OBJECTIVES: The development of External Quality Assessment Schemes (EQAS) for clinical flow cytometry (FCM) is challenging in the context of rare (immunological) diseases. Here, we introduce a novel EQAS monitoring the primary immunodeficiency Orientation Tube (PIDOT), developed by EuroFlow, in both a 'wet' and 'dry' format. This EQAS provides feedback on the quality of individual laboratories (i.e., accuracy, reproducibility and result interpretation), while eliminating the need for sample distribution. METHODS: In the wet format, marker staining intensities (MedFIs) within landmark cell populations in PIDOT analysis performed on locally collected healthy control (HC) samples, were compared to EQAS targets. In the dry format, participants analyzed centrally distributed PIDOT flow cytometry data (n=10). RESULTS: We report the results of six EQAS rounds across 20 laboratories in 11 countries. The wet format (212 HC samples) demonstrated consistent technical performance among laboratories (median %rCV on MedFIs=34.5 %; average failure rate 17.3 %) and showed improvement upon repeated participation. The dry format demonstrated effective proficiency of participants in cell count enumeration (range %rCVs 3.1-7.1 % for the major lymphoid subsets), and in identifying lymphoid abnormalities (79.3 % alignment with reference). CONCLUSIONS: The PIDOT-EQAS allows laboratories, adhering to the standardized EuroFlow approach, to monitor interlaboratory variations without the need for sample distribution, and provides them educational support to recognize rare clinically relevant immunophenotypic patterns of primary immunodeficiencies (PID). This EQAS contributes to quality improvement of PID diagnostics and can serve as an example for future flow cytometry EQAS in the context of rare diseases.
Congenital athymia is a life-limiting disorder due to rare inborn errors of immunity causing impaired thymus organogenesis or abnormal thymic stromal cell development and function. Athymic infants have a T-lymphocyte-negative, B-lymphocyte-positive, natural killer cell-positive immunophenotype with profound T-lymphocyte deficiency and are susceptible to severe infections and autoimmunity. Patients variably display syndromic features. Expanding access to newborn screening for severe combined immunodeficiency and T lymphocytopenia and broad genetic testing, including next-generation sequencing technologies, increasingly facilitate their timely identification. The recommended first-line treatment is allogeneic thymus transplantation, which is a specialized procedure available in Europe and the United States. Outcomes for athymic patients are best with early diagnosis and thymus transplantation before the development of infectious and inflammatory complications. These guidelines on behalf of the European Society for Immunodeficiencies provide a comprehensive review for clinicians who manage patients with inborn thymic stromal cell defects; they offer clinical practice recommendations focused on the diagnosis, investigation, risk stratification, and management of congenital athymia with the aim of improving patient outcomes.
- MeSH
- lidé MeSH
- management nemoci MeSH
- novorozenec MeSH
- syndromy imunologické nedostatečnosti * terapie diagnóza imunologie MeSH
- thymus imunologie MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- směrnice pro lékařskou praxi MeSH
- Geografické názvy
- Evropa MeSH
Schimke immuno-osseous dysplasia is a rare multisystemic disorder caused by biallelic loss of function of the SMARCAL1 gene that plays a pivotal role in replication fork stabilization and thus DNA repair. Individuals affected from this disease suffer from disproportionate growth failure, steroid resistant nephrotic syndrome leading to renal failure and primary immunodeficiency mediated by T cell lymphopenia. With infectious complications being the leading cause of death in this disease, researching the nature of the immunodeficiency is crucial, particularly as the state is exacerbated by loss of antibodies due to nephrotic syndrome or immunosuppressive treatment. Building on previous findings that identified the loss of IL-7 receptor expression as a possible cause of the immunodeficiency and increased sensitivity to radiation-induced damage, we have employed spectral cytometry and multiplex RNA-sequencing to assess the phenotype and function of T cells ex-vivo and to study changes induced by in-vitro UV irradiation and reaction of cells to the presence of IL-7. Our findings highlight the mature phenotype of T cells with proinflammatory Th1 skew and signs of exhaustion and lack of response to IL-7. UV light irradiation caused a severe increase in the apoptosis of T cells, however the expression of the genes related to immune response and regulation remained surprisingly similar to healthy cells. Due to the disease's rarity, more studies will be necessary for complete understanding of this unique immunodeficiency.
