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Dive into the research topics where M. D. Rickard is active.

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Featured researches published by M. D. Rickard.


Parasitology | 1988

Excretory–secretory products of helminth parasites: effects on host immune responses

Marshall W. Lightowlers; M. D. Rickard

Parasitic helminths excrete or secrete (ES) a variety of molecules into their mammalian hosts. The effects of these ES products on the hosts immune responses are reviewed. Investigations into the source of antigenic or immunoregulatory ES products have identified the cuticular and tegumental surfaces of some nematodes and trematodes respectively as being important sources of ES products; other ES molecules are released through specialized excretory or secretory organs. It is proposed that the active shedding of surface antigens may serve as an important source of parasite antigens available to the immune system in a form in which they can be taken up and processed by antigen-presenting dendritic cells, macrophages and certain B cells for presentation to T helper cells. The ES products of nematodes, trematodes and cestodes contribute to immune evasion strategies of the parasites through mechanisms including shedding of surface-bound ligands and cells, alteration of lymphocyte, macrophage and granulocyte functions and modulation of complement and other host inflammatory responses. Immunopathology may be induced by ES products as in the development of granulomas around entrapped schistosome eggs. In some host-parasite systems ES antigens may induce host-protective immune responses and this source of protective antigens has been utilized in the successful vaccination against helminth infections, particularly against infection with trichurid nematodes and the metacestode stage of cestode parasites. The use of ES antigens in immunodiagnosis of helminth infection is also briefly discussed.


Molecular and Biochemical Parasitology | 1989

Subunit composition and specificity of the major cyst fluid antigens of Echinococcus granulosus

Marshall W. Lightowlers; Dongyou Liu; Anna Haralambous; M. D. Rickard

The subunit composition and specificity of the major Echinococcus granulosus cyst fluid antigens were determined by immunochemical analysis using murine monoclonal antibodies against Antigen 5 and Antigen B and human sera. Immune complexes cut out from immunoelectrophoresis gels and murine hybridomas were used as a source of specific anti-Antigen 5 and anti-Antigen B antibodies. Immunoprecipitation and Western blot analyses in sodium dodecyl sulphate-polyacrylamide gels using these reagents identified Antigen 5 to be a heterodimer composed of 24-kDa and 38-kDa subunits linked by disulphide bonding. Antigen B comprised a regularly spaced group of molecules with the smallest subunit estimated to be 8 kDa and the other components each differing in size by approximately 8 kDa, i.e., 16 kDa, 24 kDa, 32 kDa etc.; all possibly derived from the 8-kDa monomer. The relative abundance of the Antigen B subunits decreased asymptotically with increasing molecular weight. Neither the Antigen 5 nor the Antigen B subunit was specific for E. granulosus. Both antigens generated readily detectable levels of specific antibody in the sera of patients with Echinococcus multilocularis, Echinococcus vogeli or E. granulosus infection. Relatively high levels of antibody to Antigen 5 were also detected in the sera of patients infected with Taenia solium. The presence of phosphorylcholine epitope(s) on Antigen 5 was confirmed.


Parasitology Research | 1975

The developmental cycle of a species of Sarcocystis occurring in dogs and sheep, with observations on pathogenicity in the intermediate host

B. L. Munday; I. K. Barker; M. D. Rickard

SummaryTwelve dogs were fed mutton containing small sarcocysts, and killed 1, 3, 4, 6, 7, 10, 15, 16, 17 days after infection (DAI). Beginning 13–15 DAI sporocysts 14.7×9.0 μm were passed in the faeces of the dogs killed 15–17 DAI. Histological examination showed that developing stages were most numerous in the subepithelial tissue at the tips of villi in the proximal third of the small intestine. Macrogametes containing tiny PAS+ granules, and microgametocytes with peripheral developing microgametes were present 1 DAI. By 4 DAI oocysts, with a small nucleus and vacuolate cytoplasm were seen. Sporulation was observed 7–10 DAI. The first nuclear division resulted in 2 polar nuclei which divided laterally, resulting in 2 sporocysts each with 2 polar nuclei. This process was repeated once more to produce 4 nucleated sporozoites in each of 2 sporocysts. PAS+ granules were seen at the periphery of sporulating oocysts and sporocysts. There was a large PAS+ granule in the mid zone of sporozoites, with a smaller granule at one tip. Numerous sporulated sporocyst pairs were present beneath the epithelium at the tips of villi in dogs killed during patency.Four lambs were inoculated orally with sporocysts passed by dogs following feeding of infected mutton. Fifteen DAI schizonts were seen in the endothelium of arteries and arterioles in many organs, but not brain. Twenty-four DAI, smaller schizonts were seen in capillary endothelium in many organs, including brain. The two other lambs died 42 and 104 DAI, after an illness characterized by anaemia and ill-thrift. Mature schizonts were found in cells in the brain 42 DAI, associated with nonsuppurative meningoencephalitis. Developing sarcocysts were found in muscle, associated with myositis. Sarcocysts in muscle 104 DAI were mature. In the brain there were degenerate cysts and mature sarcocysts, and nonsuppurative meningoencephalitis.


