This study revises our understanding of the effectiveness of cell-mediated adaptive immunity and treatment against microsporidia using molecular detection and quantification of microsporidia in immunocompetent C57Bl/6 and immunodeficient CD4-/-and CD8-/-mice for the first time. We demonstrate an intense dissemination of microsporidia into most organs within the first weeks post-infection in all strains of mice, followed by a chronic infection characterized by microsporidia persistence in CD4-/-and C57Bl/6 mice and a lethal outcome for CD8-/-mice. Albendazole application reduces microsporidia burden in C57Bl/6 and CD4-/-mice, whereas CD8-/-mice experience only a temporary effect of the treatment. Surprisingly, treated CD8-/-mice survived the entire experimental duration despite enormous microsporidia burden. On the basis of our results, we conclude that microsporidia survive despite the presence of immune mechanisms and treatments that are currently considered to be effective and therefore that CD8 T lymphocytes represent a major, but not sole effector mechanism controlling microsporidiosis. Furthermore, the survival of mice does not correspond to spore burden, which provides new insight into latent microsporidiosis from an epidemiological point of view.
- MeSH
- adaptivní imunita imunologie MeSH
- albendazol terapeutické užití MeSH
- anthelmintika terapeutické užití MeSH
- buněčná imunita imunologie MeSH
- CD4-pozitivní T-lymfocyty imunologie MeSH
- CD8-pozitivní T-lymfocyty imunologie MeSH
- Encephalitozoon cuniculi růst a vývoj imunologie MeSH
- encephalitozoonóza imunologie mikrobiologie MeSH
- lymfopenie imunologie MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Encephalitozoon cuniculi is probably the most common microsporidia which infects a wide range of vertebrates, including human. So far, four genotypes of this parasite have been identified based on the rRNA internal transcribed spacer variations. The course of infection caused by E. cuniculi III had very massive onset in immunocompetent host characterized by the presence of this parasite in all organs and tissues within one week after peroral infection. Encephalitozoonosis caused by E. cuniculi III had very progressive spreading into all organs within first week post inoculation in immunocompromised SCID mice and led to the death of the host. The experimental treatment with albendazole of immunocompetent BALB/c mice infected with E. cuniculi III have shown very weak effect. Our findings clearly showed that the different course of infection and response to treatment depends not only on the immunological status of the host, but also on the genotype of microsporidia. It could be very important especially for individuals under chemotherapy and transplant recipients of organs originating from infected donors.
- MeSH
- albendazol farmakologie terapeutické užití MeSH
- antifungální látky farmakologie terapeutické užití MeSH
- Encephalitozoon cuniculi účinky léků genetika imunologie fyziologie MeSH
- encephalitozoonóza farmakoterapie imunologie parazitologie MeSH
- feces parazitologie MeSH
- genotyp MeSH
- imunokompetence * MeSH
- imunokompromitovaný pacient * MeSH
- myši inbrední BALB C MeSH
- myši SCID MeSH
- myši MeSH
- spory hub MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The breeding of domestic rabbit (Oryctolagus cuniculus) for human consumption has a long tradition mainly in European and Asian countries. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animals health. This study aimed to collect sera from rabbits bred in different conditions and test the presence of Toxoplasma gondii and Encephalitozoon cuniculi antibodies. Whether infections were active or latent was assessed by determining the occurrence of IgM or IgM together with IgG antibodies which indicated active infection whereas latent infection was characterized by finding IgG antibodies only. An ELISA test was performed with 1883 sera samples collected throughout the Czech and Slovak Republics. The seroprevalence of T. gondii in 902 samples from 6 commercial farms (CF) was very low with only 4 rabbits (0.4%) being positive. In total 99 (10.1%) individuals out of 981 samples from 29 household farms (HF) were positive for T. gondii antibodies. Only 2 (50%) of the T. gondii positive CF rabbits had active infections while the rest were latently infected. The serological results showed that 35 (35.4%) rabbits from the T. gondii positive HF group suffered from active infection. Out of CF samples 185 (20.5%) were positive for E. cuniculi. Antibodies of E. cuniculi were detected in 497 (50.7%) HF rabbits. Active E. cuniculi infections were determined in 85.9% of CF and 56.3% of HF rabbits; respectively. Interestingly, the E. cuniculi positive rabbits were significantly more often positive for anti-T. gondii antibodies in comparison to E. cuniculi negative individuals. Prevalence of T. gondii in CF rabbits was negligible. According to our results meat of HF rabbits still poses a risk of T. gondii infection. Nevertheless, the risk is on its lowest level in 20 years which is apparently caused due to changes in feeding practices. The occurrence of E. cuniculi antibodies was significantly lower in rabbits from commercial farms, apparently because of better hygiene conditions.