- MeSH
- apoptóza genetika MeSH
- arterioskleróza genetika etiologie imunologie MeSH
- dítě MeSH
- DNA-helikasy genetika MeSH
- lidé MeSH
- metabolické nemoci kostí etiologie genetika MeSH
- nefrotický syndrom etiologie genetika MeSH
- oprava DNA * genetika MeSH
- osteochondrodysplazie * genetika imunologie MeSH
- plicní embolie genetika etiologie MeSH
- poruchy růstu genetika etiologie MeSH
- primární imunodeficience * genetika diagnóza imunologie MeSH
- syndromy imunologické nedostatečnosti genetika imunologie MeSH
- T-lymfocyty imunologie MeSH
- ultrafialové záření škodlivé účinky MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Termín s cirhózou asociovaná imunitná dysfunkcia predstavuje široké spektrum zmien v lokálnej a systémovej imunitnej odpovedi. CAID je charakterizovaná súčasnou prítomnosťou systémového zápalu a imunitného deficitu, ktorých intenzita prejavu závisí od stupňa pokročilosti ochorenia, predisponujúcich a precipitujúcich faktorov. Kľúčovú úlohu v patogenéze CAID zohráva spojenie črevo–pečeň. Porucha ich vzájomnej súhry, ako aj zmeny v zložení mikrobiómu, vedú k poškodeniu črevnej bariéry a následnej permanentnej antigénnej stimulácii, ktorá vedie k novému prozápalovému nastaveniu imunitného systému. Táto zmena charakterizuje nízkozápalový fenotyp CAID. Opakované zosilnenia mikrobiálnej translokácie a opakované pôsobenie precipitujúceho faktora vedú k vzplanutiu systémového zápalu a prelomeniu imunitnej tolerancie. Výsledkom je dysregulovaná hyperinflamačná imunitná odpoveď, ktorá reprezentuje vysokozápalový fenotyp CAID. Zmena fenotypu je podkladom pre progresiu cirhózy do štádia dekompenzácie. Miera systémového zápalu zodpovedá jednotlivým subtypom dekompenzácie, pričom najvyšší stupeň dosahuje pri ACLF, ktoré reprezentuje fulminantný imunofenotyp CAID. K rozvoju dysfunkcie jednotlivých orgánových systémov prispievajú okrem orgánovo špecifických mechanizmov aj hyperinflamačná imunitná odpoveď, imunopatologické a imunometabolické mechanizmy. Trvajúca dysregulovaná prozápalová odpoveď vedie k postupnému vyčerpaniu imunity a preprogramovaniu vrodených a získaných imunitných buniek. Navodená imunoparalýza je príčinou problémových infekcií a predstavuje indolentný imunofenotyp CAID.
The term cirrhosis-associated immune dysfunction represents a wide spectrum of alterations in the local and systematic immune response. CAID is characterised by parallel ongoing systematic inflammation and immune deficiency, the intensity of manifestation of which depends on the stage of the disease, predisposing and precipitating factors. The gut-liver axis plays a key role in the pathogenesis of CAID. Impaired interplay between the gut and liver and changes in microbiome composition are responsible for damage to the intestinal barrier and continuous antigen stimulation, leading to the new proinflammatory setting of the immune system, which represents the low-grade inflammation phenotype CAID. Episodic aggravation of microbial translocation and episodic effect of precipitating factors cause a burst of systematic inflammation and breakthrough of the immune tolerance. The result is dysregulated hyperinflammatory immune response, which represents the high-grade inflammation phenotype of CAID. The change of the phenotype stays in the background of transition to decompensated cirrhosis. The systemic inflammation increases across the subtypes of decompensation with the highest degree reached in the ACLF, which represents the fulminant immunophenotype of CAID. Hyperinflammatory immune response, immunopathologic and immunometabolic changes act synergistically with organ-specific mechanisms in the development of organ dysfunctions. Lasting dysregulated pro-inflammatory reaction leads to immunity exhaustion and innate and adaptive immune cells reprogramming. This immune paralysis is the cause of problematic infections and represents the indolent immunophenotype of CAID.
