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1.
Parasitol Res ; 123(4): 196, 2024 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-38662084

RESUMO

Many pathogens are related to carcinogenesis. Chronic inflammation, as a result of persistent infection, leads to DNA damage, higher expression of oncogenes, decreased apoptosis and immunosuppression, which are some of the reasons for cancer induction. Among parasites, Schistosoma, Opistorchis and Clonorchis are recognised as infectious agents which contribute to cancer. A relationship between Anisakis and cancer was hypothesised because cellular responses to Anisakis products could result in inflammation and DNA damage. Previous research has shown a decrease in CD8+ γδ T-cells and an increase in αß and γδ T-cell apoptosis in colon cancer (CC) samples. Ninety-two CC patients and 60 healthy subjects were recruited. γδ and αß T-cells were analysed, and their apoptosis was evaluated. Anti-Anisakis antibodies were tested in sera from CC patients and controls. Anti-Anisakis IgG, IgM, IgA and IgE antibodies were significantly higher in CC patients. A significant increase in anti-Anisakis IgA levels was observed in patients with angiolymphatic invasion. The number of all γδ T-cells, as well as CD3+ CD4+ αß T-cells, was significantly lower in CC patients. The apoptosis of all T-cells was significantly increased in patients with CC. We observed a significantly higher percentage of anti-Anisakis IgE positive patients having a deficit of CD3+ γδ T-cells. Our results suggest a relationship between Anisakis and CC.


Assuntos
Anisakis , Anticorpos Anti-Helmínticos , Neoplasias do Colo , Humanos , Masculino , Pessoa de Meia-Idade , Anticorpos Anti-Helmínticos/sangue , Anticorpos Anti-Helmínticos/imunologia , Feminino , Neoplasias do Colo/imunologia , Neoplasias do Colo/parasitologia , Idoso , Animais , Anisakis/imunologia , Adulto , Apoptose , Idoso de 80 Anos ou mais , Subpopulações de Linfócitos T/imunologia , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Linfócitos T/imunologia
2.
Int Immunopharmacol ; 100: 108120, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34537480

RESUMO

AIMS: The objective of this investigation is to evaluate the mechanisms Anisakis simplex employs to modify its host immune system, regarding the larval antigens interactions with Toll-Like-Receptors (TLRs). METHODS AND RESULTS: In a previous study, we described that the stimulation of bone marrow derived dendritic cells (BMDCs) with A. simplex larval antigens drive an acute inflammatory response in BALB/c mice, but a more discrete and longer response in C57BL/6J. Moreover, when A. simplex larval antigens were combined with TLR agonists (TLR 1/2-9), they modified mainly TLR2, TLR4 and TLR9 agonists responses in both mice strains, and also TLR3, TLR5 and TLR7 in BALB/c. Antigen-presenting ability was analyzed by the detection of CD11c + cells expressing surface markers (CD80-86, MHC I-II), intracellular cytokines (IL-10, IL-12, TNF-α) and intracellular proteins (Myd88, NF-κß) by Flow Cytometry. Secreted IL-10 was measured by ELISA. CONCLUSION: Our findings confirm not only that the host genetic basis plays a role in the development of a Th2/Th1/Treg response, but also it states A. simplex larval antigens present specific mechanisms to modify the innate response of the host. As allergies share common pathways with the immune response against this particular helminth, our results provide a better understanding into the specific mechanisms of A. simplex allergy related diseases.


Assuntos
Anisakis/imunologia , Antígenos/imunologia , Imunomodulação , Larva/imunologia , Receptores Toll-Like/imunologia , Animais , Feminino , Antígenos H-2 , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Receptor 2 Toll-Like , Receptores Toll-Like/agonistas
3.
Int Immunopharmacol ; 90: 107241, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33321294

RESUMO

The impact of immunization with Anisakis simplex larval antigen on the occurrence and progression of experimental autoimmune encephalomyelitis (EAE) induced in mice was studied. C57BL/6J mice were immunized with the MOG35-55 peptide and one batch was treated with A. simplex total larval antigen on days 1, 8, 10 and 12 after EAE induction. Significantly higher values were obtained in the EAE clinical parameters of the antigen-treated group. Likewise, there was a significant decrease in the weights of the animals. Anisakis-treatment produced a significant decrease in anti-MOG35-55 specific IgG1 on day 21. On day 14 there was an increase in serum IL-2, IL-6, IL-10, IL-17A, and TGF-ß in the treated group. On day 21, a decrease in IL-4, IL-6, TNF-α, TGF-ß was observed. All brain determinations were made on day 21. The treatment decreased values of IL-6, IL-10, IL-17A and TNF-α. A. simplex antigen caused a significantly higher incidence of EAE and an advance in the appearance of the disease manifestations. However, treatment with the antigen was able to cause a decrease in proinflammatory cytokines (IL-6, IL-17A, and TNF-α) in nervous tissue that could establish a future preventive scenario for myelin damage.