- MeSH
- ELISA veterinární MeSH
- Encephalitozoon cuniculi imunologie izolace a purifikace MeSH
- encephalitozoonóza veterinární MeSH
- králíci imunologie parazitologie MeSH
- prevalence MeSH
- protilátky protozoální krev MeSH
- séroepidemiologické studie MeSH
- Toxoplasma imunologie izolace a purifikace MeSH
- toxoplazmóza zvířat epidemiologie MeSH
- veřejné zdravotnictví MeSH
- zvířata MeSH
- Check Tag
- králíci imunologie parazitologie MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Slovenská republika MeSH
A total of 9 (8 stallions and 1 mare) 1 year old ponies were used for the experimental infection caused by Encephalitozoon cuniculi genotype II (10(7) spores per animal). Subsequently, individual horses were slaughtered 7, 14, 21, 28, 35, 42, 49, 56, and 63 days post infection. Immediately after slaughter, tissues samples of stomach, duodenum, jejunum, ileum, caecum, colon, spleen, liver, kidney, bladder, heart, lungs, and brain were sampled. In addition, urine, feces and blood specimens were collected. Enzyme-linked immunosorbent assay was used for determination of humoral immune response and nested PCR targeting 16S rDNA, whole ITS, and 5.8S rDNA was used for detection of E. cuniculi in collected organs, blood, feces and urine. No clinical signs of microsporidiosis including diarrhea or colic, neurological signs and fever were observed in any horses during whole experiment. Acute microsporidiosis in ponies was characterized by the dissemination of microsporidia into almost all organs and significant increase of concentration of specific antibodies in blood was observed from 28 to 42 DPI. After this acute stage microsporidia disappeared from most organs with the exception of the kidney, which was positive up to 63 DPI when the experiment was terminated. No pathological changes were observed in any organs with exception of one mare's brain, where E. cuniculi-positive cavity measuring 5 cm × 3 cm in diameter formed in the lobus piriformis.
