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Biomedical subjects

J Huntley

Publications and source records attributed to J Huntley.

At least 19 recordsLinked to original sources

Phenotypic and functional characterisation of follicle-associated epithelium of rectal lymphoid tissue.

Lymphoid follicles cluster in the terminal rectum of various animal species and of man and hence this site may be important in the development of immune responses to pathogens. For the induction of immune responses at mucosal sites, interplay is required between various cell types performing functions ranging from antigen-sampling cells via antigen-presenting cells to antigen-specific lymphocytes. Therefore, we have characterised the cell populations and relevant functioning of follicle-associated epithelium (FAE) and associated follicles in the terminal portion of rectum in cattle as a representative mammal. Immunohistochemical studies of this region identified immune cell subsets (CD4+, CD8+, WC 1+gammadelta, CD2+, CD 21+ and CD 40+ cells) characteristic of an immune-inductive site. Examination of FAE identified a subset of cells with structural and functional features of antigen-sampling M-cells. Cells of the FAE and adjacent follicle-associated crypts expressed vimentin and a subset of these cells internalised microparticles, a further attribute of M-cells. The FAE cells were phenotypically heterogeneous and therefore the function and phenotype of these cell subsets requires further characterisation, particularly with respect to their potentially important role in the interaction of hosts with pathogens and the development of immune responses.

Animals↗

The effect of dietary polyunsaturated fatty acids (PUFA) on infection with the nematodes Ostertagia ostertagi and Cooperia oncophora in calves.

Diet-induced changes in the polyunsaturated fatty acid (PUFA) content of immune cells influences the immune phenotype that develops following infection. The aim of this study was to examine the effect of manipulating dietary PUFA supply on tissue fatty acids composition and immunity to a mixed infection with an abomasal and an intestinal nematode parasite in calves. Calves (n=24) were allocated into two treatment groups and fed 25 g/day of either fish oil (n-3 group) or a binary mixture of palm/rapeseed oil (normal group) as a supplement in milk replacer. Within each treatment group eight calves were infected with 2000 L3 Ostertagia ostertagi and Cooperia oncophora, three times per week for 8 weeks, the remaining calves were pair-fed uninfected controls. Faecal egg counts (FEC) were carried out twice weekly. At slaughter, the whole gut was removed intact for worm counts and tissue samples were taken for fatty acid analysis. Samples of abomasum, duodenum and mid-gut were also collected for immunohistological analysis. FEC were not significantly influenced by oil supplement but tended to remain higher in the palm/rapeseed oil-fed group (normal infected). The number of intestinal immature worms was significantly (p<0.05) higher in the n-3 group. Mucosal mast cell (MMC) and eosinophil numbers were significantly increased (p<0.05) by infection and were significantly lower (p<0.05) in the intestinal tissue of the fish oil supplemented and infected group (n-3 infected group). These results suggest that feeding an n-3 PUFA-rich supplement (fish oil) can influence cellular mediators of immunity to nematode infection. This is the first report of the establishment of patency and the subsequent development of immunity to a mixed infection with O. ostertagi and C. oncophora in calves undergoing early rumen development. The trend in the FEC, MMC and eosinophil numbers in the n-3 group suggests that decreasing the dietary n-6/n-3 PUFA ratio may be a worthwhile immunonutritional strategy for potentiating the immune response to nematode parasite infection in the calf.

Animals↗

Nonatopic allergen-independent mast cell activation in parasitized eosinophilic athymic rats.

It is possible to induce eosinophilia in congenitally athymic rats by infection with the parasite Ascaris suum. Athymic bronchial eosinophilia is associated with increased expression of IL-5 and eotaxin mRNA, and with the presence of residual T cells and mast cells. Anamnestic mastocytosis is particularly pronounced and in this study we examine the relationship between mast cell degranulation and IgE production in athymic rats following infection. Incubation of peritoneal mast cells from athymic rats with anti-IgE induced dose-dependent degranulation, as measured by histamine release. However, the failure of mast cells from infected athymic rats to degranulate following incubation with all but one of the parasite antigens selected confirms the absence of a specific IgE response. In contrast, all agonists induced degranulation in euthymic rats. The only parasite-derived factor to induce histamine release in all mast cells was Ascaris body fluid, which contains an element capable of inducing IgE-independent degranulation. Furthermore, increased levels of rat mast cell protease II in athymic bronchoalveolar lavage fluid suggest degranulation of mast cells by IgE and allergen independent means in vivo. We believe that the development of eosinophilic lung inflammation in the absence of specific IgE makes this a prototype for investigating the immunological mechanisms underlying nonatopic asthma.