Assuntos
Anisakis/imunologia , Anti-Inflamatórios não Esteroides/uso terapêutico , Antígenos/uso terapêutico , Encefalomielite Autoimune Experimental/tratamento farmacológico , Animais , Peso Corporal/efeitos dos fármacos , Química Encefálica/efeitos dos fármacos , Citocinas/sangue , Encefalomielite Autoimune Experimental/imunologia , Feminino , Imunoglobulina G/imunologia , Larva/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Glicoproteína Mielina-Oligodendrócito/imunologia , Fragmentos de Peptídeos/imunologia
4.
J Sci Food Agric ; 101(9): 3908-3916, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33348457

RESUMO

BACKGROUND: Freezing is considered the most suitable technological treatment to avoid Anisakis infection from eating raw or undercooked fish but modifications of their cuticles upon freezing may reduce their resistance to gastric fluids, provoking a greater release of allergens. This work aimed to study the relationship between freezing-induced modifications of Anisakis simplex s.l., antigen recognition, and resistance to oral and gastric digestion in spiked fish mince. RESULTS: (i) Differences between non-treated larvae and larvae that survived freezing / thawing were studied in terms of respiratory capacity, survival in simulated gastric fluid (SGF), recognition of antigens and allergens. (ii) Untreated (i.e. chilled) mince containing live larvae, mince frozen at two freezing rates, with a negative (uninfected) mince and a positive mince (infected with broken larvae) as controls, were subjected to the oral and gastric phases of a simulated digestion process. Anisakis able to survive freezing showed lower resistance to gastric fluid (i.e. faster mortality as compared to controls). Untreated larvae released significantly more antigens than freeze-surviving larvae but only after 96 h in SGF. In treatments rendering complete larvae mortality, the highest loss of larvae integrity was found upon fast freezing. There was a positive correlation between antigen release and the number of ruptures of larvae after the oral digestion phase, whereas a more complex trend was observed after oral plus gastric digestion phases. CONCLUSION: These results suggest a new factor to consider for sensitized patients and suggest that the numbers of L3 should be reduced before industrial freezing to minimize risk. © 2020 Society of Chemical Industry.


Assuntos
Anisaquíase/metabolismo , Anisakis/metabolismo , Antígenos de Helmintos/metabolismo , Contaminação de Alimentos/análise , Gadiformes/parasitologia , Suco Gástrico/enzimologia , Animais , Anisaquíase/parasitologia , Anisakis/classificação , Anisakis/genética , Anisakis/imunologia , Antígenos de Helmintos/análise , Manipulação de Alimentos , Congelamento , Humanos , Larva/classificação , Larva/genética , Larva/imunologia , Larva/metabolismo , Modelos Biológicos
5.
Parasitol Int ; 80: 102223, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33137497

RESUMO

To better understand the molecular mechanisms underlying allergens and parasite immunity and discover the stage-enriched gene expression of fish-borne zoonotic nematodes in the stomach, we used RNA-seq to study the transcriptome profiles of Anisakis pegreffii (Nematoda: Anisakidae, AP) in simulated gastric juice. Mobile L3 larvae were incubated in simulated medium at 37 °C in 5% CO2 (AP-GJ) and the control group larvae were collected in PBS under the same conditions (AP-PBS). We found that the sequences of A. pegreffii were highly similar to Toxocara canis sequences. Among the transcripts, there would be 138 up-regulated putative genes and 251 down-regulated putative genes in AP-GJ group. Several lipid binging-related genes were more highly expressed in AP-GJ larvae. Moreover, 17 allergen genes were up-regulated and 29 were down-regulated in AP-GJ larvae. Eleven allergen genes belonged to one or more of the following three categories: biological process, cellular component, and molecular function. According to KEGG analysis, the main pathways that were represented included protein processing in transcription, immune system, cancer, and infectious disease. In particular, the most significant changes in the expression of parasite-derived allergen products occurred in AP-GJ larvae. This study helps us to extend our understanding of the biology of the fish-borne zoonotic parasite A. pegreffii and could be helpful for more precise risk assessment and providing guidelines for allergic consumers.


