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Strongyloides stercoralis mesenteric lymphadenopathy: clue to the etiopathogenesis of intestinal pseudo-obstruction in HIV-infected patients.

Mesenteric lymph node involvement in Strongyloides stercoralis hyperinfective states, described as an autopsy finding, remains a relatively poorly recognized and possibly underreported, antemortem phenomenon. Furthermore, the occurrence of S stercoralis mesenteric lymphadenopathy as a tocsin of bowel strongyloidiasis and the clue to the cause of intestinal pseudo-obstruction are undescribed. We report S stercoralis mesenteric lymphadenopathy and intestinal pseudo-obstruction in 5 HIV seropositive male patients, 21 to 42 years, who presented with abdominal pain and variable vomiting, diarrhea, and constipation. All were pale, pyrexial, and emaciated with abdominal distension. The preoperative diagnosis was intestinal obstruction. Poor clinical response on conservative therapy necessitated laparotomy. Dilated small bowel loops, ascites, and mesenteric lymphadenopathy were consistently noted; a diagnosis of pseudo-obstruction due to underlying tuberculosis or lymphoma was made. The mesenteric lymph nodes were biopsied. The pertinent nodal features were a dense infiltrate of eosinophils, eosinophil microabscesses and degranulation, a focal Splendore-Hoeppli phenomenon, and randomly disposed, but elusive, S stercoralis filariform larvae. Clinical deterioration confirmed intestinal complications at repeat laparotomy. Intestinal resections were performed in 4 patients; histopathologic appraisal confirmed intestinal strongyloidiasis. All patients died within 3 to 7 days after surgery. Heightened awareness of S stercoralis mesenteric lymphadenopathy as a sentinel of intestinal strongyloidiasis and etiopathogenetic clue of intestinal pseudo-obstruction may allow timely diagnosis and medical treatment and avoidance of further surgery, potentially reducing the long-term morbidity associated with S stercoralis hyperinfection.

AIDS-Related Opportunistic Infections↗

Strongyloides stercoralis: Amphidial neuron pair ASJ triggers significant resumption of development by infective larvae under host-mimicking in vitro conditions.

Resumption of development by infective larvae (L3i) of parasitic nematodes upon entering a host is a critical first step in establishing a parasitic relationship with a definitive host. It is also considered equivalent to exit from the dauer stage by the free-living nematode Caenorhabditis elegans. Initiation of feeding, an early event in this process, is induced in vitro in L3i of Strongyloides stercoralis, a parasite of humans, other primates and dogs, by culturing the larvae in DMEM with 10% canine serum and 5mM glutathione at 37 degrees C with 5% CO(2). Based on the developmental neurobiology of C. elegans, resumption of development by S. stercoralis L3i should be mediated, in part at least, by neurons homologous to the ASJ pair of C. elegans. To test this hypothesis, the ASJ neurons in S. stercoralis first-stage larvae (L1) were ablated with a laser microbeam. This resulted in a statistically significant (33%) reduction in the number of L3i that resumed feeding in culture. In a second expanded investigation, the thermosensitive ALD neurons, along with the ASJ neurons, were ablated, but there was no further decrease in the initiation of feeding by these worms compared to those in which only the ASJ pair was ablated.

Animals↗

The fork head transcription factor FKTF-1b from Strongyloides stercoralis restores DAF-16 developmental function to mutant Caenorhabditis elegans.

