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PubMed2002

Protective immunity against Taenia crassiceps murine cysticercosis induced by DNA vaccination with a Taenia saginata tegument antigen.

This study investigated the protective capacity of the recombinant Taenia saginata Tso18 antigen administered as a DNA vaccine in the Taenia crassiceps murine model of cysticercosis. This Tso18 DNA sequence, isolated from a T. saginata oncosphere cDNA library, has homologies with Taenia solium and Echinococcus sp. It was cloned in the pcDNA3.1 plasmid and injected once intramuscularly into mice. Compared to saline-vaccinated control mice, immunization reduced the parasite burden by 57.3-81.4%, while lower levels of non-specific protection were induced in control mice injected with the plasmid pcDNA3.1 (18.8-33.1%) or a plasmid with irrelevant construct, pcDNA3.1/3D15 (33.4-38.8%). Importantly, significant levels of protection were observed between the pcDNA3.1/Tso18 plasmid and pcDNA3.1/3D15 plasmid immunized mice. Mice immunized with pTso18 synthesized low levels of, primarily IgG1 sub-class, antibodies. These antibodies were shown to recognize a 66 kDa antigen fraction of T. crassiceps and T. solium. Splenocytes enriched in both CD4+CD8- and CD4-CD8+ T cells from these vaccinated mice proliferated in vitro when exposed to antigens from both T. solium and T. crassiceps cestodes. Immunolocalization studies revealed the Tso18 antigen in oncospheres of T. saginata and T. solium, in the adult tapeworm and in the tegument of T. solium cysticerci. The protective capacity of this antigen and its extensive distribution in different stages, species and genera of cestodes points to the potential of Tso18 antigen for the possible design of a vaccine against cestodes.

Animals↗
PubMed2006

Molecular and functional characterization of a Taenia adhesion gene family (TAF) encoding potential protective antigens of Taenia saginata oncospheres.

Two clones from an activated Taenia saginata oncosphere cDNA library, Ts45W and Ts45S, were isolated and sequenced. Both of these genes belong to the Taenia ovis 45W gene family. The Ts45W and Ts45S cDNAs are 997- and 1,004-bp-long, each corresponding to 255 amino acids and with theoretical molecular masses of 27.8 and 27.7 kDa, respectively. Southern blot profiles obtained with Ts45W cDNA as a probe suggest that these two genes are members of a multigene family with tandem organization. The full genomic sequence was determined for the Ts45W gene and a new family member, the Ts45W/2 gene. The genomic sequences of the T. saginata Ts45W and Ts45W/2 genes were at least 2.2 kb in length with four exons separated by three introns. Exons 1 and 4 coded for hydrophobic domains, while, importantly, exons 2 and 3 coded for fibronectin homologous domains. These domains are presumably responsible for the demonstrated cell adhesion and, perhaps, the protective nature of this family of molecules and the acronym TAF (Taenia adhesion family) is proposed for this group of genes. We hypothesize that these TAF proteins and another T. saginata-protective antigen, HP6, have evolved the dual functions of facilitating tissue invasion and stimulating protective immunity to first ensure primary infection and subsequently to establish a concomitant protective immunity to protect the host from death or debilitation through superinfection by subsequent infections and thus help ensure parasite survival.

Amino Acid Sequence↗
PubMed2003

Helminth antigens (Taenia solium, Taenia crassiceps, Toxocara canis, Schistosoma mansoni and Echinococcus granulosus) and cross-reactivities in human infections and immunized animals.