- MeSH
- jaterní cirhóza * imunologie terapie MeSH
- lidé MeSH
- přirozená imunita MeSH
- syndromy imunologické nedostatečnosti * imunologie terapie MeSH
- zánět MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
Most primary immunodeficiency diseases, and select secondary immunodeficiency diseases, are treated with immunoglobulin (IG) therapy, administered intravenously or subcutaneously (SCIG). The first instance of IG replacement for primary immunodeficiency disease was a 16.5% formulation administered subcutaneously in 1952. While most SCIG products are now a 10 or 20% concentration, this review will focus on SCIG 16.5% products with a historical overview of development, including the early pioneers who initiated and refined IG replacement therapy, as well as key characteristics, manufacturing and clinical studies. In determining an appropriate IG regimen, one must consider specific patient needs, characteristics and preferences. There are advantages to SCIG, such as stable serum immunoglobulin G levels, high tolerability and the flexibility of self-administered home treatment.
- MeSH
- injekce subkutánní MeSH
- intravenózní imunoglobuliny aplikace a dávkování imunologie terapeutické užití MeSH
- lidé MeSH
- pasivní imunizace metody MeSH
- subkutánní infuze MeSH
- syndromy imunologické nedostatečnosti farmakoterapie imunologie MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- MeSH
- antibakteriální látky terapeutické užití MeSH
- antivirové látky terapeutické užití MeSH
- biologická terapie MeSH
- Crohnova nemoc * chirurgie diagnóza komplikace MeSH
- imunologické testy MeSH
- lidé MeSH
- mladý dospělý MeSH
- nežádoucí účinky léčiv MeSH
- pasivní imunizace MeSH
- syndromy imunologické nedostatečnosti * diagnóza farmakoterapie imunologie MeSH
- tonzilektomie MeSH
- tonzilitida diagnóza farmakoterapie komplikace MeSH
- ustekinumab terapeutické užití MeSH
- Check Tag
- lidé MeSH
- mladý dospělý MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- MeSH
- humorální imunita imunologie MeSH
- imunita MeSH
- imunosupresivní léčba škodlivé účinky MeSH
- interakce mikroorganismu a hostitele * fyziologie imunologie MeSH
- lidé MeSH
- oportunní infekce imunologie mikrobiologie MeSH
- syndromy imunologické nedostatečnosti imunologie patologie MeSH
- transplantace škodlivé účinky MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
B-cell precursors (BCP) arise from hematopoietic stem cells in bone marrow (BM). Identification and characterization of the different BCP subsets has contributed to the understanding of normal B-cell development. BCP first rearrange their immunoglobulin (Ig) heavy chain (IGH) genes to form the pre-B-cell receptor (pre-BCR) complex together with surrogate light chains. Appropriate signaling via this pre-BCR complex is followed by rearrangement of the Ig light chain genes, resulting in the formation, and selection of functional BCR molecules. Consecutive production, expression, and functional selection of the pre-BCR and BCR complexes guide the BCP differentiation process that coincides with corresponding immunophenotypic changes. We studied BCP differentiation in human BM samples from healthy controls and patients with a known genetic defect in V(D)J recombination or pre-BCR signaling to unravel normal immunophenotypic changes and to determine the effect of differentiation blocks caused by the specific genetic defects. Accordingly, we designed a 10-color antibody panel to study human BCP development in BM by flow cytometry, which allows identification of classical preB-I, preB-II, and mature B-cells as defined via BCR-related markers with further characterization by additional markers. We observed heterogeneous phenotypes associated with more than one B-cell maturation pathway, particularly for the preB-I and preB-II stages in which V(D)J recombination takes place, with asynchronous marker expression patterns. Next Generation Sequencing of complete IGH gene rearrangements in sorted BCP subsets unraveled their rearrangement status, indicating that BCP differentiation does not follow a single linear pathway. In conclusion, B-cell development in human BM is not a linear process, but a rather complex network of parallel pathways dictated by V(D)J-recombination-driven checkpoints and pre-BCR/BCR mediated-signaling occurring during B-cell production and selection. It can also be described as asynchronous, because precursor B-cells do not differentiate as full population between the different stages, but rather transit as a continuum, which seems influenced (in part) by V-D-J recombination-driven checkpoints.
