Harnessing immunomodulatory mechanisms of Trichinella spiralis to design novel nanomedical approaches for restoring self-tolerance in autoimmunity.

The rapid increase in the prevalence of autoimmune diseases in recent decades, especially in developed countries, coincided with improved living conditions and healthcare. Part of this increase could be ascribed to the lack of exposure to infectious agents like helminths that co-evolved with us and display potent immune regulatory actions. In this review we discussed many investigations, including our own, showing that Trichinella spiralis via its excretory-secretory products attenuate Th1/Th17 immunopathological response in autoimmunity and potentiate the protective Th2 and or regulatory T cell response, acting as an effective induction of tolerogenic dendritic cells (DCs), and probably mimicking the autoantigen in some diseases. A recent discovery of T. spiralis extracellular vesicles (TsEVs) suggested that inducing a complex regulation of the immune response requires simultaneous delivery of different signals in nano-sized packages. Indeed, different artificial nanomedical approaches discussed here suggested that co-delivery of multiple signals via nanoparticles is the most promising strategy for the treatment of autoimmune diseases. Although a long way is ahead of us before we could completely replicate natural nano-delivery systems which are both safe and potent in restoring self-tolerance, a clear path is being opened from a careful examination of parasite-host interactions.

Murine hepatoma treatment with mature dendritic cells stimulated by Trichinella spiralis excretory/secretory products.

Excretory/Secretory Products (ESPs) of the nematode Trichinella spiralis contain antitumor-active substances that inhibit tumor growth. Mature dendritic cells (DCs) play a critical role in the antitumor immunity of the organism. As pathogen-derived products, it ought to be discussed whether T. spiralis ESPs will reduce the antitumor effect of mature DCs from the host before it is applied to patients' tumors. Therefore, the aim of this work was to evaluate the immunological effect of DCs stimulated by T. spiralis ESPs in H22 tumor-bearing mice. H22 tumor model mice in this study were randomly divided into four groups according to the treatment: PBS control group, ESP group, DCs group, and DCs stimulated with T. spiralis ESP (ESP+DCs group). The antitumor effect was evaluated by tumor inhibition rate and cytokine detection using ELISA. The results showed significant inhibition in tumor growth in the ESP+DCs, DCs and ESP groups when compared with the PBS control group (p < 0.01, p < 0.01, and p < 0.05, respectively). However, no significant difference was observed on tumor inhibition rates between the ESP+DCs and DCs groups. The decrease in IL-4, IL-6, and IL-10, and the increase in IFN-γ between the DCs and ESP+DCs groups were also not significant. Therefore, DCs stimulated by ESP did not reduce the antitumor effect of mature DCs, which demonstrated that the T. spiralis ESP would not affect the antitumor effect of mature DCs by modulating the immune response of the host, and that ESPs are safe in antitumor immunology when applied in a tumor model mice.

TITLE: Traitement des hépatomes murins avec des cellules dendritiques mûres stimulées par des produits d'excrétion-sécrétion de Trichinella spiralis. ABSTRACT: Les produits d'excrétion-sécrétion (PES) du nématode Trichinella spiralis contiennent des substances actives antitumorales qui inhibent la croissance tumorale. Les cellules dendritiques (CD) mûres jouent un rôle essentiel dans l'immunité anti-tumorale de l'organisme. En tant que produits dérivés d'agents pathogènes, il convient de déterminer si les PES de T. spiralis réduisent l'effet antitumoral des CD mûrs de l'hôte, avant leur application aux tumeurs des patients. Par conséquent, l'objectif de ce travail était d'évaluer l'effet immunologique des CD stimulées par les PES de T. spiralis chez des souris porteuses de tumeurs H22. Les souris modèles à tumeurs H22 dans cette étude ont été réparties au hasard en quatre groupes selon le traitement : groupe témoin PBS, groupe PES, groupe CD et groupe CD stimulés par les PES de T. spiralis (CD+PES). L'effet antitumoral a été évalué par le taux d'inhibition des tumeurs et la détection des cytokines en utilisant un dosage ELISA. Les résultats ont montré une inhibition significative de la croissance tumorale dans les groupes CD+PES, CD et PES par rapport au groupe témoin PBS (p < 0,01, p < 0,01 et p < 0,05, respectivement). Cependant, aucune différence significative n'a été observée sur le taux d'inhibition de la tumeur entre les groupes CD+PES et CD. La diminution de l'IL-4, de l'IL-6, de l'IL-10 et l'augmentation de l'IFN-γ entre les groupes CD et CD+PES n'étaient pas non plus significatives. Ainsi, les CD stimulées par les PES n'ont pas réduit l'effet antitumoral des CD mûrs, ce qui a démontré que les PES de T. spiralis n'affectaient pas l'effet antitumoral des CD mûrs en modulant la réponse immunitaire de l'hôte et que les PES sont sûrs en immunologie antitumorale quand ils sont appliqués à des souris modèles tumorales.

