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At least 19 recordsLinked to original sources
PubMed1994

A novel type of glutathione S-transferase in Onchocerca volvulus.

Onchocerca volvulus is a pathogenic human filarial parasite which, like other helminth parasites, is capable of evading the host's immune responses by a variety of defense mechanisms which are likely to include the detoxification and repair mechanisms of the enzyme glutathione S-transferase (GST). In this study, we show that one of the previously described GSTs from O. volvulus appears to possess the characteristics of a secreted enzyme. When the complete O. volvulus GST1 (OvGST1) sequence presented here is compared with those of other GSTs, 50 additional residues at the N terminus are observed, the first 25 showing characteristics of a signal peptide. This is consistent with the N-terminal sequence data on the native mature enzyme which begins at amino acid 26, based on the deduced protein sequence from the cDNA. The native protein, without the signal peptide sequence, possesses a 24-amino-acid extension not present in other GSTs. The deduced amino acid sequence of the OvGST1 cDNA clone was shown to possess four potential N-glycosylation sites. Digestion of O. volvulus homogenate with endoglycosidase, followed by detection of OvGST1 with specific antibody, indicated that the enzyme possesses at least two N-linked oligosaccharide chains. Gel filtration of the Escherichia coli-produced recombinant OvGST1 showed that it is enzymatically active as a nonglycosylated dimer. OvGST1 is found in the media surrounding adult worms maintained in culture, indicating that, in vitro, this enzyme is released from the worm. The strongest immunostaining for OvGST1 was observed in the outer cellular covering of the adult worm body, the syncytial hypodermis, especially in the interchordal hypodermis, where the peripheral membrane forms a series of lamellae which run into the outer zone of the hypodermal cytoplasm.

Amino Acid Sequence↗
PubMed1992

Effects of three-month doses of ivermectin on adult Onchocerca volvulus.

Onchocerca volvulus worms in nodules from Guatemalan patients treated with four, eight, or 11 single doses of ivermectin (150 micrograms/kg of body weight) that were given once every three months were examined by routine histologic techniques and compared with worms in control nodules from untreated persons living in the same location over the same time periods. All treated nodules were removed four months after the last dose of ivermectin, i.e., 13, 25, or 34 months after the start of the trial. At the 25th and 34th months, i.e., after the eighth or eleventh doses of ivermectin, there were excess mortalities in female worms of 25.5% and 32.6%, respectively, over and above the levels in controls. Furthermore, the proportions of live females still producing scanty embryos up to the gastrula stage were only 7.7% and 18.2%, and no females were producing microfilariae. Ivermectin given at 3-month intervals also reduced significantly the mean numbers of live male worms in nodules, as well as the proportions of inseminated females. This regimen was effective in preventing embryogenesis to the microfilarial stage while, at the same time, it caused a slow but steady attrition of the adult worms.

Adolescent↗
PubMed1990

On the reproductive activity of the female Onchocerca volvulus.

Onchocerca volvulus worms, extracted from nodules by collagenase digestion, stained with haematoxylin and cleared in glycerol, were unravelled for longitudinal examination and later embedded in brain blocks for study of serial transverse sections. A classification system for female worms is proposed, based on the reproductive status of 446 worms from Guatemala, 94 from Liberia and 125 from Mali. They were categorized into fecund, inseminated specimens; uninseminated, but potentially fertile specimens, shedding ova destined to degenerate; worms changing from the uninseminated to the inseminated state and vice versa, which were few in number; old worms, with degenerate ovaries, whose genital tracts were either empty or had disappeared; and moribund or dead worms, characterized by loss of turgor, collapse and degeneration, calcification, or invasion by polymorphic, basophilic cells. Potentially fertile worms shed oocytes continuously and, when they were inseminated, embryonic development ensured. No evidence was found of a periodic cycle of reproduction. Inseminated worms were found in nodules without a male worm, and uninseminated worms in nodules harbouring male worms. Measurements are recorded of portions of the female reproductive tract and of the length of uterus occupied by the various embryonic stages in fully fecund worms. A significant difference in the length of the body behind the first and second ovaries was observed as between worms from West African savanna (Mali) and forest (Liberia). Limited observations were also made on meiosis in the oocyte, penetration of the oocyte by sperm, formation of the ovum, syngamy and zygote formation.

Animals↗
PubMed1986

Identification of antigens of Onchocerca volvulus and Onchocerca gibsoni for diagnostic use.