- MeSH
- buněčná diferenciace genetika imunologie MeSH
- dítě MeSH
- lidé MeSH
- prekurzorové B-lymfoidní buňky imunologie MeSH
- průtoková cytometrie metody MeSH
- receptory antigenů B-buněk genetika imunologie MeSH
- syndromy imunologické nedostatečnosti genetika imunologie MeSH
- těžké řetězce imunoglobulinů genetika MeSH
- V(D)J rekombinace genetika imunologie MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Postzygotic de novo mutations lead to the phenomenon of gene mosaicism. The 3 main types are called somatic, gonadal, and gonosomal mosaicism, which differ in terms of the body distribution of postzygotic mutations. Mosaicism has been reported occasionally in patients with primary immunodeficiency diseases (PIDs) since the early 1990s, but its real involvement has not been systematically addressed. OBJECTIVE: We sought to investigate the incidence of gene mosaicism in patients with PIDs. METHODS: The amplicon-based deep sequencing method was used in the 3 parts of the study that establish (1) the allele frequency of germline variants (n = 100), (2) the incidence of parental gonosomal mosaicism in families with PIDs with de novo mutations (n = 92), and (3) the incidence of mosaicism in families with PIDs with moderate-to-high suspicion of gene mosaicism (n = 36). Additional investigations evaluated body distribution of postzygotic mutations, their stability over time, and their characteristics. RESULTS: The range of allele frequency (44.1% to 55.6%) was established for germline variants. Those with minor allele frequencies of less than 44.1% were assumed to be postzygotic. Mosaicism was detected in 30 (23.4%) of 128 families with PIDs, with a variable minor allele frequency (0.8% to 40.5%). Parental gonosomal mosaicism was detected in 6 (6.5%) of 92 families with de novo mutations, and a high incidence of mosaicism (63.9%) was detected among families with moderate-to-high suspicion of gene mosaicism. In most analyzed cases mosaicism was found to be both uniformly distributed and stable over time. CONCLUSION: This study represents the largest performed to date to investigate mosaicism in patients with PIDs, revealing that it affects approximately 25% of enrolled families. Our results might have serious consequences regarding treatment and genetic counseling and reinforce the use of next-generation sequencing-based methods in the routine analyses of PIDs.
- MeSH
- alely * MeSH
- frekvence genu * MeSH
- lidé MeSH
- mozaicismus * MeSH
- rodina MeSH
- syndromy imunologické nedostatečnosti genetika imunologie MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky MeSH
- práce podpořená grantem MeSH
- MeSH
- adjuvancia imunologická terapeutické užití MeSH
- alergie farmakoterapie imunologie MeSH
- autoimunitní nemoci farmakoterapie imunologie MeSH
- gynekologická onemocnění * farmakoterapie imunologie MeSH
- imunitní systém účinky léků MeSH
- imunologické faktory terapeutické užití MeSH
- imunomodulace * imunologie MeSH
- imunosupresiva terapeutické užití MeSH
- komplikace těhotenství * farmakoterapie imunologie MeSH
- lidé MeSH
- syndromy imunologické nedostatečnosti farmakoterapie imunologie patofyziologie MeSH
- těhotenství imunologie MeSH
- zánět farmakoterapie imunologie MeSH
- ženská infertilita * imunologie MeSH
- Check Tag
- lidé MeSH
- těhotenství imunologie MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