Animals↗

Studies using Teladorsagia circumcincta in an in vitro direct challenge method using abomasal tissue explants.

An in vitro direct challenge (IVDC) method in which abomasal tissue explants maintained at 37 degrees C in Hanks/Hepes solution (pH 7.6) in a high oxygen concentration (incubator gassed with pure oxygen and sealed prior to use) were-challenged with exsheathed third stage larvae of Teladorsagia circumcincta has been used to investigate the tissue association phase of larval establishment. Studies using naïve and recently challenged immune sheep and goat abomasal tissues have shown significant reductions (P < 0.05) in the numbers of tissue associated larvae in material from immune animals. The mechanisms that affect the tissue association process appear to be labile since more larvae were recovered from the tissue digests of previously infected sheep that had not been recently exposed to larval challenge in comparison to those that had been recently challenged (P < 0.05). The method has also been used to demonstrate the influence of protein nutrition on the establishment of larvae in abomasal tissues and region specific differences in the efficacy of exclusion mechanisms. The technique appears to have potential as a means of investigating the crucial first phase of the process of establishment.

Abomasum↗

A comparison of larval development and mucosal mast cell responses in worm-naïve goat yearlings, kids and lambs undergoing primary and secondary challenge with Teladorsagia circumcincta.

Larval development, mucosal mast cell (MMC) and eosinophil responses in worm-nai;ve lambs, yearling goats and goat kids were compared using two different experimental challenge regimes involving oral administration of infective Teladorsagia circumcincta L(3). Experimental challenge regimes enabled primary and secondary immune responses in the two species to be compared. Goats carried higher worm burdens than lambs and there were significant differences in the stages of development attained by the larval challenge that established in the two species. Possible physiological reasons for these differences are discussed. There were also differences in the establishment and development of larvae in individual yearlings which may indicate the development of a weak age-related immune response. Quantitative analysis of MMC and globule leukocyte (GL) recruitment and functional activity in the form of mast cell-specific proteinase (MCP) production demonstrated differences between the species with goat tissues containing significantly higher numbers of GL and lower concentrations of MCP than the lambs. Quantitative analysis of blood and tissue eosinophil responses failed to demonstrate any significant differences in either species under the two challenge regimes.

Aging↗

Approaches to vaccines for Psoroptes ovis (sheep scab).

A brief outline of the history of sheep scab in the UK is presented together with the current chemical methods used for its control and problems associated with these. Possible alternative approaches to control are discussed, as are selected aspects of the physiology of Psoroptes ovis and the pathogenesis of the scab lesion from the perspective of control through immunisation. Evidence is provided that immunity to the disease can indeed be acquired, both naturally after a previous infection and following inoculation of mite proteins in adjuvant. These results support the view that control by vaccination may be possible, although little is known to date about the antigens involved or the mechanism of protection.

Animal Feed↗

Susceptibility of high and low producer dairy goats to single experimental infection with Trichostrongylus colubriformis.

Culled dry dairy goats, which differed in their level of production in previous lactations, received a single infection with Trichostrongylus colubriformis. The objectives of the study were twofold. First, the study aimed at examining the ability of dairy goats to develop an immune response to trichostrongyle infection and the associated cellular changes developing within the intestinal mucosa. Second, a comparison between animals differing in their level of production was assessed, in order to determine whether these differed in their susceptibility to infection. No difference occurred in egg excretion, worm burden and local inflammatory cell responses between high (HP) and low (LP) producer dairy goats, in contrast to observations in previous studies. Because their nutrition was controlled and milk production absent in the goats employed in the present study, these results suggest that any genetic component associated with the selection of HP and LP goats had little influence on the development of acquired resistance to T. colubriformis. The analysis of the relationship between different cell types in the mucosa and some characteristics of the worm population show that eosinophils are negatively related to worm burden. Also, a role is suggested for mast cells and globule leukocytes in the modulation of egg excretion.

Animals↗

The effect of truncated infections with Ostertagia ostertagi on the development of acquired resistance in calves.

