas/efectos de los fármacos , Fenotipo , Análisis de Supervivencia , Adulto JovenAsunto(s)
Síndrome de Bernard-Soulier/genética , Proteínas Motoras Moleculares/genética , Mutación , Cadenas Pesadas de Miosina/genética , Síndrome de Bernard-Soulier/patología , Plaquetas/metabolismo , Plaquetas/patología , Preescolar , Femenino , Humanos , Linaje , Glicoproteínas de Membrana Plaquetaria/metabolismoRESUMEN
A pilot study was conducted to examine the magnitude of cell-cycle delay and apoptosis in bone marrow (BM) cells collected at 18, 42 and 66 hr from radiosensitive CBA/CaJ mice and radioresistant C57BL/6J mice following a whole-body in vivo exposure to 1 GeV/amu (56)Fe ions or (137)Cs gamma rays. At each sacrifice, BM cells were collected from three mice of each strain per dose of (56)Fe ions (0, 10 and 100 cGy) and two mice of each strain per dose of (137)Cs gamma rays (0, 100 and 300 cGy). A significant G1-arrest (ANOVA, p < 0.05) was observed at 18 hr after exposure of mice to 100 cGy of (56)Fe ions or 300 cGy of (137)Cs gamma rays, relative to their corresponding sham-controls, resulting in a significant decrease in the percentage of cells cycling into S-phase in both strains. The percentage of S-phase cells subsequently increased and persisted up to 66 hr post-irradiation. Significant numbers of G2/M cells were found at 18 and 66 (but not at 42) hr post-irradiation, regardless of radiation-type or mouse-strain. It is likely that BM cells have undergone at least one cell cycle at 66 hr after exposure of mice to either 100 cGy (56)Fe ions or 300 cGy (137)Cs gamma rays. Our study is the first to investigate the in vivo effects of (56)Fe ions (1 GeV/amu) on the cell cycle of mouse BM cells using flow cytometry. The cell-cycle distribution (but not the number of apoptotic cells) was dependent on radiation-dose and harvest-time.
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Células de la Médula Ósea/citología , Células de la Médula Ósea/efectos de la radiación , Ciclo Celular/fisiología , Iones Pesados , Radioisótopos de Hierro , Animales , Ciclo Celular/efectos de la radiación , Células Cultivadas , Relación Dosis-Respuesta en la Radiación , Masculino , Ratones , Ratones Endogámicos C57BL , Dosis de RadiaciónRESUMEN
Tick-borne diseases are the most common vector-borne diseases in the United States, with serology being the primary method of diagnosis. We developed the first multiplex, array-based assay for serodiagnosis of tick-borne diseases called the TBD-Serochip. The TBD-Serochip was designed to discriminate antibody responses to 8 major tick-borne pathogens present in the United States, including Anaplasma phagocytophilum, Babesia microti, Borrelia burgdorferi, Borrelia miyamotoi, Ehrlichia chaffeensis, Rickettsia rickettsii, Heartland virus and Powassan virus. Each assay contains approximately 170,000 12-mer linear peptides that tile along the protein sequence of the major antigens from each agent with 11 amino acid overlap. This permits accurate identification of a wide range of specific immunodominant IgG and IgM epitopes that can then be used to enhance diagnostic accuracy and integrate differential diagnosis into a single assay. To test the performance of the TBD-Serochip, we examined sera from patients with confirmed Lyme disease, babesiosis, anaplasmosis, and Powassan virus disease. We identified a wide range of specific discriminatory epitopes that facilitated accurate diagnosis of each disease. We also identified previously undiagnosed infections. Our results indicate that the TBD-Serochip is a promising tool for a differential diagnosis not available with currently employed serologic assays for TBDs.
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Pruebas Serológicas , Enfermedades por Picaduras de Garrapatas/diagnóstico , Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Humanos , Enfermedades por Picaduras de Garrapatas/sangre , Enfermedades por Picaduras de Garrapatas/virologíaRESUMEN
INTRODUCTION: T cells occupy a central role in MS and CIDP pathogenesis. High dose cyclophosphamide's in-vivo cytotoxic-effect on circulating memory and naïve T cells is unknown. METHOD: Three MS and five CIDP patients received cyclophosphamide (200 mg/kg) for refractory disease. Before and after chemotherapy administration, peripheral blood T-cell subsets were determined. Patients underwent serial neurologic evaluations quarterly. RESULTS: Cyclophosphamide uniformly decreased clinical disease activity. Compared to memory T cells, naïve T cells were preferentially eradicated. DISCUSSION: Cyclophosphamide effectiveness in autoimmune illness may result from Naïve T-cell destruction, as this compartment may be the source of autoreactive lymphocytes.
