Association between CD4 lymphocyte count and the incidence of comorbidities in Human immunodeficiency virus positive patients with virological suppression after antiretroviral treatment.

BACKGROUND: The incidence of comorbidities is higher in HIV-positive patients than in the general population due to factors, such as HIV-related chronic inflammation. There is no consensus on whether a low CD4 lymphocyte count after virological suppression at long-term follow-up increases the risk of comorbidities. This study evaluates the association between CD4 lymphocyte count and the incidence of comorbidities during the first 5 years of virological suppression after highly active antiretroviral treatment. METHODS: We conducted a cohort study of HIV-positive adults who achieved virological suppression in an HIV program between 2002 and 2016 in Colombia. A generalized equation estimation model was used to estimate the association between CD4 lymphocyte count and the incidence of comorbidities. RESULTS: A follow-up period of at least 1 year was completed in 921 HIV-positive patients with virological suppression. We found 71 comorbidities during a maximum of 5 years of follow-up; 41 (59%) were AIDS-defining comorbidities and 19 (46%) of them occurred during the first semester. Thirty cases of non-AIDS- defining comorbidities were diagnosed.We did not find any association between CD4 lymphocyte count and the incidence of comorbidities (OR 0.92, CI 95% 0.45 -1.91 for CD4 201-499 cells/µL vs CD4 ≤200 cells/µL, and OR 0.55, 95% CI 0.21-1.44 for CD4 ≥500 cells/µL vs CD4 ≤200 cells/µL). CONCLUSION: No association was found between CD4 lymphocyte count and the incidence of AIDS-defining or non-AIDS-defining comorbidities in patients with virological suppression. Further studies are needed to assess the risk of comorbidities in this population to design interventions aimed at improving their prognosis.

An Extensive Quality Control and Quality Assurance (QC/QA) Program Significantly Improves Inter-Laboratory Concordance Rates of Flow-Cytometric Minimal Residual Disease Assessment in Acute Lymphoblastic Leukemia: An I-BFM-FLOW-Network Report.

Monitoring of minimal residual disease (MRD) by flow cytometry (FCM) is a powerful prognostic tool for predicting outcomes in acute lymphoblastic leukemia (ALL). To apply FCM-MRD in large, collaborative trials, dedicated laboratory staff must be educated to concordantly high levels of expertise and their performance quality should be continuously monitored. We sought to install a unique and comprehensive training and quality control (QC) program involving a large number of reference laboratories within the international Berlin-Frankfurt-Münster (I-BFM) consortium, in order to complement the standardization of the methodology with an educational component and persistent quality control measures. Our QC and quality assurance (QA) program is based on four major cornerstones: (i) a twinning maturation program, (ii) obligatory participation in external QA programs (spiked sample send around, United Kingdom National External Quality Assessment Service (UK NEQAS)), (iii) regular participation in list-mode-data (LMD) file ring trials (FCM data file send arounds), and (iv) surveys of independent data derived from trial results. We demonstrate that the training of laboratories using experienced twinning partners, along with continuous educational feedback significantly improves the performance of laboratories in detecting and quantifying MRD in pediatric ALL patients. Overall, our extensive education and quality control program improved inter-laboratory concordance rates of FCM-MRD assessments and ultimately led to a very high conformity of risk estimates in independent patient cohorts.

Comparison of the QuantiFERON-TB and tuberculin skin test for detection of latent tuberculosis infection in cancer patients in a developing country.

Cancer patients have an increased risk of reactivation of latent tuberculosis infection. It is unknown which strategy on screening should be used in this population in developing countries. We aimed to determine the concordance between the tuberculin skin test (TST) and QuantiFERON®-TB (QFT) assay in order to diagnose latent tuberculosis infection in cancer patients. We conducted a cross-sectional study of the agreement of diagnostic tests. Prevalence and agreement between tests were calculated. A logistic regression to assess predictors of discordance was performed. The accuracy of the TST to predict QFT results by a receiver operating characteristic (ROC) curve was evaluated. We included 149 adults with cancer without active tuberculosis. Prevalence of latent tuberculosis infection was 21.5% (n=32), defined as positive results on either test. Test agreement was moderate for the diagnosis of latent tuberculosis infection (κ=0.43, 90% CI 0.26-0.6). No predictor was associated with the chance of discordant results. Agreement improved slightly using a cut-off point ≥8 mm (κ=0.5, 90% CI 0.35-0.66). In a moderate-incidence setting, a moderate agreement was found between tests in cancer patients. Modification of the cut-off points of test results achieved marginally better agreement between the TST and QFT.

