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SUMMARY: The toxic effects of thioacetamide (TAA) and carbon tetrachloride on the human body are well recognized. In this study, we examined whether TAA intoxication can induce kidney leukocyte infiltration (measured as leukocyte common antigen CD45) associated with the augmentation of the reactive oxygen species (ROS)/tumor necrosis factor-alpha (TNF-α) axis, as well as biomarkers of kidney injury with and without metformin treatment. Rats were either injected with TAA (200 mg/kg; twice a week for 8 weeks) before being sacrificed after 10 weeks (experimental group) or were pre-treated with metformin (200 mg/kg) daily for two weeks prior to TAA injections and continued receiving both agents until the end of the experiment, at week 10 (protective group). Using basic histology staining, immunohistochemistry methods, and blood chemistry analysis, we observed profound kidney tissue injury such as glomerular and tubular damage in the experimental group, which were substantially ameliorated by metformin. Metformin also significantly (p0.05) increase in kidney expression of CD45 positive immunostaining cells. In conclusion, we found that TAA induces kidney injury in association with the augmentation of ROS/TNF-α axis, independent of leukocyte infiltration, which is protected by metformin.
Son bien conocidosos los efectos tóxicos de la tioacetamida (TAA) y el tetracloruro de carbono en el cuerpo humano. En este estudio, examinamos si la intoxicación por TAA puede inducir la infiltración de leucocitos renales (medida como antígeno leucocitario común CD45) asociada con el aumento de las especies reactivas de oxígeno (ROS)/factor de necrosis tumoral-alfa (TNF-α), así como biomarcadores de daño renal con y sin tratamiento con metformina. A las ratas se les inyectó TAA (200 mg/kg; dos veces por semana durante 8 semanas) antes de sacrificarlas a las 10 semanas (grupo experimental) o se les pretrató con metformina (200 mg/kg) diariamente durante dos semanas antes de las inyecciones de TAA y continuaron recibiendo ambos agentes hasta el final del experimento, en la semana 10 (grupo protector). Usando tinción histológica básica, métodos de inmunohistoquímica y análisis químico de la sangre, observamos una lesión profunda del tejido renal, como daño glomerular y tubular en el grupo experimental, que mejoraron sustancialmente con la metformina. La metformina también inhibió significativamente (p0,05) en la expresión renal de células de inmunotinción positivas para CD45. En conclusión, encontramos que el TAA induce la lesión renal en asociación con el aumento del eje ROS/TNF-α, independientemente de la infiltración de leucocitos, que está protegida por metformina.
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Animais , Masculino , Ratos , Tioacetamida/toxicidade , Injúria Renal Aguda/tratamento farmacológico , Hipoglicemiantes/uso terapêutico , Metformina/uso terapêutico , Imuno-Histoquímica , Biomarcadores , Fator de Necrose Tumoral alfa , Espécies Reativas de Oxigênio , Antígenos Comuns de Leucócito , Injúria Renal Aguda/induzido quimicamente , InflamaçãoRESUMO
Abstract Objective: The prognostic importance of Tumor-Infiltrating Lymphocytes (TILs) in the tumor microenvironment of various cancers is increasingly recognized. In the present study, we aimed to investigate the prognostic value of CD3+, CD4+, CD8+, and CD45RO + TILs and their relation to histopathological features in larynx squamous cell carcinoma. Methods: Formalin-Fixed and Paraffin-Embedded (FFPE) samples from 63 primary larynx squamous cell carcinoma patients were immunostained for CD3, CD4, CD8, and CD45RO expression. Positive cells in micrographs from Invasive Margin (IM) and Tumor Center (CT) of tissue specimens counted by ImageJ software and their correlation with disease outcome were analyzed. Results: The expression level of TILs subpopulations was associated with clinicopathological markers as well as Overall Survival (OS) and Disease-Free Survival (DFS). In multivariate analysis, high frequency of CD45RO + cells in IM were confirmed as an independent prognostic marker for DFS (p = 0.007, HR = 4.968) and OS (p = 0.007, HR = 4.957). Similar findings were observed in the multivariate analysis of the combined frequency of CD45RO+cells in IM and CT. Conclusion: TILs are associated with patients clinicopathological features. Also, our findings indicate that CD45RO + TILs are a valuable marker for risk prediction in larynx SCC and could predict patients' outcomes.
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SUMMARY: Diabetes and hypertension account for the majority of chronic kidney injury cases that can lead to renal failure. The link between the leukocytes common antigen (CD45) and diabetic kidney disease (DKD) with and without metformin incorporation in an animal model has not been investigated before. Therefore, we sought to assess the extent of leukocytes infiltration into kidney tissues 10 weeks following the induction of diabetes in rats treated with metformin. In addition, we monitored blood and urine parameters associated with diabetes. The model group of rats received streptozotocin (STZ; 50 mg/kg) injection after being fed for 14 days on a high-fat diet (HFD) and continuously fed a HFD until they were culled, at week 12. The protective group was treated in the same way except that these animals were put from day 1 on metformin (200 mg/kg) until being culled, on week 12. Kidneys were immunostained with CD45 as a marker of leukocytes infiltration and examined by light microscopy. Urine samples were tested for urine albumin and collected blood was analyzed for sugar, urea, creatinine, and oxidative stress and antioxidants biomarkers. Kidney injury secondary to diabetes was developed as demonstrated by (i) increased blood glucose, urea, and malondialdehyde (MDA) as a marker of lipid peroxidation; and (ii) kidney tissue damage and marked increase in kidney tissues expressing CD45 positive cells. The above markers were inhibited (p0.0006) by metformin. Also, a significant correlation was observed between CD45 score and glycemia, urea, MDA, and the antioxidant superoxide dismutase (SOD). Thus, our data demonstrate an association between the infiltration of CD45+ inflammatory cells into kidney tissues and biomarkers of kidney damage in a rat model of DKD, which was effectively protected by metformin.
