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D Wakelin

Publications and source records attributed to D Wakelin.

At least 91 records · Page 5Linked to original sources
PubMed1993

ELISAs for parasitologists: or lies, damned lies and ELISAs.

The introduction in 1971, by Engvoll and Perlmann, of enzyme labels in immunoassays represented a significant technical advance. Their enzyme-linked immunosorbent assay (ELISA) proved to be as sensitive as radioimmunoossoys but safer to use. Since then, ELISAs have been widely used in the assay o f anti-bodies and antigens. In this article, Pradhib Venkateson and Derek Wokelin focus on some o f the problems associated with ELISAs when applied to parasitic infections, which can modify antibody responses, by immunosuppression or polyclonal activation. The article also considers optimization, which is an essential step in the establishment o f on ELISA.

Journal Article↗
PubMed1993

Failure to demonstrate suppressor T cell activity in the lamina propria of Hymenolepis diminuta-infected rats.

The aim of this study was to examine whether there was an increase of suppressor T cells, relative to helper T cells, in the intestinal lamina propria of Hymenolepis diminuta-infected rats, a condition which might allow the parasite to survive for the life-span of the host. Lamina propria cells were isolated by enzymatic procedure. All lymphocytes were passed over nylon wool columns in order to remove B cells; then the T cells were purified by a panning technique using monoclonal anti-T cell antibodies. Changes in the mitogen-stimulated synthesis of IgM, IgG and IgA by normal peripheral blood indicator lymphocytes, as measured by sandwich ELISA, was used as an index of help and/or suppression. No significant suppressor T cell activity was observed in the cultures, either from control or infected intestine.

Animals↗
PubMed1993

Influence of variation in host strain and parasite isolate on inflammatory and antibody responses to Trichinella spiralis in mice.

Variation in the immunogenicity of 3 isolates of Trichinella spiralis was assessed by the parameters of adult worm recovery, mast cell, eosinophil and antibody responses in mice of defined response phenotype. The levels of the protective, inflammatory and immune responses induced by infection differed between the isolates. Isolates showed considerable variation in the capacity to elicit mast cell and eosinophil responses. All induced increases in parasite-specific antibody, levels of total (IgGAM) antibody and of IgM and IgG isotypes rose steadily after infection, but there were significant differences in levels of response. The IgGAM response was correlated with the number of worms present, i.e. the greatest response was seen in low responder (C57BL/10) mice infected with the longest-surviving isolates. All isolates elicited specific IgG1 and IgG2a antibodies after infection, although, again, there were isolate-specific differences in the levels and kinetics of response. Levels of these isotypes were always higher, although not significantly so, in high-responder NIH mice. Low-responder mice showed higher IgE serum levels than high-responder mice after infection, one isolate giving much higher IgE values than the other two.

Animals↗
PubMed1993

Response of inbred mice to infection with a new isolate of Trypanosoma musculi.

A new isolate of Trypanosoma musculi has been derived from organisms recovered from Mus spretus captured in Lisbon, Portugal. The time-course and profiles of infection with this isolate in inbred mice have been compared with those obtained with the existing Partinico II isolate. Infections with the Portuguese isolate are less intense, and controlled more quickly than those with the Partinico isolate. As with the latter, there are marked mouse strain-dependent influences on infection with the Portuguese isolate, but these strain-dependent characteristics differ considerably with each isolate. For example, NIH mice were the most susceptible to the Partinico II isolate, but virtually refractory to the Portuguese isolate. Mice exposed to infection with one isolate show complete immunity to both homologous and heterologous challenge infections. These striking interactions between host and parasite genotype are discussed in terms of immunological influences on infection.

Animals↗
PubMed1993

Vaccination against Trichinella spiralis in mice using antigens from different isolates.

Antigen preparations from three isolates of Trichinella spiralis (sensu-stricto) have been tested for their ability to immunize mice against homologous and heterologous challenge infections. Immunization was measured by accelerated expulsion of adult worms, reduction in female worm fecundity, stunting of growth, and level of antibody responses before and after challenge. Antigens of two isolates, from London (L) and Poland (P), immunized effectively against homologous challenge, adult worm recoveries being reduced by 64% and 51% respectively within 6 days. Antigen from a Spanish (S) isolate gave no homologous protection in terms of worm recovery at day 6 and only 43% reduction at day 8. L- and P-isolate antigens immunized well against heterologous challenge (day 6 worm recoveries reduced by 66% and 59%) but similar protection against S-isolate challenge was not seen until day 8. Immunization with S antigen gave better protection at day 6 against L-isolate challenge (77%) than against challenge with P-isolate (37%). These results imply considerable, but not complete, antigen cross-reactivity between the isolates and this was reflected in antibody responses. These variations in immunogenicity and immunization potential are discussed in terms of constraints on the use of vaccines against parasites that are widely distributed geographically.