Secondary erythrocytosis due to hypoxemia as prognosis in exacerbated chronic pulmonary diseases / Eritrocitosis secundaria a hipoxemia como pronóstico en neumopatías crónicas exacerbadas

Abstract Introduction: Even though exacerbations are the main cause of emergency consultation in patients suffering from lung diseases, erythrocyte parameters are not assessed in their prognosis. Thus, determining the implications of erythrocyte parameters might contribute to define the usefulness of phlebotomy or red blood cells transfusion in these patients. Objective: To establish a possible relationship between the different hematocrit levels with a 30-day prognosis in patients admitted with exacerbated chronic lung disease and hypoxemia. Materials and methods: A study based on a 30- day follow-up was conducted. Variables were described using an additional categorization by hematocrit levels and an adjustment in a multivariate model through logistic regression. Results: Follow-up was completed for 110 Patients. The frequency of anemia was 7.3% and of erythrocytosis, 14.5%. A significant association to the outcome using Anthonisen score (OR=10.45, 95%CI: 1.11-98.48, p=0.04), hypertension (OR=11.02, 95%CI: 1.32-91.75, p=0.026) and heart failure (OR=0.09, 95%CI: 0.01-0.82, p=0.032) was found. Conclusion: This research could not determine any relationship between erythrocyte parameters and prognosis of patients suffering from pulmonary diseases; nevertheless, extreme values of hematocrits tended to have adverse outcomes.

Resumen Introducción. Aunque las exacerbaciones de las neumopatías crónicas son las principales causas de consulta a urgencias de los pacientes que las padecen, los parámetros eritrocitarios no son evaluados en su pronóstico. Por tanto, determinar las implicaciones de los parámetros eritrocitarios podría ayudar a definir la utilidad de la flebotomía o la transfusión de eritrocitos en estos pacientes. Objetivo. Establecer si hay relación entre los distintos niveles de hematocrito con pronóstico a 30 días en pacientes con neumopatía crónica exacerbada e hipoxemia. Materiales y métodos. Estudio de seguimiento a 30 días. Se realizó la descripción de las variables con una categorización adicional por niveles de hematocrito y un ajuste en un modelo multivariado por regresión logística. Resultados. Se completó el seguimiento en 110 pacientes. La frecuencia de anemia fue de 7.3% y de eritrocitosis de 14.5%. Se encontró asociación significativa al desenlace con la clasificación Anthonisen (OR=10.45, IC95%: 1.11-98.48; p=0.04), hipertensión arterial (OR=11.02, IC95%: 1.32-91.75; p=0.026) y falla cardiaca (OR=0.09, IC95%: 0.01-0.82; p=0.032). Conclusión. Este estudio no pudo determinar relación alguna entre los parámetros eritrocitarios y el pronóstico de pacientes con enfermedades pulmonares crónicas; sin embargo, hubo una tendencia a que los valores extremos del hematocrito presentaran desenlaces adversos.

Niveles séricos del factor de crecimiento semejante a la insulina tipo I como posible marcador de la evolución de pacientes con leucemia linfática crónica / Growth factor similar insulin like 1 serum levels as prognostic factor of patients with chronic lymphotic leukemia evolution

La leucemia linfática crónica (LLC) es una enfermedad caracterizada por la acumulación de linfocitos B,usualmente CD5+, que tienen una larga vida y se encuentran detenidos en la fase GO/1 temprana del ciclo celular debido aun defecto en su apoptosis. Como el factor de crecimiento semejante a la insulina tipo I (IGF-I) es un conocido factor antiapoptótico en diferentes tipos celulares, hemos investigado su posible participación autocrina/paracrina en las células leucémicas. Observamos que los niveles de IGF-I séricos en pacientes con LLC fueron elevados mientras que la hormona de crecimiento (HC) se mantuvo normal. Las células LLC expresaron el ARNm del IGF-I y fueron capaces de secretar el factor de crecimiento in vitro. Por lo tanto, la producción local del IGF-I puede ser responsable del aumento de los niveles séricos de IGF-I, de forma independiente de la HC y podría estar relacionada al control autocrino/paracrino de la supervivencia de los linfocitos. Más aun, los pacientes estables que tuvieron IGF-I sérico elevado, mostraron una sobrevida más corta en un seguimiento de 4 años. Nuestros hallazgos indican que los niveles séricos de IGF-I en pacientes estables podrían ser utilizados como un marcador pronóstico en LLC.