Helminth antigens were investigated in the search for accessible heterologous antigens capable to discriminate different helminthiases, by the enzyme linked immunosorbent assay (ELISA) and the immunoblot assay (IB). Antigens used were: Taenia solium cysticercus total saline (Tso); Taenia crassiceps cysticercus vesicular fluid (Tcra-VF); T. crassiceps cysticercus glycoproteins (Tcra-GP and Tcra-(18-14)-GP); Toxocara canis larva excretory-secretory (TES); Schistosoma mansoni adult total saline (Sm) and Echinococcus granulosus hydatid fluid (Eg). The assayed sera were from patients with: cysticercosis (n = 18); toxocariasis (n = 40); schistosomiasis (n = 19) and hydatidosis (n = 50) with proven clinical and laboratory diagnosis, and sera from rabbits immunized with Tso, Tcra-VF, TES and Eg. Cross-reactivity occurred mostly between infections caused by Taenia and Echinococcus or in immunized rabbits, by ELISA. Moreover, the cross-reactivity among helminthiases was found with the use of antigens belonging to phylogenetically related parasite species, Eg, Tso and Tcra-VF, by sharing same antigenic components. Lower cross-reactivities were obtained by IB technique, when only peptides were considered as antigens, and the use of T. crassiceps purified glycoproteins demonstrated high sensitivity and specificity in the diagnosis of human cysticercosis, similarly to that using homologous antigen (Tso) by the same technique.

Animals↗
PubMed2001

Alternative splicing and sequence diversity of transcripts from the oncosphere stage of Taenia solium with homology to the 45W antigen of Taenia ovis.

Genes and transcripts which show homology to the host-protective 45W antigen of Taenia ovis have been cloned from the human parasite Taenia solium. The T. solium genes cloned in this study (TSO45) show conserved genomic structural features which are also features of the T. ovis 45W gene family. The TSO45 genes consist of a four exon and three intron structure. Eight TSO45 transcripts, encoded by at least five genes, were cloned from T. solium oncospheres and comparison of their DNA sequence indicates that some transcripts have arisen by alternative splicing, the first demonstration of exon inclusion/exon skipping in cestodes. Alternative splicing occurred with respect to both exons II and III with three splice variants identified from the TSO45-1 gene and two splice variants from TSO45-5. The proteins encoded by this family of genes contain putative N-linked glycosylation sites, an amino terminal secretory signal, a hydrophobic carboxy terminal sequence characteristic of GPI-anchored proteins and fibronectin type III motifs. These features are common to their T. ovis and Taenia saginata homologues. The similarities of the TSO45 genes cloned in this study with genes encoding host-protective antigens of T. ovis and T. saginata indicates that the encoded T. solium proteins are quite possibly antigenic and have potential use as a vaccine to prevent T. solium infection in the parasite's intermediate host. In this respect, the generation of sequence diversity and hence potential antigenic diversity through alternative splicing of TSO45 genes may have implications for the use of these proteins in vaccines against T. solium cysticercosis.

Alternative Splicing↗
PubMed2000

Specific Taenia crassiceps and Taenia solium antigenic peptides for neurocysticercosis immunodiagnosis using serum samples.

Neurocysticercosis (NC), i.e., the presence of the larval form of Taenia solium in tissues, is the most frequent and severe infection involving the central nervous system. Paired serum and cerebrospinal fluid (CSF) samples from patients with NC, CSF and serum samples from a control group, and serum samples from patients with other parasitoses were studied by enzyme-linked immunosorbent assay (ELISA) and by immunoblotting with Taenia crassiceps vesicular fluid antigen (Tcra) and Taenia solium total saline antigen (Tso) for the detection of immunoglobulin G antibodies. ELISAs carried out with the Tso and Tcra antigens showed 94.1 and 95.6% sensitivities, respectively, for the detection of antibodies in CSF and 70.6% and 91.2% sensitivities, respectively, for the detection of antibodies in serum, with 100% specificity for the detection of antibodies in CSF and 80% specificity for the detection of antibodies in serum for both antigens. On the basis of the reactivities of the peptides in the samples analyzed, the peptides of </=23, 39, 85 to 77, and 97 kDa were found to be Tso specific by immunoblotting and the peptides of </=62, 74, 109, 121, and 131 kDa were found to be Tcra specific. Tests with Tcra extract had higher sensitivities and more homogeneous results and permitted us to obtain the parasites easily. We suggest the use of Tcra ELISA for the study of serum and confirmation of the results for sera positive by an immunoblotting analysis in which specific peptides (e.g., peptides of 19 to 13 kDa) are detected.