RESUMEN: La diabetes y la hipertensión representan la mayoría de los casos de lesión renal crónica que pueden provocar insuficiencia renal. El vínculo entre el antígeno común de los leucocitos (CD45) y la enfermedad renal diabética (DKD) con y sin incorporación de metformina en un modelo animal no se había anteriormente investigado. El objetivo fue evaluar el grado de infiltración de leucocitos en los tejidos renales 10 semanas después de la inducción de diabetes en ratas tratadas con metformina. Además, monitoreamos los parámetros de sangre y orina asociados con la diabetes. El grupo modelo de ratas recibió una inyección de estreptozotocina (STZ; 50 mg/kg) después de ser alimentadas durante 14 días con una dieta alta en grasas (HFD) y continuamente alimentadas con un HFD hasta que fueron sacrificadas, en la semana 12. El grupo protector fue tratado de la misma manera excepto que estos animales fueron recibieron desde el día 1 metformina (200 mg/kg) hasta ser sacrificados, en la semana 12. Los riñones fueron inmunoteñidos con CD45 como marcador de infiltración de leucocitos y examinados por microscopía óptica. Las muestras de orina se analizaron en busca de albúmina y la sangre recolectada se analizó en busca de glucosa, urea, creatinina y biomarcadores de estrés oxidativo y antioxidantes. La lesión renal secundaria a la diabetes se desarrolló como lo demuestra (i) el aumento de la glucosa en sangre, la urea y el malondialdehído (MDA) como marcador de la peroxidación lipídica; y (ii) daño del tejido renal y marcado aumento en los tejidos renales que expresan células positivas para CD45. Los marcadores anteriores fueron inhibidos (p≤0.0006) por metformina. Además, se observó una correlación significativa entre la puntuación de CD45 y la glucemia, la urea, la MDA y la superóxido dismutasa antioxidante (SOD). Por lo tanto, nuestros datos demuestran una asociación entre la infiltración de células inflamatorias CD45+ en los tejidos renales y biomarcadores de daño renal en un modelo de rata con DKD, que fue protegido de manera efectiva por metformina.
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Animais , Ratos , Diabetes Mellitus , Injúria Renal Aguda/prevenção & controle , Hipoglicemiantes/administração & dosagem , Metformina/administração & dosagem , Biomarcadores , Antígenos Comuns de Leucócito , Estresse Oxidativo/efeitos dos fármacos , Modelos Animais de Doenças , Hipoglicemiantes/uso terapêutico , Inflamação , Rim/efeitos dos fármacos , Metformina/uso terapêuticoRESUMO
SUMMARY: Ingestion of an overdose of paracetamol (also called acetaminophen, or APAP) induces hepatotoxicity that can lead to liver failure. The link between the pro-inflammatory microRNA-155 (miR-155) and leukocyte infiltration (CD45) in APAP- antioxidant depletion and liver toxicity with and without the natural polyphenolic compounds, quercetin (QUR) plus resveratrol (RES) has not been previously studied. Therefore, acute hepatic injury was induced in rats by 2 g/kg APAP (single dose, orally) and another group started QUR (50 mg/kg) plus RES (30 mg/kg) treatment one week prior to APAP ingestion. Animals were culled 24 hours post the paracetamol treatment. APAP overdose induced hepatic and blood levels of miR-155 expression, CD45 (leukocyte common antigen) immunostaining, degenerated hepatocytes, and hepatic injury enzymes; alanine aminotransferase (ALT) and aspartate aminotransferase (AST), which were markedly decreased by QUR+RES. Whereas, APAP intoxication ameliorated liver tissue levels of the antioxidants, glutathione peroxidase and superoxide dismutase that were augmented by QUR+RES. Moreover, a significant (p<0.05) correlation between miR-155/CD45 axis and liver tissue injury was observed. These findings show that paracetamol intoxication augments miR- 155/CD45 axis-mediated modulation of antioxidants and liver injury in rats, and is protected by QUR+RES.
RESUMEN: La ingestión de una sobredosis de paracetamol (también llamado acetaminofeno o APAP) induce hepatotoxicidad que puede provocar insuficiencia hepática. El vínculo entre el microARN-155 proinflamatorio (miR-155) y la infiltración de leucocitos (CD45) en el agotamiento de APAP- antioxidante y la toxicidad hepática con y sin los compuestos polifenólicos naturales, quercetina (QUR) más resveratrol (RES) no ha sido previamente investigado. En este estudio, se indujo daño hepático agudo en ratas con 2 g/kg de APAP (dosis única, por vía oral) y otro grupo comenzó el tratamiento con QUR (50 mg/ kg) más RES (30 mg/kg) una semana antes de la ingestión de APAP. Los animales se sacrificaron 24 horas después del tratamiento con paracetamol. La sobredosis de APAP indujo niveles hepáticos y sanguíneos de expresión de miR-155, inmunotinción de CD45 (antígeno leucocitario común), degeneración de los hepatocitos y daño hepático enzimático; alanina aminotransferasa (ALT) y aspartato aminotransferasa (AST), disminuyeron notablemente con QUR+RES. Mientras que la intoxicación con APAP mejoró los niveles de antioxidantes, glutatión peroxidasa y superóxido dismutasa en el tejido hepático los que aumentaron con QUR+RES. Además, se observó una correlación significativa (p<0,05) entre el eje miR-155/CD45 y la lesión del tejido hepático. Estos hallazgos muestran que la intoxicación por paracetamol aumenta la modulación mediada por el eje miR-155/CD45 de los antioxidantes y la lesión hepática en ratas, y está protegida por QUR+RES.