Niveles séricos del factor de crecimiento semejante a la insulina tipo I como posible marcador de la evolución de pacientes con leucemia linfática crónica / Growth factor similar insulin like 1 serum levels as prognostic factor of patients with chronic lymphotic leukemia evolution

La leucemia linfática crónica (LLC) es una enfermedad caracterizada por la acumulación de linfocitos B,usualmente CD5+, que tienen una larga vida y se encuentran detenidos en la fase GO/1 temprana del ciclo celular debido aun defecto en su apoptosis. Como el factor de crecimiento semejante a la insulina tipo I (IGF-I) es un conocido factor antiapoptótico en diferentes tipos celulares, hemos investigado su posible participación autocrina/paracrina en las células leucémicas. Observamos que los niveles de IGF-I séricos en pacientes con LLC fueron elevados mientras que la hormona de crecimiento (HC) se mantuvo normal. Las células LLC expresaron el ARNm del IGF-I y fueron capaces de secretar el factor de crecimiento in vitro. Por lo tanto, la producción local del IGF-I puede ser responsable del aumento de los niveles séricos de IGF-I, de forma independiente de la HC y podría estar relacionada al control autocrino/paracrino de la supervivencia de los linfocitos. Más aun, los pacientes estables que tuvieron IGF-I sérico elevado, mostraron una sobrevida más corta en un seguimiento de 4 años. Nuestros hallazgos indican que los niveles séricos de IGF-I en pacientes estables podrían ser utilizados como un marcador pronóstico en LLC.(AU)

Autocrine/paracrine involvement of insulin-like growth factor-I and its receptor in chronic lymphocytic leukaemia.

Chronic lymphocytic leukaemia (CLL) is characterized by the accumulation of long-lived B lymphocytes blocked in G(0/1) by impaired apoptosis. As insulin-like growth factor-I (IGF-I) is known for its antiapoptotic effects in different cell types, we investigated whether IGF-I and its receptor (IGF-IR) participate in autocrine/paracrine loops affecting the survival of CLL cells. IGF-IR protein and mRNA was present in CLL cells in 44% and 59% of patients respectively. IGF-IR expression in CLL patients was positively correlated with the expression of the antiapoptotic protein Bcl-2 and was involved in CLL cell survival in vitro. Serum IGF-I was elevated in CLL patients, but growth hormone (GH) was normal. CLL cells expressed IGF-I mRNA and secreted the growth factor in vitro. Therefore, local production of IGF-I can account for the increased levels of serum IGF-I, independently of GH, and may be related to autocrine/paracrine control of lymphocyte survival acting at IGF-IR. This is the first demonstration of IGF-IR expression in a subgroup of CLL patients and of its antiapoptotic effects in vitro, highlighting the importance of this growth factor receptor as a possible therapeutic target in CLL.

Insulin-like growth factor-1 receptor regulation in activated human T lymphocytes.

OBJECTIVE: To investigate the kinetics of insulin-like growth factor-1 receptor (IGF-1R) expression in PHA-stimulated T lymphocytes. METHODS: IGF-1R protein and mRNA were detected by flow cytometry and RT-PCR respectively, between 0 and 48 h after cell activation. RESULTS: Few minutes after T lymphocytes were activated, internalization of the IGF-1R from the cell membrane was observed, achieving the lower level between 1 and 6 h and was accompanied by a reduction in its mRNA. This was followed by re-expression of IGF-1R on the cell surface and an increase in IGF-1R mRNA levels in the cytoplasm, reaching levels higher than those recorded initially after 48 h activation. CONCLUSION: This down- and up-regulation suggests that restoration of IGF-1R would be the result of receptor recycling and de novo synthesis and highlights its importance for T lymphocyte proliferation.

Primer estudio multicéntrico realizado por el Grupo Rioplatense de Citometría de Flujo: control de calidad en las mediciones de subpoblaciones de linfocitos T / First multicentric study by Grupo Rioplatense de Citometría de Flujo : quality control of lymphocyte subsets

El control de calidad se efectuó sobre los valores obtenidos, relativos y absolutos, de linfocitos T y de sus subpoblaciones CD4+ y CD8+ en muestras de sangre de pacientes infectados con el virus de la inmunodeficiencia humana (HIV). El estudio incluyó dieciocho centros: diez utilizaron citómetros de flujo de Becton Dickinson, tres de Coulter y 5 de Ortho que representan a 17 laboratorios de Argentina y a uno de Uruguay. Los siguientes programas se utilizaron para analizar los datos : SimulSET, Paint a Gate (Becton Dickinson), Profile II, XL System (Coulter), ImmunoCount Trio y Combo Cytoron (Ortho). Se obtuvieron muestras de sangre periférica en horas de la mañana (8 a 10 hs) de 10 voluntarios normales (por serología y hemograma) y de 10 pacientes HIV positivos con valores previos de CD4 que variaron entre 200-350 células por microlito y fueron procesadas dentro de las 12 horas. Cada centro obtuvo los valores relativos con el procedimiento técnico habitual y el de los valores absolutos utilizando el hemograma propio. Además, en un contador hematológico Cell-Dyn 3500 se obtuvo para cada muestra el hemograma correspondiente considerado de referencia. Los valores absolutos medios, obtenidos en cada centro con el hemograma propio, para los linfocitos T y los de sus subpoblaciones fueron significativamente diferentes. No hubo diferencias significativas para los valores porcentuales entre los diferentes centros ni para los valores absolutos obtenidos con el hemograma de referencia. Concluimos que las diferencias en los valores absolutos de los linfocitos T y sus subpoblaciones dependen del recuento hematológico empleado.

