Biomedical subjects

S J Prowse

Publications and source records attributed to S J Prowse.

At least 19 recordsLinked to original sources

PubMed2005

A long-term study in Angora goats experimentally infected with Mycobacterium avium subsp. paratuberculosis: clinical disease, faecal culture and immunological studies.

Two longitudinal experiments involving Angora goats challenged with either bovine or ovine strains of Mycobacterium avium subspecies paratuberculosis (Map) have been conducted over a period of 54 and 35 months, respectively. Blood samples for the interferon-gamma (IFN-gamma) test and the absorbed ELISA and faecal samples for bacteriological culture were taken pre-challenge and monthly post-challenge. Persistent shedding, IFN-gamma production, seroconversion and clinical disease occurred earlier with the bovine Map gut mucosal tissue challenge inoculum than with cultured bacteria. The IFN-gamma responses of the gut mucosal tissue and bacterial challenge groups were substantially and consistently higher than those of the control group. The in vivo and cultured cattle strains were much more pathogenic for goats than the sheep strains with persistent faecal shedding, seroconversion and clinical disease occurring in the majority of bovine Map challenged goats. With the ovine Map, 3 goats developed persistent antibody responses but only one of these goats developed persistent faecal shedding and clinical disease. However, there was no significant difference between the IFN-gamma responses of the tissue challenged, bacterial challenged and control groups. Compared with sheep, the ELISA appeared to have higher sensitivity and the IFN-gamma test lower specificity.

Animals↗

PubMed2004

A long-term study in Merino sheep experimentally infected with Mycobacterium avium subsp. paratuberculosis: clinical disease, faecal culture and immunological studies.

Two longitudinal experiments involving Merino sheep challenged with either bovine or ovine strains of Mycobacterium avium subsp. paratuberculosis (Map) have been conducted over a period of 54 and 35 months, respectively. Blood samples for the interferon-gamma test, the absorbed ELISA and faecal samples for bacteriological culture were taken pre-challenge and monthly post-challenge. Infections were induced with either a bovine or ovine strain of Map in separate experiments with infections being more easily established, in terms of faecal bacterial shedding and clinical disease when the challenge inoculum was prepared from gut mucosal tissue than cultured bacteria. The patterns of response for shedding and clinical disease were similar. Cell-mediated immune responses were proportionally elevated by at least an order of magnitude in all sheep dosed with either a bovine or ovine strain of Map. Conversely, antibody responses were only elevated in a relatively small proportion of infected sheep. Neither of the clinically affected tissue challenged sheep developed an antibody response despite the presence of persistent shedding and the development and decline in cell-mediated immunity. The results indicated that for sheep the interferon-gamma test may be useful for determining if a flock has been exposed to ovine Johne's disease.

Animals↗

PubMed1997

In vivo effects of chicken interferon-gamma during infection with Eimeria.

Newly hatched chickens are highly susceptible to infection by opportunistic pathogens during the first 1 or 2 weeks of life. The use of cytokines as therapeutic agents has been studied in animal models as well as in immunosuppressed patients. This approach has become more feasible in livestock animals, in particular poultry, with the recent cloning of cytokine genes and the development of new technologies, such as live delivery vectors. We have recently cloned the gene for chicken interferon-gamma (Ch-IFN-gamma). Poly-HIS-tagged recombinant Ch-IFN-gamma was expressed in Escherichia coli, was purified by Ni chromatography, and was found to be stable at 4 degrees C and an ambient temperature for at least several months and Several weeks, respectively. Ch-IFN-gamma was capable of protecting chick fibroblasts from undergoing virus-mediated lysis, induced nitrite secretion from chicken macrophages in vitro, and enhanced MHC class II expression on macrophages. Administration of recombinant Ch-IFN-gamma to chickens resulted in enhanced weight gain over a 12-day period. Furthermore, the therapeutic potential of Ch-IFN-gamma was assessed using a coccidial challenge model. Birds were treated with Ch-IFN-gamma or a diluent control and then infected with Eimeria acervulina. Infected birds treated with Ch-IFN-gamma showed improved weight gain relative to noninfected birds. The ability of Ch-IFN-gamma to enhance weight gain in the face of coccidial infection makes it an excellent candidate as a therapeutic agent.

Animals↗

PubMed1994

Purification and characterisation of a serine-type protease from Eimeria tenella oocysts.