- MeSH
- Encephalitozoon cuniculi imunologie MeSH
- encephalitozoonóza imunologie veterinární MeSH
- humorální imunita fyziologie MeSH
- koně MeSH
- nemoci koní imunologie mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cieľ práce: Zistiť výskyt protilátok proti intracelulárnym patogénom (Toxoplasma gondii, Encephalitozoon cuniculi a Encephalitozoon intestinalis) u slovenských žien. Materiál a metodika: Na detekciu špecifických IgG protilátok proti Toxoplasma gondii, Encephalitozoon cuniculi a Encephalitozoon intestinalis bola použitá imunoenzýmová analýza. Špecifické protilátky IgG boli detegované v krvných sérach 118 slovenských žien s rozličnou diagnózou pomocou ELISA. Výsledky: Ženy pochádzajúce zo Slovenska boli rozdelené do troch skupín na základe veku. V skupine 1 (n = 26, vek > 51 rokov) bolo pozitívnych 9 žien na prítomnosť špecifických protilátok proti T. gondii, 2 na E. cuniculi a 1 na E. intestinalis. V skupine 2 (n = 53, vek 36–50 rokov) bolo 18 žien pozitívnych na T. gondii, 3 na E. cuniculi a 7 na E. intestinalis. V skupine 3 (n = 39, vek 20–35 rokov) bolo 13 žien pozitívnych na prítomnosť protilátok proti T. gondii, 2 na E. cuniculi a 4 na E. intestinalis. V rámci tejto štúdie sme sa pokúsili zistiť súvislosť medzi klinickou diagnózou a výskytom jednotlivých typov infekcií. Ženy boli preto rozdelené do štyroch skupín na základe diagnóz (diagnózy súvisiace s nervovým, respiratórnym a imunitným systémom a gravidné ženy). V našej štúdii bol potvrdený štatisticky významný vzťah medzi diagnózou a výskytom infekcie (p < 0.001). U žien s výskytom ochorení respiratórneho systému bola detegované v najväčšej miere toxoplazmóza. U žien s ochoreniami nervového systému sa najčastejšie vyskytovala infekcia s E. intestinalis a u žien s ochoreniami imunitného systému sa vyskytovala prevažne infekcia E. cuniculi. Záver: V rámci našej štúdie sme zistili, že prevalencia infekcií u gravidných žien a jednotlivých infekcií bola veľmi podobná. V skupine žien s respiratórnymi ochoreniami bola najčastejšia infekcia s T. gondii. Naše výsledky potvrdzujú skutočnosť, že u ľudí môže toxoplazmóza postihnúť rozdielne orgánové systémy a môže byť spojená s rozličnými klinickými príznakmi. Respiratórny systém býva v prípade toxoplazmózy postihnutý veľmi často a môže vyústiť až do pneumónie. Medzi veľmi často sa vyskytujúce klinické príznaky patria príznaky podobné chrípke. V skupine s diagnózami, ktoré súviseli s imunitným systémom, sa najčastejšie vyskytovala infekcia s E. cuniculi a druhou najčastejšou infekciou bola toxoplazmóza. Oba patogény patria medzi oportúnne patogény, ktoré môžu spôsobiť vážne zdravotné problémy v dôsledku imunodeficiencií (HIV, transplantácia, chemoterapia). V skupine s diagnózami spojenými s nervovým systémom sa najčastejšie vyskytovala infekcia s E. intestinalis. V prípade E. intestinalis nie je úplne jasné, ktoré patologické zmeny môže spôsobiť. Infekcia veľmi často prebieha diseminovane a veľmi často postihuje tráviaci trakt. Ale keďže je E. intestinalis veľmi blízky s E. cuniculi, ktorý postihuje nervový systém, je možné, že E. intestinalis môže spôsobiť poškodenie nervového systému.
Aim of the study: To determine the prevalence of antibodies to intracellular pathogens (Toxoplasma gondii, Encephalitozoon cuniculi, and Encephalitozoon intestinalis) in sera of Slovak women. Material and methods: Enzyme-linked immunosorbent assay (ELISA) was used for the detection of specific IgG antibodies to Toxoplasma gondii, Encephalitozoon cuniculi, and Encephalitozoon intestinalis in 118 sera samples from Slovak women with different diagnoses. Results: Women were divided into three groups based on the age. In group 1 (n = 26, age > 51), nine women tested positive for T. gondii, two for E. cuniculi, and one for E. intestinalis. The respective numbers in group 2 (n = 53, age range 36–50) were 18, three, and seven, and in group 3 (n = 39, age range 20–35) 13, two, and four. In an attempt to find the relationship between the clinical diagnosis and the prevalence of the three pathogens, we divided the study subjects into four groups based on the condition (diseases of the nervous, respiratory, and immune systems and pregnancy). A statistically significant relationship between the diagnosis and the prevalence of a pathogen was found (p < 0.001). In women with diseases of the respiratory system, antibodies against Toxoplasma gondii were most often detected, while infection with E. intestinalis was most commonly associated with nervous system diseases and the highest seroprevalence of E. cuniculi was found in women diagnosed with immune system disorders. Conclusions: In our study, the seroprevalence rates of the three infections in pregnant women were similar, showing no significant difference. Women with diseases of the respiratory system were most often seropositive for T. gondii. Our result is consistent with the known fact that in humans, toxoplasmosis may affect various organs of the body, causing diverse clinical signs. However, the respiratory system is commonly involved and pneumonia may result. The most common finding is a mild, flu-like illness that lasts a few days. The women with different types of immunodeficiency disorders had the highest seropositivity rates for E. cuniculi and the second most common infection in this group was toxoplasmosis. Both of these parasites are opportunistic and can cause serious problems in immunocompromised individuals (HIV-positives, organ transplant recipients, chemotherapy patients). The women diagnosed with the nervous system disorders were most commonly infected with E. intestinalis. The relationship between the disease and this pathogen is not clear, but E. intestinalis is responsible for various pathologies; it causes mainly disseminated infections and damage to the digestive tract. E. intestinalis is a parasite very close to E. cuniculi affecting either the nervous system or the respiratory tract depending on the type of host.