Assuntos
Alérgenos/genética , Anisakis/fisiologia , Imunidade Inata/genética , Transcriptoma/imunologia , Alérgenos/imunologia , Animais , Anisakis/genética , Anisakis/crescimento & desenvolvimento , Anisakis/imunologia , Suco Gástrico/química , Imunidade Inata/imunologia , Larva/genética , Larva/crescimento & desenvolvimento , Larva/imunologia , Larva/fisiologia
6.
Sci Rep ; 10(1): 11275, 2020 07 09.
Artigo em Inglês | MEDLINE | ID: mdl-32647149

RESUMO

The high frequency of infection by Anisakis simplex (A. simplex) has led to an increase in IgE sensitization, turning allergy to this parasite a relevant contemporary health problem. Improving the lack of conventional diagnosis test specificity is crucial to better understand these clinical scenarios. Specific IgE (sIgE) to A. simplex extract by ImmunoCAP (Anisakis-sIgE) was determined in sera from 403 blood donors (BD) from Cantabria (North of Spain) of which 51 subjects resulted sensitized. Among these latter, 47 were asymptomatic (sABD). The values of total IgE, prick-test, Anisakis-sIgE, and sIgE to Ani s 1 (anti-rAni s 1) and Ani s 7 (anti-rAni s 7) were compared between 46 sABD and 49 A. simplex allergic patients. The IgE seroprevalence by ImmunoCAP among BD was 12.65%. Allergic patients and sABD showed significant differences in all serum biomarkers evaluated. The area under the curve was assessed for Anisakis-sIgE (0.892), sIgE-rAni s 1 (0.672) and sIgE-rAni s 7 (0.668). After a severe reaction, significantly higher levels of Anisakis-sIgE and sIgE anti-rAni s 1 were detected. Determinations of sIgE by ImmunoCAP, Ani s 1 and Ani s 7 presented different sensitization patterns between allergic and asymptomatic individuals. The Ani s 1 allergen arises as a possible biomarker to detect patients at risk of suffering severe allergic reactions.


Assuntos
Alérgenos/imunologia , Anisaquíase/imunologia , Antígenos de Helmintos/imunologia , Biomarcadores/sangue , Proteínas de Ligação ao Cálcio/imunologia , Proteínas de Helminto/imunologia , Hipersensibilidade/parasitologia , Adulto , Idoso , Animais , Anisakis/imunologia , Estudos Transversais , Dermatophagoides pteronyssinus/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina E/imunologia , Masculino , Pessoa de Meia-Idade , Penaeidae/imunologia , Prevalência , Estudos Prospectivos , Curva ROC , Estudos Soroepidemiológicos
7.
Genes (Basel) ; 11(6)2020 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-32585969

RESUMO

The view of the nucleolus as a mere ribosomal factory has been recently expanded, highlighting its essential role in immune and stress-related signalling and orchestrating. It has been shown that the nucleolus structure, formed around nucleolus organiser regions (NORs) and attributed Cajal bodies, is prone to disassembly and reassembly correlated to various physiological and pathological stimuli. To evaluate the effect of parasite stimulus on the structure of the leukocyte nucleolus, we exposed rat peripheral blood mononuclear cells (PBMC) to the crude extract of the nematode A. pegreffii (Anisakidae), and compared the observed changes to the effect of control (RPMI-1640 media), immunosuppressive (MPA) and immunostimulant treatment (bacterial lipopolysaccharide (LPS) and viral analogue polyinosinic:polycytidylic acid (poly I:C)) by confocal microscopy. Poly I:C triggered the most accentuated changes such as nucleolar fragmentation and structural unravelling, LPS induced nucleolus thickening reminiscent of cell activation, while MPA induced disassembly of dense fibrillar and granular components. A. pegreffii crude extract triggered nucleolar segregation, expectedly more enhanced in treatment with a higher dose. This is the first evidence that leukocyte nucleoli already undergo structural changes 12 h post-parasitic stimuli, although these are likely to subside after successful cell activation.