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Ciclofosfamida/farmacología , Inmunosupresores/farmacología , Esclerosis Múltiple/tratamiento farmacológico , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/tratamiento farmacológico , Linfocitos T/efectos de los fármacos , Adulto , Recuento de Linfocito CD4 , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/inmunología , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Memoria Inmunológica/efectos de los fármacos , Memoria Inmunológica/inmunología , Antígenos Comunes de Leucocito/inmunología , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/inmunología , Esclerosis Múltiple/fisiopatología , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/inmunología , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/fisiopatología , Recuperación de la Función/efectos de los fármacos , Recuperación de la Función/inmunología , Linfocitos T/inmunología , Resultado del TratamientoRESUMEN
Peripheral T-cell lymphomas (PTCLS) comprise a diverse group of difficult to treat, very aggressive non-Hodgkin's lymphomas (NHLS) with poor prognoses and dismal patient outlook. Despite the fact that PTCLs comprise the majority of T-cell malignancies, the standard of care is poorly established. Chimeric antigen receptor (CAR) immunotherapy has shown in B-cell malignancies to be an effective curative option and this extends promise into treating T-cell malignancies. Because PTCLS frequently develop from mature T-cells, CD3 is similarly strongly and uniformly expressed in many PTCL malignancies, with expression specific to the hematological compartment thus making it an attractive target for CAR design. We engineered a robust 3rd generation anti-CD3 CAR construct (CD3CAR) into an NK cell line (NK-92). We found that CD3CAR NK-92 cells specifically and potently lysed diverse CD3+ human PTCL primary samples as well as T-cell leukemia cells lines ex vivo. Furthermore, CD3CAR NK-92 cells effectively controlled and suppressed Jurkat tumor cell growth in vivo and significantly prolonged survival. In this study, we present the CAR directed targeting of a novel target - CD3 using CAR modified NK-92 cells with an emphasis on efficacy, specificity, and potential for new therapeutic approaches that could improve the current standard of care for PTCLs.
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Complejo CD3/inmunología , Inmunoterapia Adoptiva/métodos , Células Asesinas Naturales/inmunología , Linfoma de Células T Periférico/terapia , Receptores de Antígenos de Linfocitos T/inmunología , Animales , Complejo CD3/metabolismo , Técnicas de Cocultivo , Humanos , Células Jurkat , Estimación de Kaplan-Meier , Células Asesinas Naturales/trasplante , Linfoma de Células T Periférico/inmunología , Linfoma de Células T Periférico/mortalidad , Masculino , Ratones , Ratones SCID , Receptores de Antígenos de Linfocitos T/uso terapéutico , Proteínas Recombinantes de Fusión/inmunología , Proteínas Recombinantes de Fusión/uso terapéutico , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: Endothelial dysfunction is emerging as a common denominator for diverse and highly prevalent cardiovascular diseases. Increased level of plasminogen activator inhibitor-1 (PAI-1) and procoagulant activity have been recognized as hallmarks of endothelial dysfunction. This study was aimed at investigating cellular actions of PAI-1 and a potential link between PAI-1 and procoagulant state. METHODS AND RESULTS: Human umbilical vein endothelial cells treated with PAI-1 were subjected to laser confocal fluorescence microscopy, immunoprecipitation and Western blotting, and FACS analysis for isolation and identification of endothelial microparticles. PAI-1 treatment resulted in a reduced expression of uPAR, its colocalization with caveolin, and the concomitant increase of uPAR abundance in the culture medium. FACS analysis revealed that PAI-1 rapidly and dose-dependently increased the number of endothelial microparticles expressing uPAR and alpha(V)beta3 integrin. This process was attenuated by pretreatment with neutralizing anti-uPAR antibodies. PAI-1 knockout mice showed a significantly decreased number of circulating endothelial microparticles than wild-type mice; however, PAI-1-deficient animals responded to infusion of PAI-1 with a more pronounced rise in the number of microparticles. PAI-1 treatment increased the number of microparticles stained with Annexin V, evidence for the expression of anionic phospholipids. This was accompanied by the accelerated generation of thrombin. CONCLUSIONS: The data disclose a novel effect of PAI-1 to dose-dependently promote formation of endothelial microparticles with the reduced transmembrane asymmetry of phospholipids. This phenomenon may be responsible for the observed increase in in vitro thrombin generation. These findings could potentially link these hallmarks of endothelial dysfunction-elevated levels of PAI-1 and propensity toward thrombosis.
