Biomedical subjects

D Wakelin

Publications and source records attributed to D Wakelin.

At least 37 records · Page 2Linked to original sources

PubMed2001

Mucosal responses to infection with Trichinella spiralis in mice.

Infections with T. spiralis in mice elicit strong inflammatory responses. The nature and control of these responses, and their relationship to the process of worm expulsion, have been debated for many years. Many components of inflammation are, like worm expulsion, T cell-dependent, but some are not. The paper describes novel observations on Paneth cell responses to infection in immunologically normal mice and in a variety of T cell-deficient mice. Responses occurred normally in nu/nu and scid/scid mice but not in beta/delta knock out mice incapable of generating cells with functional TCR. However all of these mice failed to expel worms in the pattern seen in immunologically normal controls. These data are incorporated into a discussion of the causal relevance of intestinal inflammatory changes to the process of worm expulsion.

Animals↗

PubMed2000

Murine model study of the practical implication of trypanosome-induced immunosuppression in vaccine-based disease control programmes.

The relevance of trypanosome-induced immunosuppression in relation to the efficacy of vaccine-induced immunity was studied in mice. Mice were immunised with crude Trichinella spiralis muscle larvae homogenate vaccine and infected with T. spiralis and/or Trypanosoma brucei. Vaccination significantly decreased adult worm burden (p<0. 05) and accelerated worm expulsion in mice infected with T. spiralis only. T. brucei superinfection resulted in monocytosis, suppressed eosinophilia, significant decrease in PCV (p<0.001), higher numbers of adult worms (p<0.001) and failure to expel all adult worms by Day 12 post infection (p.i.). Regardless, they produced anti-Trichinella IgG(1) responses similar to those of the vaccinated non-T. brucei-infected group. T. brucei also suppressed the proliferative responses of spleen cells to stimulation with Con A and T. spiralis antigen, and induced strong production of interferon-gamma (IFN-gamma) in culture supernatants of antigen stimulated spleen and mesenteric lymph node cells. Interleukin-5 (IL-5) production was suppressed by T. brucei in supernatants of Con A- and antigen-stimulated spleen cells. It was concluded that trypanosome infections and the associated immunosuppression are of great practical significance in trypanosome endemic areas, especially with regards to disease control programmes involving vaccine-induced herd immunity.

Animals↗

PubMed2000

Immunopathology of intestinal helminth infection.

The relationship between intestinal pathology and immune expulsion of gastrointestinal nematodes remains controversial. Parasite expulsion is associated with intestinal pathology in several model systems and both of these phenomena are T cell dependent. However, while immune expulsion of gastrointestinal helminth parasites is usually associated with Th2 responses, the effector mechanisms directly responsible for parasite loss have not been elucidated. In contrast, the intestinal pathology observed in many other disease models closely resembles that seen in helminth infections, but has been attributed to Th1 cytokines. We have used infection with the nematode Trichinella spiralis in mice defective for cytokines to demonstrate that although parasite expulsion is indeed IL-4 dependent, contrary to expectations, the enteropathy is also regulated by IL-4. Furthermore, abrogation of severe pathology in iNOS deficient and TNF receptor defective animals does not prevent parasite expulsion. TNF and iNOS are therefore involved in intestinal pathology in nematode infections, apparently under regulation by IL-4 and Th2 mediated responses. Therefore, it appears that the IL-4-dependent protective response against the parasite operates by a mechanism other than merely the gross degradation of the parasite's environment brought about by the immune enteropathy. However, it remains important to elucidate the protective mechanisms involved in parasite expulsion, which are still unclear.

Animals↗

PubMed1999

Cellular immune responses in mice infected with the intestinal nematode Trichuris muris.