Antibodies, Helminth↗
PubMed2005

Apoptosis patterns in experimental Taenia solium and Taenia crassiceps strobilae from golden hamsters.

Apoptosis or programmed cell death (PCD) patterns of two taeniid species, Taenia solium and Taenia crassiceps, were explored in adult tapeworms grown in golden hamsters. Animals were fed either ten viable T. solium cysticerci from naturally infected pigs or from T. crassiceps WFU strain maintained in Balb/c mice. Adult strobilae were recovered from the intestine at different times after infection and either frozen at -70 degrees C or fixed in paraformaldehyde-glutaraldehyde. Frozen sections were processed using the DNA fragmentation fluorescent TUNEL reagents and examined in an epifluorescent microscope. Fixed tissues were processed for light and electron microscopy. Typical apoptotic cells were found in the central core of scolex and strobilar tissues, mainly in the germinal tissue and subtegumentary areas. By the TUNEL technique, cells exhibited the characteristic fluorescent images of condensed nuclear chromatin. By light microscopy of thick sections stained with toluidine blue, we found a number of small rounded cells which had lost their cytoplasmic bridges and had shrunken nuclei with aggregated chromatin, cells which were found interspersed with normal syncytial cells. Similar cell morphology was confirmed by electron microscopy. Stunted viable worms, recovered with longer mature specimens, had very short strobilae and exhibited a large number of apoptotic cells in the germinal neck tissues. The results are consistent with the syncytial nature of these parasites, and strongly suggest that cell proliferation and PCD in these adult cestodes are continuous processes of the germinal tissue and tegumentary cytons.

Animals↗
PubMed2004

Differential diagnosis of Taenia saginata and Taenia saginata asiatica taeniasis through PCR.

New multiplex-PCR and PCR-linked restriction fragment length polymorphism protocols, derived from Taenia saginata HDP2 DNA sequence, have been designed that allow the simultaneous and specific identification of T. saginata and Taenia saginata asiatica. Proglottids expelled from 20 different Spanish taeniasis patients, previously diagnosed as T. saginata by both morphological identification and multiplex HDP2-PCR, were also examined by the newly developed PCR protocols, and the original diagnosis of T. saginata infection was confirmed. All of the 20 T. saginata samples were negative in the T. saginata asiatica-specific PCR. Three authentic T. saginata asiatica samples were unambiguously identified as such in the T. saginata asiatica PCR. These new protocols have immediate potential for the specific, sensitive, and rapid identification of T. saginata asiatica and may assist in taxonomic studies.

Animals↗
PubMed2006

Taenia asiatica and Taenia saginata: genetic divergence estimated from their mitochondrial genomes.

We conducted a differential identification of Taenia asiatica and Taenia saginata, through the mapping of mitochondrial genomes and the sequencing of the cox1 and cob genes. The entire mitochondrial genomes of T. asiatica and T. saginata were amplified by long-extension PCR and cloned; each was approximately 14 kb in size. Restriction maps of T. asiatica and T. saginata mitochondrial genomes were then constructed using 13 restriction enzymes. The resulting restriction patterns enable us to estimate their genetic divergence at 4.8%. The actual sequence divergence was computed 4.5% from the cox1 gene, and 4.1% from the cob gene. These results support the designation of T. asiatica as a separate species from T. saginata.

Animals↗
PubMed2006

Taenia solium and Taenia ovis: stage-specific expression of the vaccine antigen genes, TSOL18, TSOL16, and homologues, in oncospheres.

Recombinant antigens that have been cloned from Taenia solium and Taenia ovis have been shown to be highly effective when used as vaccines against cysticercosis in the intermediate hosts. This study investigated the presence of mRNA encoding the TSOL18 and TSOL16 antigens in different life-cycle stages of T. solium, and their related homologues in T. ovis. Reverse transcription-PCR and Southern blotting demonstrated that the antigens are stage-specifically expressed in the oncosphere. The apparent absence of expression of TSOL18 in the metacestode life-cycle stage suggests that the vaccine based on this antigen targets exclusively the early stages in the development of the parasite.