Animals↗
PubMed1993

Immunity to coccidiosis: genetic influences on lymphocyte and cytokine responses to infection with Eimeria vermiformis in inbred mice.

Cellular and cytokine responses to infection with Eimeria vermiformis were compared in BALB/c (resistant) and C57BL/6 (B6-susceptible) inbred mice. Cellular responses in the mesenteric lymph node (MLN) occurred sooner after primary infection in the resistant BALB/c strain. In contrast, proliferative responses occurred earlier after challenge in B6 mice. Resting levels of CD4 + ve and CD8 + ve T-lymphocytes in the MLN differed between the two strains but the relative numbers of each subset remained relatively constant throughout primary infection. MLN cells taken at intervals after infection were assayed for release of the cytokines IFN-gamma, IL-5 and IL-10 after culture in vitro with the mitogen Concanavalin A (Con-A) or with parasite antigen. With either stimulus cells from resistant BALB/c mice released IFN-gamma and IL-5 earlier after infection than did B6 cells. The strains had a comparable absolute ability to produce IFN-gamma but BALB/c cells released more IL-5 than did B6, levels declining, rather than increasing, during primary infection in the latter. Only cells from BALB/c mice released IL-10 during infection. Cells taken after a secondary infection released relatively little cytokine after pulsing in vitro. These data suggest that the difference in response phenotype between the two strains when infected with E. vermiformis reflect a kinetic, rather than a qualitative, difference in ability to mount protective T-helper (Th) cell subset responses. No evidence was found for a Th2-mediated interference with ability to release IFN-gamma, the cytokine most closely associated with protective immunity.

Animals↗
PubMed1993

Trichinella spiralis: immunity, ecology, and evolution.

Parasites exploit the environment provided by their hosts in the same way that free-living animals exploit their physical and biotic environments. There is, however, 1 important difference: the host environment can respond adaptively to infection in ways that may be disadvantageous to the parasite. Immunity provides the most effective adaptive host response and exerts a major selective pressure in parasite evolution. Among parasites many ways of maintaining the ability to exploit the host in the face of adaptive responses have evolved; such responses additionally may promote survival by enhancing transmission. The nematode Trichinella spiralis provides an excellent model with which to explore immuno-parasite-ecological ideas. Its biology is well known, and host immunity is described in detail. Immunity acts to limit parasite survival, but it also helps to ensure transmission by allowing the host to survive until infective stages mature. Host and parasite engage in a complex interplay, mediated by immunity, in which the genetic variability of each partner plays an important and necessary role.

Animals↗
PubMed1992

Inflammatory responses in the intestine during tapeworm infections. Mucosal mast cells and mucosal mast cell proteases in Sprague-Dawley rats infected with Hymenolepis diminuta.

Comparative studies were made of two populations of Sprague-Dawley rats infected with Hymenolepis diminuta. The time course of infection, the development of mucosal mastocytosis and the levels of rat mucosal mast cell (MMC) protease (RMCP II) in serum and in jejunal mucosal tissues were monitored at intervals after infection with 40 cysticercoids of the tapeworm. Worm expulsion patterns differed markedly between the two populations, rats of New Zealand origin showing an abrupt and clear-cut loss of worms, rats of English origin showing a more gradual decline over a longer time period. In both populations, however, numbers of MMC and levels of tissue RMCP II were positively correlated with time after infection and negatively correlated with worm numbers. In only one of the three experiments (using English strain rats over a short time period) did levels of serum RMCP II change with time. In the other two experiments, in which English-strain and New Zealand-strain rats were used, there were no correlations between serum RMCP II and time, numbers of MMC, numbers of worms or levels of tissue RMCP II. The absence of correlation between serum RMCP II and worm loss in these experiments implies that MMC have no direct role in expulsion of H. diminuta. The data do show, nevertheless, that this purely luminal tapeworm is fully capable of activating the mucosal T lymphocyte-MMC precursor axis to elicit a mucosal mastocytosis.

Animals↗
PubMed1992

Susceptibility of adult Heligmosomoides polygyrus to intestinal inflammatory responses induced by heterologous infection.

