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D A Denham

Publications and source records attributed to D A Denham.

At least 55 records · Page 3Linked to original sources
PubMed1985

Secreted and circulating antigens of the filarial parasite Brugia pahangi: analysis of in vitro released components and detection of parasite products in vivo.

A range of excretory-secretory (ES) antigens have been characterised following in vitro culture of adult Brugia pahangi filarial nematodes in serum-free medium. Analysis by radioiodination, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and immunoprecipitation of purified macromolecules with antibodies from human and experimental animal infections reveals both host and parasite components. Two host molecules appear by molecular weight and immunoprecipitation analysis to be immunoglobulin and serum albumin, presumed to be taken up from the jird host from which adult worms were recovered. A further prominent component, of 19 kDa, reacts with neither anti-host nor anti-filarial antibodies, and may represent a non-immunogenic parasite product. Three additional bands, although less intensely radiolabelled, did prove to be consistently antigenic, with apparent molecular weights of 15, 29 and 40 kDa. A further ES antigen, which does not take up radio-iodine or lend itself to electrophoretic analysis, has also been detected. This molecule reacts in a immunoradiometric assay in which monoclonal antibody directed against a repetitive epitope acts both to capture and indicate antigen presence. The same antibody, Bp-1, may also be employed to detect circulating antigen in the serum of animals experimentally infected with Brugia pahangi, and in the serum of patients with each of the three species of human lymphatic filariasis, Brugia malayi, Brugia timori and Wuchereria bancrofti.

Animals↗
PubMed1985

Surface antigens of a filarial nematode: analysis of adult Brugia pahangi surface components and their use in monoclonal antibody production.

The surface antigens of adult worms of the filarial nematode Brugia pahangi have been investigated further by surface radioiodination and detergent solubilisation techniques. In addition to yielding new information on the distribution of antigenic components of this stage, detergent-solubilised molecules were used in both radiometric and enzyme-linked assays for human and mouse antibody. These assays were subsequently used in screening for monoclonal antibodies from hybrid cells derived from animals infected with living parasites and boosted with detergent-extracted antigen. Three monoclonal antibody-producing cell lines were isolated, with differing antigenic specificities: Bp-1, which binds a non-iodinatable antigen with high ELISA activity; Bp-2, which reacts with a determinant found on but not unique to the major surface Iodogen-labelled 29 kDa antigen; and Bp-3, which is specific for a minor antigen of 20 kDa revealed by Iodogen labelling.

Animals↗
PubMed1985

The induction of functional mononuclear and multinuclear macrophages in murine brugian filariasis: morphological and immunological properties.

This present study describes the ontogeny and morphology of mononuclear and multinuclear macrophages in murine Brugia pahangi infections. Intraperitoneal infection with L 3 resulted in the development of adult worms and the production of microfilariae in many mice; however, all worms were usually removed by 100 days post infection (p.i.). The infection induced predominantly mononuclear and multinuclear macrophage responses in the peritoneal cavity, with maximum numbers of peritoneal cells being achieved by 8-12 weeks p.i. and at this time some 20% of the macrophages were multinucleated. Lymphocyte numbers were also increased during infection. The reactions around the filariae involved macrophages (both single and multinucleated), eosinophils, fibroblasts and the laying down of collagen. Most worms were involved in these cellular reactions by 5-6 weeks p.i., with the multinuclear macrophages generally seen lying close to the parasites. All stages of parasites developed vacuolar internal changes early in their degeneration with calcification being a later change. The destruction and removal of the sheath was not apparently a prerequisite for degeneration of the body of microfilariae. Rosetting techniques showed that 75% of peritoneal cells (macrophages) in infected animals possessed Fc receptors compared with 32% of those in uninfected mice, and for C3 receptors the respective results were 35% and 5%. The multinuclear cells possessed both types of receptors at levels similar to those of mononuclear macrophages. Likewise both cell types were found to readily phagocytose yeast and latex particles, as well as staining positively for non-specific esterases.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Specificity of surface molecules of adult Brugia parasites: cross-reactivity with antibody from Wuchereria, Onchocerca and other human filarial infections.