The purpose of this study was to determine whether Strongyloides stercoralis FKTF-1, a transcription factor of the FOXO/FKH family and the likely output of insulin/IGF signal transduction in that parasite, has the same or similar developmental regulatory capabilities as DAF-16, its structural ortholog in Caenorhabditis elegans. To this end, both splice variants of the fktf-1 message were expressed under the control of the daf-16alpha promoter in C. elegans carrying loss of function mutations in both daf-2 (the insulin/IGF receptor kinase) and daf-16. Under well-fed culture conditions the majority (91%) of untransformed daf-2; daf-16 double mutants developed via the continuous reproductive cycle, whereas under the same conditions 100% of daf-2 single mutants formed dauers. Transgenic daf-2; daf-16 individuals expressing fktf-1b showed a reversal of the double mutant phenotype with 75% of the population forming dauers under well-fed conditions. This phenotype was even more pronounced than that of daf-2; daf-16 mutants transformed with a homologous rescuing construct, daf-16alpha::daf-16a (56% dauers under well fed conditions), indicating that S. stercoralis fktf-1b can almost fully rescue loss-of-function mutants in C. elegans daf-16. By contrast, daf-2; daf-16 mutants expressing S. stercoralis fktf-1a, encoding the second splice variant of FKTF-1, showed a predominantly continuous pattern of development identical to that of the parental double mutant stock. This indicates that, unlike FKTF-1b, the S. stercoralis transcription factor FKTF-1a cannot trigger the shift to dauer-specific gene expression in C. elegans.

Animals↗

Successful transgenesis of the parasitic nematode Strongyloides stercoralis requires endogenous non-coding control elements.

Critical investigations into the cellular and molecular biology of parasitic nematodes have been hindered by a lack of modern molecular genetic techniques for these organisms. One such technique is transgenesis. To our knowledge, the findings reported here demonstrate the first heritable DNA transformation and transgene expression in the intestinal parasite Strongyloides stercoralis. When microinjected into the syncitial gonads of free-living S. stercoralis females, a construct fusing the S. stercoralis era-1 promoter, the coding region for green fluorescent protein (gfp) and the S. stercoralis era-1 3' untranslated region was expressed in intestinal cells of normally developing F1 transgenic larvae. The frequency of transformation and GFP expression among F1 larvae was 5.3%. By contrast, expression of several promoter::gfp fusions incorporating only Caenorhabditis elegans regulatory elements was restricted to abortively developing F1 embryos of S. stercoralis. Despite its lack of regulated expression, PCR revealed that one of these C. elegans-based vector constructs, the sur-5::gfp fusion, is incorporated into F1 larval progeny of microinjected female worms and then transmitted to the F2 through F5 generations during two host passages conducted without selection and punctuated by free-living generations reared in culture. Heritable DNA transformation and regulated transgene expression, as demonstrated here for S. stercoralis, constitute the essential components of a practical system for transgenesis in this parasite. This system has the potential to significantly advance the molecular and cellular biological study of S. stercoralis and of parasitic nematodes generally.

Animals↗

Eosinophilic oophoritis: association with positive Strongyloides stercoralis serology and clinical response to ivermectin.

BACKGROUND: Localized eosinophilic infiltration causing ovarian dysfunction is an unusual clinical problem. CASE: We report a 16-year-old girl with abdominal pain, ovarian cysts, and biopsy-proven eosinophilic oophoritis, which resulted in right oophorectomy. Subsequently, a large cyst developed in the left ovary, causing abdominal pain that was worse at the time of her menses, and biopsy again showed numerous eosinophilic microabscesses. Unexpectedly, a parasite serology test to Strongyloides stercoralis was positive, although stool tests for ova and parasites were negative and the total IgE and total eosinophil count were normal. After treatment with ivermectin, the patient's abdominal pain resolved, the serologic antibody titers to S stercoralis returned to normal, and subsequent ultrasonographic evaluations showed involution of the large cyst in the remaining ovary. CONCLUSION: Eosinophilic oophoritis is a new disorder of localized tissue eosinophilia.

Adolescent↗

Toll-like receptor 4 (TLR4) is required for protective immunity to larval Strongyloides stercoralis in mice.

