58 research outputs found
EuroFlow-based flowcytometric diagnostic screening and classification of primary immunodeficiencies of the lymphoid system
Guidelines for screening for primary immunodeficiencies (PID) are well-defined and several consensus diagnostic strategies have been proposed. These consensus proposals have only partially been implemented due to lack of standardization in laboratory procedures, particularly in flow cytometry. The main objectives of the EuroFlow Consortium were to innovate and thoroughly standardize the flowcytometric techniques and strategies for reliable and reproducible diagnosis and classification of PID of the lymphoid system. The proposed EuroFlow antibody panels comprise one orientation tube and seven classification tubes and corresponding databases of normal and PID samples. The 8-color 12-antibody PID Orientation tube (PIDOT) aims at identification and enumeration of the main lymphocyte and leukocyte subsets; this includes naive pre-germinal center (GC) and antigen-experienced post-GC memory B-cells and plasmablasts. The seven additional 8(-12)-color tubes can be used according to the EuroFlow PID algorithm in parallel or subsequently to the PIDOT for more detailed analysis of B-cell and T-cell subsets to further classify PID of the lymphoid system. The Pre-GC, Post-GC, and immunoglobulin heavy chain (IgH)-isotype B-cell tubes aim at identification and enumeration of B-cell subsets for evaluation of B-cell maturation blocks and specific defects in IgH-subclass production. The severe combined immunodeficiency (SCID) tube and T-cell memory/effector subset tube aim at identification and enumeration of T-cell subsets for assessment of T-cell defects, such as SCID. In case of suspicion of antibody deficiency, PIDOT is preferably directly combined with the IgH isotype tube(s) and in case of SCID suspicion (e.g., in newborn screening programs) the PIDOT is preferably directly combined with the SCID T-cell tube. The proposed >= 8-color antibody panels and corresponding reference databases combined with the EuroFlow PID algorithm are designed to provide fast, sensitive and cost-effective flowcytometric diagnosis of PID of the lymphoid system, easily applicable in multicenter diagnostic settings world-wide
The EuroFlow PID orientation tube for flow cytometric diagnostic screening of primary immunodeficiencies of the lymphoid system
Copyright © 2019 van der Burg, Kalina, Perez-Andres, Vlkova, Lopez-Granados, Blanco, Bonroy, Sousa, Kienzler, Wentink, MejstrĂkovĂĄ, Ć inkorova, Stuchly, van Zelm, Orfao and van Dongen. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.In the rapidly evolving field of primary immunodeficiencies (PID), the EuroFlow consortium decided to develop a PID orientation and screening tube that facilitates fast, standardized, and validated immunophenotypic diagnosis of lymphoid PID, and allows full exchange of data between centers. Our aim was to develop a tool that would be universal for all lymphoid PIDs and offer high sensitivity to identify a lymphoid PID (without a need for specificity to diagnose particular PID) and to guide and prioritize further diagnostic modalities and clinical management. The tube composition has been defined in a stepwise manner through several cycles of design-testing-evaluation-redesign in a multicenter setting. Equally important appeared to be the standardized pre-analytical procedures (sample preparation and instrument setup), analytical procedures (immunostaining and data acquisition), the software analysis (a multidimensional view based on a reference database in Infinicyt software), and data interpretation. This standardized EuroFlow concept has been tested on 250 healthy controls and 99 PID patients with defined genetic defects. In addition, an application of new EuroFlow software tools with multidimensional pattern recognition was designed with inclusion of maturation pathways in multidimensional patterns (APS plots). The major advantage of the EuroFlow approach is that data can be fully exchanged between different laboratories in any country of the world, which is especially of interest for the PID field, with generally low numbers of cases per center.The coordination and innovation processes of this study were supported by the EuroFlow Consortium (Chairmen: MvdB and AO). MvZ is supported by Senior Research Fellowship GNT1117687 from the Australian National Health and Medical Research Council. TK and EM were supported by projects 15-28541A from Ministry of Health, LO1604 from Ministry of Education, Youth and Sports and GBP302/12/G101 from Grant Agency of the Czech Republic. MP-A, EB, and AO were supported by a grant from the Junta de Castilla y LeĂłn (Fondo Social Europeo, ORDEN EDU/346/2013, Valladolid, Spain) and the CB16/12/00400 grant (CIBER/ONC, Instituto de Salud Carlos III, Ministerio de EconomĂa y Competitividad, - Madrid, Spain- and FONDOS FEDER), the FIS PI12/00905-FEDER grant (Fondo de InvestigaciĂłn Sanitaria of Instituto de Salud Carlos III, Madrid, Spain) and AP119882013 grant (FundaciĂłn Mutua Madrileña, Madrid, Spain). Publishing costs for this article were covered by the International Union of Immunological Societies (IUIS).info:eu-repo/semantics/publishedVersio