Adults of Onchocerca volvulus and Onchocerca gibsoni were identically fractionated into a surface-enriched fraction, a phosphate buffered saline (PBS) extract and a PBS insoluble-detergent soluble fraction. Glycoproteins were prepared from these extracts and all fractions were examined by the Western blot technique using sera from individuals infected with a variety of filarial and non-filarial nematode worms. Using antisera to O. volvulus, a number of antigens were demonstrated in all of the extracts, with some antigens of each extract being unique. Many antigens were glycoproteins, and a high cross-reactivity was observed between O. volvulus and O. gibsoni. The different fractions of both species were also analysed using a panel of different sera in order to identify Onchocerca-specific antigens. The studies revealed that the lower molecular weight antigens showed greater Onchocerca specificity in all of the extracts examined. The surface-enriched fraction, however, clearly contained less widely cross-reacting components than the somatic and glycoprotein fractions. Finally, using surface labelling and coprecipitation techniques, O. gibsoni was shown to possess a 20 kDa Onchocerca-specific antigen, previously described for O. volvulus. The findings indicate a number of Onchocerca-specific antigens which may have potential in diagnosis of human onchocerciasis. They also show that the related bovine parasite O. gibsoni, may be an alternative source of material.

Animals↗
PubMed1991

Comparison of the effects of a single dose and of four six-monthly doses of ivermectin on adult Onchocerca volvulus.

Adult Onchocerca volvulus worms, extracted from nodules of Guatemalans by collagenase digestion, were examined whole and by histological techniques. One group of persons received a single 150 micrograms/kg dose of ivermectin; two other groups (one with older and one with younger nodules) received four similar doses of ivermectin at 6-month intervals. For each group, there were comparable untreated controls. All nodules were removed six months after the last dose. After a single dose, the only significant difference from the controls was in the decreased proportion of female worms producing live microfilariae. After four doses, there were significant increases in the proportions of moribund/dead female worms and of live uninseminated females, when compared with the corresponding controls. There were also fewer male worms present, but this difference was not significant. Six months after the conclusion of the 4-dose regimen, the proportion of female worms producing live microfilariae was significantly lower than in the groups that had received a single dose.

Animals↗
PubMed1995

Immunity to onchocerciasis: putative immune persons produce a Th1-like response to Onchocerca volvulus.

Immunity to Onchocerca volvulus (Ov) infection is suggested by the presence of putatively immune (PI) subjects in a region of Ecuador in which Ov is endemic. PI subjects were identified by traditional diagnostic methods combined with a polymerase chain reaction-based assay for Ov DNA in skin snips. Responses of peripheral blood mononuclear cells (PBMC) from the PI group (n = 16) were compared with those of persons with active infection (microfiladermic [MF] subjects; n = 51). PBMC of PI subjects proliferated significantly more to Ov antigen (OvAg; P < .009) than did PBMC of MF persons but less to streptolysin-O (P < .001). Cytokine analysis of PBMC culture supernatants revealed that PI subjects (n = 11) produced significantly more interferon-gamma to OvAg than did those in the MF group (n = 18; P = .018), less interleukin (IL)-5 to nonparasite antigen (P = .003) and mitogen (P = .012), and less IL-10 spontaneously (P = .016). Thus, immunity to Ov may in part be mediated by an antigen-specific Th1-type response.

Adolescent↗
PubMed1986

Detection of protein kinase substrates in extracts of Onchocerca volvulus.

Extracts of Onchocerca volvulus were phosphorylated in the presence of (gamma 32P)ATP and Mg2+ by endogenous protein kinase activity and exogenous rabbit muscle catalytic sub-unit of the adenosine 3'5' monophosphate dependent protein kinase (E.C. 2.7.1.37). Sodium dodecylsulfate polyacrylamide gel electrophoretic analysis of the 32P-labelled extracts revealed at least seven (32P)-phosphoproteins with apparent Mr of 92,000; 86,000; 40,000; 27,000; 26,000; 23,000 and 17,000. The phosphorylation of the components with apparent Mr of 23,000 and 17,000 was catalysed by both endogenous and exogenous protein kinases, whereas the other components required exogenous protein kinase for their phosphorylation. The endogenous protein kinase activity was inhibited by suramin and the heat-stable protein inhibitor of the adenosine 3'5' monophosphate dependent protein kinase. The (32P)phosphoproteins identified in this investigation are probably candidate regulatory molecules in O. volvulus; though their physiological functions remain to be determined.