TLR4 is important for immunity to various unicellular organisms and has been implicated in the immune responses to helminth parasites. The immune response against helminths is generally Th2-mediated and studies have shown that TLR4 is required for the development of a Th2 response against allergens and helminth antigens in mice. C3H/HeJ mice, which have a point mutation in the Tlr4 gene, were used in this study to determine the role of TLR4 in protective immunity to the nematode Strongyloides stercoralis. It was demonstrated that TLR4 was not required for killing larval S. stercoralis during the innate immune response, but was required for killing the parasites during the adaptive immune response. No differences were seen in the IL-5 and IFN-gamma responses, antibody responses or cell recruitment between wild type and C3H/HeJ mice after immunization. Protective immunity was restored in immunized C3H/HeJ mice by the addition of wild type peritoneal exudate cells in the environment of the larvae. It was therefore concluded that the inability of TLR4-mutant mice to kill larval S. stercoralis during the adaptive immune response is due to a defect in the effector cells recruited to the microenvironment of the larvae.

Animals↗

daf-7 and the development of Strongyloides ratti and Parastrongyloides trichosuri.

daf-7 is a key ligand in one of the three pathways that control dauer larva development in Caenorhabditis elegans. Given the similarities between dauer larvae of free-living nematodes and third stage infective larvae of animal parasitic nematodes, we hypothesised that daf-7 may be involved in the development of these infective larvae. To investigate this, we cloned daf-7 orthologues from Strongyloides ratti and Parastrongyloides trichosuri and analysed their RNA level by semi-quantitative RT-PCR during the S. ratti and P. trichosuri life cycles and in a range of in vitro and in vivo conditions. We found that, in both species, the RNA level of daf-7 was low in free-living stages but peaked in the infective L3 (iL3) stage with little or no expression in the parasitic stages. This contrasts with the daf-7 RNA level in C. elegans, which peaks in L1, decreases thereafter, and is absent in dauer larvae. The RNA level of daf-7 in infective larvae was reduced by larval penetration of host skin or development in the host, but not by a shift to the body temperature of the host.

Amino Acid Sequence↗

An expressed sequence tag analysis of the life-cycle of the parasitic nematode Strongyloides ratti.

14,761 expressed sequence tags (ESTs) were generated, representing five stages during the parasitic and free-living phases of the life-cycle of the parasitic nematode Strongyloides ratti. These ESTs formed 4152 clusters, of which 97% contained 10 or fewer ESTs and 66% were singletons. These 4152 clusters are likely to represent approximately 20% of S. ratti's genes. The clusters' consensus sequences were used to assign each cluster to one of three databases: (i) Caenorhabditis elegans and C. briggsae sequences; (ii) other nematode sequences; (iii) non-nematode sequences. This approach has identified putative nematode-specific genes, that may be targets for developing approaches for parasitic nematode control. Approximately 25% of the clusters have no significant alignments and may therefore represent novel genes. The EST representation between the libraries was used to analyse stage-specific or -biased expression in silico. This showed that 81% of clusters are present in only one library and 12% are present in any two libraries, indicating substantial stage-specificity of gene expression. The 30-most abundantly expressed clusters were analysed in further detail. Many of these have significantly different parasitic- or free-living-specific or -biased expression. Many of the parasitic-specific genes are, as yet, uncharacterised: one of these represents 25% of all ESTs obtained from the parasitic stage.

Animals↗

Strongyloides ratti: chemokinesis of glycolytic enzyme- and lectin-treated third-stage infective larvae in vitro.

The infective third-stage larvae (L3s) of Strongyloides ratti, a parasitic nematode in rodents, showed two types of chemokinesis on a gradient of sodium chloride (NaCl) in an in vitro agarose tracking assay. The types were a consistent directional avoidance behavior under unfavorable environmental conditions and a reduced avoidance behavior under favorable conditions. We examined the effects of treatments with glycolytic enzymes and lectins by analyzing the avoidance behavior. L-Fucose dehydrogenase, hyaluronidase, beta-glucosidase, alpha-mannosidase, beta-galactosidase, concanavalin A, wheat germ agglutinin and soybean agglutinin exhibited inhibitory or enhancive effects on chemokinesis. We also confirmed the sites of the amphids of L3s aside from the mouth at the anterior end by scanning electron microscopy, and that concanavalin A-binding sites existed in the vicinity of the amphids using lectin-histochemistry. The carbohydrate moieties in the amphids of S. ratti L3s may play an important role as chemosensors in perceiving environmental cues.