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Coagulación Sanguínea/efectos de los fármacos , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Inhibidor 1 de Activador Plasminogénico/farmacología , Trombosis/etiología , Animales , Anticuerpos/farmacología , Coagulación Sanguínea/fisiología , Western Blotting , Caveolas/metabolismo , Caveolina 1 , Caveolinas/metabolismo , Células Cultivadas , Medios de Cultivo Condicionados/metabolismo , Relación Dosis-Respuesta a Droga , Endotelio Vascular/citología , Citometría de Flujo , Humanos , Sustancias Macromoleculares , Ratones , Ratones Noqueados , Microscopía Confocal , Tamaño de la Partícula , Fosfolípidos/biosíntesis , Inhibidor 1 de Activador Plasminogénico/deficiencia , Inhibidor 1 de Activador Plasminogénico/genética , Pruebas de Precipitina , Receptores de Superficie Celular/antagonistas & inhibidores , Receptores de Superficie Celular/metabolismo , Receptores del Activador de Plasminógeno Tipo Uroquinasa , Receptores de Vitronectina/metabolismo , Trombina/biosíntesisRESUMEN
Single-cell suspensions of primary keratinocytes comprise a heterogeneous cell population that consists of basal cells (stem cells, transient amplifying cells, and post-mitotic basal cells) and suprabasal cells at different stages of differentiation. Quantitative data for the differential expression of epitopes on single cells can be obtained using a flow cytometer. Simultaneous analysis of two intracellular epitopes, keratin 14 and connexin 43, using flow cytometry after keratinocyte isolation, fixation, permeabilization, and fluorescent immunolabeling is described. Three subsets of cells could be distinguished: stem cells (basal cells [keratin 14 positive] that lack connexin 43 expression); suprabasal cells (connexin 43-positive, keratin 14-negative cells); and basal cells (keratin 14 positive) that express connexin 43. The last population of keratinocytes includes both transient amplifying cells and postmitotic basal cells. The scatter characteristics of each cell population are also described.
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Diferenciación Celular/fisiología , Conexina 43/metabolismo , Queratinocitos/citología , Queratinas/metabolismo , Animales , Células Epidérmicas , Epidermis/metabolismo , Citometría de Flujo/métodos , Humanos , Inmunohistoquímica/métodos , Queratina-14 , Queratinocitos/metabolismoRESUMEN
OBJECTIVES: Determine whether surface dust grab samples taken from a large military base in Iraq are toxic and respirable. METHODS: X-ray diffraction for mineral content, x-ray fluorescence for elemental content, in vivo mouse dust challenges for assessment of histological changes, bronchoalveolar lavage for cytokines, polarizing light microscopy for crystals in lung tissue, and Fluorescence Activated Cell Sorting for cell surface and intracellular markers were utilized. RESULTS: Camp Victory, Iraq dust taken during wartime contains respirable particles 2.5 microns in size, constituting particulate matter air pollution. Dust particles are angular and have sharp edges. Trace metals (including titanium) calcium and silicon are present. Mice with airway instillation of dust have polarizable crystals in lung and septate inflammation. Regulatory T cells (CD4âºCD25âºFOXP3âº) are decreased in thymus and spleen. Interleukin-2 (IL-2) is upregulated in bronchoalveolar lavage. CONCLUSIONS: Respirable Iraq dust leads to lung inflammation in mice similar to that seen in patients with polarizable crystals, which seem to be titanium.