The Disease Ecology, Epidemiology, Clinical Manifestations, and Management of Trichinellosis Linked to Consumption of Wild Animal Meat.

Historically, human trichinellosis was caused by Trichinella spiralis and transmitted to humans by consumption of undercooked domestic pork. Today, most cases of trichinellosis are caused by other Trichinella species and transmitted by consumption of raw or undercooked wild game meats. Given the increasing global prevalence of wild animal meat-linked trichinellosis, the objectives of this review are: 1) to describe the life cycle and global distribution of Trichinella worms; 2) to describe the changing epidemiology of trichinellosis; 3) to describe the clinical phases of trichinellosis; 4) to recommend the latest diagnostic tests; and 5) to recommend treatment and prevention strategies. Internet search engines were queried with keywords as subject headings to meet the objectives of this review. Although trichinellosis surveillance systems and laws regulating commercial pork production have limited T spiralis-caused trichinellosis in Europe and the United States, trichinellosis due to consumption of raw and undercooked wild boar and feral hog meat continues to occur throughout Southeast Asia. Trichinellosis due to consumption of raw or undercooked meats of other infected game, such as bear, deer, moose, and walrus, continues to occur worldwide. Only adherence to hygienic practices when preparing wild game meats and cooking wild game meats to recommended internal temperatures can prevent transmission of trichinellosis to humans. Wilderness medicine clinicians should be prepared to advise hunters and the public on the risks of game meat-linked trichinellosis and on how to diagnose and treat trichinellosis to prevent fatal complications.

Compromiso neurológicoy cardiovascularen pacientes con triquinosis / Neurological and cardiovascular involventin patients with trichinosis

Introducción: La triquinosis es una infección parasitaria causada por nematodos del género Trichinella. El compromiso cardíaco no es habitual, pero representa la causa más frecuente de muerte por triquinosis, mientras que la afectación neurológica ocurre principalmente en pacientes severamente enfermos. Se sabe que a mayor cantidad de larvas ingeridas menor es el tiempo de incubación y mayor es la severidad de la enfermedad. Se presentan dos pacientes con compromiso del sistema nervioso central, uno de ellos cardiovascular, ambos pertenecientes a un brote de triquinosis ocurrido en la ciudad de Bahía Blanca.

Background: Trichinosis is a parasitic infection caused by nematodes of the genus Trichinella. Cardiac involvement is not usual but represents the most frequent cause of death by trichinosis, while neurological affectation occurs mainly in severely ill patients. A greater number of larvae ingested, the shorter the incubation time and the greater the severity of the disease. We present two patients with compromise of the central nervous system and one of them cardiovascular, both belonging to an outbreak of trichinosis in the city of Bahía Blanca

Effect of probiotic bacteria on phagocytosis and respiratory burst activity of blood polymorphonuclear leukocytes (PMNL) in mice infected with Trichinella spiralis.