CBA and B10.BR mice show variation in immune response to the intestinal nematode Trichuris muris. CBA mice develop strong resistance, eliminating worms from the intestine; B10BR mice are permissive and develop chronic infections. It is already known that resistance and permissiveness reflect differential T helper responses. The data reported here show that resistant CBA mice express good antigen-specific lymphocyte proliferative responses to infection, whereas cells from B10.BR mice are relatively anergic, although still responsive to Concanavalin A (ConA). The possibility that the altered proliferative responsiveness seen in infected B10.BR mice reflected quantitative or qualitative changes in T helper cells was examined by comparing cytokine production and expression of cell surface markers (CD4, CD8, and CD28) in mesenteric lymph node cells and spleen cells from both strains and comparing these with the characteristics of cells from resistant CBA mice and from CBA mice that had been rendered permissive to infection by a combination of irradiation and corticosteroid treatment. As expected, cells from B10.BR mice produced high levels of interferon-gamma (IFN-gamma), whereas those from CBA mice released high levels of IL-5, whether stimulated with adult worm somatic antigens, excretory/secretory antigens, or ConA. Immunosuppressed CBA mice produced high levels of both IFN-gamma and IL-5 throughout the experiment. FACS analysis revealed a decrease of CD4+ and an initial increase in CD8+ cells in all infected mice. No major changes occurred in the relative proportion of CD28(+) cells. Further evaluation of the CD28 costimulatory molecule, measured as mean fluorescence intensity, displayed down-regulation in permissive and immunosuppressed mice. The data obtained suggest that lymphocyte unresponsiveness and permissiveness to T. muris infection may be associated with a down-regulation or an initially reduced expression of costimulatory CD28 molecules.

Animals↗

PubMed1999

Trypanosome-induced suppression of responses to Trichinella spiralis in vaccinated mice.

Mice vaccinated against the gastro-intestinal (GI) nematode Trichinella spiralis by injection of muscle larval homogenate antigen express a strong immunity to subsequent infection, reflected in earlier expulsion of adult worms from the intestine and reduced female worm fecundity. Infection with Trypanosoma brucei at the time of vaccination, or at the time of infection with T. spiralis, significantly reduced the level of immunity expressed, the effect being greatest when vaccination and T. brucei infection were given together. Trypanosome infection reduced T. spiralis-specific antibody responses in vaccinated mice, the effect being most apparent against IgM, IgG1 and IgG2b, and ablated the eosinophil response to T. spiralis. In vaccinated mice infected with both trypanosomes and T. spiralis, the proliferative responses of lymphocytes to the mitogen Con A or to T. spiralis antigen were much lower than in vaccinated mice infected only with the nematode. Whereas cells from mice infected only with T. spiralis produced the cytokine IL-4 and little or no IFNgamma when stimulated in vitro, cells from animals infected with T. spiralis and with trypanosomes released large amounts of IFNgamma but no IL-4. These observations are consistent with the known, IFNgamma-dependent, nitric-oxide-mediated suppressive effects of trypanosomes on lymphocyte function and the Th1 bias associated with these infections, both of which reduce the effectiveness of the Th2-mediated responses involved in immunity against GI nematode infections. The data are discussed in the context of the possible use of vaccines against GI nematodes in ruminants in countries where concurrent trypanosome-GI nematode infections are widespread.

Animals↗

PubMed1999

Primary infections with Babesia microti are not prolonged by concurrent Heligmosomoides polygyrus.

Seven experiments were carried out to test the hypothesis that concurrent infection with the chronic and immunomodulatory intestinal nematode parasites, Heligmosomoides polygyrus, and the piroplasm, Babesia microti, would result in more intense and long-lasting infections with the hemoprotozoan. However, despite variations in the experimental protocols (different mouse strains, varying levels of infection and different intervals between infection with the two species) a significantly higher B. microti parasitaemia was detected on only one occasion, and a significantly lower parasitaemia on two occasions, relative to control mice. In none of our experiments was the duration of infection prolonged. We conclude that the presence of H. polygyrus does not interfere markedly with the host's ability to mount a protective response against B. microti and hence season-dependent peaks of abundance of H. polygyrus in wild rodents are unlikely to present a particular threat to human communities by providing a greater reservoir of infection with B. microti in wild rodents than at other times of the year.

Animals↗

PubMed1999

The response of Nigerian West African Dwarf goats to experimental infections with Haemonchus contortus.

One option for controlling haemonchosis in warm pastoral regions is improvement of resistance by selective breeding. Variation in acquired immunity to H. contortus and immunological correlates of infection were studied in West African Dwarf (WAD) goats. Following exposure to 5000 L3, 63 per cent of the inoculum established but 77 per cent of established worms were expelled by week 5. All infected animals were anaemic (day 14). When exposed to 2000L3, 36 per cent of the inoculum was still present (day 35) with no loss by day 49. Persisting primary infection worms survived a superimposed challenge (day 35), but their growth was slowed and resistance to challenge was significant. Most goats showed eosinophilia and parasite-specific IgG responses to primary infection, but only eosinophilia increased after challenge. No consistent associations were found between parasite burden and any immunological measures of infection, but parasite egg counts showed considerable variation. Overall, our results suggest that resistant genotypes exist among the WAD goat population.