Animals↗
PubMed2006

Metabolism of steroid hormones by Taenia solium and Taenia crassiceps cysticerci.

Previous in vitro experiments showed that both, Taenia crassiceps and Taenia solium cysticerci have the ability to metabolize exogenous androstenedione to testosterone. Here we evaluate on the capacity of both cysticerci to synthesize several sex steroid hormones, using different hormonal precursors. Experiments using thin layer chromatography (TLC) showed that both cysticerci were able to produce (3)H-hydroxyprogesterone, (3)H-androstenedione and (3)H-testosterone when (3)H-progesterone was used as the precursor. They also synthesized (3)H-androstenediol and (3)H-testosterone when (3)H-dehydroepiandrosterone was the precursor. In addition, both cysticerci interconverted (3)H-estradiol and (3)H-estrone. These results, strongly suggest the presence and activity of the Delta4 and Delta5 steroid pathway enzymes, 3beta-hydroxysteroid dehydrogenase/Delta(5-4) isomerase-like enzyme (3beta-HSD), that converts androstenediol into testosterone; and the 17beta-hydroxysteroid dehydrogenase that interconverts estradiol and estrone, in both types of cysticerci.

Androgens↗
PubMed2003

Steroid hormone production by parasites: the case of Taenia crassiceps and Taenia solium cysticerci.

Many examples of reciprocal endocrine interactions between parasites and hosts have been found in insects, arthropods and mammals. Cysticercosis produced by Taenia solium metacestodes is a widely distributed parasite infection that affects the human and the pig. Taenia crassiceps experimental murine cysticercosis has been used to explore the role of biological factors involved in host-parasite interactions. We had shown that T. crassiceps cysticercosis affects the serum concentration of steroid hormones and the reproduction behavior of the male mice host. In an effort to understand the biology of the parasite, we had investigated the parasite capacity to produce sex steroids. For this purpose, T. crassiceps cysticerci were incubated in the presence of different steroid precursors. TLC and recrystallization procedures showed that testosterone is produced from 3H-androstenedione in cysticerci. The conversion of 3H-testosterone to androstenedione, although present is much less significant. In addition, we had studied the production of testosterone by T. solium cysticerci. For this purpose, cysticerci were dissected from pork meat and incubated as above described. The results showed that T. solium cysticerci also produce testosterone. We have speculated about the importance of androgens in the growth of T. crassiceps cysticerci and found that the addition of the antiandrogen flutamide to the culture media of the parasites significantly decreased 3H-thymidine incorporation. We therefore hypothesized, that the ability of cysticerci to produce testosterone from steroid precursors might be important for the parasite growth and development.

Androgens↗
PubMed1995

Transmission dynamics of Echinococcus granulosus, Taenia hydatigena and Taenia ovis in sheep in Uruguay.

A base-line survey was carried out on the transmission dynamics of Echinococcus granulosus, Taenia hydatigena and Taenia ovis in sheep in the Department of Florida, Uruguay. Mean life expectancy at birth of the sheep population in this Department was estimated at 3.5 and 4.8 years for male and female sheep, respectively. Both intensity and prevalence of E. granulosus infection increased with age providing no evidence that immunity was acquired in the population. This revealed that E. granulosus was only relatively stable and in an equilibrium endemic steady state with an estimate of the basic reproduction ratio (Ro) of about 1.2. The mature larval population in sheep was over-dispersed with parameter k, from fitting a negative binomial, being 0.199 and 0.556 in 42- and 54-month-old sheep, respectively. The mean number of cysts increased by 1.05 cysts per year and the infection pressure was calculated as 0.174 infections per year, implying that each infection consisted of 693.6 eggs to produce 6.03 cysts. Neither intensity nor prevalence of infection with T. hydatigena or T. ovis increased with age. This indicated that immunity was acquired to reinfection by both parasites and they were in an equilibrium hyperenedemic steady state with an estimated Ro of 4.2-5.8. Viability and fertility of E. granulosus, and viability of T. hydatigena cysts in the older age groups were low. The effects of the different transmission levels of the parasites on potential control strategies are discussed.