The relative contribution of the third (L3), fourth (L4) and adult stages of Ostertagia ostertagi to the development of immunity was assessed in calves which were either continuously infected during 21 weeks or subjected to infections truncated by anthelmintic treatment at the L3 or L4 stage. A fourth group remained uninfected (control group). Faecal samples and blood samples were collected weekly for faecal egg counts and determination of pepsinogen and antibody levels. Only the continuously infected animals showed positive egg counts, which fell towards the end of the primary infection period. Pepsinogen and antibody levels remained high in the continuously infected group until the end of the primary infection period. At that time, they were significantly higher compared to the control calves, with intermediate values in the truncated infection groups. After the 21 weeks primary infection period all animals were dewormed. To evaluate the protection provided by the different immunisation protocols, all animals were challenged 1 week later with 156000 Ostertagia L3, spread over 12 consecutive days. The marked reduction in egg counts following challenge infection indicated a certain degree of immunity in the continuously infected calves, which was confirmed at necropsy by the reduced worm burdens, the high percentage of inhibited early L4 larvae, the reduced size of the adult worms and the higher numbers of mucosal mast cells in this group. Numbers of globule leucocytes and eosinophils were not significantly different from the control group. Infections truncated by anthelmintic treatment elicited poor development of immunity as shown by the egg output after the challenge infection and the percentages of arrested larvae and the lengths of adult worms which were intermediate to those of the continuously infected calves and control animals.

Animals↗

Analysis of the expression and secretion of isotypes of sheep B cell immunoglobulins with a panel of isotype-specific monoclonal antibodies.

Monoclonal antibodies to sheep light chain, IgM and IgG were produced and used to assess total immunoglobulin (Ig) synthesis by sheep B cells in culture and antibodies to specific antigens. By using these antibodies in a dual fluorescence-activated cell sorting analysis of sheep efferent lymph B lymphocytes the percentage change in surface Ig isotype of B lymphoblasts from IgM to IgG after the antigenic stimulation of the local lymph node was measured. An extension of this analysis to paired blood and afferent or efferent lymph B cells made it possible to investigate the recirculation characteristics of B cells expressing different Ig isotypes.

Animals↗

Intracellular pH in lizards after hypercapnia.

We used the transmembrane distribution of 5,5-[2-14C]dimethyloxazolidine-2,4-dione ([14C]DMO) and 31P magnetic resonance spectroscopy (NMR) to investigate the effects of hypercapnia on intracellular pH (pHi) in brain and skeletal muscle of two lizard species: Anolis equestris and Dipsosaurus dorsalis. In control animals (normocapnic), plasma PCO2 (3.3 +/- 0.1 kPa) and plasma pH (7.52 +/- 0.01) for D. dorsalis were not significantly different from the values for A. equestris (2.8 +/- 0.2 kPa and 7.59 +/- 0.02, respectively). Furthermore 60 min of 5% CO2 increased plasma PCO2 and decreased plasma pH by the same amounts in both species. Brain pHi values determined with the DMO method were not significantly different from values determined with NMR. Control values of brain pHi (DMO, 7.16 +/- 0.01; NMR, 7.11 +/- 0.02) and muscle pHi were significantly higher for D. dorsalis (DMO, 7.15 +/- 0.03) than for A. equestris (DMO, 6.99 +/- 0.03; NMR, 7.02 +/- 0.02 for brain; DMO, 6.97 +/- 0.03 for muscle). In addition, changes in tissue pHi after 60 min of 5% CO2 were significantly different for the two species. In D. dorsalis muscle and brain pHi decreased significantly after hypercapnia, whereas in A. equestris muscle pHi decreased significantly but brain pHi was unchanged. Our findings were independent of the methods used to determine pHi. The smaller change in brain and muscle pHi than in plasma pH for A. equestris is consistent with the view that pHi regulation involves active processes such as transmembrane ion transport.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Mast cell recovery following chronic treatment with compound 48/80.