The extent of structural and immunological similarity between surface antigens of three species of Brugia filarial parasites was studied by lodogen-mediated surface labelling of adult worms of B. malayi, B. pahangi and B. timori. The close homology and cross-reactivity between these antigens reported in previous surface labelling studies with Bolton-Hunter reagent, was verified in this system. The surface antigens of adult B. pahangi are also recognised by antibody from patients with Wuchereria bancrofti, the major human lymphatic filariae, and by antibody to Loa loa and Mansonella perstans. Further experiments have begun to establish the boundaries of these cross-reactions: antibodies to nonfilarial nematodes such as Trichinella, Necator and Strongyloides does not recognise the adult surface antigens; however, although most anti-Onchocerca sera show little or no reaction to the major (29 kDa) surface antigen, there is consistent reactivity to the secondary 20 kDa antigen, and extensive recognition of a minor antigen of 15 kDa.

Animals↗
PubMed1984

The effect of humidity on the transmission of Brugia pahangi infective larvae to mammalian hosts by Aedes aegypti.

The transmission of Brugia pahangi by Aedes aegypti to the mammalian host was compared at low and high humidity. There was no statistical difference between the number of infected mosquitoes feeding or the egress of infective larvae from these mosquitoes at high or low humidity. The penetration of the host by the infective larvae was however significantly greater (p less than 0.05) at high than at a low humidity.

Aedes↗
PubMed1984

Induction of host resistance to Brugia pahangi in jirds (Meriones unguiculatus) protected by chemoprophylaxis.

Jirds were given a chemoprophylactic inoculation of flubendazole (FMBZ) and then five injections of infective larvae of Brugia pahangi whilst still protected by the FMBZ. When the drug was thought to be non-effective the jirds (and controls) were given a challenge infection of B. pahangi larvae. By comparison with control jirds the treated-infected-challenged jirds had 40% fewer adult worms. The control treated-challenged jirds contained mostly sterile female worms showing that they were still partially protected by FMBZ but worms numbers were not significantly reduced as compared with untreated controls.

Animals↗
PubMed1984

Biochemical surface components of Brugia pahangi microfilariae.

The sheath and cuticle of microfilariae of Brugia pahangi were examined by electron microscopy and the presence of various proteins, carbohydrate and enzymes sought. The epicuticle of microfilariae consists of a pentalaminate structure (24.0 +/- 1.4 nm), a cortex (13.7 +/- 3.6 nm) and a basal zone (27.8 +/- 4.8 nm) which is often banded in appearance. The pentalaminate layers are not continuous at the base of the interannular grooves. The sheath and the epicuticle of B. pahangi stained positively with concanavalin A and saccharated iron oxide. The sheath of approximately 50% of microfilariae showed activity for acid phosphatase, 5' nucleotidase and peroxidase, but not for ATPases, alkaline phosphatase or esterase. No enzymes were detected in the epicuticle although the cortex and basal layers of the cuticle did show enzymic activity. Structures beneath the cuticle in the main body of the worms contained considerable enzymic activity. Microfilariae directly isolated from the blood of infected cats were found by immunochemical means to carry serum proteins on their sheaths but not on their cuticles. These studies extend the definition of the outer structures of microfilariae and confirm that they significantly differ in morphology and enzyme content from typical mammalian cell membranes.

Acid Phosphatase↗
PubMed1984

Increased activity of macrophages from mice infected with Brugia pahangi: in vitro adherence to microfilariae.

After the inoculation of infective larvae of Brugia pahangi into the peritoneal cavities of CBA/Ca mice adult worms developed, but by 12 weeks post-infection the parasites were usually dead and surrounded by granulomatous tissue. Macrophages were the most common cell type in these granulomas and were also found in increasing numbers free in the peritoneal cavity. We have investigated the ability of macrophages to damage microfilariae in vitro, in a system where microfilariae were cultured together with cells and serum taken from either uninfected female CBA/Ca mice or at various intervals after infection. Macrophages adhered to and killed microfilariae in this system and there was an increase in vitro adherence of these cells during the course of infection. The peak level of in vitro activity of these cells coincided with the phase of parasite killing in vivo. The presence of serum also affected the degree of macrophage adherence and subsequent death of the parasite. Analysis of serum components using EGTA, zymosan or heat inactivation suggested that complement and possibly heat labile antibody were involved. Immunoglobulins were shown by immunofluorescence to be present on the surface of microfilariae cultured in serum from infected mice. It is concluded from this study that macrophages are actively involved in the termination of murine filarisis.