Homogenates of sporulated oocysts of E. tenella have detectable proteolytic activity which is completely inhibited by phenylmethylsulfonyl fluoride and L-trans-epoxysuccinyl-leucyl-amido-(4-guanidino)-butane, indicating the presence of both serine and cysteine-type proteases in sporulated oocysts. A serine-type protease has been purified from the homogenate using immobilised-bacitracin affinity chromatography. The monomeric enzyme had an apparent M(r) of 20,000 and a pI of 8.6. The maximum proteolytic activity with azocasein and gelatin was observed at pH 8.0. Antibodies raised in rabbits and chickens against purified protease recognised this protein on blots of sporozoite homogenates.

Animals↗

PubMed1993

Molecular characterisation of peanut agglutinin-binding glycoproteins from Eimeria tenella.

A group of glycoproteins, which strongly bind peanut agglutinin (PNA) was found in Eimeria tenella. Two major antigenic glycoproteins, Et110gp and Et35gp, were identified in sporulated oocysts and sporozoites. Molecular characterisation of carbohydrate moieties (lectin binding, enzymic hydrolysis and monosaccharide composition) revealed that both glycoproteins are rich in galactose and N-acetylgalactosamine, and appear to be sialylated. Both glycoproteins were susceptible to treatment with neuraminidase followed by O-glycosidase, suggesting that the oligosaccharide chains are attached to the protein by an O-glycosidic linkage to serine and/or threonine. Purified Et35gp contained a large number of serine (14) and threonine (33) residues, and was rich in glycine. This protein aggregated after repetitive lyophilisation and migrated on SDS-PAGE gels as an 85,000 protein. Sera against purified Et35gp raised in chickens and rabbits, and anti-E. tenella immune chicken serum recognised both antigens on blots and on the surface of sporozoites. Chickens immunised with purified Et35gp were not protected against coccidial infection.

Animals↗

PubMed1992

Mannitol metabolism in Eimeria tenella.

Unsporulated oocysts of Eimeria tenella contain large quantities of carbohydrates, namely amylopectin, mannitol and glucose. Analysis of carbohydrate content of sporulating oocysts revealed that mannitol content increased markedly during early stages of sporogony (first 4-6h) but slowly diminished during the next 40h of sporulation. Accumulation of mannitol was accompanied by a rapid decrease in amylopectin and free glucose, suggesting that mannitol might be synthesized from glucose released from amylopectin. Mannitol was also detected in sporozoite and merozoite extracts. All four mannitol cycle enzymes were detected in oocysts. Sporozoites excysted in vitro had lower activities of all four enzymes. Mannitol-1-phosphatase and mannitol dehydrogenase activity was also detected in merozoites obtained from the second stage schizonts. Sporozoites incubated with 14C-glucose accumulated radioactively labelled precursor continuously for over 12h and some of the 14C-glucose was converted into 14C-mannitol. These results indicate that mannitol plays an important role in the metabolism and development of the intracellular stages of the parasite.

Amylopectin↗

PubMed1992

Enhanced H2O2 release from immune chicken leucocytes following infection with Eimeria tenella.

Cell-mediated immunity is thought to be important in the resistance of chickens to infection by coccidia, and it has been demonstrated that sporozoites of Eimeria tenella are very sensitive to superoxide ions. Therefore an investigation into the cellular responses in naive specific pathogen-free and hyperimmune birds was carried out with particular attention to their ability to produce reactive derivatives of oxygen. Leucocytes were isolated from the blood, spleen and caecal mucosa of chickens infected with E. tenella and assessed for their ability to release H2O2. Leucocytes obtained from the blood and spleen of hyperimmune birds 1 day after challenge showed an elevated ability to produce reactive oxygen intermediates. In contrast, the ability of leucocytes from naive chickens to produce these molecules was transiently depressed after challenge. Prior to challenge, mucosal leucocytes from immune chickens were also able to release heightened levels of H2O2 when compared with cells from naive chickens.

Animals↗

PubMed1991

Cell-mediated immunity to Eimeria in the fowl: the absence of cross-species protection is not due to the lack of cross-reactive T cells.