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Animais , Ratos , Quercetina/farmacologia , Doença Hepática Induzida por Substâncias e Drogas , Resveratrol/farmacologia , Acetaminofen/toxicidade , Antioxidantes/farmacologia , Ratos Sprague-Dawley , Antígenos Comuns de Leucócito/efeitos dos fármacos , MicroRNAs/efeitos dos fármacosRESUMO
BACKGROUND@#Lung cancer is the most common malignancy world-wide. There are a variety of immune infiltrating cells in tumor microenvironment, which is an important component of tumor immunity and has clinical significance for the prognosis of patients. CD45RO is a surface marker of memory T cells. The expression of CD45RO⁺ tumor infiltrating lymphocytes (TILs) is associated with the prognosis of many tumors. The purpose of this study was to evaluate the relationship between the density of CD45RO⁺ TILs in tumor and stromal area and the clinical characteristics of patients with non-small cell lung cancer (NSCLC) and its impact on the prognosis of patients. We aimed to explore the clinical value of CD45RO⁺ TILs and programmed cell death ligand 1 (PD-L1) as prognostic markers.@*METHODS@#Multiple fluorescent immunohistochemical staining was used to stain the tissue microarray chips of 167 patients with NSCLC, marking CD45RO, cytokeratin (CK) and PD-L1. Using artificial intelligence image recognition technology and tumor cell-specific CK staining, divide the tumor and stromal area in the tissue, evaluate the density of CD45RO⁺ TILs in the tumor and stromal area, and the expression level of PD-L1 in tumor cells. The non-parametric test was used to analyze the relationship between CD45RO⁺ TILs and the clinical characteristics of patients, and the Kaplan-Meier method and Cox risk ratio model were used to analyze the relationship between CD45RO⁺ TILs independently or in combination with PD-L1 and tumor prognosis.@*RESULTS@#The density of CD45RO⁺ TILs was significantly associated with patient age, smoking, tumor stage, and pathological type. Single-factor survival analysis showed that NSCLC (P=0.007) stromal region and lung adenocarcinoma (LUAD) (P<0.001) with CD45RO⁺ TILs high density had better OS. Multivariate survival analysis showed that the high density of CD45RO⁺ TILs in the stromal region of NSCLC (HR=0.559, 95%CI: 0.377-0.829, P=0.004) and lung adenocarcinoma (HR=0.352, 95%CI: 0.193-0.641, P=0.001) were independent prognostic factors for overall survival time (OS). Combined with PD-L1 score of tumor cells in tumor tissues and infiltration score of CD45RO⁺ TILs in all tumor tissues, the patients were divided into 4 groups: patients with PD-L1⁺/CD45RO⁺ had the longest disease-free survival (DFS) time, and patients with PD-L1⁺/CD45RO- had the shortest DFS time. Multivariate Cox regression analysis showed that PD-L1⁺/CD45RO- was an independent prognostic factor for DFS and had a higher risk of poor prognosis compared to the other three groups (HR=2.221, 95%CI: 1.258-3.919, P=0.006).@*CONCLUSIONS@#In tumor tissues, the density of CD45RO⁺ TILs, as well as the combination of CD45RO⁺ TILs and PD-L1 in tumor areas, significantly correlated with clinicopathological features and prognosis of NSCLC, which can be used as a new prognosis marker.
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Objective:To investigate the expression of IL-2/IL-15 receptor β subunit (IL-2/IL-15Rβ) on memory CD3 + CD8 + CD45RO + T cells in patients with chronic hepatitis B (CHB) receiving antiviral treatment and its significance. Methods:Sixty-eight patients with chronic active hepatitis B (CAHB) and 47 asymptomatic hepatitis B virus (HBV) carriers attending in the Department of Infectious Diseases, the First Affiliated Hospital of Wannan Medical College from March 2019 to December 2020 were enrolled in the study; and 30 health subjects were also enrolled as healthy control group. Among 60 CAHB patients there were 30 cases with positive HBeAg and 30 cases with negative HBeAg. All CAHB patients received nucleos(t)ide analogue therapy, the HBV-related markers, Alanine aminotransferase (ALT) and the expression of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were determined and compared between HBeAg-positive and negative patients, before and after treatment. Normal distribution measurement data among 3 groups were compared with One-way ANOVA; normal distribution measurement data between 2 groups were compared with paired samples t test; non-normal distribution measurement data between the two groups were compared with Mann-Whitney U test; Pearson’s correlation coefficient was performed for correlation analysis. P<0.05 was considered statistically significant. Results:The proportion of CD8 + CD45RO + T cells on PBMC CD3 + T cells in CAHB group [(8.6±3.7)%] was higher than that of asymptomatic HBV carriers group [(5.7±2.5)%] and healthy control group [(5.5±1.5)%] (all P<0.05). The expression percentage of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells in CAHB group [(6.8±4.7)%] was higher than that of asymptomatic HBV carriers group [(4.7±2.8)%] and healthy control group [(4.3±2.2)%] (all P<0.05). The MFI of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells in CAHB group (243±168) was higher than those of asymptomatic HBV carriers group (160±91) and healthy control group [160±63] (all P<0.05). The expression percentage and MFI of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells were positively correlated with the percentage of CD3 + CD8 + CD45RO + T cells in CAHB patients ( r=0.33 and 0.28, all P<0.05). The proliferation percentage of PBMC CD3 + CD8 + CD45RO + T cells in CAHB group[ (43.7±16.0)%] was higher than that of asymptomatic HBV carriers group [(29.1±9.4)%] and healthy control group [(26.8±9.6)%] after stimulation with Anti-CD3+ super-2 (all P<0.05). After the expression of IL-2/IL-15Rβ was blocked, the proliferation percentage of CD3 + CD8 + CD45RO + T cells was decreased [(11.2±6.3)%] compared with the untreated CAHB group ( P<0.05). The percentages of PBMC CD3 + CD8 + CD45RO + T cells secreting IFN-γ, IL-2 and TNF-α in CAHB group were (13.8±5.4)%, (14.0±4.3)% and (12.3±4.6)% respectively, which were higher than those of asymptomatic HBV carriers [(8.4±2.6)%, (9.4±3.2)% and (6.8±3.3)%] and healthy control group [(6.9±2.7)%, (9.9±3.0)% and (7.7±3.8)%] after stimulation with Anti-CD3+ super-2 (all P<0.05). After the expression of IL-2/IL-15Rβ was blocked, the percentages of PBMC CD3 + CD8 + CD45RO + T cells secreting IFN-γ [(2.4±1.6)%], IL-2 [(4.1±1.9)%] and TNF-α [(4.1±1.8)%] were decreased compared with the untreated CAHB group (all P<0.05). HBeAg, ALT, the expression percentage and MFI of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were 521.4 (68.9, 1 339.0) COI, 292 (160, 528) U/L, (6.4±3.2)% and (239±136) in 30 HBeAg-positive CAHB patients before treatment, which were higher than those after treatment [3.5(1.5, 17.5)COI、20(14, 31) U/L, (4.1±2.4)% and (134±58)] ( Z=5.337 and 6.403, t=3.229 and 3.892, all P<0.05). HBsAg, ALT, the expression percentage and MFI of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were (5 310±2 851) COI, (328±207) U/L, (7.1±5.8)% and (252±110) in 30 HBeAg-negative CAHB patients before treatment, which were higher than those after 48 weeks of treatment [(3 811±2 495) COI, (33±14) U/L, (4.6±2.9)% and (154±73)] ( t=2.167, 5.595, 2.116 and 2.383, all P<0.05). Conclusion:The study suggests that up-regulated expression of IL-2/IL-15Rβ is associated with elevated frequency, proliferation and secretion function of memory CD3 + CD8 + CD45RO + T cells in CAHB patients.