Animals↗
PubMed1984

Differentiation of species and life cycle stages of Brugia spp. by isoenzyme analysis.

The isoenzyme patterns of glucose phosphate isomerase and phosphoglucomutase of 3 species of Brugia, B. pahangi, subperiodic B. malayi, and B. patei, and 3 life cycle stages, adult, third-stage larva, and microfilaria were compared using the technique of isoelectricfocusing on polyacrylamide gels. The results demonstrated that the adults of all 3 species could be identified from one another and that differences existed between the sexes of any one species. Hybridization between B. pahangi and B. patei could be detected in the progeny of the cross. Both the third-stage larvae and microfilariae of B. malayi and B. pahangi were differentiated and the epidemiological significance and the application of these findings to arthropod-borne filarial infections were discussed.

Animals↗
PubMed1984

The action of diethylcarbamazine on the skin-dwelling microfilariae of Monanema globulosa (Nematoda: Filarioidea) in rodents.

Diethylcarbamazine (DEC) administered orally to jirds and striped mice (300 mg/kg for five days) was effective against the microfilariae of Monanema globulosa in experimental infections. Based on the small numbers of animals used in this trial, it is questionable whether the drug had significant activity against the adult worms in pulmonary arteries. Densities of microfilariae in the ears (mf/mg) fell by 80-100% of pretreatment levels over a four week period following treatment. There was no apparent action of the developing embryos in utero, and in some rodents densities of microfilariae in the skin returned to 73-88% of pretreatment levels within 70 days. No microfilariae were detected in the blood or urine during treatment. Twenty-four hours after the first dose of DEC, many of the microfilariae in the dermis became surrounded by a local infiltration of neutrophils, eosinophils and histiocytes. Cellular reactions were accompanied by an acute pruritus in rodents, as indicated by scratching and distress exhibited soon after treatment: No reactions to microfilariae were seen in untreated rodents. Similarities between this response and the Mazzotti reaction induced by DEC in human onchocerciasis indicate that M. globulosa may be of value as a rodent model to study adverse drug reactions in Onchocerca volvulus infections.

Animals↗
PubMed1984

Parasitological observations on Meriones unguiculatus singly or multiply infected with Brugia pahangi.

When jirds were infected with a single inoculum of 25-50 infective larvae of Brugia pahangi an overall mean recovery of adult worms of 44.5% (n = 41) was obtained. There was no difference in recoveries between male and female jirds. If jirds were repeatedly inoculated with larvae into the peritoneal cavity yields were only slightly reduced. Yields were 30.5% for 5 infections (n = 10), 26.7% for 10 infections (n = 8), 34.4% for 15 infections (n = 10) and 28.5% for 20 infections (n = 7). Twice as many worms were recovered from intraperitoneally inoculated jirds than from subcutaneously inoculated jirds.

Animals↗
PubMed1983

Cross-reactive surface antigens on three stages of Brugia malayi, B. pahangi and B. timori.

Surface antigens of three stages of three species of the filarial nematode genus Brugia have been analysed by radio-iodination and immunoprecipitation. These surface antigens have been shown to be characteristic for each stage by polyacrylamide gel electrophoresis. For example, infective larvae and adult worms have relatively complex patterns while microfilariae have few bands which are not found when other stages are radio-isotope labelled by the same technique. The surface antigens of Brugia malayi, B. timori and B. pahangi adult worms are all closely homologous, as are the surface antigens of infective larvae of the same three species, and of microfilariae of B. malayi and B. pahangi. Immunoprecipitation revealed that antibody raised in mice against one stage or species reacted with surface antigens from other stages and species. For example, sera raised against B. pahangi male adults reacted strongly with surface antigens from all three species. This cross-reactivity was dominant despite the apparent stage-specificity of the surface pattern seen on SDS-PAGE analysis. Moreover, in cross-immunization experiments, infective larvae were able to stimulate a secondary antibody response in mice previously primed with microfilarial surface antigens. The major microfilarial surface antigens (of mol. wt 65-70 000 Daltons) were recognized by serum antibody from microfilariae-, infective larvae- or adult-infected animals. Thus, although the dominant antigens from each stage are of different molecular weight, cross-reactions with stage-specific antisera suggest that there must be shared epitopes on Brugia surface antigens from each stage. Such shared antigenic determinants dominate the immune response, although other evidence, including the differences in molecular weight, indicates the existence of stage- and species-specific components.