- Klíčová slova
- Encephalitozoon intestinalis,
- MeSH
- dospělí MeSH
- ELISA MeSH
- Encephalitozoon cuniculi imunologie patogenita MeSH
- Encephalitozoon imunologie patogenita MeSH
- encephalitozoonóza epidemiologie imunologie MeSH
- financování organizované MeSH
- imunokompromitovaný pacient imunologie MeSH
- kohortové studie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikrosporidióza epidemiologie imunologie krev MeSH
- nemoci dýchací soustavy imunologie MeSH
- nemoci imunitního systému imunologie MeSH
- nemoci nervového systému imunologie MeSH
- prevalence MeSH
- protilátky fungální imunologie krev MeSH
- séroepidemiologické studie MeSH
- těhotenství imunologie MeSH
- toxoplazmóza epidemiologie imunologie krev MeSH
- věkové rozložení MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- těhotenství imunologie MeSH
- ženské pohlaví MeSH
- Geografické názvy
- Slovenská republika MeSH
Encephalitozoon cuniculi is an obligate intracellular pathogen that has a wide host distribution, but primarily affects rabbits. The aim of this study was to characterize both the cell-mediated and the antibody response in rabbits after experimental infection using 2 different infection routes: oral and ocular. SPF rabbits were infected with low (10³ spores) and high (10⁷ spores) infection doses. Monitored parameters included clinical signs, detection of spores in urine, antibody response detected with ELISA, and cell-mediated immunity detected by antigen-driven lymphocyte proliferation. At week 13 post-infection, half of the rabbits in each group were suppressed by intramuscular administration of dexamethasone. At week 18 post-infection, animals were euthanized. Clinical signs were mild with exacerbation after immunosuppression. Spores in urine and antigen-specific cell-mediated immunity were detected from weeks 5 and 4 post-infection, respectively. Specific IgM was detected 1 week after infection, and IgG antibodies followed 1 week later in rabbits infected with the high dose. Immunological responses were dose dependent. The authors can conclude that both oral and ocular experimental infection with E. cuniculi resulted in an immune response of the infected animals. Rabbits could be used as an experimental model for the study of ocular microsporidiosis.
