Out of three genotypes of Encephalitozoon cuniculi (I-III) available for experimental studies, E. cuniculi genotype I remains the less characterized. This study describes for the first time individual phases of microsporidiosis caused by E. cuniculi genotype I and efficacy of albendazole treatment in immunocompetent BALB/c and C57Bl/6 mice and immunodeficient SCID, CD4-/- and CD8-/- mice using molecular detection and quantification methods. We demonstrate asymptomatic infection despite an intense dissemination of microsporidia into most organs within the first weeks post infection, followed by a chronic infection characterized by significant microsporidia persistence in immunocompetent, CD4-/- and CD8-/- mice and a lethal outcome for SCID mice. Albendazole application led to loss E. cuniculi genotype I infection in immunocompetent mouse strains, decreased spore burden by half in CD4-/- and CD8-/- mice, and prolongation of survival of SCID mice. These results showed Encephalitozoon cuniculi genotype I infection extend and albendazole sensitivity was comparable to E. cuniculi genotype II, but the infection onset speed and mortality rate was similar to E. cuniculi genotype III. These imply that differences in the course of infection and the response to treatment depend not only on immunological status of the host, but also on the genotype causing the infection.
- MeSH
- albendazol aplikace a dávkování MeSH
- antigeny CD4 genetika MeSH
- antigeny CD8 genetika MeSH
- antiinfekční látky aplikace a dávkování MeSH
- Encephalitozoon cuniculi klasifikace genetika MeSH
- encephalitozoonóza imunologie parazitologie MeSH
- genotyp MeSH
- imunokompetence MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši SCID MeSH
- myši MeSH
- polymerázová řetězová reakce MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Encephalitozoon cuniculi genotype III disseminated intensively into most of the organs in all strains of mice, followed by a chronic infection with massive microsporidia persistence in immunodeficient mice and a partial decrease in C57Bl/6 mice. Treatment with 0.2 mg Albendazole/mouse/day temporarily reduces the number of affected organs in immunocompetent C57Bl/6 mice, but not in CD4-/- and CD8-/- mice. The application of medication temporarily decreased the spore burden at least by one order of magnitude in all groups. These results demonstrate that the E. cuniculi genotype III infection had a progressive course and surprisingly, Albendazole treatment had only a minimal effect. The E. cuniculi genotype III spore burden in individual organs reached up to 108 or 109 in immunocompetent or immunodeficient mice, respectively; however, these mice did not demonstrate any obvious clinical signs of microsporidiosis, and the immunodeficient mice survived longer. Our findings clearly show that the survival of mice does not correspond to spore burden, which provides new insight into latent microsporidiosis from an epidemiological point of view.
- MeSH
- albendazol terapeutické užití MeSH
- antigeny CD4 genetika MeSH
- antigeny CD8 genetika MeSH
- Cercopithecus aethiops MeSH
- Encephalitozoon cuniculi genetika MeSH
- encephalitozoonóza farmakoterapie mikrobiologie patologie MeSH
- genotyp * MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- Vero buňky MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Of four genotypes of Encephalitozoon cuniculi, E. cuniculi genotype II is considered to represent a parasite that occurs in many host species in a latent asymptomatic form, whereas E. cuniculi genotype III seems to be more aggressive, and infections caused by this strain can lead to the death of even immunocompetent hosts. Although albendazole has been considered suitable for treatment of Encephalitozoon species, its failure in control of E. cuniculi genotype III infection has been reported. This study determined the effect of a 100× recommended daily dose of albendazole on an Encephalitozoon cuniculi genotype III course of infection in immunocompetent and immunodeficient mice and compared the results with those from experiments performed with a lower dose of albendazole and E. cuniculi genotype II. The administration of the regular dose of abendazole during the acute phase of infection reduced the number of affected organs in all strains of mice and absolute counts of spores in screened organs. However, the effect on genotype III was minor. Surprisingly, no substantial effect was recorded after the use of a 100× dose of albendazole, with larger reductions seen only in the number of affected organs and absolute counts of spores in all strains of mice, implying variations in albendazole resistance between these Encephalitozoon cuniculi genotypes. These results imply that differences in the course of infection and the response to treatment depend not only on the immunological status of the host but also on the genotype causing the infection. Understanding how microsporidia survive in hosts despite targeted antimicrosporidial treatment could significantly contribute to research related to human health.
- MeSH
- albendazol aplikace a dávkování farmakologie MeSH
- antifungální látky aplikace a dávkování farmakologie MeSH
- antigeny CD4 genetika MeSH
- antigeny CD8 genetika MeSH
- buněčné linie MeSH
- Cercopithecus aethiops MeSH
- Encephalitozoon cuniculi účinky léků genetika izolace a purifikace MeSH
- encephalitozoonóza farmakoterapie MeSH
- genotyp MeSH
- hostitel s imunodeficiencí imunologie MeSH
- mikrobiální testy citlivosti MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši SCID MeSH
- myši MeSH
- počet mikrobiálních kolonií MeSH
- Vero buňky MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Quantitative approaches for characterizing molecular organization of cell membrane molecules under physiological and pathological conditions profit from recently developed super-resolution imaging techniques. Current tools employ statistical algorithms to determine clusters of molecules based on single-molecule localization microscopy (SMLM) data. These approaches are limited by the ability of SMLM techniques to identify and localize molecules in densely populated areas and experimental conditions of sample preparation and image acquisition. We have developed a robust, model-free, quantitative clustering analysis to determine the distribution of membrane molecules that excels in densely labeled areas and is tolerant to various experimental conditions, i.e. multiple-blinking or high blinking rates. The method is based on a TIRF microscope followed by a super-resolution optical fluctuation imaging (SOFI) analysis. The effectiveness and robustness of the method is validated using simulated and experimental data investigating nanoscale distribution of CD4 glycoprotein mutants in the plasma membrane of T cells.