This study focusses on the effect of probiotic (bacteriocinogenic) strains on parasite infection and innate immunity - phagocytosis and oxidative burst of blood monocytes and polymorphonuclear leukocytes (PMNL) in mice infected with Trichinella spiralis. Bacteriocinogenic and probiotic strains of different origin (Enterococcus faecium AL41=CCM8558, Enterococcus durans ED26E/7, Lactobacillus fermentum AD1=CCM7421, Lactobacillus plantarum 17L/1) were administered daily in dose of 109CFU/ml in 100µl and mice were infected with 400 larvae of T. spiralis on 7th day of treatment. Phagocytic activity of blood leukocytes was inhibited at week 3 and 4 post infection (p.i.), i.e. in the time of massive muscle invasion with larvae T. spiralis. Administration of bacterial strains to mice prior to T. spiralis infection elevated and prolonged phagocytic activity of blood leukocytes and their ingestion capability from week 1 to 3 of the infection and the phagocytosis was inhibited only at week 4 p.i. The highest stimulative effect on phagocytosis was induced by strains E. durans ED26E/7, L. fermentum AD1=CCM7421, and L. plantarum 17L/1. The percentage of cells with respiratory burst and their enzymatic activity was increased after T. spiralis infection with the exception of week 3 p.i. In contrast, in all mice treated with bacterial strains the enzymatic stimulation was observed after the infection, with the highest intensity caused by strains E. durans ED26E/7, L. fermentum AD1=CCM7421 and L. plantarum 17L/1. The administration of probiotic strains stimulated phagocytosis and respiratory burst of blood PMNL that could contribute to a decreased larval migration and a destruction of muscle larvae and then reduced parasite burden in the host. The protective effect against T. spiralis infection was induced by all strains, but the highest reduction was recorded by E. faecium AL41=CCM8558.

Nematode infections: soil-transmitted helminths and trichinella.

Infection with soil-transmitted helminths occurs via ingestion of nematode eggs with contaminated food and water, via hands, or inhalation of dust, or by penetration of larvae through the skin. Trichinella infections are caused by the ingestion of larvae contained in undercooked meat. In highly endemic areas, preventive chemotherapy (ie, regular administration of anthelmintic drugs to at-risk populations) is the key strategy against soil-transmitted helminthiasis. Integrated control approaches, including improved hygiene, sanitation, and water, are required for lasting effects. Because of growing tourism, travel, and migration, clinicians and specialized travel clinics must remain aware of the diagnosis, prevention, and treatment of soil-transmitted helminth and Trichinella infections.

Evaluación de la infección de Trichinella spiralis en cerdos gonadectomizados, Zacatecas, México / An assessment of Trichinella spiralis infection in gonadectomized pigs in Zacatecas, Mexico

Objetivo: Evaluar la infección de Trichinella spiralis en cerdos gonadectomizados. Material y método: El presente trabajo se realizó utilizando 9 cerdos de la raza York de 4 meses de edad divididos en tres grupos: 3 cerdos control sanos, 3 cerdos infectados con Trichinella spiralis y 3 cerdos gonadectomizados y posteriormente infectados con Trichinella spiralis, todos los animales fueron sangrados pretratamiento y postratamiento para detección de anticuepos anti T. spiralis por las técnicas inmunologicas, Wester Blot y Inmunofluorescencia indirecta. Los animales fueron sacrificados a las 6 semanas de infección por Trichinella spiralis, se obtuvo tejido para las técnicas directas de compresión en placa y digestión artificial para evaluación de carga parasitaria. Resultados: en los cerdos gonadectomizados e infectados se presentó un ataque agudo de Trichinellosis, la carga parasitaria fue mayor estadísticamente significativa en relación a los cerdos infectados con P < 0.01), por el método de ANOVA y sin gonadectomizar, las técnicas inmunológicas fueron positivas en ambos grupos infectados y gonadectomizados. Conclusión: El diagnóstico oportuno de Trichinella spiralis mediante la evaluación directa y aletoria en camadas de cerdos a través de la observación de larva infectante en tejido cerebral, detección de anticuerpos anti - T. spiralis. por inmunofluorecescia indirecta e inmunoelectrotransferencia resultan eficaces y eficientes para hacer un diagnóstico oportuno en el animal para consumo humano.