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Introducción: El CD45 se expresa en las células hematopoyéticas, su determinación es indispensable para la clasificación inmunofenotípica de la leucemia linfoide aguda (LLA). Objetivo: Evaluar la expresión del antígeno CD45 en los blastos de pacientes pediátricos con LLA y su relación con las características biológicas, morfológicas y clínicas al inicio de la enfermedad, la respuesta al tratamiento y la supervivencia global (SG) de los enfermos. Métodos: Se estudiaron 160 pacientes con LLA entre diciembre del 2012 y diciembre del 2017, tratados con el protocolo ALL-IC BFM-SG 2009. El inmunofenotipaje celular de la médula ósea se realizó por citometría de flujo. Resultados: El fenotipo B CD45+ predominó en los menores de seis años de edad y en los mayores de diez, el fenotipo T CD45+. Se encontró diferencia significativa entre la ausencia de adenopatías mediastínicas, el fenotipo leucémico y la ausencia de CD45 (p=0.004); entre la respuesta a la prednisona en sangre periférica al día ocho, el fenotipo leucémico y la ausencia de CD45 (p=0.001). Se encontraron diferencias significativas entre la respuesta a la prednisona en sangre periférica el día ocho y la respuesta en médula ósea el día 33, según fenotipo leucémico (p=0.009) y la presencia en los blastos del antígeno CD45 (p=0.02). Se encontró diferencia significativa entre la SG de los enfermos, según fenotipo leucémico y la ausencia del antígeno CD45 (p=0.017). Conclusión: La expresión o ausencia del antígeno de CD45 en los blastos tiene relación con la respuesta al tratamiento y la SG de pacientes pediátricos con LLA(AU)
Introduction: CD45 is expressed in hematopoietic cells, its determination is essential for the immunophenotypic classification of acute lymphoid leukemia (ALL). Objective: To evaluate the expression of the CD45 antigen in the blasts of pediatric patients with ALL and its relationship with the biological, morphological and clinical characteristics at the onset of the disease, the response to treatment and the overall survival (OS) of the patients. Methods: 160 patients with ALL were studied between December 2012 and December 2017, treated with the ALL-IC BFM-SG 2009 protocol. Bone marrow cellular immunophenotyping was performed by flow cytometry. Results: Patients with the CD45 + B phenotype predominated in those under six years of age, while those with a CD45 + T phenotype in those older than ten. A significant difference was found between the absence of mediastinal lymph nodes, the leukemic phenotype and the absence of CD45 (p = 0.004). A significant difference was found between the response to prednisone in peripheral blood at day eight, the leukemic phenotype and the absence of CD45, p = 0.001. Significant differences were found between the response to prednisone in peripheral blood on day eight and the response in bone marrow on day 33, according to leukemic phenotype and the presence in blasts of the CD45 antigen (p = 0.009 and p = 0.02, respectively). A significant difference was found between the OS of patients, according to leukemic phenotype and the absence of the CD45 antigen, p = 0.017. Conclusion: The expression or absence of the CD45 antigen in blasts is related to the response to treatment and OS of pediatric patients with ALL(AU)
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Humanos , Feminino , Lactente , Pré-Escolar , Criança , Adolescente , Imunofenotipagem/métodos , Antígenos Comuns de Leucócito/uso terapêutico , Leucemia-Linfoma Linfoblástico de Células Precursoras/epidemiologia , Citometria de Fluxo/métodos , Fenótipo , Análise de SobrevidaRESUMO
Background: The most fundamental trait of cancer cells involves their ability to sustain chronic proliferation. Tumors have a complex cellular ecology that establishes the malignant potential of the tumor. In these ecosystems, innate immune cells are highly represented. Many contradictory reports have been published regarding the impact of tumor-infiltrating immune cells on proliferation of the tumors. Aim: This study aims to assess the impact of CD45RO+ve immune cells on proliferation and dedifferentiation of node-negative squamous cell carcinomas of cheek mucosa (SCC-CM). Materials and Methods: Thirty formalin-fixed paraffin-embedded tissue blocks of previously diagnosed node-negative SCC-CM subclassified as Grade I SCC – 10 cases; Grade II SCC – 10 cases; and Grade III SCC – 10 cases (Broders' classification – 1927). Immunohistochemistry performed on each selected tissue section using anti-p53 and anti-CD45RO as primary antibodies. Semi-quantitative analyses performed for all the tissue sections to assess the p53 and CD45RO expression. p53:CD45RO expression ratio calculated. The data were statistically analyzed using GraphPad Prism 5 for Windows. Results: Our results showed statistically significant increase (P = 0.0006) in p53 expression and decrease (P = 0.0044) in CD45RO+ immune cell response with the decrease in differentiation of SCC-CMs using Fisher's exact test and statistically significant increase (P < 0.001) in p53:CD45RO expression ratio with decrease in differentiation using one-way ANOVA. Conclusion: Based on all these findings from the present study, we perceive the following findings. In node-negative SCC-CMs, CD45RO+ immune cells play a possible role in controlling the dedifferentiation of the tumor and in limiting the proliferative potential of the tumor cells which are tumor antagonistic in nature
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We present a case of B-acute lymphoblastic leukemiain an elderly patient who presented with severe weakness and pancytopenia. The patient was a 75 year old Female whose blasts had an unusual morphology in form of coarse azurophilic granules and cytoplasmic blebs and on flow cytometry the blasts were present in the bright CD45 zone with a high side scatter. Bone marrow aspirate sample was subjected to multicolour flow cytometry using Beckman Coulter Navios® which is an 8 colour flow cytometer.Flow cytometricanalysis of the bone marrow aspirate showed blasts in the monocytic zone with a precursor B cell immunophenotype. Complete blood counts showed pancytopenia with peripheral blood film not showingany blasts. Bone marrow aspirate smears showed 20% blasts with coarse azurophilic granules and cytoplasmic blebs.The position of the blasts in this case which were in monocytic zone giving them a bright expression of CD45 and a high side scatter on the CD45 side scatter. This is not the usual positionfor blasts in B-acute lymphoblastic leukemia as these blasts are less complex. A bright expression of CD45 by blasts in B-acute lymphoblastic leukemia is known to be associated with a poor prognosis but the clinical significance of blasts being bright CD45 with a high side scatter is a very rare occurrence and more number of cases with a similar presentation are required to determine a prognostic significance.