Adult H. polygyrus are capable of surviving for many months after primary exposure of mice to infective larvae, raising the possibility that worms of this species have inherent resistance to intestinal immune responses. Accordingly experiments were carried out to determine whether H. polygyrus are resistant to the inflammatory changes elicited during the acute phase of the intestinal response to Trichinella spiralis. Adult worms were expelled from mice when their presence coincided with the most intense phase of inflammation elicited by T. spiralis. The effect was dose-dependent with more intense T. spiralis challenge resulting in a correspondingly greater loss of H. polygyrus. Even the less pathogenic species T. pseudospiralis elicited a response of sufficient intensity in NIH mice to cause the expulsion of H. polygyrus from concurrently infected animals. Tissue larval stages of H. polygyrus were protected from expulsion by their location deep in the intestinal walls and the maximum detrimental effect against H. polygyrus was observed during the adult phase or during the establishment of L3 larvae. Acceleration of the response to T. spiralis in immune challenged mice resulted in earlier loss of H. polygyrus. When the expulsion of T. spiralis was delayed (e.g. from slow responder C57BL/10 mice) the loss of H. polygyrus took place correspondingly later. These experiments demonstrate unequivocally that mouse strains which normally tolerate chronic infections with H. polygyrus have the capacity to mount intestinal inflammatory responses of sufficient vigour to remove the worms but that this potential is not normally realized. However, the observation that some H. polygyrus always survived even when the response induced by T. spiralis was of the rapid secondary type suggests that the parasites are resilient in the face of the inflammatory response capable of removing most of the worms. It is suggested that in addition to the immunomodulatory strategy employed by adult worms to prevent the intestinal response being elicited, the worms have a second line of defence which is reflected in their resilience to responses which they have been unable to prevent.

Animals↗
PubMed1992

Immunization against geographical isolates of Trichinella spiralis in mice.

Partially purified antigen preparations from six isolates of Trichinella spiralis were used to immunize mice. Immunogenicity of the antigens was assessed in terms of antibody and lymphocyte responses and ability to stimulate protective immunity against challenge. Isolate antigens showed considerable cross-reactivity, and all elicited protective responses. Two major patterns of immunizing ability could be distinguished: (a) isolates that immunized well against heterologous challenge and elicited good immunity in London isolate-immunized mice (C-76 and Laso), and (b) isolates that immunized poorly against heterologous challenge and were least effective in London isolate-immunized mice (GM-1 and Mad-83). The immunogenicity of one isolate (C-76) was markedly greater than the others, inducing rapid loss in unvaccinated mice, almost complete protection in London isolate-immunized mice, and immunity in London isolate-challenged mice equivalent to the homologous antigen. These variations in immunogenicity, cross-reactivity and immunizing ability are discussed in terms of constraints that may operate against the development and use of vaccines against parasites that are widely distributed geographically.

Animals↗
PubMed1992

Genetic variation in resistance to parasitic infection: experimental approaches and practical applications.

Parasitic infections remain a major threat to the successful maintenance of domestic livestock. Control is still centered around management and husbandry, and the use of chemotherapeutic compounds. However, the spread of drug resistance in many important protozoal and helminth infections is placing an increasing emphasis on the development of alternative forms of control. At the present time vaccination is available for only a limited number of parasites, and may never be practical or cost-effective for some of the most important infections. This review examines current views of the mechanisms, control and likely value of genetically determined differences in resistance to parasitic infection. There is an extensive literature dealing with experimental analyses of genetic variation to parasitic infection in both laboratory and domestic species, and a growing interest in the application of this information in the field of practical parasite control.

Animals↗
PubMed1992

Genetic influences upon eosinophilia and resistance in mice infected with Trichinella spiralis.

Genetic influences upon host variation in eosinophilia and resistance to helminth infection, and the relationship between these parameters, were investigated in 7 inbred and 1 hybrid strains of mice infected with Trichinella spiralis. Clear strain-dependent variations were observed in the maximum peripheral blood, bone marrow and spleen eosinophilia attained in infected animals. SWR, NIH and SJL strains of mice all gave high responses to infection; four congenic strains sharing the B10 background (C57BL10 [B10], B10.S, B10.G and B10.BR) were low responders. Some of the genes for high responsiveness appeared to be dominant, as F1 hybrids from high- and low-response phenotype parental strains showed intermediate to high responses to infection. Intestinal eosinophilia showed no correlation with either peripheral blood or bone marrow responses (NIH and B10 strains having similar levels of eosinophil response in gut tissue) and was unrelated to the level of resistance to infection. Whereas NIH were highly resistant, with adult worm burdens at 13 days post-infection and muscle larval burdens at 35 days post-infection significantly lower than all other strains, B10 were quite susceptible, retaining substantial worm burdens at day 13 and harbouring large numbers of muscle larvae. Measurements of the level of the eosinophilopoietic cytokine IL-5 in sera during infection showed that the two strains differed in the kinetics of release but not in their absolute capacity to produce this cytokine. NIH mice released high levels during a primary infection, B10 released high levels during a secondary infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Immune control of murine coccidiosis: CD4+ and CD8+ T lymphocytes contribute differentially in resistance to primary and secondary infections.