Objective: To assess the prevalence of Trichinella spiralis in gonadectomized pigs. Material and methods: The study was performed using 9 four-month old pigs, York race, which were divided in three groups: 3 healthy control animals, 3 pigs infected with Trichinella spiralis, and 3 gonadectomized animals that later were infected with T. spiralis. Blood samples were taken from every animal before and after therapy for the detection of anti-T. spiralis antibodies using Western Blot and indirect immunofluorescence techniques. All animals were killed after 6 weeks of being infected with T. spiralis, and tissue samples were obtained for performing assessments using direct methods, such as plate compression and artificial digestion in order to look for the parasite load. Results: Gonadectomized and infected pigs developed an acute presentation of trichinellosis, their parasite load was statistically higher compared to infected non-gonadectomized pigs using ANOVA (P<0.01), and immune methods yielded positive results in both infected groups of animals. Conclusion: The timely diagnosis of Trichinella spiralis in pigs using direct and random assessments by observing the presence of infective worms in brain tissue, together with the detection of anti-T. spiralis antibodies using indirect immunofluorescence and immunoelectrotransference are both efficacious and efficient for the early diagnosis of this infection in animals whose meat is about to be consumed.

The intraperitoneal inoculation of Lactobacillus casei in mice induces total protection against Trichinella spiralis infection at low challenge doses.

The following effects of Lactobacillus casei in NIH mice were evaluated: the establishment of Trichinella spiralis adult worms in the intestine (AWI), larvae per gram of muscle tissue (LPG), levels of IgG and IgA, and levels of IL-4 and IFN-γ. One hundred and eight mice were allocated at random into 18 groups of six mice each. Each mouse in treated or non-treated groups was inoculated intraperitoneally once a week during 6 weeks with L. casei or phosphate-buffered solution. Later each mouse was challenged either with 200, 50, or 25 T. spiralis infective larvae. When the infection dose was 200 T. spiralis infective larvae, the reductions in AWI were 78.6% at 4 days after infection (dai) and 76.7% at 10 dai; while the reduction of LPG was 80.9% with respect to control groups. When the infection dose was 50 or 25 T. spiralis infective larvae, the reductions of AWI were 100% both at 4 and 10 dai; while the reduction of LPG at 30 dai was also 100% with respect to control groups. The levels of IgG and IgA anti-T. spiralis and IL-4 were significantly higher (P < 0.01) at 4 and 10 dai in mice from groups treated with L. casei than in animals in control groups; while at 10 dai, the levels of IFN-γ were higher in control mice (P < 0.01) than in L. casei-treated animals. The results suggest that frequent treatment of mice with L. casei induces a total protection against infection with low doses of T. spiralis.

Reviews on trichinellosis (II): neurological involvement.

Neurological involvement may occur in 0.2%-52% of cases with trichinellosis, generally in the most severely affected patients. This review focuses on neurotrichinellosis and includes a brief overview of selected cases reported in the literature. Our primary goal was to increase the awareness of infectious diseases specialists, neurologists, and general practitioners about these major complications with possible fatal outcome. Seventy seven of the cases, for which enough details were available, have been pooled for statistical analysis. The mean age of the investigated group was 34.6 ± 16.8 years. Patients with both focal and diffuse manifestations predominated (55.8%), and they were significantly older (40 ± 15.5 years old) than those who presented solely focal (28.9 ± 17.8 years old; p = 0.03) or diffuse lesions (27.9 ± 15.3 years old; p = 0.007). In most of the cases (59.7%), complete recovery was reported, whereas 23.4% of cases had sequelae and 16.9% of the patients died. Patients who died had significantly lower eosinophil counts (13.8% ± 14%) when compared with those who made complete recovery (28.7% ± 18%; p = 0.015) and the cases with sequelae (35% ± 17.9%; p = 0.006). To sum up, trichinellosis must be considered in the differential diagnosis of any patient with encephalitis or other central nervous system malady of ambiguous etiology.