Parasite Immunology | 1995

Cellular responses during liver fluke infection in sheep and its evasion by the parasite

Els N.T. Meeusen; C.S. Lee; M. D. Rickard; Malcolm R. Brandon

The cellular immune response in sheep to an acute and chronic primary and an acute secondary liver fluke infection were examined by immunohistology of liver tissue and flowcytometry of lymphocytes from the draining hepatic lymph nodes. Ten days after primary infection, portal tract areas surrounding migratory tunnels were infiltrated with CD4+ and CD8+ lymphocytes with fewer B cells and T19+ T cells. Micro abscesses were distributed sporadically in the liver parenchyma and young flukes could be easily observed in the liver tissue free from inflammatory cells. More intensive infiltration of the portal tract areas was observed during a secondary liver fluke infection characterized by a pronounced increase in eosinophils, B cells and CD4+ T cells. In addition, there was an increase in MHC class II+ fibroblastic‐like cells surrounding the migratory tracts. In contrast to the primary infection, no young flukes were observed in the same tissue areas during the secondary infection. Chronic primary infections were characterized by perilobular fibrosis and a predominance of CD8+ and γδ‐TCR+ T19‐ T cells distributed within fibrotic strands. Distinct B cell follicles were observed in the fibrotic strands and near major bile ducts and necrotic patches. Pronounced lymphocyte infiltration could occasionally be observed surrounding liver fluke eggs lodged in liver tissue. A progressive increase in lymph node weight, cell number and CD4/CD8 ratio was observed in the acute and chronic primary infections. The role of the infiltrating cell populations and possible mechanisms of immune evasion by the parasite are discussed.


International Journal for Parasitology | 1984

Immunization against Taenia taeniaeformis in mice: Studies on the characterization of antigens from oncospheres

Marshall W. Lightowlers; Graham F. Mitchell; David Bowtell; Robin F. Anders; M. D. Rickard

Abstract Mature eggs of Taenia taeniaeformis hatched readily in the presence of sodium hypochlorite and no loss in infectivity of oncospheres for mice was observed after hatching. Crude and sodium deoxycholate-solubilized antigens (termed TtO-DOC) prepared from such oncospheres stimulated high levels of protection against T. taeniaeformis infection in immunized mice similar to those described previously for oncospheres prepared by other methods. Mice immunized with TtO-DOC antigens that had been exposed to potassium metaperiodate remained significantly protected against infection. Exposure of TtO-DOC antigens to pronase and thermolysin, or to trypsin, significantly reduced the ability of these antigens to protect mice against infection. These data suggest that the antigens which immunize mice against infection include protein components. 125 I-labelled TtO-DOC antigens were immunoprecipitated with sera from mice infected with T. taeniaeformis and the immunoprecipitates analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Immunoprecipitation with sera from C3H/He mice infected for 28 days revealed a single major labelled protein antigen having a relative molecular mass ( M r ) of 31,000. Sera from 5-month infected C3H/He mice immunoprecipitated at least thirteen labelled antigens, including one at M r 31,000. Attempts to use SDS-PAGE separated proteins to immunize mice showed that oncosphere antigens exposed to the reducing conditions prior to SDS-PAGE lost their ability to protect mice against infection. It was concluded that SDS-PAGE was an unsatisfactory technique for the isolation of a host protective fraction of TtO-DOC antigens. TtO-DOC proteins were resolved by PAGE performed in the presence of sodium deoxycholate (DOC-PAGE) and mice were vaccinated with cut-outs from the gel. A fraction of the DOC-polyacrylamide gel was found to be effective in immunizing mice against infection. Thus, although the characteristics of the protein antigens in this DOC-PAGE fraction have yet to be determined, an important fractionation technique has been identified. It was shown that partial removal of DOC from oncosphere antigen preparations solubilized in 1% DOC was required for the antigen to stimulate protective immunity. These findings will facilitate further antigen characterization studies towards the development of a defined-antigen vaccine in murine cysticercosis. This is particularly so as attempts to raise anti-oncospheral monoclonal antibodies capable of passively transferring protection to mice by using crude antigen preparations to immunize donor mice have not been successful.