Animals↗
PubMed1983

The resistance to re-infection of cats repeatedly inoculated with infective larvae of Brugia pahangi.

Seven microfilaraemic and five amicrofilaraemic cats which had been repeatedly infected with Brugia pahangi were challenged along with normal cats 28, 14 and 1 day before autopsy. The lymphatics of the amicrofilaraemic cats contained no female adult worms originating from the repeat infections and only two adult males (both from the same cat). Only 5.2% of the worms in the control cats were recovered from the amicrofilaraemic cats. Most of the challenge worms were killed in the first 24 h. The microfilaraemic cats all contained fertile adult male and female worms derived from the repeated infection but in such low numbers as to indicate considerable resistance to infection. Compared to their controls 26.4% of the challenge worms were recovered. Analysis of the life-cycle stages recovered showed that in both groups there was attrition of all stages and that although a number of worms reached L5 these were all killed later in the amicrofilaraemic cats.

Animals↗
PubMed1983

Effect of immunosuppressive or inflammatory agents on lymphatic dilatation in cats infected with Brugia pahangi.

Cats infected with Brugia pahangi were immunosuppressed and the development of lymphatic pathology, as measured xeroradiographically by dilatation of the vessels, in these animals was compared with that of normally infected cats. Individual cats showed wide variations in response. Niridazole and antilymphocyte serum both reduced dilatation, niridazole being particularly effective in the early stages of infection. When silica, used as an inflammatory agent, was injected subcutaneously in conjunction with worm infection lymphatic dilatation developed, this being potentiated in degree and rate of development as compared with control animals. The possible role of a non-filarial inflammatory response in the development of lymphatic dilatation is discussed.

Animals↗
PubMed1983

Infection of inbred and nude (athymic) rats with Brugia spp.

Infective larvae of Brugia pahangi were injected subcutaneously into inbred PVG (-RTIc) rats, and 'nude' (PVG-rnu/rnu) (athymic) rats. Adult worms or circulating microfilariae were recovered from 20/34 (59%) of PVG-RTIc rats and from 30/30 (100%) of 'nude' rats. Fertile worms were regularly found in the lumbar lymphatics and hearts of both strains of rat. Blood eosinophilia first developed in PVG-RTIc rats about 17 days, and in all such animals by 6 weeks. High circulating eosinophil counts persisted only in patent animals, proving a useful hallmark for the presence of microfilariae. Nude rats despite patency, developed eosinophilia only latterly and then to a lesser extent. Specific anti-B. pahangi IgG antibody was first detected at 7 days in all infected PVG-RTIc rats, with levels rising until 8 weeks and remaining high only in microfilaraemic animals; total IgE showed a similar response. Specific IgE rose in all the eight patent rats inconsistently and only to low levels in eight non-patent infected rats. IgG and IgE were undetectable in nude rats. Other strains of inbred rats of different RTI haplotype were also successfully infected with B. pahangi and the human parasite B. malayi, a total of 10/23 (43%) and 5/15 (33%) becoming patent respectively. In the small numbers tested no major influence of RTI haplotype was detected. Infection by the intraperitoneal route did not result in the development of microfilariae. The difference in patency rates between 'nude' and normal PVG rats supports the contention that the development of filarial infections is T lymphocyte dependent. Inbred and 'nude' rats provide a valuable model of human filariasis, in which many features of filarial immunopathology can be studied.

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