Dissection of the pre-germinal center B-cell maturation pathway in common variable immunodeficiency based on standardized flow cytometric EuroFlow tools
Copyright © 2021 del Pino-Molina, López-Granados, Lecrevisse, Torres Canizales, Pérez-Andrés, Blanco, Wentink, Bonroy, Nechvatalova, Milota, Kienzler, Philippé, Sousa, van der Burg, Kalina, van Dongen and Orfao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Introduction: Common Variable Immunodeficiency (CVID) is characterized by defective antibody production and hypogammaglobulinemia. Flow cytometry immunophenotyping of blood lymphocytes has become of great relevance for the diagnosis and classification of CVID, due to an impaired differentiation of mature post-germinal-center (GC) class-switched memory B-cells (MBC) and severely decreased plasmablast/plasma cell (Pb) counts. Here, we investigated in detail the pre-GC B-cell maturation compartment in blood of CVID patients.
Methods: In this collaborative multicentric study the EuroFlow PID 8-color Pre-GC B-cell tube, standardized sample preparation procedures (SOPs) and innovative data analysis tools, were used to characterize the maturation profile of pre-GC B-cells in 100 CVID patients, vs 62 age-matched healthy donors (HD).
Results: The Pre-GC B-cell tube allowed identification within pre-GC B-cells of three subsets of maturation associated immature B-cells and three subpopulations of mature naïve B-lymphocytes. CVID patients showed overall reduced median absolute counts (vs HD) of the two more advanced stages of maturation of both CD5+ CD38+/++ CD21het CD24++ (2.7 vs 5.6 cells/”l, p=0.0004) and CD5+ CD38het CD21+ CD24+ (6.5 vs 17 cells/”l, p1 (CD38, CD5, CD19, CD21, CD24, and/or smIgM) phenotypic marker (57/88 patients; 65%) for a total of 3 distinct CVID patient profiles (group 1: 42/88 patients, 48%; group 2: 8/88, 9%; and group 3: 7/88, 8%) and ii) CVID patients with a clearly altered pre-GC B cell maturation pathway in blood (group 4: 31/88 cases, 35%).
Conclusion: Our results show that maturation of pre-GC B-cells in blood of CVID is systematically altered with up to four distinctly altered maturation profiles. Further studies, are necessary to better understand the impact of such alterations on the post-GC defects and the clinical heterogeneity of CVID.The coordination and innovation processes of this study were supported by the EuroFlow Consortium (Chairmen: MB and AO). LP-M was supported by FIS PI16/01605 and JTC by FIS PI13/02296 (Fondo de Investigación Sanitaria Instituto de Salud Carlos III, Madrid, Spain). The work was partially supported by grant PI20/01712-FEDER (Fondo de Investigación Sanitaria Instituto de Salud Carlos III, Madrid, Spain) and a grant from Fundación Mutua Madrileña (MMA, Madrid, Spain).info:eu-repo/semantics/publishedVersio
Age-associated distribution of normal B-cell and plasma cell subsets in peripheral blood
Background: Humoral immunocompetence develops stepwise throughout life and contributes to individual susceptibility to infection, immunodeficiency, autoimmunity, and neoplasia. Immunoglobulin heavy chain (IgH) isotype serum levels can partly explain such age-related differences, but their relationship with the IgH isotype distribution within memory B-cell (MBC) and plasma cell (PCs) compartments remains to be investigated. Objective: We studied the age-related distribution of MBCs and PCs expressing different IgH isotypes in addition to the immature/transitional and naive B-cell compartments. Methods: B-cell and PC subsets and plasma IgH isotype levels were studied in cord blood (n = 19) and peripheral blood (n = 215) from healthy donors aged 0 to 90 years by using flow cytometry and nephelometry, respectively. Results: IgH-switched MBCs expressing IgG1, IgG2, IgG3, IgA1, and IgA2 were already detected in cord blood and newborns at very low counts, whereas CD27+IgM++IgD+ MBCs only became detectable at 1 to 5 months and remained stable until 2 to 4 years, and IgD MBCs peaked at 2 to 4 years, with both populations decreasing thereafter. MBCs expressing IgH isotypes of the second immunoglobulin heavy chain constant region (IGHC) gene block (IgG1, IgG3, and IgA1) peaked later during childhood (2-4 years), whereas MBCs expressing third IGHC gene block immunoglobulin isotypes (IgG2, IgG4, and IgA2) reached maximum values during adulthood. PCs were already detected in newborns, increasing in number until 6 to 11 months for IgM, IgG1, IgG2, IgG3, IgA1, and IgA2; until 2 to 4 years for IgD; and until 5 to 9 years for IgG4 and decreasing thereafter. For most IgH isotypes (except IgD and IgG4), maximum plasma levels were reached after PC and MBC counts peaked. Conclusions: PC counts reach maximum values early in life, followed by MBC counts and plasma IgH isotypes. Importantly, IgH isotypes from different IGHC gene blocks show different patterns, probably reflecting consecutive cycles of IgH isotype switch recombination through life
EuroFlow-based flowcytometric diagnostic screening and classification of primary immunodeficiencies of the lymphoid system
Guidelines for screening for primary immunodeficiencies (PID) are well-defined and several consensus diagnostic strategies have been proposed. These consensus proposals have only partially been implemented due to lack of standardization in laboratory procedures, particularly in flow cytometry. The main objectives of the EuroFlow Consortium were to innovate and thoroughly standardize the flowcytometric techniques and strategies for reliable and reproducible diagnosis and classification of PID of the lymphoid system. The proposed EuroFlow antibody panels comprise one orientation tube and seven classification tubes and corresponding databases of normal and PID samples. The 8-color 12-antibody PID Orientation tube (PIDOT) aims at identification and enumeration of the main lymphocyte and leukocyte subsets; this includes naĂŻve pre-germinal center (GC) and antigen-experienced post-GC memory B-cells and plasmablasts. The seven additional 8(-12)-color tubes can be used according to the Eu
The EuroFlow PID Orientation Tube for Flow Cytometric Diagnostic Screening of Primary Immunodeficiencies of the Lymphoid System
In the rapidly evolving field of primary immunodeficiencies (PID), the EuroFlow
consortium decided to develop a PID orientation and screening tube that facilitates
fast, standardized, and validated immunophenotypic diagnosis of lymphoid PID, and
allows full exchange of data between centers. Our aim was to develop a tool that would
be universal for all lymphoid PIDs and offer high sensitivity to identify a lymphoid PID
(without a need for specificity to diagnose particular PID) and to guide and prioritize
further diagnostic modalities and clinical management. The tube composition has been
defined in a stepwise manner through several cycles of design-testing-evaluationredesign in a multicenter setting. Equally important appeared to be the standardized preanalytical procedures (sample preparation and instrument setup), analytical procedures
(immunostaining and data acquisition), the software analysis (a multidimensional view
based on a reference database in Infinicyt software), and data interpretation. This
standardized EuroFlow concept has been tested on 250 healthy controls and 99 PID
patients with defined genetic defects. In addition, an application of new EuroFlow
software tools with multidimensional pattern recognition was designed with inclusion of
maturation pathways in multidimensional patterns (APS plots). The major advantage of
the EuroFlow approach is that data can be fully exchanged between different laboratories
in any country of the world, which is especially of interest for the PID field, with generally
low numbers of cases per center
Defects in memory B-cell and plasma cell subsets expressing different immunoglobulin-subclasses in patients with CVID and immunoglobulin subclass deficiencies
Background: Predominantly antibody deficiencies (PADs) are
the most prevalent primary immunodeficiencies, but their B-cell
defects and underlying genetic alterations remain largely
unknown.
Objective: We investigated patients with PADs for the
distribution of 41 blood B-cell and plasma cell (PC) subsets,
including subsets defined by expression of distinct
immunoglobulin heavy chain subclasses.
Methods: Blood samples from 139 patients with PADs, 61
patients with common variable immunodeficiency (CVID), 68
patients with selective IgA deficiency (IgAdef), 10 patients with
IgG subclass deficiency with IgA deficiency, and 223 agematched control subjects were studied by using flow cytometry
with EuroFlow immunoglobulin isotype staining. Patients were
classified according to their B-cell and PC immune profile, and
the obtained patient clusters were correlated with clinical
manifestations of PADs.
Results: Decreased counts of blood PCs, memory B cells
(MBCs), or both expressing distinct IgA and IgG subclasses
were identified in all patients with PADs. In patients with
IgAdef, B-cell defects were mainly restricted to surface
membrane (sm)IgA1 PCs and MBCs, with 2 clear subgroups
showing strongly decreased numbers of smIgA1 PCs with
mild versus severe smIgA1 MBC defects and higher
frequencies of nonrespiratory tract infections, autoimmunity,
and affected family members. Patients with IgG subclass
deficiency with IgA deficiency and those with CVID showed
defects in both smIgA1 and smIgG1 MBCs and PCs.
Reduced numbers of switched PCs were systematically found
in patients with CVID (absent in 98%), with 6 different
defective MBC (and clinical) profiles: (1) profound decrease
in MBC numbers; (2) defective CD271 MBCs with almost
normal IgG3
1 MBCs; (3) absence of switched MBCs; and (4)
presence of both unswitched and switched MBCs without
and; (5) with IgG2
1 MBCs; and (6) with IgA
Search for eccentric black hole coalescences during the third observing run of LIGO and Virgo
Despite the growing number of binary black hole coalescences confidently observed through gravitational waves so far, the astrophysical origin of these binaries remains uncertain. Orbital eccentricity is one of the clearest tracers of binary formation channels. Identifying binary eccentricity, however, remains challenging due to the limited availability of gravitational waveforms that include the effects of eccentricity. Here, we present observational results for a waveform-independent search sensitive to eccentric black hole coalescences, covering the third observing run (O3) of the LIGO and Virgo detectors. We identified no new high-significance candidates beyond those that have already been identified with searches focusing on quasi-circular binaries. We determine the sensitivity of our search to high-mass (total source-frame mass M > 70 Mâ) binaries covering eccentricities up to 0.3 at 15 Hz emitted gravitational-wave frequency, and use this to compare model predictions to search results. Assuming all detections are indeed quasi-circular, for our fiducial population model, we place a conservative upper limit for the merger rate density of high-mass binaries with eccentricities 0 < e †0.3 at 16.9 Gpcâ3 yrâ1 at the 90% confidence level
Moving from understanding to action: Breaking barriers for transformation
No abstract available
The importance of training in the initial assessment of clinical interviews: relevant aspects
La ValoraciĂłn, primera fase del Proceso Enfermero1, consiste en la recogida, interpretaciĂłn y
organizaciĂłn de los datos sobre el paciente, la familia y el entorno. Proporciona datos que
constituyen la base para las decisiones y actuaciones enfermeras posteriores.2
La bibliografĂa consultada sobre la prĂĄctica enfermera2-3-4 recomienda como mĂ©todos para la
recogida de datos la observaciĂłn, la entrevista clĂnica y el examen fĂsico, destacando la entrevista
como el método mås importante ya que si tenemos en cuenta las fuentes que utilizamos, paciente,
historia clĂnica y familia, es el paciente la fuente primaria o directa, pasando a constituir fuentes
secundarias o indirectas la familia y la historia clĂnica.
La realizaciĂłn de la entrevista clĂnica es el primer paso para empezar a cuidar con calidad 2. No
obstante, la calidad y cantidad de datos que obtengamos a travĂ©s de la entrevista clĂnica va a
depender de la habilidad que tengamos las enfermeras para establecer una relaciĂłn de confianza,
para observar, escuchar y preguntar.ABSTRACT
Assessment, the first stage in the nursing process, consists of the collection, interpretation and
organization of particulars about a patient, his or her family and their environment. This will provide us
with basic data for future decisions and interventions.
The bibliography on nursing practice consulted recommends some methods for this data collection,
such as observation, physical evaluation and clinical interview, the latter being the most important. If
we consider our sources of information, i.e. patient, medical record and family, it is the patient the one
who constitutes the primary or direct source, medical record and family being secondary or indirect. The clinical interview is the first step in quality nursing care. Nevertheless, both the quality and the
amount of those data obtained will depend on the ability on the part of the nurse to establish a nursepatient relation based on confidence, on our ability as regards observing, listening to and asking
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