International Journal for Parasitology | 1996

Identification and cDNA cloning of two novel low molecular weight host-protective antigens from Taenia ovis oncospheres

G.B.L. Harrison; D.D. Heath; R.P. Dempster; Charles G. Gauci; Susan E. Newton; W.G. Cameron; C.M. Robinson; S.B. Lawrence; Marshall W. Lightowlers; M. D. Rickard

Oncosphere antigens of Taenia ovis were solubilised in sodium dodecyl sulphate and separated by electrophoresis in polyacrylamide gels (SDS-PAGE). Antigen-containing gel fractions cut from the region covering 18-12 kDa were shown to be highly immunogenic in sheep challenge experiments. Specific antisera against 2 candidate antigens at 18 and 16 kDa were used to screen a cDNA library prepared from T. ovis oncosphere mRNA. Recombinant proteins selected with antibody to the 16 and 18 kDa native antigens were expressed as GST fusion proteins. Vaccination trials using either of the 2 fusion proteins To16.17-GST and To18-GST, revealed that each was capable of inducing high levels of immunity in sheep against challenge infection with T. ovis eggs. Antibodies induced by vaccination with the recombinant antigens reacted specifically with their respective 18 or 16 kDa native oncosphere antigens. There was no apparent homology between the T. ovis cDNA coding for To18 and To16.17, or with another host-protective antigen, To45W, described previously. These additional host-protective antigens should prove a valuable adjunct to To45W and permit the development of effective vaccination strategies.


International Journal for Parasitology | 1980

Taenia taeniaeformis in mice: Protective immunization with oncospheres and their products

G.R. Rajasekariah; Graham F. Mitchell; M. D. Rickard

Abstract Taenia taeniaeformis oncospheres, injected parenterally with or without adjuvants, stimulated a high degree of protective immunity in mice against challenge infection with eggs. Furthermore, the supernatant from a centrifuged (twice for 30 min at 4500 g ) preparation of oncospheres which had been frozen, thawed and sonicated, induced more than 90% protection when used as a vaccine with adjuvant. By contrast, centrifuged (1 h at 3500 g ) supernatant medium collected during 72 h incubation of activated T. taeniaeformis oncospheres in vitro in serum-free culture medium was only marginally host-protective. The results indicate that materials from disrupted oncospheres should be a suitable starting preparation for identification and purification of ‘host-protective’ antigens.


Parasitology | 1995

Taenia ovis recombinant vaccine--'quo vadit'.

M. D. Rickard; G.B.L. Harrison; D.D. Heath; Marshall W. Lightowlers

Several years have elapsed since the publication by Johnson et al. (1989) of the cloning of a recombinant antigen from the cestode parasite Taenia ovis which stimulated high levels of protective immunity in sheep. A great deal of subsequent research and development was necessary to bring the fledgling vaccine to the point of being a registered commercial product. The results of these subsequent studies are dealt with briefly in this paper, including the results of field trials. The T. ovis vaccine was registered by the New Zealand Animal Remedies Board in February 1994. Where then is the commercial product? This paper gives a background to market problems which have emerged through the politics (and realities) of the NZ T. ovis control campaign. It serves as notice that the best science dedicated to producing vaccines or products for parasitic, or other, diseases often faces significant hurdles in the real world of commerce and politics.


Veterinary Parasitology | 1976

Vaccination of calves against Taenia saginata infection using a “parasite-free” vaccine

M. D. Rickard; A.J. Adolph

Abstract Calves were vaccinated with antigens collected during in vitro cultivation of the larval stages of Taenia ovis, T. hydatigena or T. saginata , and challenged 4 weeks later with 4 000 T. saginata eggs. Calves vaccinated with T. saginata antigen were highly resistant to the challenge infection and those groups vaccinated with T. ovis and T. hydatigena were also significantly resistant.


Parasitology Research | 1975

The location of Taenia pisiformis, Taenia ovis and Taenia hydatigena in the gut of the dog and its effect on net environmental contamination with ova.

Brian J. Coman; M. D. Rickard

SummaryAutopsy of dogs 56 days after infection with either T. pisiformis, T. ovis or T. hydatigena showed that these worms could be found attached at any point along the length of the small intestine, but were most commonly in the anterior half. The mean relaxed lengths of T. pisiformis, T. ovis and T. hydatigena were 107 cm, 156 cm and 177 cm respectively. Attached gravid proglottides contained a mean of 41000 eggs each in T. pisiformis, 31000 eggs in T. hydatigena and 95000 eggs in T. ovis, whereas proglottides free in the gut contained means of only 1370, 500 and 1400 eggs respectively; therefore, the majority of eggs were released into the gut before segments passed out into the faeces. It was shown that eggs of all 3 species of worms hatched and activated in the small intestine of the dog, especially in the anterior half. Eggs of T. pisiformis which had been passaged through the intestine of the dog and stored in the faeces for 5 days were poorly infective for rabbits compared with eggs only stored in faeces. It was concluded, therefore, that during taeniid infections of dogs the point of apolysis in the gut plays a significant role in determining environmental contamination with eggs. Puppies which had been fed 10000 T. ovis eggs daily for 6 weeks prior to infection with T. ovis cysticerci showed no difference in susceptibility to the infection when compared with untreated puppies.

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Graham F. Mitchell

Walter and Eliza Hall Institute of Medical Research

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D. Liu

University of Melbourne

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