Animals↗

PubMed1998

Early cytokine responses during intestinal parasitic infections.

Infections with gastro-intestinal nematodes elicit immune and inflammatory responses mediated by cytokines released from T-helper type-2 (Th2) cells. In vitro assays of cells from the mesenteric lymph nodes (MLN) of experimentally infected rodents confirm that, after about 1 week, the dominant cytokine responses to mitogens and antigens are those associated with this Th-cell subset. Polarization of the Th response in this way implies an initial local cytokine environment that favours Th2 development. However, experimental infections with Trichinella spiralis and Nippostrongylus brasiliensis show that, within 2 days of worms reaching the intestine, MLN cells (MLNC) respond with a Th1 rather than a Th2 response [i.e. there is an increase in mRNA for the type 1 cytokine interferon-gamma (IFN-gamma), and mitogen-stimulated MLNC release IFN-gamma rather than interleukin-5 (IL-5)]. Antigen stimulation at this time does not elicit IFN-gamma release and the MLNC cannot adoptively transfer immunity. Within a few days the MLNC phenotype changes. There is a Th2 response (IL-5 release) to both mitogen and antigen stimulation and MLNC can adoptively transfer immunity. Early release of IFN-gamma is T-cell dependent, with CD4+ T cells playing the major role. The data are discussed in relation to factors regulating the mucosal response to invasion by parasites.

Animals↗

PubMed1998

Immunodepression induced by Trypanosoma cruzi and mouse hepatitis virus type 3 is associated with thymus apoptosis.

Trypanosoma cruzi-infected mice show disturbance in the peripheral immune system such as polyclonal lymphocyte activation, autoantibody production, and immunosuppression of T lymphocytes. Previous observations in our laboratory showed that some stocks of T. cruzi can be contaminated with mouse hepatitis virus type 3 (MHV-3). Literature has shown that MHV-3 infection induces immunologic disorders characterized by thymic involution with marked cell depletion. However, the effects of interactions between MHV-3 and the parasite on the immune system are not well understood. In the present study specific-pathogen-free CBA mice were inoculated with MHV-3, alone or associated with different stocks of T. cruzi. Concurrent murine virus infection resulted in increased pathogenicity of T. cruzi infection shown by profound thymic atrophy; loss of cortical thymocytes; depletion of Thy1.2+, CD4+, and CD8+ cells; enhancement of in situ labeling of nuclear DNA fragmentation; and eventually, death of the animals. Such lines of evidence show that the mechanism underlying this thymic atrophy is associated with apoptosis. These results also suggest that MHV-3 can account for the increased immunosuppression observed during experimental infection with the parasite.

Animals↗

PubMed1998

Production of proinflammatory cytokines and inflammatory mediators in human intestinal epithelial cells after invasion by Trichinella spiralis.

Epithelial cells are the first point of host contact for invasive intestinal pathogens and may initiate mucosal inflammatory responses via production of proinflammatory cytokines and mediators. The aim of the present study was to investigate in vitro the initial invasion of a parasitic nematode (Trichinella spiralis), to measure the early production of specific epithelial cytokines and inflammatory mediators after invasion, and to compare these responses with those to invasive bacteria. Monolayers of human colonic epithelial cell lines (HT29, T84, and Caco-2) were infected by T. spiralis or Listeria monocytogenes. Bile-activated infective larvae of T. spiralis invaded and migrated into the epithelial cell monolayers, leaving trails of dead cells. Transmission electron microscopy studies of damaged cells along the trail showed a progressive increase in size, disruption of cell membranes, loss or dilution of cytoplasmic proteins, and swelling of mitochondria and nuclei. However, no nuclear fragmentation was observed. With reverse transcription-PCR and an enzyme-linked oligonucleotide chemiluminescent assay, mRNA transcripts of interleukin-1beta (IL-1beta), IL-8, and epithelial neutrophil-activating peptide 78 were shown to increase in epithelial cells invaded by T. spiralis or L. monocytogenes, but only L. monocytogenes elicited increased inducible nitric oxide synthase (iNOS) mRNA. No increase in tumor necrosis factor alpha or transforming growth factor beta mRNA was seen after T. spiralis invasion. Increased levels of IL-8 were also released from the basolateral surfaces of infected monolayers as detected by sandwich enzyme-linked immunosorbent assay. Induction and secretion of proinflammatory cytokines in epithelial cells after nematode or bacterial invasion may initiate the acute inflammatory response of the small intestine. The upregulation of iNOS in bacterial infections may contribute to mucosal defense and may also be associated with subsequent cell death, whereas different mechanisms appear to operate after nematode invasion.