Primer estudio multicéntrico realizado por el Grupo Rioplatense de Citometría de Flujo: control de calidad en las mediciones de subpoblaciones de linfocitos T / First multicentric study by Grupo Rioplatense de Citometría de Flujo : quality control of lymphocyte subsets

El control de calidad se efectuó sobre los valores obtenidos, relativos y absolutos, de linfocitos T y de sus subpoblaciones CD4+ y CD8+ en muestras de sangre de pacientes infectados con el virus de la inmunodeficiencia humana (HIV). El estudio incluyó dieciocho centros: diez utilizaron citómetros de flujo de Becton Dickinson, tres de Coulter y 5 de Ortho que representan a 17 laboratorios de Argentina y a uno de Uruguay. Los siguientes programas se utilizaron para analizar los datos : SimulSET, Paint a Gate (Becton Dickinson), Profile II, XL System (Coulter), ImmunoCount Trio y Combo Cytoron (Ortho). Se obtuvieron muestras de sangre periférica en horas de la mañana (8 a 10 hs) de 10 voluntarios normales (por serología y hemograma) y de 10 pacientes HIV positivos con valores previos de CD4 que variaron entre 200-350 células por microlito y fueron procesadas dentro de las 12 horas. Cada centro obtuvo los valores relativos con el procedimiento técnico habitual y el de los valores absolutos utilizando el hemograma propio. Además, en un contador hematológico Cell-Dyn 3500 se obtuvo para cada muestra el hemograma correspondiente considerado de referencia. Los valores absolutos medios, obtenidos en cada centro con el hemograma propio, para los linfocitos T y los de sus subpoblaciones fueron significativamente diferentes. No hubo diferencias significativas para los valores porcentuales entre los diferentes centros ni para los valores absolutos obtenidos con el hemograma de referencia. Concluimos que las diferencias en los valores absolutos de los linfocitos T y sus subpoblaciones dependen del recuento hematológico empleado. (AU)

Fine needle aspiration for flow cytometry immunophenotyping of non Hodgkin lymphoma

The aim was to evaluate the usefulness of lymph node biopsies obtained by fine needle aspiration (FNA) for immunophenotyping of non Hodgkin lymphoma (NHL). Seventeen superficial and deep lymph node samples were fractioned for conventional cytological examination and immunophenotyping studies. Out of ten NHL, nine were readily detected by flow cytometry (FC), while failure on the remaining case was due to selective loss of large cell population, which is liable to occur with this procedure. A single case, which proved negative for all markers employed, was finally diagnosed by immunohistochemistry as germ cell tumor. The other six cases, presenting lymphoid population without phenotypic abnormalities, were diagnosed by cytology and/or histology as Hodgkin disease or hyperpiasic disorders. To conclude, FC immunophenotyping seems to improve the efficacy of FNA in NHL diagnosis, whereas for Hodgkin disease and hyperplasic disorders, classic morphological criteria are more useful for differential diagnosis. Although FNA for FC immunophenotyping cannot replace histopathological examination for NHL diagnosis, it proves to be a useful tool for staging and follow up, making surgical procedures for sample collection unnecesary.

Fine needle aspiration for flow cytometry immunophenotyping of non Hodgkin lymphoma

The aim was to evaluate the usefulness of lymph node biopsies obtained by fine needle aspiration (FNA) for immunophenotyping of non Hodgkin lymphoma (NHL). Seventeen superficial and deep lymph node samples were fractioned for conventional cytological examination and immunophenotyping studies. Out of ten NHL, nine were readily detected by flow cytometry (FC), while failure on the remaining case was due to selective loss of large cell population, which is liable to occur with this procedure. A single case, which proved negative for all markers employed, was finally diagnosed by immunohistochemistry as germ cell tumor. The other six cases, presenting lymphoid population without phenotypic abnormalities, were diagnosed by cytology and/or histology as Hodgkin disease or hyperpiasic disorders. To conclude, FC immunophenotyping seems to improve the efficacy of FNA in NHL diagnosis, whereas for Hodgkin disease and hyperplasic disorders, classic morphological criteria are more useful for differential diagnosis. Although FNA for FC immunophenotyping cannot replace histopathological examination for NHL diagnosis, it proves to be a useful tool for staging and follow up, making surgical procedures for sample collection unnecesary.(AU)