Animals↗
PubMed2000

Two Taenia species found in Japan, with new distribution record of Taenia polyacantha Leuckart, 1856 (Cestoda: Taeniidae).

In an epidemiological survey for Echinococcus multilocularis in rodents and insectivores from the northernmost part of the central mainland of Japan (Honshu), two taeniid species, Taenia crassiceps and Taenia polyacantha, were found in Microtus montebelli and Apodemus argenteus, respectively. The latter is the first record of distribution in Japan, and the former is the second after its first recovery from the central part of Japan. Although we have found neither larval nor strobilar stage of E. multilocularis there, discovery of these taeniid species, having overlapping global distribution with E. multilocularis in red foxes Vulpes vulpes as well as multiple occurrences of hydatid patients having no history of visits to the endemic areas shows the possibility that the life-cycle of E. multilocularis might be maintained at least in the northernmost part of Honshu.

Animals↗
PubMed2004

Differential expression of AP-1 transcription factor genes c-fos and c-jun in the helminth parasites Taenia crassiceps and Taenia solium.

Homologues of c-fos and c-jun from total DNA of Taenia crassiceps and Taenia solium were cloned and sequenced. The amino acid alignment analysis revealed that c-fos DNAs from T. crassiceps and T. solium were highly homologous (96%), and both have high homology compared to several mammalian c-fos proteins (93% to mouse, 96% to rat and 86% to human). The c-jun protein alignment showed higher homology (T. crassiceps and T. solium have 98%), when compared with mouse, rat and human, being 92%, 98% and 93% respectively. RT-PCR amplification of the parasite's total RNA, showed that T. crassiceps expressed both AP-1 complex genes, while T. solium only expressed c-fos. Southern blot hybridization analysis confirmed the true origin of each amplified gene. AP-1 transcription gene expression is regulated by oestradiol in the same fashion as their mammalian counterparts only in T. crassiceps. To study if AP-1 genes are involved in a physiological function of the cyst, reproduction was studied in vitro. Oestradiol treatment stimulated reproduction in T. crassiceps but not in T. solium cysticerci. This is the first report of the detection and functionality of AP-1 transcription factor genes in any species of helminth parasite.

Amino Acid Sequence↗
PubMed2001

Detection of IgG in cerebrospinal fluid for diagnosis of neurocysticercosis: evaluation of saline and SDS extracts from Taenia solium and Taenia crassiceps metacestodes by ELISA and immunoblot assay.

We compared saline (S) and sodium dodecyl sulphate (SDS) extracts from Taenia solium (homologous species - HO) and Taenia crassiceps (heterologous species - HE) metacestodes in order to detect IgG by ELISA and immunoblot assay (IBA) in cerebrospinal fluid (CSF) for the diagnosis of human neurocysticercosis (NC). CSF samples were obtained from 93 patients. Of these, 40 had NC, five had a diagnosis of probable NC, nine had central nervous system schistosomiasis or strongyloidiasis and 39 had other neurological alterations. Samples were analysed by ELISA and the results were compared with IBA in all samples with confirmed and probable NC diagnosis, in all samples with other central nervous system parasitic infection, and in 10 of those with another neurological alterations. ELISA sensitivity was 100%, 85%, 95% and 87.5% for the S-HO, S-HE, SDS-HO and SDS-HE extracts, respectively, and ELISA specificity was 100% for S-HO, S-HE, SDS-HO extracts and 97.9% for SDS-HE antigen. Immunodominant peptides detected by IBA were, by decreasing percentage of recognition: 64-68 and 45 kDa for S-HO; 108-114, 92-95, 64-68, 83 and 88 kDa for S-HE; 64-68, 108-114, 77 and 86 kDa for SDS-HO; and 108-114, 88 and 92-95 kDa for SDS-HE. Overall the homologous antigenic extracts showed higher sensitivity than the heterologous extracts in the diagnosis of NC in CSF samples. The heterologous extracts contained most of the immunodominant peptides presented in the homologous extracts, which are recognized by IgG antibodies in CSF samples.