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[Absract] Objective This paper was designed to reveal the new mechanism on ASI Ⅱ triggered CD4+T cells activation via regulating CD45 molecular and provide a basis for the theoretical foundation of antitumor immunotherapy of Astragalus.Methods The CD4+T cells were randomly divided into negative group,stimulated control group,ASI Ⅱ group,CD45 inhibitor group,and the combination of ASI Ⅱ and CD45 inhibitor group.Besides negative group,the cells from other groups were activated by anti-CD3/CD28 antibody.ASI Ⅱ group was treated with 10 nmol/L ASI Ⅱ,CD45 inhibitor group was treated with 0.8 μmol/L CD45 inhibitor,and the combination group were treated with 10 nmol/L ASI Ⅱ and 0.8 iμmol/L CD45 inhibitor.After 36h culture,the proliferation of CD4+T cells was detected by Ki-67 intracellular staining assay.Cytokine production of Th1 and Th2 were examined ELISA method.The proportion of surface marker (CD44 and CD25)and Th1 intracellular cytokines (IFN-γ) were detected by flow cytometry.Results Compared with stimulated group,Astragaloside Ⅱ group in CD4+Ki67+T positive proportion (5.37% ± 0.92% vs.1.19% ± 0.23%),in CD4+CD25+ positive proportion (50.23% ± 4.65 % vs.15.89% ± 1.13%),in CD4+CD44+ positive proportion (33.16% ± 6.08% vs.15.36% 4 1.45%),in CD4+IFN-γ+ positive proportion (1.42% ± 0.44 % vs.0.38% ± 0.06%) were significntly increased.And the secretion of IFN-γ,IL-4 and IL-2 in ASI Ⅱ group were higher than stimulated group.The anti-mouse CD45 Ab treatment markedly blocked the proliferation and Th1 cytokines production which induced by ASI Ⅱ.Furthermore,the anti-mouse CD45 Ab treatment significantly decreased the expression of surface marker (CD44 and CD25).Conclusions Activating CD45 protein tyrosine phosphatase may be involved in ASI Ⅱ triggered CD4+T cells activation.This study will provide a basis for antitumor immunotherapy of Astragalus.
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Objective To investigate the expressions of CD4+CD45RA+ T cells and CD4+CD45RO+ T cells in peripheral blood of patients with acute coronary syndrome (ACS) and their significance. Methods A case-control study was conducted. Ninety-four patients receiving coronary angiography (CAG) admitted to Tianjin Chest Hospital from March 5th to April 27th in 2018 were enrolled. They were divided into non-coronary heart disease (CHD) group (n = 12), unstable angina pectoris (UAP) group (n = 27), acute non-ST elevation myocardial infarction (NSTEMI) group (n = 27) and acute ST elevation myocardial infarction (STEMI) group (n = 28) according to the patients' symptoms, electrocardiogram, troponin test and angiographic results. General data, blood routine parameters, and biochemical indicators were collected. The ratios of CD4+CD45RA+ T cells and CD4+CD45RO+ T cells were determined by flow cytometry. Multivariate Logistic regression was used to evaluate whether CD4+CD45RA+ T cells and CD4+CD45RO+ T cells were associated with STEMI. Results Ninety-four patients were included initially. After excluding the patients who died during the intervention, 93 patients were enrolled in the data analysis finally, with 12 patients in the non-CHD group, 27 patients in the UAP group, and the same as the NSTEMI group and the STEMI group. Compared with the non-CHD group, white blood cell count (WBC) was decreased (×109/L: 6.03±1.30 vs. 6.60±1.30, P > 0.05), and lymphocyte ratio was increased (0.273±0.059 vs. 0.269±0.070, P > 0.05) in patients of the UAP group; however, in the NSTEMI group and STEMI group, WBC was increased (×109/L: 8.29±2.28, 9.86±2.76 vs. 6.60±1.30, both P < 0.05), and lymphocyte ratio was decreased (0.236±0.076, 0.173±0.094 vs. 0.269±0.070, P > 0.05 and P < 0.05), especially in the STEMI group [WBC (×109/L): 9.86±2.76 vs. 6.60±1.30, lymphocyte ratio: 0.173±0.094 vs. 0.269±0.070, both P < 0.05]. There was no significant difference in biochemical indicators among all of the groups. Flow cytometry results showed that the ratios of CD4+CD45RO+ T cells in the UAP group and NSTEMI group were higher than those in the non-CHD group (0.323±0.074, 0.319±0.078 vs. 0.314±0.058, both P > 0.05); however, the ratio of CD4+CD45RO+ T cells in the STEMI group showed a decreased tendency (0.270±0.057 vs. 0.314±0.058, P > 0.05), and it was significantly lower than that in the UAP group and the NSTEMI group (0.270±0.057 vs. 0.323±0.074, 0.319±0.078, both P < 0.05). There was no significant difference in the ratio of CD4+CD45RA+ T cells among all of the groups. Multivariate Logistic regression analysis showed that CD4+CD45RA+ T cells ratio was not significantly correlated with the occurrence of STEMI [odds ratio (OR) = 0.976, 95% confidence interval (95%CI) was 0.907-1.050, P = 0.518], but CD4+CD45RO+ T cells ratio was significantly correlated with the occurrence of STEMI (OR = 0.888, 95%CI was 0.821-0.961, P = 0.003). Conclusions There was no significant difference in the ratio of CD4+CD45RA+ T cells among UAP, NSTEMI and STEMI patients, and CD4+CD45RO+ T cells ratio in the STEMI group was significantly lower than that in the UAP group and NSTEMI group. CD4+CD45RO+ T cells ratio may be risk factor of STEMI.