Animals↗

[Strongyloides stercoralis infection simulating polyarteritis nodosa].

INTRODUCTION: Parasitic infection can present with many different clinical manifestations. EXEGESIS: A 77 year-old Russian woman, who's been living in France since 50 years was admitted for polyarthritis, myalgia, fever, abdominal pain, and eosinophilia simulating polyarterisis nodosa. Before admission, she was treated by steroids for polymyalgia rheumatica. The diagnosis of Strongyloides stercoralis was performed by parasitologic analysis of feces and colic biopsies. The outcome was favourable under treatment by ivermectine and steroid withdrawal. CONCLUSION: S. stercoralis can be associated with reactive arthritis. Case reports of S. stercoralis infection mimicking systemic vasculitis are exceptionnal.

Adrenal Cortex Hormones↗

Distribution of human Oesophagostomum bifurcum, hookworm and Strongyloides stercoralis infections in northern Ghana.

A cross-sectional study was carried out in 216 randomly selected, representative rural villages in the northeastern part of Ghana from March 1995 to May 1998. Inhabitants of randomly selected households, stratified by age and gender, were included. The geographical position of villages was recorded with a global positioning system (GPS). The prevalence of Oesophagostomum, hookworm and Strongyloides stercoralis infections in a study population of 20250 people was determined by microscopic examination of larvae in stool cultures. The overall prevalence was 10.2, 50.6 and 11.6% for the three nematodes, respectively. Hookworm infections were seen in all but one (99.5%) and S. stercoralis in 88.4% of the 216 villages, while Oesophagostomum infections were found to be common in a limited area with prevalences varying from 0 to 75%. An association was found between Oesophagostomum and hookworm infection, both at the individual and at the village level. Spatial analysis of the prevalence data indicated that the endemic area is relatively clearly demarcated to the south of the study area.

Adolescent↗

Enhanced survival of rats concurrently infected with Trypanosoma brucei and Strongyloides ratti.

The interaction between the blood protozoan parasite, Trypanosoma brucei and the gastrointestinal nematode parasite, Strongyloides ratti was studied in outbred white albino rats. Rats were grouped and given either single infection with T. brucei or S. ratti or concurrently infected with both parasites. Blood parasitaemia and packed cell volume, faecal egg/larva output, adult worm burden and survivability were monitored in order to assess the interactive effects of the infections. All trypanosome-infected rats became parasitaemic within 1 week of infection but surprisingly parasitaemia was higher in the single than concurrently infected group of rats. In addition all animals with single T. brucei infection had died by 14 days after the infection, whereas animals with concurrent infection were still alive by day 28 after the infection when the experiment was terminated. Concurrent infection resulted in significant increase in daily S. ratti egg/larval output in faeces (P < 0.01), but lesser number of adult worms were recovered from the intestine of sacrificed rats on day 8 post-infection. Taken together these results suggest that T. brucei and S. ratti interact in a manner that ameliorates their pathogenic effects resulting in a decrease in the level of parasitaemia and intestinal worm burden and in increased life span of the infected rats. These results differ from the classical immunosuppressive attributes of T. brucei and the results are discussed in the context of the possible immune responses that might have contributed to this outcome and the potential significance of the findings in alternative control method of trypanosomosis.

Animals↗

Chemoattraction and chemorepulsion of Strongyloides stercoralis infective larvae on a sodium chloride gradient is mediated by amphidial neuron pairs ASE and ASH, respectively.