Assuntos
Anisaquíase/imunologia , Anisakis/imunologia , Nucléolo Celular/imunologia , Região Organizadora do Nucléolo/imunologia , Animais , Anisaquíase/genética , Anisaquíase/patologia , Anisakis/patogenicidade , Nucléolo Celular/efeitos dos fármacos , Humanos , Imunossupressores/farmacologia , Células Intersticiais de Cajal/efeitos dos fármacos , Células Intersticiais de Cajal/imunologia , Leucócitos/imunologia , Leucócitos/metabolismo , Leucócitos Mononucleares/imunologia , Lipopolissacarídeos/farmacologia , Região Organizadora do Nucléolo/efeitos dos fármacos , Região Organizadora do Nucléolo/genética , Poli I-C/farmacologia
8.
Genes (Basel) ; 11(6)2020 06 22.
Artigo em Inglês | MEDLINE | ID: mdl-32580523

RESUMO

In plant and animal nematode parasites, proteins derived from esophageal gland cells have been shown to be important in the host-nematodes relationship but little is known about the allergenic potential of these proteins in the genus Anisakis. Taking into account the increase of anisakiasis and allergies related to these nematodes, immunoreactive properties of gland cell proteins were investigated. Two hundred ventricles were manually dissected from L3 stage larvae of Aniskakis simplex s.s. to allow direct protein analysis. Denaturing gel electrophoresis followed by monochromatic silver staining which revealed the presence of differential (enriched) proteins when compared to total nematode extracts. Such comparison was performed by means of 1D and 2D electrophoresis. Pooled antisera from Anisakis spp.-allergic patients were used in western blots revealing the presence of 13 immunoreactive bands in the ventricular extracts in 1D, with 82 spots revealed in 2D. The corresponding protein bands and spots were excised from the silver-stained gel and protein assignation was made by MALDI-TOF/TOF. A total of 13 (including proteoforms) were unambiguously identified. The majority of these proteins are known to be secreted by nematodes into the external environment, of which three are described as being major allergens in other organisms with different phylogenetic origin and one is an Anisakis simplex allergen.


Assuntos
Anisaquíase/imunologia , Anisakis/imunologia , Doenças dos Peixes/imunologia , Interações Hospedeiro-Parasita/imunologia , Alérgenos/imunologia , Alérgenos/isolamento & purificação , Animais , Anisaquíase/genética , Anisaquíase/parasitologia , Anisakis/patogenicidade , Esôfago/imunologia , Esôfago/parasitologia , Doenças dos Peixes/genética , Doenças dos Peixes/parasitologia , Humanos , Larva/genética , Larva/imunologia , Larva/patogenicidade , Filogenia , Proteínas/imunologia , Alimentos Marinhos/parasitologia , Especificidade da Espécie , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
9.
Int Arch Allergy Immunol ; 181(7): 507-511, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32392567

RESUMO

INTRODUCTION: Anisakiasis is a zoonosis of parasitic origin whose diffusion seems to be continuously increasing. OBJECTIVE: The aim of this study was to evaluate the benefits of a fish-free diet in patients allergic to Anisakis simplex as well as underlining the importance of awareness and prevention. Furthermore, we aimed to investigate the critical issues related to the spread of anisakiasis in relation to eating habits. METHODS: Patients were assessed by means of skin prick tests (SPTs) and targeted laboratory testing, with an 18-month-long fish-free diet being recommended in cases of severe sensitization. The degree of awareness about anisakiasis was evaluated from interviews. Patients were subjected to follow-up visits after 18 months. RESULTS: A total of 70 cases of sensitization to A. simplex were evaluated. The Interview answers highlighted a general state of misinformation among patients and healthy subjects along with a remarkable underestimation of anisakiasis-related risks. An overall lack of care regarding eating habits and diet plans also emerged. In 21 patients affected by severe sensitization, clinical and laboratory evaluations were repeated after 18 months of the subjects being on a fish-free diet. There was a remarkable improvement in serum IgE levels and clinical symptoms. CONCLUSION: Data analysis proved the need to implement new and more effective awareness-raising and prevention campaigns in order to reduce the incidence of anisakiasis. It is crucial to establish an adequate diet therapy for sensitized patients. Evaluation of cytokine patterns suggests how a polyphenol-rich regime can activate regulatory T cell function and possibly reduce the allergic and inflammatory components of the disease.


Assuntos
Anisaquíase/dietoterapia , Anisaquíase/prevenção & controle , Hipersensibilidade/etiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Anisakis/imunologia , Feminino , Humanos , Hipersensibilidade/dietoterapia , Hipersensibilidade/prevenção & controle , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Adulto Jovem
10.
PLoS Negl Trop Dis ; 14(4): e0008177, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32243436

RESUMO

Urticaria remains a major problem in terms of aetiology, investigation, and management, and although parasitic diseases are considered potential causes, the absence of a consistent link between parasitic infections and skin allergy symptoms leads to the need for a deeper study of parameters that support this association. The objectives of this study were to analyse a possible relationship between parasitism by Ascarididae (Toxocara canis and Anisakis simplex) and the clinical expression of urticaria and to identify possible parasitic molecular markers for improving the diagnosis of unknown urticaria aetiology. The prevalence of Toxocara and Anisakis infestations was evaluated by measuring the levels of specific IgG (sIgG) and IgE (sIgE) antibodies against crude extracts and isolated components from whole larvae of Anisakis simplex (Ani s 1, Ani s 3 and Ani s 7) and Toxocara canis (TES-120, TES-70, TES-32 and TES-26) using immunologic and molecular diagnostic methods. A cross-sectional study was performed in a group of 400 individuals. The study group consisted of 95 patients diagnosed with urticaria (55 with chronic urticaria and 40 with acute urticaria). A control group consisted of 305 subjects without urticaria (182 diagnosed with respiratory allergy and 123 without allergy). Statistically significant differences were demonstrated in the seroprevalence of specific IgG and IgE antibodies between the urticaria patients and the healthy general population when isolated ascarid antigens were evaluated. The prevalence of IgG antibodies against Ani s 1, IgE antibodies against TES-120 and IgE antibodies against TES-70 were significantly different between the control individuals (healthy general population) and patients with urticaria. Moreover, the urticaria patient group demonstrated a higher seroprevalence of antibodies (sIgE and sIgG) against Anisakis simplex larva whole extract than the control group but just with statistically diferences when sIgE was evaluated. The presence of IgE and/or IgG antibodies against Ani s 3 (tropomyosin) can help to discriminate between patients with and without urticaria. Both ascarids seem to be associated with urticaria, although in our region, Anisakis seems to have greater involvement than Toxocara in this relationship. Molecular diagnostics can be used to associate urticaria with parasite infestations. Tropomyosin and Ani s 1 were the most relevant markers to demonstrate the association between urticaria and the most relevant Ascarididae parasites in our region.


Assuntos
Anticorpos Anti-Helmínticos/imunologia , Antígenos de Helmintos/imunologia , Ascaridoidea/patogenicidade , Urticária/diagnóstico , Urticária/imunologia , Urticária/parasitologia , Adolescente , Adulto , Alérgenos/imunologia , Animais , Anisaquíase/imunologia , Anisakis/imunologia , Estudos Transversais , Feminino , Proteínas de Helminto/imunologia , Humanos , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Larva/imunologia , Masculino , Pessoa de Meia-Idade , Estudos Soroepidemiológicos , Pele/imunologia , Toxocara canis/imunologia , Adulto Jovem
12.
PLoS Negl Trop Dis ; 14(1): e0008038, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31986138

RESUMO

We undertook the first study systematically evaluating the risk of Anisakis-sensitization in Croatian fish-processing workers and potential genetic susceptibility to anisakiasis. Anti-Anisakis IgE seroprevalence and risk factors for 600 employees of Croatian fish processing facilities and 466 blood donor controls, were assessed by indirect ELISA targeted with: recombinant Ani s 1 and Ani s 7 allergens, an Anisakis crude extract, the commercial ImmunoCAP kit, and questionnaires. Genetic susceptibility to anisakiasis was evaluated by genotypisation of human leukocytes alleles (HLA). Anti-Anisakis seropositive and a fraction of negative subjects were also assessed by ELISA and Western Blot (WB) for IgG seroprevalence to Trichinella spp. Overall, the observed anti-Anisakis seroprevalence inferred by indirect ELISA was significantly higher in fish processing workers (1.8%, 95% CI 0.9-3.3%) compared to the controls (0%, 0-0.8%). Seven out of 11 Ani s 1 and Ani s 7-positives and none of selected 65 negative sera, tested positive on whole-Anisakis extract (ImmunoCAP), whereas Anisakis crude extract ELISA detected 3.9% (2.4-6.0%) seropositives in fish processing workers, three (14%) of which showed IgE reactivity to milk proteins. The highest risk associated with Anisakis-sensitization among workers was fishing in the free time, rather than any of attributes related to the occupational exposure. Although no association was observed between anti-Anisakis seropositivity and wearing gloves or protective goggles, the majority of workers (92%) wore protective gloves, minimizing the risk for Anisakis sensitization via skin contact. Six HLA alleles within DRB1 gene were significantly associated with seropositivity under dominant, allelic or recessive models. All sera confirmed negative for anti-Trichinella spp. IgG. The study exhaustively covered almost all marine fish processing workers in Croatia, reflecting real-time Anisakis sensitization status within the industry, already under the influence of wide array of allergens.


Assuntos
Anisakis/imunologia , Peixes/parasitologia , Manipulação de Alimentos , Hipersensibilidade , Exposição Ocupacional , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos , Croácia , Dispositivos de Proteção dos Olhos , Luvas Protetoras , Proteínas de Helminto , Humanos , Fatores de Risco , Trichinella/imunologia
14.
Parasite ; 26: 52, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31441776

RESUMO

Anisakis pegreffii, a recognised etiological agent of human anisakiasis, is a parasite of homeothermic hosts at the adult stage and of ectothermic hosts at the third larval stage. Among distinct factors, temperature appears to be crucial in affecting parasite hatching, moulting and to modulate parasite-host interaction. In the present study, we investigated the gene transcripts of proteins having an antigenic role among excretory secretory products (ESPs) (i.e., a Kunitz-type trypsin inhibitor, A.peg-1; a glycoprotein, A.peg-7; and the myoglobin, A.peg-13) after 24 h, in A. pegreffii larvae maintained in vitro, under controlled temperature conditions. Temperatures were 37 °C and 20 °C, resembling respectively homeothermic and ectothermic hosts conditions, and 7 °C, the cold stress condition post mortem of the fish host. Primers of genes coding for these ESPs to be used in quantitative real-time PCR were newly designed, and qRT-PCR conditions developed. Expression profiles of the genes A.peg-1 and A.peg-13 were significantly up-regulated at 20 °C and 37 °C, with respect to the control (larvae kept at 2 °C for 24 h). Conversely, transcript profiles of A.peg-7 did not significantly change among the chosen temperature conditions. In accordance with the observed transcript profiles, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed the presence of the three target ESPs at 37 °C, while only A.peg-13 was observed at 7 °C. The results suggest that temperature conditions do regulate the gene expression profiles of A.peg-1 and A.peg-13 in A. pegreffii larvae. However, regulation of the glycoprotein A.peg-7 is likely to be related to other factors such as the host's immune response.


Assuntos
Anisaquíase/veterinária , Anisakis/genética , Antígenos de Helmintos/genética , Proteínas de Helminto/genética , Temperatura , Animais , Anisaquíase/imunologia , Anisakis/imunologia , Antígenos de Helmintos/imunologia , Peixes/parasitologia , Interações Hospedeiro-Parasita , Larva/genética , Reação em Cadeia da Polimerase em Tempo Real , Transcriptoma
15.
Parasite Immunol ; 41(4): e12616, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30719721

RESUMO

AIMS: The objective of this work is to investigate whether Anisakis simplex larval antigens present immunomodulatory properties by the induction of tolerogenic dendritic cells (DCs) from two strains of mice (BALB/c and C57BL/6J). METHODS AND RESULTS: We used mouse bone marrow-derived DCs. We determined their antigen-presenting ability by expression of membrane markers (MHC I and MHC II, CD80, CD86) and intracellular expression levels of IL-10 and IL-12 cytokines. We also analysed whether stimulation with A simplex larval antigens is enhanced by the co-administration of the TLR4 and TLR9 agonists [LPS E coli 026B6 and CpG (ODN1826), respectively]. Two differential types of responses were found in the two mouse strains studied: the BALB/c strain showed an acute and inflammatory response, whereas the C57BL/6J mice developed a more discrete and resistant response. This suggests the coexistence of two opposing responses generated by A simplex larval antigens and confirms that the host genetic basis plays a role in the development of a Th2 or Treg response. CONCLUSION: The study of the mechanisms by which Anisakis manipulates the immune response through anti-inflammatory molecules is of interest not only for the direct application on the development of anthelmintic strategies, but also for the development of new anti-inflammatory products.


Assuntos
Anisakis/imunologia , Antígenos de Helmintos/imunologia , Células Dendríticas/imunologia , Larva/imunologia , Linfócitos T Reguladores/imunologia , Células Th2/imunologia , Animais , Anisakis/embriologia , Antígeno B7-1 , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Larva/metabolismo , Lipopolissacarídeos/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Oligodesoxirribonucleotídeos/farmacologia , Transdução de Sinais/imunologia , Receptor 4 Toll-Like/agonistas , Receptor Toll-Like 9/agonistas
16.
Exp Parasitol ; 197: 9-15, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30584924