Animals↗
PubMed2005

Sero-prevalence of Taenia solium cysticercosis and Taenia solium taeniasis in California, USA.

OBJECTIVES: Taenia solium Cysticercosis is a leading cause of epilepsy and neurological disability in the developing world. It is caused by ingestion of the eggs of the tapeworm, T. solium Taeniasis. The prevalence of either T. solium Cysticercosis or T. solium Taeniasis in the United States in populations at risk is poorly understood. The primary objectives of this study are to perform the first study of the sero-prevalence of T. solium Cysticercosis and T. solium Taeniasis in an at-risk community in the USA, specifically rural Southern California; identify T. solium Taeniasis positive individuals, and treat positive individuals for the tapeworm T. solium Taeniasis. METHODS: Community based sero-prevalence study of antibodies to T. solium Cysticercosis and T. solium Taeniasis in 449 subjects living in a federally funded, predominantly Hispanic residential community; and in two migrant farm worker camps in rural Ventura County, California, USA. For this study, fingerstick blood samples were obtained. Serum immunoblots for both T. solium Cysticercosis and T. solium Taeniasis were performed. RESULTS: The sero-prevalence of T. solium Cysticercosis was 1.8% and the sero-prevalence of T. solium Taeniasis by serum immunoblot was 1.1%. Taenia solium Cysticercosis and T. solium Taeniasis antibodies were not detected in children. The sero-prevalence of T. solium Taeniasis was highest in the migrant farm worker community. Handwashing frequency was correlated with T. solium Taeniasis sero-positivity. CONCLUSION: The sero-prevalence of T. solium Cysticercosis and T. solium Taeniasis in this population, as detected by serum immunoblot, approximates the prevalence in some endemic areas of Latin America. Importantly, most patients likely had prior exposure, not active infection. This study establishes for the first time, the relative sero-prevalence of T. solium Cysticercosis and T. solium Taeniasis in at-risk populations in the United States.

Adolescent↗
PubMed2002

Use of Taenia crassiceps cysticercus antigen preparations for detection of antibodies in cerebrospinal fluid samples from patients with neurocysticercosis (Taenia solium).

Antigen extracts obtained from the vesicular fluid of Taenia crassiceps cysticerci and from fractions purified by affinity chromatography with the lectin concanavalin A and the glycoprotein antigen separated by electrophoresis were used for the detection of Taenia solium anticysticercus antibodies. The sensitivity and specificity obtained for all antigens were 100% in enzyme-linked immunosorbent assay with good reproducibility. Using immunoblotting of the three antigens, low-molecular-mass peptides (18 and 14 kDa) were characterized only in cerebrospinal fluid samples from patients with neurocysticercosis. The results confirm that antigen fractions purified from T. crassiceps cisticerci are important sources of specific peptides and proved to be efficient in detecting anti-T. solium antibodies.

Animals↗
PubMed2001

Two epitopes shared by Taenia crassiceps and Taenia solium confer protection against murine T. crassiceps cysticercosis along with a prominent T1 response.

Taenia crassiceps recombinant antigens KETc1 and KETc12 have been shown to induce high level of protection against experimental murine T. crassiceps cysticercosis, an experimental model successfully used to test candidate antigens for use in vaccination against porcine Taenia solium cysticercosis. Based on the deduced amino acid sequence, KETc1 and KETc12 were chemically synthesized in linear form. Immunization with KETc1 induced 66.7 to 100% protection against murine cysticercosis, and immunization with KETc12 induced 52.7 to 88.1% protection. The elicited immune response indicated that both peptides contain at least one B-cell epitope (as demonstrated by their ability to induce specific antibodies) and one T-cell epitope that strongly stimulated the proliferation of T cells primed with either the free peptide or total cysticercal T. crassiceps antigens. The high percentage of spleen cells expressing inflammatory cytokines points to the likelihood of a T1 response being involved in protection. The protective capacity of the peptides and their presence in all developmental stages of T. solium point to these two epitopes as strong candidates for inclusion in a polyepitopic synthetic vaccine against T. solium pig cysticercosis.

Animals↗