Animals↗

PubMed1998

Absence of lysozyme (muramidase) in the intestinal Paneth cells of newborn infants with necrotising enterocolitis.

AIM: To determine immunocytochemically whether preterm and newborn infants with necrotising enterocolitis (NEC) show differences in numbers of lysozyme positive Paneth cells compared with normal controls, and to relate the findings to the possibility that lysozyme deficiency may facilitate the bacterial infections thought to be associated with this condition. METHODS: Tissues from 10 infants with NEC and from 11 matched controls were sectioned and stained immunocytochemically for lysozyme. Differences in the numbers of Paneth cells and degree of lysozyme positivity in the tissues were assessed. RESULTS: Tissues from NEC patients showed no, or very few, lysozyme positive Paneth cells, whereas controls showed strong positive staining. CONCLUSIONS: A deficiency or developmental defect in Paneth cells, resulting in an absence of lysozyme, may render the intestine more susceptible to bacterial infection, allowing organisms to adhere and translocate across the mucosa. Such enhancement of infection may contribute to the pathogenesis of NEC.

Bacterial Infections↗

PubMed1998

Genetic resistance to parasitic infection.

The ability of animals to resist infections with parasites is genetically determined and therefore variable between individuals or breeds of a given host species. Such variation may involve innate (non-immunological) and acquired (immunologically mediated) resistance mechanisms, and is determined by both major histocompatibility complex (MHC)-linked and non-MHC genes. Resistance is inherited as a dominant trait, with heritability often exceeding 0.3. Genetic variation can be exploited to improve the capacity of domestic animals to resist parasitic infection. Methods to achieve this exploitation are discussed in relation to gastrointestinal nematode infections in sheep.

Animals↗

PubMed1997

Immune response to the cestode Hymenolepis nana: cytokine production during infection with eggs or cysts.

Analysis of cytokine production (IFN-gamma, IL-2, IL-3, IL-4, IL-5) by in vitro Con A-stimulated mesenteric lymph node cells measured daily after egg or cyst infection of mice with Hymenolepis nana showed that cytokine production varies during parasite development and between different host strains (BALB/c and C3H/He mice). Egg infection stimulates a rapid increase in IFN-gamma, independent of mouse strain. In addition, in BALB/c mice a Th2-like response (IL-4, IL-5 secretion) was stimulated 4-5 days p.i., when the parasites are thought to begin their lumenal phase. After infection with cysts significant increases in IFN-gamma, IL-2, IL-4 and IL-5 were observed at the time when autoinfection with eggs is thought to occur. The level of IFN-gamma paralleled that seen after a primary egg infection. This suggests that there is a predominantly Th1-type response during the tissue phase of H. nana development and that, in BALB/c mice, a Th2 polarization occurs during the first few days of the lumenal phase. The cytokine patterns observed are discussed in relation to host responses during chronic helminth infection.

Animals↗

PubMed1997

Cytokine production during infection with Hymenolepis diminuta in BALB/c mice.

Levels of the cytokines IFN-gamma, IL-2, IL-3, IL-4 and IL-5 were monitored in cells from the mesenteric lymph nodes (MLN) of BALB/c mice infected with Hymenolepis diminuta after stimulation in vitro with Con-A. Infection was associated with an increased production of IL-3, IL-4 and IL-5 and a low production of IFN-gamma, indicating the preferential activation of a Th2 response. It is suggested that this reflects the purely lumenal development of this worm. The results are discussed in relation to effector mechanisms known to be involved in immunity to intestinal helminths.