The effect of treatment with monoclonal antibodies (Mabs) which deplete CD4+ or CD8+ T lymphocytes, on infections with Eimeria spp. was examined in NIH mice. Treatment with anti-CD4 Mab increased susceptibility to primary infections with E. vermiformis or E. pragensis and reduced the subsequent resistance of the mice to homologous challenge. Similar treatment of immune mice did not affect their resistance to re-infection but this was reduced in mice depleted of CD8+ T lymphocytes. In mice immunized with E. vermiformis the effect of CD8(+)-depletion was very slight, apparent only as the presence of small numbers of oocysts in the faeces of some mice; in mice immunized with E. pragensis there was a small, though significant, increase in oocyst production, compared with controls and anti-CD4-treated groups. These results confirm the importance of mechanisms involving the function of CD4+ T lymphocytes in the control of primary infections with Eimeria spp. and indicate that CD8+ cells play some part in the expression of resistance to reinfection. They also show that a major part of this resistance was not affected by either of the treatments given.

Animals↗
PubMed1991

Acanthocheilonema viteae (Dipetalonema viteae) in mice: differences in the relative binding of microfilarial surface-specific antibody may explain the contrasting response phenotypes of BALB/c and C57BL/10.

Experiments were carried out to obtain additional data concerning the role of IgM antibodies, specific for the cuticular surface of the microfilariae (mf) of A. viteae, in clearing microfilaraemia from high- and low-responder mice infected by transplanted adult worms. Although BALB/c mice, which sustain a chronic microfilaraemia, produced IgM mf surface-specific antibodies, the binding to target mf was weak when compared to that of antibodies from the serum of the resistant C57BL/10 mice. Furthermore, antibodies from BALB/c mice were not as efficient as those from C57BL/10 mice in promoting the adherence of immune or control leukocytes to mf in vitro. Evidence is provided to show that mf shed surface bound antibody. Although the results do not establish conclusively the mechanism underlying the contrasting response phenotypes of C57BL/10 and BALB/c mice, they provide support for the involvement of antibody in controlling microfilaraemia and suggest that quantitative and qualitative differences in the amount and affinity of IgM antibody specific for the mf surface, together with the natural tendency of the mf to shed surface bound antibody at 37 degrees C, may combine to allow the former strain to clear microfilaraemia efficiently whilst the latter sustains a chronic infection.

Animals↗
PubMed1991

Trichuris muris: antigen recognition and transfer of immunity in mice by IgA monoclonal antibodies.

Mesenteric node lymphocytes from mice that had been infected with the nematode Trichuris muris, and then boosted with adult worm excretory-secretory antigens were fused with myeloma cells to produce a panel of 9 monoclonal antibodies (MoAbs). Five of the MoAbs were of the IgA isotype. The antigen recognition profiles of these MoAbs were studied using SDS-PAGE and immunoblotting; three major profile patterns were identified. Five MoAbs recognized a major band in the MW range 43-48 kD; all recognized a range of antigens. Three MoAbs were used to localize antigens in the bodies of adult worms. Granules within the anterior stichocytes were recognized strongly, as was material within the eggs and pseudocoelom. Two MoAbs stained the cuticle. Although the phosphorylcholine (PC) determinant was widely distributed within worm tissues none of the MoAbs tested recognized PC. Passive transfer of immunity was achieved using two of the IgA monoclonals; no immunity was transferred by the IgM and IgG MoAbs used. The limited recognition profiles of these IgA MoAbs, and the ability to stain stichocyte granules, suggest that their protective activity results from an interaction with ES antigens.

Animals↗
PubMed1991

Interferon-gamma-mediated effects upon immunity to coccidial infections in the mouse.

The effect of treatment with a monoclonal antibody (MoAb) capable of neutralising interferon-gamma (IFN-gamma) on the course of coccidial infections in mice (C57BL/6 and NIH infected with Eimeria vermiformis or E. pragensis, and BALB/c infected with E. pragensis) was examined. The results differed with the species of parasite, the strain of mouse, the measure of infection and whether the infection was a primary or secondary one. The replication of E. vermiformis in primary infections was enhanced in both C57BL/6 and NIH mice, but less MoAb was required in NIH than in C57BL/6 to produce similar effects. In neither strain did treatment prevent priming or interfere with the complete immunity to challenge normally induced by moderate infection with E. vermiformis. The replication of E. pragensis in primary infections was not affected in any of the strains of mouse but the clinical effects were exacerbated. Priming with E. pragensis was unaffected by treatment but the partial immunity to challenge, normally induced by infection with this species, was reduced when MoAb was given 2 h before challenge. This reduction was evident as an increased faecal output of oocysts and loss of body weight. These results confirm the role of IFN-gamma in resistance to coccidiosis and further emphasise the complexity of the immune response in this disease.

Animals↗