- MeSH
- algoritmy MeSH
- antigeny CD4 genetika metabolismus MeSH
- buněčná membrána metabolismus MeSH
- fluorescenční barviva MeSH
- fluorescenční mikroskopie metody statistika a číselné údaje MeSH
- Jurkat buňky MeSH
- lidé MeSH
- membránové proteiny genetika metabolismus MeSH
- mutantní proteiny genetika metabolismus MeSH
- optické zobrazování metody statistika a číselné údaje MeSH
- shluková analýza MeSH
- T-lymfocyty imunologie metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- validační studie MeSH
Hyper-IgM syndrom (HIGM) je skupina vzácných a závažných vrozených imunodeficiencí, pro které jsou typické velmi nízké hladiny imunoglobulinu tříd IgG, IgA a IgE s normální nebo zvýšenou hladinou IgM. Příčinou onemocnění je porucha mechanismu izotypového přesmyku. Defekt postihuje signalizační dráhu molekuly CD40 nebo vnitřní signalizaci B-lymfocytů, případně buněčný DNA reparační mechanismus. Nejčastější poruchou je poškození ligandu pro CD40 (X-vázaný hyper-IgM syndrom). Kromě humorální imunity je postižena i složka buněčná, takže pacienti jsou ohroženi oportunními infekcemi (především Pneumocystis jirovecii, CMV, kryptokoky nebo mykobakteriemi). Typické jsou Časté recidivující respirační infekce a chronický průjem doprovázený neprospíváním. Více než u poloviny pacientů nalézáme chronickou neu"''^penii, dále se vyskytují artritidy, trombocytopenie, hemolytické anemie a nefritidy. Pacienti s deficitem CD40L jsou také zvýšeně ohroženi vznikem maligních onemocnění. Základem léčby jsou režimová opatření, antibiotická profylaxe a substituční léčba imunoglobuliny. Jediná kauzální terapie je alogenní transplantace hematopoetických buněk. V letech 1993 až 2011 bylo v České republice diagnostikováno pět pacientů s vrozeným deficitem CD40 ligandu. Tři pacienti byli po stanovení diagnózy indikováni k transplantaci hematopoetických buněk. V článku předkládáme přehled průběhu onemocnění, diagnostiky a léčby těchto pacientů.
Hyper-IgM syndrome (HIGM) is a group of rare and serious primary immunodeficiencies. It is characterised by very low levels of IgG, IgA and IgE immunoglobulins whereas the level of IgM is normal or elevated. It is caused by impairment of immunoglobulin isotype sw itching. Either the signalling pathway of CD40 molecule, intrinsic B-cell signalling or DNA reparation mechanism are defective. The most frequent defect lies in the gene coding for CD40 ligand (X-linked Hyper-IgM syndrome). Apart from humoral immunity also the T-cell funct ion is affected. Thus, the patients suffer from infections caused by opportunistic pathogens (most frequently Pneumocystic jirovecii , CMV, Crypto- cocci, Mycobacteria). Recurrent respiratory tract infections are frequently seen. Chronic diarrhoea may be present as well and may associate with failure to thrive. More than a half of the patients suffer from chronic neutropenia. Arthritis, thrombocytopenia, haemolyt ic anaemia and nephritis are documented in some patients. Patients with CD40 ligand deficiency have increased risk of malignant diseases. The cornerstones of the therapy are preventive measures, antibiotic prophylaxis and immunoglobulin substitution. The haematopoietic stem cell tr ansplantation is the only curative therapy currently available. Between 1993 and 2011 there were 5 patients with CD40 ligand deficiency diagnosed and treated in the Czech Republic. Three pati ents were indicated for haematopoietic stem cell transplantation. The authors present an overview of the disease’s course, diagnostic pro cess and treatment of these patients.
- Klíčová slova
- deficit CD40L,
- MeSH
- antigeny CD4 genetika chemie MeSH
- B-lymfocyty metabolismus MeSH
- buněčná membrána metabolismus MeSH
- dítě MeSH
- dysgamaglobulinemie MeSH
- financování organizované MeSH
- imunodeficience s hyper-IgM epidemiologie genetika imunologie MeSH
- intravenózní imunoglobuliny aplikace a dávkování MeSH
- lidé MeSH
- ligand CD40 metabolismus MeSH
- mladý dospělý MeSH
- posunová mutace genetika imunologie MeSH
- předškolní dítě MeSH
- průtoková cytometrie MeSH
- transplantace hematopoetických kmenových buněk využití MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- Publikační typ
- kazuistiky MeSH
- MeSH
- akutní lymfatická leukemie diagnóza etiologie genetika MeSH
- algoritmy MeSH
- antigeny CD27 genetika MeSH
- antigeny CD4 genetika MeSH
- exprese genu genetika imunologie MeSH
- genetické markery genetika MeSH
- genotyp MeSH
- lidé MeSH
- mikročipová analýza metody využití MeSH
- prognóza MeSH
- Check Tag
- lidé MeSH
- MeSH
- antigenní modulace * genetika imunologie MeSH
- antigeny CD4 genetika imunologie MeSH
- B-lymfocyty MeSH
- buněčná imunita genetika imunologie MeSH
- exprese genu * genetika imunologie MeSH
- histokompatibilita - antigeny třídy II * imunologie účinky léků MeSH
- HLA-D antigeny genetika imunologie MeSH
- humorální imunita genetika imunologie MeSH
- interferony genetika imunologie MeSH
- interleukiny genetika imunologie MeSH
- Langerhansovy buňky MeSH
- lidé MeSH
- makrofágy imunologie MeSH
- membránové glykoproteiny genetika imunologie MeSH
- slezina MeSH
- T-lymfocyty MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH