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Helminth regulation of host IL-4Ralpha/Stat6 signaling: mechanism underlying NOS-2 inhibition by Trichinella spiralis.

Gastrointestinal nematode infection is known to alter host T cell activation and has been used to study immune and inflammatory reactions in which nitric oxide (NO) is a versatile player. We previously demonstrated that Trichinella spiralis infection inhibits host inducible NO synthase (NOS-2) expression. We now demonstrate that (i) an IL-4 receptor alpha-subunit (IL-4Ralpha)/Stat6-dependent but T cell-independent pathway is the key for the nematode-induced host NOS-2 inhibition; (ii) endogenous IL-4 and IL-13, the only known IL-4Ralpha ligands, are not required for activating the pathway; and (iii) treatment of RAW264.7 cells with parasite-cultured medium inhibits NOS-2 expression but not cyclooxygenase 2 expression. We propose that a yet-unidentified substance is released by the nematode during the host-parasite interaction.

Animals↗

Further potential limitations of the undifferentiated faecal egg count reduction test for the detection of anthelmintic resistance in sheep.

Fifteen (36%) of 42 mixed gastro-intestinal nematode infections in sheep, identified as drench-susceptible by the undifferentiated faecal egg count reduction test, were found to harbour anthelmintic-resistant worms when analysed on the basis of changes in the egg counts of individual nematode genera. Most of these cases involved resistance in a single nematode genus, with Ostertagia and Trichostrongylus being implicated most frequently. The possible contributory role of larval culture results in helping to reduce the chances of the faecal egg count reduction test producing these and similar types of errors is discussed.

Journal Article↗

Evaluation of the treatment of intestinal helminthiases with albendazole in Djohong (North Cameroon).

246 inhabitants of Djohong, a township located in North-east Cameroon, presented with single or mixed nematode infections. They were treated by 400 mg albendazole in a single dose. The results were evaluated either with the Kato thick-smear technique and the Ritchie technique on stools and/or the Graham test: albendazole has proved to be 100% efficacious in pinworm and roundworm treatment, 63% to 84% in hookworm treatment (difference due to the type of technique used). Albendazole showed a mean efficacy in whipworm treatment (about 50% cure rate). In case of residual worm infection, the egg count is reduced from 80% to 90% which is of the utmost importance. The relative treatment failures occurring with large worm load. This systematic treatment of a whole population considerably reduces the spread of nematode eggs over the soil (18-fold for Nector americanus, 10-fold for Trichuris trichiura). This broad spectrum anthelmintic is strongly recommended in mass treatments for its efficacy and excellent tolerance, as its ovicide action reduces the probability of fecal pollution of the environment by treated patients who have residual parasites.

Adolescent↗

Cloning and characterisation of a prohibitin gene from infective larvae of the parasitic nematode Toxocara canis.

Infective larvae of the parasitic nematode Toxocara canis express an mRNA (Tc-pro-1) encoding a predicted protein that shares significant homology with prohibitin, a protein involved in inhibition of cell proliferation - The closest homologues of Tc-pro-1 include an expressed sequence tag (EST) from Caenorhabditis elegans and Drosophila L2Cc, a protein thought to be essential for larval development and moulting. Other homologues include prohibitin from rat and human and an EST from Saccharomyces cerevisiae. Parasite life cycles generally include periods of developmental arrest, which in the larvae of T. canis may persist for many years without loss of metabolic activity. This report of the first full-length gene encoding prohibitin from a parasitic nematode raises interesting suggestions about the potential role of prohibitin in diapause and in the regulation of moulting in development.

Amino Acid Sequence↗

Efficacy of doramectin for treatment of experimentally induced infection with gastrointestinal nematodes in calves.

OBJECTIVE: To determine the efficacy of injectable doramectin against experimentally induced infections with adult and larval (L4) stages of gastrointestinal nematodes in calves. PROCEDURES: In each experiment, between 20 (experiments 1 and 2) and 30 (experiment 3) parasite-free yearling calves were selected and allotted to groups of 10/group. All calves received oral inoculation of the following infective larval stages of these nematode species: Cooperia onchophora, C punctata, C zurnabada, Cooperia spp, Haemonchus placei, Nematodirus helvetianus, Oesophagostomum radiatum, Ostertagia ostertagi, Trichostrongylus axei, and T colubriformis. Infections or treatments were timed so that, by day of treatment, the population of each parasite species had matured to the stage to be tested (adult or L4). On day of treatment, calves received doramectin (200 micrograms/kg of body weight, SC), and infected nonmedicated control calves received saline solution SC at the same time. All calves were slaughtered at least 14 days after treatment. RESULTS: Efficacy against the following adult and L4 species was > or = 99%: C onchophora, C punctata, C zurnabada, Cooperia spp, H placei, Oes radiatum, O ostertagi, and T colubriformis. Doramectin was also 100% efficacious against the L4 stage of T axei. Efficacy against the adult and L4 stages of N helvetianus was 9.6 and 77.4 to 83.3%, respectively. CONCLUSIONS: Doramectin, at a dosage of 200 micrograms/kg, is effective in controlling the prevalent gastrointestinal nematodes (adult and L4 stages) found in naturally infected calves. CLINICAL RELEVANCE: Doramectin, a new anthelmintic, was effective against recent field strains of the most prevalent gastrointestinal parasites found in cattle in the United States, and could provide another alternative for control of these parasites.

Animals↗

CD4+-dependent immunity to Onchocerca volvulus third-stage larvae in humans and the mouse vaccination model: common ground and distinctions.

Onchocerciasis is a major filarial disease and is the second most common cause of infectious blindness in the world. Disease development after infection with Onchocerca volvulus varies widely and is determined by the host's immune response to the parasite. Vector control and administration of ivermectin has reduced infection and disease rates significantly. However, limitations of these programmes, including ivermectin's selective activity on microfilariae, the need for 10-15 years of annual treatments, logistical obstacles and the potential emergence of drug-resistant strains demand alternative strategies. A vaccine that targets O. volvulus infective third-stage larvae (L3) could provide an additional tool to guarantee successful elimination of infection with O. volvulus. An essential step in the development of immunoprophylactic procedures and reagents is the identification of host immune responses toward antigens of O. volvulus L3 and L3 developing to the fourth-stage larvae that are associated with protection against these stages of the parasite. This review summarises the recent advancements in understanding the immune mechanisms in particular the CD4(+) responses to L3 stages in humans and in the mouse vaccination model. Comparison between the two uncovered common immunological elements in naturally exposed humans and mice vaccinated with radiation attenuated L3 or recombinant O. volvulus antigens, as well as significant differences. These studies promisingly suggest that the O. volvulus mouse model is a very useful adjunct to the studying of natural infection in humans and could provide us with the tools to identify the target molecules and the effector immune correlates of protection in humans responsible for attrition of L3 stages. Since some of these antigens may exist in other nematodes, any insight gained into the mechanisms of vaccine-induced anti-O. volvulus L3 protective immunity in both humans and mice could be applicable to the development of vaccines against other nematode infections.

Animals↗

Disruption of the murine IL-4 gene blocks Th2 cytokine responses.

Murine T-helper clones are classified into two distinct subsets (Th1 and Th2) on the basis of their patterns of lymphokine secretion. Th1 clones secrete interleukin-2 (IL-2), tumour necrosis factor-beta (TNF-beta) and interferon-gamma (IFN-gamma), whereas Th2 clones secrete IL-4, IL-5 and IL-10 (ref. 1). These subsets are reciprocally regulated by IL-4, IL-10 and IFN-gamma and differentially promote antibody or delayed-type hypersensitivity responses. To evaluate whether IL-4 is required for mounting Th2 responses, we generated IL-4-mutant mice (IL-4-/-) and assessed the cytokine secretion pattern of T cells both from naive and Nippostrongylus brasiliensis infected mice. CD4+ T cells from naive IL-4-/- mice failed to produce Th2-derived cytokines after in vitro stimulation. The levels of Th2 cytokines IL-5, IL-9 and IL-10 from CD4+ T cells obtained after nematode infection were significantly reduced. The reduced IL-5 production in IL-4-/- mice correlated with reduced helminth-induced eosinophilia, which has been shown to be dependent on IL-5 in vivo. We conclude that IL-4 is required for the generation of the Th2-derived cytokines and that immune responses dependent on these cytokines are impaired.

Alleles↗

Prolongation of allograft survival by Nippostrongylus brasiliensis is associated with decreased allospecific cytotoxic T lymphocyte activity and development of T cytotoxic cell type 2 cells.

BACKGROUND: We have demonstrated that infection with Nippostrongylus brasiliensis (Nb), which induces strong type 2 responses, prolongs kidney allograft survival in rats. Here, we confirm that this effect is not species-specific and address immune modulation in allospecific T-cell responses mediated by nematode infection. METHODS: C57BL/6 mice were injected with Nb or phosphate-buffered saline. Four days later, mice were transplanted with BALB/c hearts and graft survival was assessed. In other experiments, Nb-infected mice were immunized with BALB/c spleen cells and allospecific T-cell responses were determined in vitro. RESULTS: In this study, we show that Nb prolongs cardiac allograft survival in mice. Further, spleen T cells from Nb-infected, allo-immunized mice exhibit reduced allospecific cytotoxic T-lymphocyte activity. In contrast, allospecific proliferation of T cells in the mixed lymphocyte reaction was not reduced by Nb, ruling out immunosuppression as the mechanism of Nb-induced allograft survival. Nb infection induced IL-4 and IL-6 and inhibited IFN-gamma production by T cells in response to allo-antigen. Furthermore, anti-IL-4 treatment reduced allospecific T-cell proliferation from Nb-infected but not control mice, indicating that type 2 allospecific T cells develop in the presence of Nb. We also double-stained T cells for CD8 and IL-4 and showed that Nb induces an 8-fold increase in Tc2 cell numbers. CONCLUSIONS: These results are consistent with a hypothesis that Nb mediates prolongation of allograft survival through induction of type 2 immunity, including the development of regulatory Tc2 cells, and subsequent inhibition of allospecific cytotoxic T-lymphocyte activity.

Animals↗

Potentiation of IgE responses to third-party antigens mediated by Ascaris suum soluble products.

A reductive approach was used to examine the potentiation of IgE responses by nematode infection. Ascaris homogenized extract, Ascaris pseudocoelomic (body) fluid (ABF) and purified Ascaris allergen (ABA) were tested for their ability to act as protein carriers and as mediators of potentiated IgE responses to third-party (ovalbumin; OVA) responses. All three nematode products were excellent protein carriers for the hapten dinitrophenol and showed significantly better activity in this respect than OVA. Neither ABF nor ABA enhanced the level of the IgE response to the third-party antigen but both prolonged the response markedly. ABF, but not ABA, induced high levels of total circulating IgE when given at the same time as OVA with alum. The data suggest that the enhancement and prolongation of IgE responses by nematodes may be two separate but related activities.

Allergens↗

Comparative efficacy of levamisole, thiabendazole and fenbendazole against cattle gastrointestinal nematodes.

Efficacy of two older anthelmintics, levamisole and thiabendazole, was compared with a newer benzimidazole, fenbendazole, against naturally acquired gastrointestinal nematode infections in cattle superimposed with experimental infections of Bunostomum phlebotomum and Dictyocaulus viviparus. Twenty-four crossbred beef heifers of 7-9 months of age and 152 kg in average weight were randomly allocated to four groups of six calves. The cattle grazed on pastures contaminated with larvae of gastrointestinal nematodes and the lungworm for 2 months prior to Day 0. Treatment groups were as follows: Group 1--levamisole, topical at 10 mg kg-1; Group 2--thiabendazole paste at 110 mg kg-1; Group 3--fenbendazole paste at 10 mg kg-1; Group 4--untreated controls. All calves were necropsied for worm recovery between 8 and 10 days after treatment. Fecal egg/larval per gram counts at 18 and 42 h post-treatment indicated greatest reductions in Groups 1 and 2. By 7 days post-treatment, reduction in counts for all treated groups ranged from 99.1 to 100%, except for the 66.7% reduction of B. phlebotomum in Group 2. Seven nematode species were present in a sufficient number of untreated controls for valid efficacy assessment at necropsy. Efficacy of fenbendazole was 100% against all species, including Cooperia spp. L4 and immature (E5) D. viviparus. The overall efficacy of levamisole and thiabendazole was generally high (93.0-100% against Haemonchus placei adults, Cooperia punctata and C. spatulata adult males, Cooperia spp. adult females, Oesophogastomum radiatum, B. phlebotomum, and D. viviparus adults).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Role of specific immunoglobulin E to excretory-secretory antigen in diagnosis and prognosis of hookworm infection.

Total immunoglobulin E (IgE) and specific IgE were assayed by a radioimmunosorbent test and a reverse enzyme immunoassay in patients with hookworm infections before and after treatment. A total of 77 subjects (30 patients with hookworm infections and 47 subjects as controls) were studied. Both specific and total IgE levels in serum and jejunal juice were raised in hookworm patients. There was a significant decrease in IgE levels after therapy. Total IgE levels were raised in other nematode infections, but specific IgE levels were low. The reverse enzyme immunoassay for specific IgE was highly specific (96%) and sensitive (100%) and may be used in the serodiagnosis of hookworm infections.

Adolescent↗

Meloidogyne incognita: molecular and biochemical characterisation of a cathepsin L cysteine proteinase and the effect on parasitism following RNAi.

RNA interference has been used to investigate the function of a cathepsin L cysteine proteinase Mi-cpl-1, in the plant-parasitic nematode Meloidogyne incognita. A reduction in gene transcript was observed and the number of nematodes infecting plants was reduced by almost 60% as was the number of established females producing eggs at 21 days post-infection. The cysteine proteinase activity of M. incognita, reported by the substrate GLUpNA, was inhibited by the cysteine proteinase inhibitor Oc-IDeltaD86. A reduction in cysteine proteinase activity was also seen following RNAi of Mi-cpl-1 in J2 stage nematodes. In situ hybridization analysis in young and mature female nematodes has shown that Mi-cpl-1 is expressed in the intestine, which suggests that its product is a digestive enzyme. The effects of knocking-out Mi-cpl-1gene function were consistent with a reduction in feeding efficiency. Here, we have shown a correlation between transcript abundance proteinase activity and parasitic success of M. incognita.

Animals↗

Very low helminth infection in sheep grazed on pastures fertilised by sewage sludge or cattle slurry.

Helminth infection acquired by lambs grazing on pastures fertilised either by urban sewage sludge or cattle slurry were studied in temperate Central Western France. The aim was to assess the risk of larval cestodoses in lambs after sewage application and of digestive tract nematode infection following the slurry application. Twenty-six sheep were allocated on two paddocks of 0.7 ha, one fertilised with sludge and the other with cattle slurry. The delay between application and actual grazing was 6 weeks; grazing on these paddocks extended from mid July to beginning of November 2002. The herbage biomass was slightly increased in the sludge paddock but it did not result in an increase of lamb live weight, compared with the slurry paddock. The lambs did not acquire cysticercosis or any other larval cestodoses in the sewage sludge group and only very limited infections with Cooperia spp. and Nematodirus spp. were observed in the slurry group. It was concluded that in our conditions the helminth risk was extremely low and was not a cause of restriction of the use of these biowastes.

Animals↗

Recombinant cDNA clones for immunodiagnosis of strongyloidiasis.

Because diagnosis of strongyloidiasis by stool examination is unreliable and because of the potential for serious disease in Strongyloides infections, there is need for improved diagnostic aids to facilitate recognition and treatment of this parasitic infection. Serologic testing, when available, requires antigen preparation from infected primates or dogs that can be difficult to maintain. Several recombinant clones from a cDNA library prepared from the infective stage of Strongyloides stercoralis were characterized. Serologic results indicate that the recombinant proteins were equally or more reactive than the larval somatic antigen. No cross-reactivity with recombinant antigen 5a was found with sera from patients with filarial or intestinal nematode infections. Recombinant antigens 5a and 12a detected parasite-specific IgE and IgG4 antibodies in Strongyloides-infected patients. Sequence analysis showed these antigens to be rich in proline and charged amino acids. Lack of homology from database searches suggests that the antigens are unique. These recombinant antigens should be useful in diagnostic and epidemiologic studies of strongyloidiasis.

Amino Acid Sequence↗

'Candidatus Paenicardinium endonii', an endosymbiont of the plant-parasitic nematode Heterodera glycines (Nemata: Tylenchida), affiliated to the phylum Bacteroidetes.

Bacteria-like endosymbionts of females of the plant-parasitic nematodes Globodera rostochiensis and Heterodera goettingiana and juveniles of Heterodera glycines were first observed during transmission electron microscopy (TEM) studies conducted in the 1970s. These organisms were characterized as being rod-shaped, ranging in size from 0.3 to 0.5 microm in diameter and 1.8 to 3 microm in length and containing structures labelled as striated inclusion bodies or tubular structures. A population of H. glycines was obtained from the soybean field where infected nematodes were first discovered in order to conduct TEM studies of females and males and to determine the phylogenetic position of the H. glycines endosymbiont among bacteria by studying the 16S rRNA and gyrB gene sequences. The bacterium was observed in the pseudocoelom and intestine of juveniles, females and males, in hypodermal chords of juveniles and males, in ovary walls and in oocytes and spermatozoa. The bacterium was polymorphic, measuring 0.4-0.8 x 2.5-4.5 microm, and many specimens contained an array of microfilament-like structures similar to those observed in "Candidatus Cardinium hertigii", the endosymbiont of Encarsia spp. wasps. Phylogenetic analysis of the 16S rRNA and gyrB genes of the H. glycines-infecting bacterium revealed 93 % and 81 % sequence identity, respectively, to the homologous genes in "Candidatus C. hertigii". Thus, the name "Candidatus Paenicardinium endonii" is proposed for the bacterial endosymbiont of the plant-parasitic nematode H. glycines.

Actin Cytoskeleton↗

The antibody repertoire in infection and vaccination with Dictyocaulus viviparus: heterogeneity in infected cattle and genetic control in guinea pigs.

The antigen recognition profiles of serum antibody from calves infected or vaccinated with irradiated Dictyocaulus viviparus larvae were analysed by immunoprecipitation of radio-iodinated in vitro-released excretory-secretory materials from live adult parasites. Immunoprecipitates were analysed by SDS-PAGE and considerable heterogeneity in antigen recognition between individual animals was observed, regardless of infection regimen. This heterogeneity was also found to occur amongst outbred guinea pigs infected with the parasite and permitted an examination of the genetics of the effect using inbred guinea pigs (Strains 2 and 13). The antibody repertoires of the two strains were distinct, with only slight variation occurring between individuals within a strain. Previous work on nematode infections in rodents has demonstrated a role for the major histocompatibility complex (MHC) in the control of the immune repertoire. If this, as is probable, holds for the guinea pig, then it can be ascribed to the MHC Class II region because Strain 2 and Strain 13 bear identical Class I alleles but disparate Class II alleles. Whilst there is no evidence to date that the efficiency of vaccination of cattle is influenced by genetic factors, the operation of vaccines based on a single or a few molecularly cloned parasite antigens might be seriously compromised by the kind of genetic restriction to the immune repertoire described here.

Animals↗

Effects of interleukin 12 on immune responses and host protection in mice infected with intestinal nematode parasites.

The cytokine interleukin (IL) 12 stimulates T cell and natural killer cell production of interferon (IFN) gamma and inhibits T cell production of IL-4. We investigated the effects of IL-12 on cytokine gene expression, immunoglobulin (Ig)E, mucosal mast cell, and eosinophil responses, and the course of infection in mice inoculated with the nematode parasite Nippostrongylus brasiliensis, as well as the IFN-gamma dependence of these effects. IL-12 stimulated IFN-gamma and IL-10 gene expression during primary and secondary N. brasiliensis infections and inhibited IL-3, IL-4, IL-5, and IL-9 gene expression during primary infections but had little inhibitory effect during secondary infections. IL-12 inhibited IgE, mucosal mast cell, and blood and tissue eosinophil responses during primary infections, but only eosinophil responses during secondary infections. IL-12 enhanced adult worm survival and egg production during primary, but not secondary infections. IL-12 needed to be administered by day 4 of a primary infection to inhibit IgE and mucosal mast cell responses, and by day 6 to strongly inhibit eosinophil responses and to enhance worm survival and fecundity. Anti-IFN-gamma mAb inhibited the effects of IL-12 on IgE secretion, intestinal mucosal mastocytosis, and parasite survival and fecundity, but did not affect IL-12 inhibition of eosinophilia. These observations indicate that IL-12, if administered during the initiation of eosinophilia. These observations indicate that IL-12, if administered during the initiation of an immune response, can change the response from one that is characterized by the production of T helper (Th)2-associated cytokines to one characterized by the production of Th-1 associated cytokines. However, IL-12 treatment has less of an effect once the production of Th2-associated cytokines has become established. In addition, our results provide evidence that Th2-associated responses protect against, and/or Th1-associated responses exacerbate, nematode infections.

Animals↗

Infection by anisakid nematodes contracaecum spp. in the Mayan cichlid fish 'Cichlasoma (Nandopsis)' urophthalmus (Gunther 1862).

Larval nematodes that parasitize the Mayan cichlid fish 'Cichlasoma (Nandopsis)' urophthalmus (Günther 1862) in southern Florida were identified as Contracaecum spp. (Nematoda: Anisakidae, Anisakinae). The objective of this study was to determine whether infection intensity and prevalence of these parasites differ between a brackish water and freshwater habitat or through ontogeny in the freshwater habitat only. The nematodes were removed from the abdominal cavity of the fishes and counted. Infection intensity was compared between habitats using analysis of covariance and evaluated through ontogeny using Spearman rank order correlation. Prevalence was compared between habitats and between adults and juveniles from the freshwater habitat using a z-test. Although infection intensity did not differ between habitats, infection prevalence was greater at the freshwater site (FWS). Both the prevalence and intensity of nematode infection increased through ontogeny at the FWS, and no nematode was found in fishes that were smaller than 93 mm standard length. Thus, the parasites appear to accumulate during the lifetime of the fishes.

Abdominal Cavity↗