Trichinellosis in children and adults: a 10-year comparative study in Western Romania.

INTRODUCTION: This study aims to evaluate and compare the epidemiologic, laboratory, clinical, and therapeutic aspects of trichinellosis in child and adult populations residing in Western Romania. PATIENTS AND METHODS: We have retrospectively investigated the medical records of patients with trichinellosis hospitalized between 1996 and 2005 in 5 infectious diseases hospitals located in 4 Western Romanian counties. RESULTS: Trichinellosis was diagnosed in 1344 patients, of whom 348 were children (26%) and 996 were adults (74%). The annual average incidence was 8.4 cases per 100,000 in children and 7.7 cases per 100,000 in adults. The mean ages for the children and adults were 11 ± 4.4 years and 38.1 ± 14.2 years, respectively. Myalgias and complications of trichinellosis were detected less frequently in children who also had lower eosinophil counts, leukocyte counts, and erythrocyte sedimentation rates compared with those for adults. Albendazole and mebendazole had comparable efficacies in adult patients; however, albendazole proved to be a better choice than mebendazole for the treatment of children. CONCLUSIONS: Trichinellosis occurs commonly among inhabitants of Western Romania. Although the general trend of decline in the incidence of trichinellosis cases cannot be used as a statistical predictor of future improvement, it provides encouragement and demonstrates the need for the implementation of correct public health and educational measures to fully prevent and eradicate trichinellosis in Western Romania.

Stimulation of TH1 response by Helicobacter pylori neutrophil activating protein decreases the protective role of IgE and eosinophils in experimental trichinellosis.

Th2 responses seem to play an important role in defence against Trichinella spiralis (Ts). The neutrophil Activating protein of Helicobacter pylori (HP-NAP), that induces IL-12, and IL-23 expression and shifts to Th1 allergen-specific Th2 cells in vitro was used as an anti-Th2 agent in BALB/c mice infected with T. spiralis. The muscle larvae (ML) burden was lower (p < 0.02) in untreated infected animals than those infected treated with HP-NAP. In both groups there was an inverse relationship between ML burden of each animal and total IgE level (controls: r -0.617, p = 0.0013 and HP-NAP-treated: r -0.678, p = 0.0001) or eosinophil count, evaluated in the same mouse on day 42 (r -0.390, p = 0.0592 and r -0.803, p = 0.0001, respectively). Inflammatory response around the nurse cell-parasite complex was significantly higher in HP-NAP-treated infected animals than in those untreated infected, on the contrary the number of eosinophils, counted around each complex was significantly lower in the first animal group. This study provides evidence of a powerful anti-Th2 activity in vivo by HP-NAP and for the partial protective effect of Th2 responses in T. spiralis infection.

[Trichinellosis]. / Trichinellose une zoonose parasitaire bien contrôlée en France.

Trichinellosis is a parasitic zoonosis caused by several species of the viviparous nematode of the genus Trichinella. Trichinellosis is a food borne zoonosis and can be contracted by eating raw or rare meat. This disease can be prevented by domestic and public health measures. France has thus considerably reinforced its system of prevention at slaughterhouses and in 2002 a Trichinella National Reference Centre was implemented. Trichinellosis is considered as a re-emergent disease throughout the world and more than 2500 cases have been reported in France since 1975. The possible severe neurological and cardiovascular complications, the risk of death highlight the need for a better knowledge of the disease and of its impact on public health.

[Visual system involvement in selected zoonotic diseases. II. Trichinellosis]. / Udzial narzadu wzroku w przebiegu niektórych zoonoz pasozytniczych. II. Wlosnica.