Animals↗
PubMed1988

Isolation and in vitro translation of Onchocerca volvulus mRNA.

Adult Onchocerca volvulus recovered for excised nodules by dissection or treatment with collagenase have been used as a source of RNA for in vitro translation experiments. RNA was purified using either the hot phenol/SDS procedure or the guanidine isothiocyanate protocol. Immunoprecipitation experiments performed on in vitro products demonstrate a marked heterogeneity in responses by individed human infection sera. Further immunoprecipitation experiments demonstrate cross reactivity between O. volvulus and other filarial nematodes.

Animals↗
PubMed1986

Antigenic similarity of Onchocerca volvulus to other helminths examined by monoclonal antibodies against O. volvulus.

Antigens in Onchocerca volvulus were characterized using anti-O. vovulus monoclonal antibodies (McAb). A total of 17 McAb which were not species-specific to O. volvulus was examined for the antigenic similarity of O. volvulus with 12 parasitic helminths using indirect immunofluorescence. The range of reactivity of these McAb was wider in nematodes in contrast to species of cestoda or trematoda. The antigenic similarity within helminths examined was generally in agreement with their taxonomic relationships but was not shown in the lower taxonomic hierarchy. The helminth which holds the antigens most in common to O. volvulus was not the filarial parasites but Ascaris suum, an intestinal nematoda. The least antigenic similarity to O. volvulus was found in Schistosoma mansoni and S. japonicum, trematoda, among helminths examined. On the other hand, the existence of a common antigen shared with all the 13 helminths examined was demonstrated by a IgM McAb.

Antibodies, Monoclonal↗
PubMed1990

An improved method of examining adult Onchocerca volvulus worms.

Adult Onchocerca volvulus worms were extracted from alcohol-fixed nodules by collagenese digestion. The worms were stained in Mayer's haematoxylin and brought up into glycerol. In this viscous fluid the stained and cleared worms could be unravelled in sequential lengths of 4-6 cm and mounted in glycerol on slides for longitudinal examination of their structure. The worm lengths were then cut into small pieces, 0.5 cm long, which were implanted sequentially into slits in a brick-shaped block of formalin-fixed calf's brain (white matter) also in glycerol. The brain blocks were then processed in paraffin wax and cut as 6 microns histological sections, enabling transverse sections of the worm, stained as required, to be examined in sequential order.

Animals↗
PubMed1989

[Effects of diethylcarbamazine and ivermectin on the mobilization of microfilariae of Onchocerca volvulus].

Microfilaria of Onchocerca volvulus are localized in superficial dermis. However, they are sometimes noticed in the blood and urines, when there is an important infestation. Diethylcarbamazine (DEC) bring on an increase of microfilaria, successively in the blood and after, in the urines. This study of 30 patients, treated in double blind by placebo, DEC and ivermectin, a new molecule with spectacular action on this filariasis, allowed to compare effect of each one on apparition of microfilariae in the blood and urines. Ivermectin bring on very important increase of microfilaremia without microfilaruria, unlike diethylcarbamazine. This establishment bring to discuss very different physiologic mechanisms between them. It should have an incidence on diagnosis and treatment of this dangerous parasite.

Adult↗
PubMed1981

A diagnostic skin test for Onchocerca volvulus infection.

Onchocerca supernatant (OS) was prepared by a technique permitting live microfilariae to migrate from nodule tissue through agar gel into sterile Hanks balanced salt/Penicillin-Streptomycin solution where they metabolized. The OS, after dialysis, was passed through Seitz viral filter and either concentrated or lyophilized. Using rabbit antiserum in immunodiffusion and immunoelectrophoresis tests, microfilariae proteins and also human protein were detected in out OS. No common antigens were found between this and somatic extracts of Loa loa, O. gutturosa, O. volvulus, L. carinii, D. immittis and A. lumbricoides. 125I labelled OS was purified by passage through protein A column and then through immunosorbent column of horse anti-human serum linked to CNB-activated sepharose 4B. Autoradiography, after sodium dodecyl sulphate polyacylamide slab gel eletrophoresis of purified OS, showed 10 protein bands in the molecular range 10,000 to 125,000. Skin prick tests with OS, shown not to be contaminated with Hepatitis B antigens, elicited immediate hypersensitivity reaction. Using our criteria, positive reactions were seen in 81% of proven onchocerca cases and only occasionally in Loasis 4.5%, ascaridiasis 13.5% or healthy controls 2.4%. The poor skin reactivity to OS in loasis was not due to immunosuppression as these patients, when also infested with ascaris, reacted just as well as onchocerca patients with ascaris to skin prick test using somatic extracts of ascaris.