- MeSH
- aktivace lymfocytů MeSH
- buněčná imunita MeSH
- Encephalitozoon cuniculi imunologie patogenita MeSH
- encephalitozoonóza imunologie parazitologie patologie MeSH
- imunoglobulin G krev MeSH
- imunoglobulin M krev MeSH
- králíci MeSH
- modely nemocí na zvířatech MeSH
- nemoci úst imunologie parazitologie patologie MeSH
- oční infekce imunologie parazitologie patologie MeSH
- outbrední kmeny zvířat MeSH
- protilátky protozoální krev MeSH
- tvorba protilátek MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Microsporidia are eukaryotic, obligate, intracellular protists that are emerging pathogens in immunocompromised hosts, including AIDS patients and organ transplant recipients. The efficacy of gamma interferon (IFN-gamma) for the treatment of microsporidiosis caused by Encephalitozoon cuniculi was studied by means of adoptive transfer and IFN-gamma administration in SCID mice. While the adoptive transfer of CD4(+) T cells from immunocompetent mice prolonged survival of SCID mice infected perorally with E. cuniculi, survival was not improved by adoptive transfer of CD4(+) T lymphocytes from IFN-gamma-deficient mice. The protective effect of IFN-gamma was confirmed in cytokine therapy experiments in which SCID mice receiving IFN-gamma survived significantly longer than mice receiving mock injections. The administration of serum containing specific antibodies against E. cuniculi was found to prolong the survival of concurrently IFN-gamma-treated SCID mice. The data presented in this study suggest that IFN-gamma is potentially useful as a cytokine therapy for microsporidiosis, especially in CD4(+) T-cell-deficient patients.
- MeSH
- CD4-pozitivní T-lymfocyty imunologie MeSH
- Encephalitozoon cuniculi imunologie MeSH
- encephalitozoonóza MeSH
- financování organizované MeSH
- imunoterapie MeSH
- interferon gama aplikace a dávkování genetika terapeutické užití MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- monoklonální protilátky imunologie terapeutické užití MeSH
- myši inbrední BALB C MeSH
- myši knockoutované MeSH
- myši SCID MeSH
- myši MeSH
- převzatá imunita MeSH
- specificita protilátek MeSH
- výsledek terapie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
The role of antibodies in the immune response to microsporidiosis was studied using a novel anti-exospore monoclonal antibody (MAb) P5/H1, which recognizes surface antigens of Encephalitozoon cuniculi. The effect of the MAb on microsporidial infection in vivo was to prolong the survival of previously CD4+ reconstituted, perorally infected and intraperitoneally MAb-treated SCID mice. The MAb decreased the numbers of E. cuniculi spores in peritoneal smears obtained post mortem. These results suggest a possible role for antibodies in protection against perorally acquired E. cuniculi infection.
- MeSH
- analýza přežití MeSH
- CD4-pozitivní T-lymfocyty imunologie MeSH
- Encephalitozoon cuniculi imunologie MeSH
- encephalitozoonóza imunologie terapie MeSH
- financování organizované MeSH
- imunoterapie metody MeSH
- monoklonální protilátky farmakologie imunologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- neparametrická statistika MeSH
- spory hub imunologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
Stav imunitného systému jedinca zohráva kľúčovú úlohu pri regulácii oportúnnych infekcií. V bojiproti intracelulárnym parazitom sa uplatňujú viaceré nešpecifické ako aj špecifické imunitnémechanizmy. Dominantnú úlohu v reakcii na infekciu zástupcami rodu Encephalitozoon mábunkousprostredkovaná imunitná odpoveď. Encephalitozoon cuniculi, ako najviac preskúmaný zástupcatohto rodu, je schopný prežívať v organizme hostiteľa aj napriek jeho aktívnej imunitnej odpovedi.Latentná asymptomatická infekcia prebieha len dovtedy, kým sú množenie parazita a imunitnáodpoveď v rovnováhe. Preto je dôležité poznať imunitné mechanizmy, ktoré sa zúčastňujú nazvládnutí tejto infekcie.
The immunity systemstatus of an individual plays the key role in regulation of opportune infection.In the fight against the intracellular parasites several non-specific as well as specific immunitymechanisms are applied. The dominant role in response to infection caused by the representativesof genus Encephalitozoon plays the cell-mediated immune response. Encephalitozoon cuniculi, asthe most explored representative of this genus is able to survive in the host organism despite hisactive immunity response. Latent asymptomatic infection goes on only as long as the parasitemultiplication and immune response are balanced.