RESUMO

Serodiagnosis of human anisakidosis is presently hampered by the current lack of standardised serological assays that allow sensitive and specific detection of Anisakidae-specific antibodies in human patients. In the present study, we comparatively evaluated the diagnostic value (by IgG-ELISA) of excretory-secretory antigens (ESAgs) of Anisakis simplex, Pseudoterranova decipiens and Contracaecum osculatum, representing the most frequently found genera responsible for human infection. In addition, we tested also a mix of the three ES preparations (Mix-ESAgs) as well as two recombinant allergens of A. simplex, rAni s 1 and rAni s 7. ES antigen from C. osculatum yielded the best diagnostic performance in IgG-ELISA-based serodiagnosis of the Spanish anisakidosis patients investigated in this study (relative serodiagnostic sensitivity 100%; specificity 89%) as compared to A. simplex ES-antigen (93% versus 57%) and P. decipiens (67% versus 93%) or a mix of the three ES antigens (100% versus 44%), respectively. Cross-reactions of C. osculatum ES antigen with serum-antibodies from patients suffering from other helminth infections were rare and were exclusively found with few sera from toxocariasis, ascariasis, and filariasis patients. The two recombinant allergens rAni s 1 and rAni s 7 did not prove sufficiently sensitive and specific in order to justify a further evaluation of these antigens regarding their suitability in IgG-ELISA-based serodiagnosis of human anisakidosis. In conclusion, the C. osculatum-ESAg-ELISA remains as key candidate to be further assessed for the serodiagnosis of symptomatic anisakidosis in different endemic regions.


Assuntos
Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/imunologia , Infecções por Ascaridida/diagnóstico , Ascaridoidea/imunologia , Ensaio de Imunoadsorção Enzimática , Imunoglobulina G/sangue , Animais , Anisakis/imunologia , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática/normas , Humanos , Coelhos , Sensibilidade e Especificidade , Testes Sorológicos
17.
Ann Parasitol ; 65(4): 303-314, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-32191412

RESUMO

Nematodes of the genus Anisakis are worldwide distributed marine species parasitized many fish and cephalopod species as larvae and sea mammals as adult form. Anisakiosis as food-borne disease is an important public health problem worldwide. Human become infected by eating raw or undercooked fish or squids. Well documented are gastrointestinal response to infection but increasingly allergic symptoms were observed also after eating well cooked fish. This is because some of allergens of Anisakis are thermostable and resistant to pepsin treatment. Due to a significant increase in human mobility and global transport of fresh products like fish on ice, food-borne diseases require educational campaigns that pay attention to threats in various parts of the world.


Assuntos
Anisaquíase , Anisakis , Alimentos Marinhos , Zoonoses , Animais , Anisaquíase/parasitologia , Anisakis/química , Anisakis/imunologia , Europa (Continente) , Hipersensibilidade Alimentar/parasitologia , Humanos , Hipersensibilidade , Larva/fisiologia , Alimentos Marinhos/parasitologia , Zoonoses/parasitologia
18.
Int Marit Health ; 69(4): 264-269, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30589066

RESUMO

BACKGROUND: Anisakis is a marine nematode. Its larvae can be found encysted in several species, both in the abdominal cavity and in the adjacent musculature. The most commonly affected commercial species are hake, whiting, cod, and mackerel. The prevalence in fish varies according to the fishing area and the size of the host. MATERIALS AND METHODS: Until now only three species have been confirmed to be involved in human anisakiasis, the most common ones being A. simplex sensu stricto (s.s.) and A. pegreffii, and anecdotally, A. physeteris. Infestation in humans occurs when they eat raw or undercooked parasitized fish or cephalopods (pickled, cold-smoked, salted, semi-preserved, prepared in certain Asian styles like sushi or sashimi, ceviche). RESULTS: The majority of anisakiasis cases have been described by Japanese authors. However, over the last few years there has been an increase in the number of cases reported in other countries including Italy and Spain. It is estimated that its incidence in the European Union is 0.32/100,000, and in the Basque Country (Spain), this parasite is responsible for 10% of anaphylaxis cases and 32% of urticaria cases in adults aged 40-60 years, around 300 cases/year. Anisakis-related disease in the work environment (occupational disease) is less common. CONCLUSIONS: We present three cases of the occupational disease in Spain due to a type I hypersensitivity to Anisakis simplex in individuals who handle fish (one fishmonger, one supermarket employee, and one chef).