Immunity to Eimeria species in the fowl has been shown to be species specific and it has been proposed that the lack of cross-protection between the species of Eimeria is due to the absence of T cells which recognize antigens from a heterologous species. When this hypothesis was tested the results showed that antigens from E. tenella elicited a strong specific response by cells from birds immune to E. tenella. In contrast cells from birds which were immune to E. acervulina responded to a similar magnitude to both E. tenella and E. acervulina antigens. This indicates that the lack of cross-protection is not due to the lack of cross-reactive T cells.

Animals↗

PubMed1991

Superoxide dismutases in Eimeria tenella.

Unsporulated oocysts of Eimeria tenella have high superoxide dismutase (SOD: superoxide:superoxide oxidoreductase, EC 1.15.1.1.) activity and contain several electrophoretically distinct forms of the enzyme, including two forms of Cu/Zn-containing SOD, two forms of Fe-SOD and two forms of Mn-SOD. SOD activity remains high during 12 h of sporulation but diminishes slowly during prolonged sporulation. Oocysts sporulated for 48 h have low levels of superoxide dismutase and contain only one form of the enzyme (Mn-SOD), which was also found in sporozoites. In vitro, sporozoites are oxidant-sensitive and die within minutes of superoxide radical (O2-) generation but SOD/catalase and mannitol protect sporozoites against oxidative damage. These data suggest that E. tenella sporulated oocysts and sporozoites lack soluble cytoplasmic SOD and that this deficiency may contribute to the oxidant sensitivity of the parasite.

Animals↗

PubMed1991

Cu,Zn superoxide dismutase from chicken erythrocytes.

1. Copper, zinc superoxide dismutase (Cu,Zn SOD) has been purified to homogeneity from chicken erythrocytes by anion-exchange, immobilized metal affinity and size exclusion chromatography. 2. Molecular properties (amino acid composition, molecular mass, subunit composition and spec. act.) of the chicken enzyme are similar to those of a bovine erythrocyte Cu,Zn SOD. 3. The chicken and bovine enzymes are immunologically similar since antisera raised against each enzyme are cross-reactive.

Animals↗

PubMed1990

A molecular karyotype of Eimeria tenella as revealed by contour-clamped homogeneous electric field gel electrophoresis.

DNA from sporozoites of Eimeria tenella was resolved by pulsed field gel electrophoresis into nine chromosomal bands. Some bands of this molecular karyotype contained more than one chromosome as determined by the relative intensity of both staining with ethidium bromide and hybridisation to an E. tenella telomeric probe. Haploid forms of the parasite must be presumed to contain at least 12 chromosomes. The two smallest chromosomes were about 1.1 and 1.4 megabases. Most chromosomes were in excess of 3 Mb with the largest over 5 Mb as determined by comparison with the co-migration of chromosomes from Schizosaccharomyces pombe. A 5S ribosomal gene probe hybridised to a single chromosomal band.

Animals↗

PubMed1989

A cytotoxic monoclonal islet cell surface antibody from the NOD mouse.

Monoclonal antibody (Mab) 1.93B7 was obtained by fusion of spleen cells from a diabetic NOD mouse with P3X63Ag8.653 myeloma cells and screening for complement mediated lysis of rat insulinoma (RIN) cells. Immunofluorescence studies revealed that this Mab binds to RIN cells but not to the rat pituitary tumour line GH3. The binding of Mab 1.93B7 to RIN cells was abolished by trypsin but not by neuraminidase treatment of the cells, suggesting that the antigen recognized is a protein. Mab 1.93B7 bound to approximately 30% of mouse (BALB/c) and rat islet cells which had been subjected to trypsin digestion and incubated as a single cell suspension for 12h to allow reexpression of trypsin sensitive antigens. Since Mab 1.93B7 is potentially pathogenic, as suggested by its reactivity to primary islet cells and its complement fixing capacity, we injected it into BALB/c and NOD mice. Cytotoxic activity against RIN cells was detected in the serum of the animals injected with Mab 1.93B7, but the Mab did not exert a diabetogenic action and failed to reverse diabetes when administered at onset in NOD mice. No modification of the course of spleen cell mediated transfer of diabetes in NOD mice was observed when the Mab was administered from the time of spleen cell inoculation to the appearance of glycosuria. The implications of the lack of an effect in vivo of Mab 1.93B7 under the conditions employed are discussed.

Animals↗

PubMed1987

Islet cytotoxicity of interleukin 1. Influence of culture conditions and islet donor characteristics.