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Polvo/inmunología , Exposición por Inhalación/efectos adversos , Interleucina-2/metabolismo , Neumonía/patología , Fibrosis Pulmonar/patología , Linfocitos T Reguladores , Animales , Birrefringencia , Líquido del Lavado Bronquioalveolar/inmunología , Calcio/análisis , Polvo/análisis , Irak , Recuento de Linfocitos , Masculino , Ratones , Ratones Endogámicos C57BL , Instalaciones Militares , Material Particulado/química , Material Particulado/inmunología , Material Particulado/toxicidad , Neumonía/etiología , Fibrosis Pulmonar/etiología , Silicio/análisis , Bazo/patología , Timo/patología , Titanio/análisis , Regulación hacia ArribaRESUMEN
BACKGROUND: Children are susceptible to mercury toxicity, and mercury has immunomodulatory effects. Lower folate and B-12, and higher homocysteine may represent susceptibility cofactors. A large proportion of variability in rubella immune response is attributable to environmental factors. OBJECTIVE: This study aimed to evaluate the interaction between total blood mercury (Hg) and nutritional and homocysteine status on rubella virus antibody concentrations. DESIGN: Cross-sectional data on rubella IgG antibody concentrations, Hg, homocysteine, methylmalonic acid (MMA, an indicator of B-12 deficiency), and folate were obtained from 2003-2004 NHANES for children aged 6-11 years with rubella seropositivity (n=690). Linear regression was used to evaluate relationships between log-transformed rubella concentrations and Hg, stratified by sex, MMA ≥, folate<, and homocysteine ≥ sample medians, adjusted for demographic and nutritional cofactors. RESULTS: Hg was significantly positively associated with rubella antibody concentrations (ß=0.24; 95% confidence interval (CI)=0.11, 0.38) in children with higher MMA, lower folate and higher homocysteine (n=110), yet inversely associated among all other children (ß=-0.18; 95% CI=-0.34, -0.03) (n=580). Among the former, estimates (ß) were positive across all Hg quartiles relative to the lowest (Q1) (Hg<0.30 µg/L): Q2: ß=0.23 (-.10, 0.56); Q3: ß=0.35 (0.13, 0.57); Q4: ß=0.53 (0.21, 0.84); P(trend)<0.01. CONCLUSION: Findings are consistent with previously reported associations between Hg and measles antibody concentrations, and highlight the importance of considering dynamics between toxicant exposures, pathogens and host susceptibility.
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Anticuerpos Antivirales/sangre , Contaminantes Ambientales/sangre , Sarampión/sangre , Mercurio/sangre , Encuestas Nutricionales , Niño , Estudios Transversales , Femenino , Ácido Fólico/sangre , Homocisteína/sangre , Humanos , Inmunoglobulina G/sangre , Modelos Lineales , Masculino , Sarampión/epidemiología , Ácido Metilmalónico/sangre , Estados Unidos/epidemiología , Vitamina B 12/sangreRESUMEN
Serum antinuclear antibody positivity (ANA) has been associated with elevated serum polychlorinated biphenyls (PCBs) among residents in PCB-polluted areas; however, associations in general populations have not been reported by congener type or with adjustment for mercury. Cross-sectional data on serum PCBs, total blood mercury, ANA, and potential confounders age, race, body mass index, menopausal status, and dietary eicosapentaenoic acid (EPA) were obtained from the 2003-2004 National Health and Nutrition Examination Survey (NHANES) for males and females aged 12-85. PCB congeners were summed separately for dioxin-like and nondioxin-like PCBs; the former were weighted for toxic equivalent factors. Total PCBs by congener type and mercury were analyzed as both continuous log-transformed variables and as categorical quintiles. Logistic regression models were stratified by sex. There were no associations between nondioxin-like PCBs or mercury and ANA among males or females. Among females (n=114 affected and 518 unaffected), adjusting for potential confounders, the prevalence odds for ANA positivity were significantly elevated per incremental increase in log-transformed dioxin-like PCBs (odds ratio {OR}=1.66; 95% confidence interval {CI}=1.24, 2.23); the highest dioxin-like PCB quintile (>0.00425-0.04339ng/g) was significantly associated with 4.04 (95% CI=2.43, 6.70) greater prevalence odds for ANA positivity relative to the lowest quintile (Ptrend<0.001). We present novel findings of an association between low-level dioxin-like PCBs and ANA among women. No associations were observed between mercury and ANA at mercury levels common to the U.S. population.
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Anticuerpos Antinucleares/sangre , Dioxinas/sangre , Exposición a Riesgos Ambientales/análisis , Contaminantes Ambientales/sangre , Mercurio/sangre , Bifenilos Policlorados/sangre , Adolescente , Adulto , Anciano , Niño , Estudios Transversales , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Encuestas Nutricionales , Factores Sexuales , Adulto JovenRESUMEN
T-cell Prolymphocytic leukemia (T-PLL) is a rare post-thymic T-cell malignancy that follows an aggressive clinical course. The classical presentation includes an elevated white blood cell (WBC) count with anemia and thrombocytopenia, hepatosplenomegaly, and lymphadenopathy. T-PLL is a disease of the elderly and to our knowledge it has never been described in the pediatric age group. We report a case of T-PLL in a 9 year old male who was initially diagnosed with T-cell acute lymphoblastic lymphoma (ALL), the diagnosis was later refined to T-PLL following additional analysis of bone marrow morphology and immunophenotype. Two unusual findings in our patient included CD117 expression and an isolated chromosomal 12(p13) deletion. The patient failed to respond to standard ALL induction chemotherapy, but achieved complete remission following treatment with a fludarabine and alemtuzumab-based regimen.