Histochemical and functional properties of mast cells (MC) in Brown Norway rats recovering from chronic treatment with the MC secretagogue compound 48/80 were examined. In the skin, treatment for 5 days with compound 48/80 resulted in a marked decrease in MC subpopulations defined by differential alcian blue/safranin staining. Both safranin-positive connective tissue MC and alcian blue staining MC were reduced in number. This was accompanied by significant decreases in skin histamine and rat MC serine protease I contents and a loss of specific IgE-mediated passive cutaneous anaphylaxis (PCA) activity. The PCA reaction did not return to normal before 2 months after stopping treatment and only when the numbers of safranin-positive connective tissue MC and skin histamine content reached pretreatment levels. The subepidermal alcian blue staining MC not eliminated by the compound 48/80 treatment were formalin resistant (unlike alcian blue staining mucosal MC of the intestine) and apparently played no role in the PCA response. MC numbers, histamine levels, and rat MC serine protease I content of the tongue were similarly decreased by compound 48/80. In contrast, mucosal MC of the gut were unaffected by the secretagogue treatment.

Alcian Blue↗

Health at work in the NHS feature.

Almost 20 million people are in paid employment in England today, spending up to 60 per cent of their waking hours in their places of work, many for 40 years or more. Workplaces are, therefore, a major influence on health and provide an invaluable setting for health promotion.

Health Promotion↗

Interactions between Teladorsagia circumcincta and Trichostrongylus vitrinus infections in young lambs.

Groups containing six lambs that previously had been infected five times each week for four, eight or 12 weeks, either with 1000 Trichostrongylus vitrinus third stage larvae (L3) or 1000 T vitrinus L3 and 2500 L3 Teladorsagia (previously Ostertagia) circumcincta were challenged, along with six worm-free control lambs, with 3000 75Se-labelled T vitrinus L3. The lambs were killed 10 days after challenge. Those given a dual infection for four and eight weeks had significantly lower total T vitrinus numbers than monospecifically infected lambs. Significantly fewer radiolabelled larvae were recovered from the dual infected group killed after four weeks exposure to infection than from lambs infected monospecifically. The negative interaction between these two nematode species appears to be physiologically mediated and may be an important factor in accounting for differences in their seasonal prevalence.

Abomasum↗

The generation of ovine bone marrow-derived mast cells in culture.

Ovine mast cells generated in vitro are morphologically and biochemically similar to mast cells localized to the gastrointestinal tract in vivo. Bone marrow (BM)-derived haemopoietic precursor cells were cultured in the presence of conditioned medium (CM) prepared from in vitro-activated lymphocytes from nematode-infected sheep. Mast cells developed rapidly after 2 weeks of culture and could persist for 2-3 months. The presence of sheep mast cell proteinase (SMCP) in the cultured cells correlated with the number of morphologically recognizable mast cells by Leishman-Giemsa staining and is definitive as, in vivo, this enzyme is unique to mast cells. The content of SMCP per cell increased as the cultures developed. In vivo, increased serum concentrations of SMCP are associated with mast cell function at the site of nematode infection. The availability of the cultured cells should now facilitate mast cell biochemical and functional studies.

Animals↗

The immune response of the pig following oral immunisation with soluble protein.

The immune response of pigs fed 200 mg per day of dinitrophenylated bovine gamma globulin has been evaluated in terms of the antibody and lymphocyte responses and of the induction of tolerance and immune exclusion. Although dosing for ten days resulted in a small IgA response as indicated by splenic plaque forming cells, serum antibody could only be detected when dosing was continued for 42 days. secretory antibody was detected at any time. Antigen feeding for two weeks rendered the animals hyporesponsive to subsequent parenteral antigenic challenge but had little effect upon their ability to exclude an oral dose of antigen from the circulation.

Administration, Oral↗

The cell-mediated immune response of the pig to orally administered antigen.

The immune response of pigs to oral immunization with dinitrophenylated bovine gamma globulin (DNP-BGG) was assessed using the indirect macrophage migration assay. Two days' exposure to the antigen resulted in the inhibition of macrophage migration by supernatants of cultures from the intestine but not the mesenteric lymph node or spleen. Stimulation for 7 days resulted in macrophage inhibition by supernatants of lymphocyte cultures prepared from the intestine, mesenteric lymph node and spleen. Supernatants of splenic lymphocytes from animals stimulated for 2 or 3 days produced enhancement of macrophage migration.

Administration, Oral↗

Passive immune haemolysis -- a class specific antibody assay for use in the pig.

An antibody assay is described based on the principle of complement mediated lysis of sheep red blood cells labelled with antigen. The technique provides a sensitive class specific assay enabling antibody in all three major immunoglobulin classes to be quantitated independently. The assay may be performed in tubes allowing precise measurement of antibody concentration, or in microtitre plates which provides a rapid estimation of antibody titre.

Animals↗