Depending on its concentration, sodium chloride acts as either an attractant or a repellant to the infective larvae (L3i) of Strongyloides stercoralis. On a concentration gradient, L3i are attracted to 0.05 M NaCl, but repelled by 2.8M. To test the hypothesis that amphidial neurons ASE and ASH might mediate attraction and repulsion, respectively, these neurons, and control neurons as well, were ablated in hatchling larvae with a laser microbeam. After the larvae attained infectivity (L3i), they were tested on a NaCl gradient. When placed at low salinity, 73.5% of normal controls migrated "up" the gradient, while 26.4% crawled randomly. In contrast, only 20.6% of ASE-ablated L3i migrated "up" the gradient, while 79.4% migrated randomly. Ablation-control ASK-ablated L3i (58.8%) migrated "up" the gradient while 41.1% crawled randomly. When placed at a region of high salinity, 100% of normal control L3i migrated "down" the gradient, whereas 62.5% of ASH-ablated L3i migrated randomly, the remaining 37.5% migrating "down" the gradient. In sharp contrast with ASH-ablated L3i, 94.1% of ablation-control larvae, i.e. ASK-ablated L3i, migrated "down" the gradient. Migration behavior of ASE- and ASH-ablated L3i was significantly different (P < 0.001) from that of ASK-ablated L3i and normal controls. It is noteworthy that 87.5% of ASE-ablated L3i that failed to exhibit chemoattractive behavior were actively chemorepelled from high salinity. Also, 70.0% of ASH-ablated L3i that failed to be chemorepelled from high salinity were capable of chemoattractive behavior, indicating that the worms had retained their behavioral responses except for those associated with the targeted neurons.

Animals↗

Parasitological and serological diagnosis of Strongyloides stercoralis in domesticated dogs from southeastern Brazil.

Canine strongyloidiasis is a parasitic infection caused by the nematode Strongyloides stercoralis and presents a great zoonotic potential. Its confirmation, using coproparasitological methods, is difficult. The detection of serum specific antibodies, however, may facilitate the diagnosis. The aims of this study were to determine the presence of S. stercoralis through the use of parasitological methods and to detect specific antibodies to the parasite in serum samples from domestic dogs by using the indirect fluorescent antibody test (IFAT) on slides and the enzyme-linked immunosorbent assay (ELISA). A total of 215 dogs of various breeds, from the cities of Uberlândia, Araxá and Campo Belo in the State of Minas Gerais, were examined and distributed according to age into the following groups: (I) 19 males and 20 females of 1-2 months old; (II) 11 males and 20 females of 2-month- to 1-year-old and (III) 41 males and 104 females, from 1 to 7 years old. Coproparasitological results showed that 63/215 (29.3%) of the dogs presented some kind of parasite, with two (0.9%) dogs (one from Araxá and the other from Uberlândia) passing S. stercoralis larvae in the feces. Serological results revealed antibodies to S. stercoralis in 45/215 (20.9%) of the dogs, with seropositivity rates of 0% (0/39) in Group I, 22.6% (7/31) in Group II, and 26.2% (38/145) in Group III. No serological cross-reactivity between S. stercoralis and hookworms or Ascaridae was found. Hookworm infections were seen in 31 dogs, but only one of these dogs (infected with both hookworm and Cystoisospora spp.) was S. stercoralis seropositive by IFAT. The present study demonstrated, for the first time, natural S. stercoralis infections in dogs diagnosed by coproparasitological and serological methods. It was concluded that the detection of specific antibodies to S. stercoralis by IFAT and ELISA may contribute to the diagnosis of canine strongyloidiasis.

Age Factors↗

A cDNA encoding a pepsinogen-like, aspartic protease from the human roundworm parasite Strongyloides stercoralis.