Trichinellosis is the medical problem that is still actual in our country. The paper presents recent taxonomic data regarding Trichinella spp. as well as selected epidemiologic aspects. Pathomechanism of trichinellosis and the outline of clinical pathology are also discussed. The image of lesions involving organ of sight, diagnostic methods and treatment are described with details as well. The paper is highlighting the role of ophthalmologist in early diagnosis of trichinellosis.

IgE enhances parasite clearance and regulates mast cell responses in mice infected with Trichinella spiralis.

Trichinella spiralis infection elicits a vigorous IgE response and pronounced intestinal and splenic mastocytosis in mice. Since IgE both activates mast cells (MC) and promotes their survival in culture, we examined its role in MC responses and parasite elimination in T. spiralis-infected mice. During primary infection, wild-type but not IgE-deficient (IgE(-/-)) BALB/c mice mounted a strong IgE response peaking 14 days into infection. The splenic mastocytosis observed in BALB/c mice following infection with T. spiralis was significantly diminished in IgE(-/-) mice while eosinophil responses were not diminished in either the blood or jejunum. Similar levels of peripheral blood eosinophilia and jejunal mastocytosis occurred in wild-type and IgE-deficient animals. Despite the normal MC response in the small intestine, serum levels of mouse MC protease-1 also were lower in parasite-infected IgE(-/-) animals and these animals were slower to eliminate the adult worms from the small intestine. The number of T. spiralis larvae present in the skeletal muscle of IgE(-/-) mice 28 days after primary infection was about twice that in BALB/c controls, and the fraction of larvae that was necrotic was reduced in the IgE-deficient animals. An intense deposition of IgE in and around the muscle larvae was observed in wild-type but not in IgE null mice. We conclude that IgE promotes parasite expulsion from the gut following T. spiralis infection and participates in the response to larval stages of the parasite. Furthermore, our observations support a role for IgE in the regulation of MC homeostasis in vivo.

Effect of viable or dead Lactobacillus casei organisms administered orally to mice on resistance against Trichinella spiralis infection.

The capacity of viable, dead Lactobacillus casei and supernatant from L. casei culture, administered by oral route, to induce resistance in mice against Trichinella spiralis infection was evaluated. The percentage of adult worm reduction in the intestine five days after T. spiralis infection as compared with the worm burden in the control group fluctuated between between 53.1 and 58% in mice treated with viable L. casei, while reductions in animals treated with dead lactobacilli or supernatant from L. casei culture were of 44 and 32.5% respectively. The percentage of larvae per gram of muscle tissue reductions, as compared with controls, obtained 30 days after infection ranged from 48.4 to 70.7% in rodents which ingested viable L. casei compared with the percentage reductions of 65.9 and 24% obtained respectively in mice treated with dead lactobacilli or with L. casei supernatant. The protective response observed in the present study may be explained on the basis of 1) lactobacilli colonization of the intestine, 2) macrophage processing of dead lactobacilli in local immune tissues and presentation of L. casei antigens to Th1 cells which, in turn, produced IL-2 to activate B cells and other T cells.

Trichinellosis: human disease, diagnosis and treatment.

In this review, the pathological mechanisms of human trichinellosis are presented, including a discussion on organ pathology, with particular attention paid to intestinal and muscular invasion. The clinical pattern in the acute stage of trichinellosis is presented, together with a classification of trichinellosis relative to severity of the disease. In turn, complications and diagnostic criteria are discussed. Drugs employed in the contemporary treatment of trichinellosis are presented (mainly those of the benzimidazole group and glucocorticosteroids) as well as indications for administering them, as related to severity of the disease.

Plasmapheresis in the treatment of trichinellosis.

Four patients with severe form of trichinellosis were treated with plasmapheresis. We noticed clinical improvement and favourable alterations in laboratory profiles (morphology and immune response parameters in the blood). The effect of plasmapheresis therapy in patients with trichinellosis was considered beneficial. No major complications of plasmapheresis were observed.