Assuntos
Anisakis/imunologia , Dermatite Alérgica de Contato/etiologia , Dermatite Ocupacional/etiologia , Manipulação de Alimentos , Hipersensibilidade Imediata , Adulto , Animais , Anisaquíase/parasitologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Alimentos Marinhos/parasitologia , Espanha
19.
PLoS One ; 13(9): e0203671, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30235242

RESUMO

Anisakis simplex spp. sensitization rates have increased worldwide, with a significant impact on health-care systems. To date, no clear-cut diagnostic criteria and laboratory algorithm have been established, so anisakiasis still represents an under-reported health problem whose clinical manifestations, when present, mimic the much more common allergic and digestive disorders. Aim of the study was to systematically review the available literature on the prevalence of sensitization against Anisakis in the general population and in specific population groups, taking into account the impact of the different available diagnostic techniques on the epidemiological data. Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement, relevant papers reporting Anisakis sensitization epidemiological data were found covering a period ranging from 1996 to February 2017. Overall, 41 studies comprising 31,701 participants from eleven countries were included in the qualitative synthesis. General asymptomatic population resulted sensitized to Anisakis in 0.4 to 27.4% of cases detected by means of indirect ELISA or ImmunoCAP specific IgE detection, and between 6.6% and 19.6% of the samples by Skin prick test (SPT). Occupationally exposed workers (fishermen, fishmongers and workers of fish-processing industries) documented specific IgE between 11.7% and 50% of cases, whereas SPT positivity ranged between 8% and 46.4%. Symptomatic allergic patients to any kind of allergen were found to be positive to Anisakis specific IgE detection between 0.0% (in children with mastocytosis) to 81.3% (among adults with shellfish allergy). Results highlighted that hypersensitivity prevalence estimates varied widely according to geographical area, characteristics of the population studied, diagnostic criteria and laboratory assays. Further studies are needed to overcome the documented misdiagnosis by improving the diagnostic approach and, consequently, providing more affordable estimates in order to address public health interventions on populations at high risk of exposure to Anisakis and to tailor health services related to specific groups.


Assuntos
Anisakis/imunologia , Anticorpos Anti-Helmínticos/análise , Hipersensibilidade/epidemiologia , Animais , Ensaio de Imunoadsorção Enzimática , Humanos , Hipersensibilidade/diagnóstico , Hipersensibilidade/parasitologia , Exposição Ocupacional/estatística & dados numéricos
20.
Parasite ; 25: 23, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29637891

RESUMO

A retrospective survey was carried out over the years 2010-2014 among all Parasitology laboratories of University hospitals in France (ANOFEL network). The objective was to estimate the incidence of anisakidosis in France as new culinary habits such as the consumption of raw fish (sushi) or undercooked fish are increasing. A total of 37 cases of anisakidosis were notified by all French laboratories: 7 proven cases with evidence of a worm, 12 possible cases with abdominal pain after consumption of raw fish with detection of anti-Anisakis precipitins, and 18 allergic cases defined as acute manifestations after consumption of fish, associated with specific IgE for Anisakis. The median age of affected individuals was 42 years (11-69) and there was a significant predominance of women (67%). Compared with previous surveys in France, this study indicates a decrease in clinical cases of anisakidosis and illustrates the emerging allergic potential of anisakids.


TITLE: Étude nationale rétrospective de l'anisakidose en France (2010-2014) : diminution de l'incidence, prédominance féminine et augmentation des allergies. ABSTRACT: Une enquête rétrospective a été réalisée au cours des années 2010-2014 auprès des laboratoires de parasitologie des hôpitaux universitaires de France (réseau ANOFEL). L'objectif était d'estimer l'incidence de l'anisakidose en France depuis l'émergence de nouvelles habitudes culinaires comme la consommation de poissons crus (sushi) ou insuffisamment cuits. Les laboratoires français de Parasitologie ont ainsi notifié 37 cas d'anisakidose: 7 cas certains avec présence du ver, 12 cas possibles se caractérisant par des douleurs abdominales après consommation de poisson cru et la présence de précipitines anti-Anisakis et 18 cas allergiques définis comme des manifestations aiguës après consommation de poisson associées à des IgE spécifiques anti-Anisakis. L'âge médian des cas était de 42 ans (11-69) et il y avait une prédominance féminine significative (67 %). Par rapport aux enquêtes précédentes en France, cette étude indique une diminution des cas cliniques d'anisakidose mais illustre le potentiel allergique émergent des anisakidés.


Assuntos
Anisaquíase/epidemiologia , Anisaquíase/etiologia , Peixes/parasitologia , Hipersensibilidade/epidemiologia , Hipersensibilidade/parasitologia , Alimentos Crus/parasitologia , Dor Abdominal , Adolescente , Adulto , Idoso , Animais , Anisaquíase/complicações , Anisakis/imunologia , Anisakis/isolamento & purificação , Criança , Feminino , Parasitologia de Alimentos , França/epidemiologia , Programas Governamentais , Humanos , Hipersensibilidade/imunologia , Incidência , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores Sexuais , Inquéritos e Questionários , Adulto Jovem
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