Animals↗

PubMed1997

Immunization of mice against Trichinella spiralis and T. britovi using excretory and secretory antigens.

Immune responses to immunization and infection with Trichinella spiralis and T. britovi were studied in NIH high-responder mice. Overall it was shown that T. britovi was the more immunogenic, immunization and challenge with this species giving greater host-protective immunity. This greater immunogenicity was reflected in higher proliferative responses when mesenteric node lymphocytes (MLNC) from immunized mice were restimulated with T. britovi antigens in vitro and in higher levels of T helper 2 (Th2) lymphocyte-dependent specific IgG1 antibody responses against this species. MLNC from mice immunized against T. britovi released more IL-5 when restimulated in vitro, again suggesting a greater T helper 2 subset response, but after infection the highest levels of IL-5 were recorded from MLNC taken from T. spiralis challenged mice. These data are discussed in relation to current understanding of immunological differences between species and isolates of the genus Trichinella.

Animals↗

PubMed1997

A comparison of mucosal inflammatory responses to Giardia muris in resistant B10 and susceptible BALB/c mice.

In the first three weeks of primary Giardia muris infections B10 mice clear infection more rapidly than BALB/c mice. There is evidence that interferon-gamma contributes to the relative resistance of B10 mice. The nature of the functional contribution of interferon-gamma is unclear and does not relate to the secretory or serum antibody response. Mucosal inflammatory events in these strains have been studied. Apart from a small rise in both strains of goblet cell and mucosal mast cell numbers, associated with release of mast cell protease-1 in serum, no inflammatory infiltrate was observed at the time trophozoites were cleared from the intestinal lumen. Inhibition of mast cell products (5-hydroxytryptamine and histamine) by cyproheptadine enhanced the intensity of infection in both strains. The relative resistance of B10 mice could not be explained in terms of the mucosal inflammatory response.

Animals↗

PubMed1997

Role of T helper 2 cells in intestinal goblet cell hyperplasia in mice infected with Trichinella spiralis.

BACKGROUND & AIMS: The four principal types of intestinal epithelial cells are derived from multipotent stem cells. Currently, there is no information on factors that regulate commitment of stem cells to differentiate along one lineage vs. another. The aim of our study was to investigate the role of T cells in the regulation of small intestinal goblet cell hyperplasia in mice infected with the parasite Trichinella spiralis. METHODS: NIH mice were infected with T. spiralis, and intestinal goblet cells and cytokine response were studied. Interferon gamma and interleukin 5 were used as candidate T helper (Th)1 and Th2 cytokines, respectively. Adoptive transfer experiments were also performed. RESULTS: Small intestinal goblet cell hyperplasia occurred 8 days after infection with T. spiralis. Th1-type cells were predominant in the mesenteric lymph nodes early in the course of infection, with a switch to Th2-predominant cells around the time of goblet cell hyperplasia. Transfer of Th2-enriched mesenteric lymph node cells further enhanced goblet cell hyperplasia in recipient mice. Neutralization of interleukin 5 activity did not affect T. spiralis-induced goblet cell hyperplasia. CONCLUSIONS: Small intestinal goblet cell hyperplasia in T. spiralis-infected mice is probably regulated by Th2 cells. We postulate that Th2-derived factors (other than interleukin 5) induce stem cells to differentiate preferentially along the goblet cell lineage.

Animals↗

PubMed1997

Oral vaccination against coccidiosis: responses in strains of mice that differ in susceptibility to infection with Eimeria vermiformis.

Four strains of mice with different susceptibilities to Eimeria vermiformis were orally dosed with a crude antigen prepared from sporulated oocysts of the parasite, with or without cholera toxin as adjuvant. The effect on subsequent challenge infections depended on the resistance and susceptibility phenotypes of the host: oocyst production was reduced in susceptible C57BL/6 and NIH mice but increased in resistant BALB/c and C3H mice. Despite this contrast, no fundamental differences were detected between the immune responses of BALB/c and C57BL/6 mice, either to vaccination or after superimposed infection, but the suppressing and enhancing effects of vaccination were transmissible to naive recipients via suspensions of mesenteric lymph node cells. The results obtained are compared with those previously reported for parenterally immunized BALB/c and C57BL/6 mice.

Administration, Oral↗