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Objective@#To investigate the expressions of CD4+CD45RA+ T cells and CD4+CD45RO+ T cells in peripheral blood of patients with acute coronary syndrome (ACS) and their significance.@*Methods@#A case-control study was conducted. Ninety-four patients receiving coronary angiography (CAG) admitted to Tianjin Chest Hospital from March 5th to April 27th in 2018 were enrolled. They were divided into non-coronary heart disease (CHD) group (n = 12), unstable angina pectoris (UAP) group (n = 27), acute non-ST elevation myocardial infarction (NSTEMI) group (n = 27) and acute ST elevation myocardial infarction (STEMI) group (n = 28) according to the patients' symptoms, electrocardiogram, troponin test and angiographic results. General data, blood routine parameters, and biochemical indicators were collected. The ratios of CD4+CD45RA+ T cells and CD4+CD45RO+ T cells were determined by flow cytometry. Multivariate Logistic regression was used to evaluate whether CD4+CD45RA+ T cells and CD4+CD45RO+ T cells were associated with STEMI.@*Results@#Ninety-four patients were included initially. After excluding the patients who died during the intervention, 93 patients were enrolled in the data analysis finally, with 12 patients in the non-CHD group, 27 patients in the UAP group, and the same as the NSTEMI group and the STEMI group. Compared with the non-CHD group, white blood cell count (WBC) was decreased (×109/L: 6.03±1.30 vs. 6.60±1.30, P > 0.05), and lymphocyte ratio was increased (0.273±0.059 vs. 0.269±0.070, P > 0.05) in patients of the UAP group; however, in the NSTEMI group and STEMI group, WBC was increased (×109/L: 8.29±2.28, 9.86±2.76 vs. 6.60±1.30, both P < 0.05), and lymphocyte ratio was decreased (0.236±0.076, 0.173±0.094 vs. 0.269±0.070, P > 0.05 and P < 0.05), especially in the STEMI group [WBC (×109/L): 9.86±2.76 vs. 6.60±1.30, lymphocyte ratio: 0.173±0.094 vs. 0.269±0.070, both P < 0.05]. There was no significant difference in biochemical indicators among all of the groups. Flow cytometry results showed that the ratios of CD4+CD45RO+ T cells in the UAP group and NSTEMI group were higher than those in the non-CHD group (0.323±0.074, 0.319±0.078 vs. 0.314±0.058, both P > 0.05); however, the ratio of CD4+CD45RO+ T cells in the STEMI group showed a decreased tendency (0.270±0.057 vs. 0.314±0.058, P > 0.05), and it was significantly lower than that in the UAP group and the NSTEMI group (0.270±0.057 vs. 0.323±0.074, 0.319±0.078, both P < 0.05). There was no significant difference in the ratio of CD4+CD45RA+ T cells among all of the groups. Multivariate Logistic regression analysis showed that CD4+CD45RA+ T cells ratio was not significantly correlated with the occurrence of STEMI [odds ratio (OR) = 0.976, 95% confidence interval (95%CI) was 0.907-1.050, P = 0.518], but CD4+CD45RO+ T cells ratio was significantly correlated with the occurrence of STEMI (OR = 0.888, 95%CI was 0.821-0.961, P = 0.003).@*Conclusions@#There was no significant difference in the ratio of CD4+CD45RA+ T cells among UAP, NSTEMI and STEMI patients, and CD4+CD45RO+ T cells ratio in the STEMI group was significantly lower than that in the UAP group and NSTEMI group. CD4+CD45RO+ T cells ratio may be risk factor of STEMI.
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Objective@#To explore the expression of CD45 in newly diagnosed multiple myeloma (MM) and its relationship with clinical efficacy and prognosis.@*Methods@#This study retrospectively analyzed expression and distribution of CD45 in 130 cases of newly diagnosed MM, comparing clinical efficacy and prognosis in CD45+/CD45- groups.@*Results@#①The CD45+ group was 33 cases (25.38%) , and CD45- group was 97 cases (74.62%) . ②The objective remission rate (ORR) of CD45+ and CD45-group was 33.33% and 64.95%, respectively. The difference was statistically significant (P=0.002) . For patients in Bortezomib regimen, the ORR of CD45+ and CD45- group was 35.71% and 66.25%, respectively. The difference was statistically significant (P=0.005) . ③The median progress free survival (PFS) of CD45+ group and CD45- group was 29.8 (95%CI 10.0-59.0) months vs 34.5 (95%CI 6.0-69.0) months (χ2=14.59, P<0.001) and the median overall survival (OS) was 32.5 (95%CI 10.0-68.0) months vs 37.6 (95%CI 6.0-78.0) months (χ2=11.42, P=0.001) , respectively. Among the patients in bortezomib regimen, The median PFS and median OS of CD45 + group and CD45- group were 30.3 (95%CI 10.0-59.0) months vs 36.3 (95%CI 6.0-69.0) months (χ2=14.75, P=0.001) and 34.0 (95%CI 10.0-68.0) months vs 39.5 (95%CI 6.0-78.0) months (χ2=10.62, P=0.001) . ④Cox risk regression model analysis showed that serum creatinine≥176.8 μmol/L (HR=5.078, 95%CI 1.744-14.723, P=0.001) , CD45 positive (HR=14.504, 95%CI 0.168-0.42, P=0.001) , LDH≥220 IU/L (HR=1.308, 95%CI 1.16-2.417, P=0.015) were independent risk prognostic factors.@*Conclusion@#CD45 expression is a risk prognostic factor of MM patients. Bortezomib did not improve the poor prognosis of CD45+ MM patients.
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Objective: To investigate the cellular components and differentiation potential of cells in rabbit pericardial fluid, and to provide morphological basis for basic research and clinical application of pericardial cells. Methods: Thirty adult New Zealand white rabbits, after aseptic thoracotomy, the pericardial fluid mixture was extracted, the fluid cells were centrifuged, isolated and cultured. The pericardial cellular morphology in the different generations was observed under the inverted microscope (The immunofluorescence staining method was used in the present study in order to analyze the pericardial cells phenotypes). Their immunological phenotypes were analyzed by using immunofluorescence staining and the CD44, vimentin, CD45 and the number of cells positively expressed in the third generation cells were observed. The expression of CD44 and vimentin related molecules was detected by PCR. Results: There was the cellular population with uniform morphology in the adult rabbit pericardial fluid. The cells with immunofluorescence positive staining for CD44 and vimentin were found in the pericardial fluid of rabbit, in addition, these cells possessed the immunofluorescence negative staining for CD45. After induction, they can differentiate into osteoblasts and adipocytes. Conclusion: Rabbit pericardial fluid contains cells with multiple differentiation potentials, which may be of positive significance for myocardial repair.