Adolescent↗
PubMed1976

Preliminary identification and characterization of antigen extracts from Onchocerca volvulus.

Sequential extraction of Onchocerca volvulus from skin nodules with buffers of different ionic strength resulted in a mixture of antigens which reacted in double immunodiffusion with sera from proven cases of onchocerciasis, the number of precipitation lines varied from 1 to 5. Triton X-100 detached mainly cationic antigen proteins. It reduced the molecular size of many antigenic proteins, but did not effect their antigenicity. Labelling of the extracted proteins with high specific activity 125I did not destroy much antigenicity. Strong cross-reactivity between Onchocera volvulus extracts and Onchocerca gutturosa and Litomosoides carinii was observed, less with extracts of Necator americanus and Ascaris suum, and none with Echinococcus multilocularis. Sera from proven cases of wuchereriasis, toxoplasmosis and echinococcosis rarely contained antibodies against O. volvulus. Isoelectric focusing, immunoelectrophoresis and gel filtration of the O. volvulus extracts indicated a variety of antigens. Preliminary attempts were made to purify the antigens using a solid-phase immuno-adsorbent prepared by coupling purified onchocerciasis IgG to CNBr-activated Sepharose.

Antigens↗
PubMed1992

The actin genes of Onchocerca volvulus.

The genome of Onchocerca volvulus was found to contain 2 actin gene classes (called 1 and 2) of 2 genes each. The 4 genes are located in 2 clusters (called A and B), each containing a gene class member. Five short introns of 122-207 bp occur within each gene. The sequences of the fourth intron and the 5' and 3' untranslated regions of all the 2 gene classes are completely different even though their coding regions share 95% identity. Mature transcripts from the actin genes have the nematode spliced leader (SL) at their 5' ends. One actin cDNA was found to be derived from an actin pre-mRNA which locks both the 5 introns and the 5' SL, suggesting that in at least some transcripts cis-splicing is completed before trans-splicing occurs.

Actins↗
PubMed1985

Serum immunoglobulin E levels in onchocerciasis: the development of a radioallergosorbent test for Onchocerca volvulus infection.

The effects of Onchocerca volvulus infection on immunoglobulin levels have been studied in subjects from three localities with different onchocerciasis prevalence rates. Infestation by O. volvulus provoked a 200-500% rise in total serum IgE levels. Immunoglobulins G, M. and A were increased less dramatically. To study further the IgE response, radioallergosorbent test (RAST) systems were developed using total extract of O. volvulus and Onchocerca supernatant (excretory-secretory) antigens. Both antigen preparations reacted with more than 80% of the onchocerciasis sera from the hyperendemic village, but the reactions were not proportionate to skin microfilarial density. There were also positive reactions with sera of some individuals whose skin biopsies showed no microfilariae, indicating prior exposure to the parasite. European control sera tested under similar conditions reacted only slightly in the RAST system. The developed RAST systems may thus be used for determining prior exposure to O. volvulus.

Cameroon↗
PubMed1977

Intracytoplasmic bacteria in Onchocerca volvulus.

Ultrastructural studies on Onchocerca volvulus disclosed intracellular organisms within the lateral chords of adult worms and of the larval stages. In the females the organisms were also present in the oogonia, oocytes, developing eggs and microfilariae. The organisms, found within vesicles of host (filarid) membrane and limited to the cytoplasm of infected cells, appeared to have a developmental cycle consisting of three morphologically distinct forms: a small spheroidal form up to 0.3 micronm in size, a bacillary form up to 1.5 micron7 in length and 0.7 micronm in diameter, and a third form, intermediate in size between the former and the latter, characterized by a dense inclusion. The intravesicular location and the developmental cycle consisting of three distinct forms are the two characteristics which suggest that these organisms are more similar to the chlamydiae than to the rickettsiae, in spite of their being transovarially transmitted. The significance of these findings with respect to the host-parasite relationship and pathogenesis of onchocerciasis is presently unknown and will require further study.

Animals↗