Using degenerate oligonucleotide primers based on conserved active site residues, we have isolated a cDNA encoding an aspartic protease from the nematode parasite Strongyloides stercoralis, an important, enteric pathogen of humans. cDNAs encoding the aspartic protease were isolated from the infective, third stage larvae of the parasite as well as from free-living, rhabditiform larvae. Based on comparisons of other aspartic proteases, the cDNA encoded a short signal peptide, an enzyme pro-segment of 35 amino acid residues, and mature enzyme of 337 residues. Homology alignments using the proenzyme sequence showed that the novel S. stercoralis zymogen was 36% identical to human pepsinogen A and 36% identical to pepsinogen C (progastricin) from humans and macaques. Phylogenetic analyses using the Phylip program and analysis of Glx/Asx and Leu/Ile ratios indicated that the proenzyme was closely related to pepsinogen A-like enzymes from the free-living nematode Caenorhabditis elegans and Haemonchous contortus, a nematode parasite of the gastro-intestinal tract of sheep. We have termed this novel enzyme strongyloidespepsin.

Amino Acid Sequence↗

Immunoaffinity-isolated antigens induce protective immunity against larval Strongyloides stercoralis in mice.

The objective of this study was to identify soluble protein antigens that would induce protective immunity against infective-stage larvae (L-3) of Strongyloides stercoralis in mice. Deoxycholate (DOC)-soluble proteins derived from L-3, adsorbed to aluminum hydroxide, induced protective immunity in BALB/c mice. The immunized mice generated parasite-specific IgG that could transfer passive immunity to naïve animals. The protective antibodies bound to parasite antigens found in the muscles and nerve cords of the L-3. An IgG affinity chromatography column generated with IgG from the sera of DOC-immunized mice was used to purify specific larval antigens. Proteins were eluted from the affinity column with sizes of 80, 75, 61, 57, 43, and 32 kDa. This antigen pool stimulated both proliferation and IL-5 production by splenocytes recovered from mice immunized with live L-3. Vaccination of mice with the immunoaffinity-isolated antigens led to significant protective immunity, with 83% of challenge larvae killed. This study demonstrates that IgG-isolated proteins are candidate antigens for a vaccine against larval S. stercoralis.

Animals↗

Strongyloides venezuelensis: longitudinal distribution of adult worms in the host intestine is influenced by mucosal sulfated carbohydrates.

Mechanisms for the longitudinal distribution of parasitic females of Strongyloides venezuelensis in the host intestine were investigated in mice. Adult worms were mostly recovered from the anterior-most one-third of the small intestine throughout the infection after infective larvae inoculation. Surgically implanted adult worms established well in the small intestinal mucosa, either in the duodenum or in the ileum, whereas a few worms could establish in the large intestine. Implanted worms in the small intestine remained where they were implanted until expelled. Mucosal mast cells were induced in the whole small intestine after the worm implantation. In the large intestine, a considerable number of adult worms settled in the mucosa of mutant mice, whose goblet cell mucins were undersulfated because of a mutation in sulfate-activating enzymes. In these mice, the degree of sulfation of goblet cell mucins in the large intestine was significantly reduced to the level of normal small intestine goblet cell mucins. Our results suggest that sulfated glycoconjugates, either from mucosal mast cells or goblet cells, have important effects on the longitudinal distribution of parasitic females of S. venezuelensis.

Animals↗

PCR amplification of putative gpa-2 and gpa-3 orthologs from the (A+T)-rich genome of Strongyloides stercoralis.

Two G protein alpha subunit genes orthologous to gpa-2 and gpa-3 in Caenorhabditis elegans have been identified in the parasitic nematode, Strongyloides stercoralis. These genes mediate chemosensory signal transduction regulating dauer arrest in C. elegans. In the parasite, they represent candidate mediators for regulation of the choice between free-living and parasitic life cycles, the obligatory developmental arrest of infective larvae, and reactivation of development after infection. The (A+T) content of these genes is 72.2% for coding sequences, 90% for introns, and 84.1% for 5' and 3' flanking regions, requiring the use of low extension temperatures for long distance PCR. The possible significance of conserved structural motifs of these proteins is discussed.

Amino Acid Sequence↗