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Objective: To explore the expression of CD45 in newly diagnosed multiple myeloma (MM) and its relationship with clinical efficacy and prognosis. Methods: This study retrospectively analyzed expression and distribution of CD45 in 130 cases of newly diagnosed MM, comparing clinical efficacy and prognosis in CD45(+)/CD45(-) groups. Results: ①The CD45(+) group was 33 cases (25.38%) , and CD45(-) group was 97 cases (74.62%) . ②The objective remission rate (ORR) of CD45(+) and CD45(-)group was 33.33% and 64.95%, respectively. The difference was statistically significant (P=0.002) . For patients in Bortezomib regimen, the ORR of CD45(+) and CD45(-) group was 35.71% and 66.25%, respectively. The difference was statistically significant (P=0.005) . ③The median progress free survival (PFS) of CD45(+) group and CD45(-) group was 29.8 (95%CI 10.0-59.0) months vs 34.5 (95%CI 6.0-69.0) months (χ(2)=14.59, P<0.001) and the median overall survival (OS) was 32.5 (95%CI 10.0-68.0) months vs 37.6 (95%CI 6.0-78.0) months (χ(2)=11.42, P=0.001) , respectively. Among the patients in bortezomib regimen, The median PFS and median OS of CD45 (+) group and CD45(-) group were 30.3 (95%CI 10.0-59.0) months vs 36.3 (95%CI 6.0-69.0) months (χ(2)=14.75, P=0.001) and 34.0 (95%CI 10.0-68.0) months vs 39.5 (95%CI 6.0-78.0) months (χ(2)=10.62, P=0.001) . ④Cox risk regression model analysis showed that serum creatinine≥176.8 μmol/L (HR=5.078, 95%CI 1.744-14.723, P=0.001) , CD45 positive (HR=14.504, 95%CI 0.168-0.42, P=0.001) , LDH≥220 IU/L (HR=1.308, 95%CI 1.16-2.417, P=0.015) were independent risk prognostic factors. Conclusion: CD45 expression is a risk prognostic factor of MM patients. Bortezomib did not improve the poor prognosis of CD45(+) MM patients.
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Humanos , Protocolos de Quimioterapia Combinada Antineoplásica , Intervalo Livre de Doença , Antígenos Comuns de Leucócito/metabolismo , Mieloma Múltiplo/diagnóstico , Prognóstico , Estudos RetrospectivosRESUMO
Objective@#To observe the change and clinical significance of the peripheral blood T-lymphocytes CD45RA+ and CD45RO+ in children with Henoch-Schonlein purpura (HSP) under different clinical classification.@*Methods@#From October 2015 to July 2017, the clinical data of 80 children with HSP in the Affiliated Hospital of North Sichuan Medical College were retrospectively analyzed.According to the clinical classification, they were divided into three groups: skin involvement group (35 cases), abdominal type group (36 cases), and renal type group (9 cases). Another 80 healthy children were selected as the control group.The changes and clinical significance of peripheral blood T-lymphocytes CD45RA+ , CD45RO+ and CD45RA+ /CD45RO+ ratio were analyzed.@*Results@#Among the 80 children, 40 cases were male and 40 cases were female, with age of (7.2±2.3)years old.The CD45RA+ and CD45RO+ rates in the HSP group were (13.19±7.09)%, (12.07±3.46)%, respectively, which were significantly lower than those in the control group [(23.26±6.01)%, (21.74±3.46)%], the differences were statistically significant (t=9.69, 16.42, all P<0.05). The CD45RA+ ratio, CD45RA+ count and CD45RA+ /CD45RO+ ratio of the kidney type group were (8.02±3.63)%, (2.19±0.33)/μL, (-0.28±0.19), respectively, which were significantly lower than those of the skin group [(15.74±7.71)%, (2.55±0.33)/μL, (0.27±0.12)], the differences were statistically significant(t=3.085, 2.709, 4.013, all P<0.05). The ratio of CD45RA+ /CD45RO+ in the abdominal group was significantly lower than that in the skin group[(-0.07±0.27)vs.(0.27±0.12), t=2.989, P<0.05].@*Conclusion@#Peripheral blood T lymphocyte subsets CD45RA+ and CD45RO+ may play a role in the generation and development of immune function changes in children with HSP, and the decrease of CD45RA+ T cells may be related to the risk of HSP nephritis.
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Objective·To study the expression of transcription factor BTB and CNC homology 2 (Bach2) in CD4+CD25+CD45RA-T ceils from patients with systemic lupus erythematosus (SLE) and its effect on cell function.Methods·The CD4+CD25+CD45RA T cells from active SLE patients and healthy volunteers were sorted by flow cytometry.The expression of Bach2 in CD4+CD25+CD45RA T cells was detected by fluorescence quantitative PCR and Western blotting.The correlation between the median flourscence indensity (MFI) of Bach2 in CD4+CD25+CD45RA-T cells and the disease activity index of SLE (SLEDAI) was analyzed.The MFI of Bach2 in IL-17+CD4+CD25+CD45RA-T ceils was compared with that in IL-17-CD4+CD25+CD45RA-T cells by flow cytometry.In Bach2 overexpression system,the expression of IL-17 in CD4+CD25+CD45RA T ceils was detected by flow cytometry and the concentration of IL-17 in the culture supernants was detected by ELISA.Results·The mRNA and protein expressions of Bach2 in CD4+CD25+CD45RA-T cells from SLE patients were significantly lower than those in healthy controls (P<0.01).There was a significant negative correlation between the MFI of Bach2 and SLEDAI (R2=0.433,P=-0.001) in patients with SLE.The expression of Bach2 in IL-17+CD4+CD25+CD45RA-T cells was significantly lower than that in IL-17-CD4+CD25+CD45RA-T cells (P=-0.013).When Bach2 was overexpressed,the percentage of CD4+CD25+CD45RA-T cells from SLE patients expressing inflammatory factor IL-17 decreased significantly (P=0.032) and the IL-17 concentration in cell culture supernatants markedly decreased (P=0.008).Conclusion·The expression of Bach2 in CD4+CD25+CD45RA-T cells from SLE patients decreases,and overexpression of Bach2 in the cells leads to the falling expression of IL-17.
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BACKGROUND: Postoperative cholangitis is a common but severe complication after Kasai portoenterostomy for biliary atresia (BA). This study aimed to identify its prognostic factors. METHODS: Two sets of liver paraffin-embedded tissue samples were collected from BA patients who received Kasai portoenterostomy (n = 25 and n = 31, respectively). Patients were divided into non-cholangitis and cholangitis groups. The infiltration of CD4+, CD8+, CD45RO+, CD68+ cells and expression of Beclin1 were quantitatively evaluated in immunohistochemical analysis. RESULTS: Cholangitis group had a significantly lower CD8+ T cell infiltration but a higher CD45RO+ cell infiltration, and a lower Beclin1 level than non-cholangitis group (all P < 0.01). Multivariate logistic regression analysis indicated that infiltration of CD8+ cells (odds ratio [OR], 0.112; 95% confidence interval [CI], 0.022–0.577) and CD45RO+ cells (OR, 3.88; 95% CI, 1.37–11.03), and Beclin1 level (OR, 0.088; 95% CI, 0.018–0.452) were independent influence factors for early postoperative cholangitis. Receiver operating characteristic (ROC) analysis showed that area under ROC curve (AUROC) values for CD8+ cells, CD45RO+ cells and Beclin1 were 0.857, 0.738 and 0.900, respectively. CONCLUSION: Our findings demonstrated the CD8+ cells, CD45RO+ cells and Beclin1 level possessed the prognostic value for early postoperative cholangitis following Kasai operation, which may be helpful to develop new prevention and treatment strategies for postoperative cholangitis.
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Humanos , Atresia Biliar , Colangite , Fígado , Modelos Logísticos , Curva ROC , Linfócitos TRESUMO
Objective: To study the expression of transcription factor BTB and CNC homology 2 (Bach2) in CD4+CD25+CD45RA- T cells from patients with systemic lupus erythematosus (SLE) and its effect on cell function. Methods: The CD4+CD25+CD45RA- T cells from active SLE patients and healthy volunteers were sorted by flow cytometry. The expression of Bach2 in CD4+CD25+CD45RA- T cells was detected by fluorescence quantitative PCR and Western blotting. The correlation between the median flourscence indensity (MFI) of Bach2 in CD4+CD25+CD45RA- T cells and the disease activity index of SLE (SLEDAI) was analyzed. The MFI of Bach2 in IL-17+CD4+CD25+CD45RA- T cells was compared with that in IL-17-CD4+CD25+CD45RA- T cells by flow cytometry. In Bach2 overexpression system, the expression of IL-17 in CD4+CD25+CD45RA- T cells was detected by flow cytometry and the concentration of IL-17 in the culture supernants was detected by ELISA. Results: The mRNA and protein expressions of Bach2 in CD4+CD25+CD45RA- T cells from SLE patients were significantly lower than those in healthy controls (P<0.01). There was a significant negative correlation between the MFI of Bach2 and SLEDAI (R2=0.433, P=0.001) in patients with SLE. The expression of Bach2 in IL-17+CD4+CD25+CD45RA- T cells was significantly lower than that in IL-17-CD4+CD25+CD45RA- T cells (P=0.013). When Bach2 was overexpressed, the percentage of CD4+CD25+CD45RA- T cells from SLE patients expressing inflammatory factor IL-17 decreased significantly (P=0.032) and the IL-17 concentration in cell culture supernatants markedly decreased (P=0.008). Conclusion: The expression of Bach2 in CD4+CD25+CD45RA- T cells from SLE patients decreases, and overexpression of Bach2 in the cells leads to the falling expression of IL-17.
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Introducción: la leucemia linfoide aguda (LLA) es la neoplasia más frecuente en la infancia. La determinación del antígeno CD45 discrimina entre los blastos y las células reactivas en la médula ósea (MO). Objetivo: evaluar la expresión del antígeno CD45 sobre los blastos de pacientes con LLA, según los distintos subtipos inmunológicos, su posible relación con las características biológicas y clínicas de presentación de la enfermedad y la respuesta al tratamiento antileucémico. Métodos: se estudiaron 150 pacientes con LLA procedentes de varios servicios oncohematológicos del país, entre enero del 2008 y mayo del 2015. El inmunofenotipaje celular de la MO se realizó por citometría de flujo. Resultados: el antígeno CD45 mostró una gran heterogeneidad de expresión sobre los linfoblastos. Del total de enfermos estudiados, 19,3 por ciento no expresaron sobre los blastos el antígeno CD45, 36,7 por ciento presentaron una expresión moderada y 44 por ciento mostraron una alta densidad de expresión. Se encontró diferencia significativa al comparar el fenotipo leucémico con la expresión del antígeno CD45 sobre los blastos (p = 0,000). Ningún enfermo presentó adenopatías mediastinales, con diferencias significativas (p = 0,000), según el fenotipo y la expresión de CD45. Los pacientes con LLA-T cuyos blastos no expresaron CD45 tuvieron una mala respuesta al tratamiento anti-leucémico los días 8 y 15 en sangre periférica y MO, respectivamente. Conclusión: la expresión de CD45 sobre los blastos, pudiera ser considerada como un factor pronóstico adicional para la estratificación en diferentes grupos de riesgos, de la LLA en el niño(AU)
Introduction: Acute lymphoblastic leukemia (ALL) is the most frequent neoplasia in infancy. Determination of CD45 antigen discriminates between blasts and reactive cells in the bone marrow (MO). Objective: To evaluate the expression of the CD45 antigen on the blasts of patients with ALL, according to the different immunological subtypes, their possible relation with the biological and clinical characteristics of the disease and the response to antileukemic treatment. Methods : 150 patients with ALL were studied from various onco-hematological services of the country, between January 2008 and May 2015. The cellular immunophenotyping of the MO was performed by flow cytometry. Results : The CD45 antigen showed a great heterogeneity of expression on the lymphoblasts. Of the total number of patients studied, 19.3 percent did not express the CD45 antigen on the blasts, 36.7 percent presented moderate expression and 44 percent showed a high expression density of it.A significant difference was found when comparing the leukemic phenotype with the expression of the CD45 antigen on the blasts (p = 0.000). No patient had mediastinal lymphadenopathy, with significant differences (p = 0.000), according to the phenotype and CD45 expression. Patients with T-ALL whose blasts did not express CD45 had a poor response to anti-leukemic treatment on days 8 and 15 in peripheral blood and MO, respectively. Conclusion: CD45 expression on blasts could be considered as an additional prognostic factor for stratification in different risk groups of ALL in children(AU)