We recently demonstrated that the macrophage product interleukin 1 (IL-1) is cytotoxic to isolated pancreatic islets and hypothesized that IL-1 is responsible for beta-cell destruction in insulin-dependent diabetes mellitus (IDDM). We studied whether the variation in IDDM preponderance with age, sex, and genetic background in vivo is reflected in different susceptibility to IL-1 toxicity of islets in vitro. In addition, we studied the effect of preculture conditions that support endocrine islet cell function and decrease nonendocrine passenger-cell survival on the susceptibility of beta-cells to IL-1 because it is unknown whether IL-1 acts directly on beta-cells or via passenger cells. No differences in susceptibility to various doses of IL-1-containing mononuclear cell supernatants were found between islets isolated from newborn or adult rats, male or female rats, or rats of four inbred strains, indicating that age, sex, and genetic background do not influence the susceptibility of the beta-cell to IL-1. Preculture of islets for 1-7 days in normal atmosphere and preculture of islet clusters in 95% O2 to delete passenger cells did not affect IL-1-mediated cytotoxicity, suggesting that IL-1 acts directly on beta-cells. Increasing the glucose concentration (22 mM) in the culture medium, which is known to protect beta-cells against alloxan toxicity, reduced IL-1 toxicity. Five or 25% normal human serum as well as 5% normal rat serum, but not equivalent concentrations of human serum albumin, inhibited IL-1 toxicity, indicating the presence of IL-1 inhibitors, IL-1 antagonists, or beta-cell-protecting factors in normal serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

PubMed1987

Circulating lymphocyte populations and autoantibodies in non-obese diabetic (NOD) mice: a longitudinal study.

Several previous observations indicate a role for the immune system in the pathogenesis of insulin-dependent diabetes mellitus (IDDM) in non-obese diabetic (NOD) mice. In order to assess the status of the immune system in this model of spontaneous diabetes we studied the phenotype of circulating lymphocytes and the humoral autoimmunity to islet cells in non-diabetic NOD mice at various ages. Lymphocyte numbers were low in young NOD mice (age less than 160 days) as compared with other strains of mice and increased later to reach values in or above the range of controls. The percentages of circulating T lymphocytes and their L3T4+ and Lyt2+ subsets were higher in NOD mice of all ages and both sexes than in controls; however, no imbalance of the L3T4+ and Lyt2+ subpopulations was found. Anti-insulin autoantibodies were detected by an ELISA assay in all the NOD mice studied throughout the entire period of observation. Autoantibodies reacting with the cytoplasm of islet cells in Bouin's fixed pancreas sections, likely to be anti-insulin antibodies, were found in 47 to 58% of the samples from NOD mice aged 75 to 150 days. Antibodies to surface antigens of rat insulinoma cells were virtually absent in young NOD mice (75-100 days) and appeared in 33 to 43% of the samples from 150 to 185 days old NOD mice. The autoantibodies and the quantitative lymphocyte abnormalities reported here, although not predictive of the appearance of overt diabetes, are likely to be involved in the pathogenesis of the disease and therefore may indicate directions for future investigations.

Aging↗

PubMed1986

The allograft response.

The allograft response is a response by host T lymphocytes reacting to transplantation antigens that are carried on allogeneic lymphoreticuler cells. In vivo this response usually leads to graft rejection. It is possible to circumvent this response by the elimination of the lymphoreticular cells from the grafts prior to transplantation. The paradox of the strong response to transplantation antigen on lymphoreticular cells and the weak response to the same antigen on graft parenchymal cells can be explained by the signaling requirements for T lymphocyte activation.

Animals↗

PubMed1986

Islet allografts are destroyed by disease occurrence in the spontaneously diabetic BB rat.

Transplantation of cultured islet and pituitary tissue from PVG (RT1c) donors to major histocompatibility complex-incompatible BB/D recipients (RT1u) results in tissue-specific destruction of the grafted islets but not of the pituitary. We interpret this response as disease occurrence in the MHC-incompatible islet graft. Islet damage is associated with eosinophil and mast cell accumulation in and around the grafted tissue. Antibody deposition is also present in tissues of the BB rat. This is suggestive of an antibody-mediated allergic reaction.

Animals↗

PubMed1986

Combined treatment with nicotinamide and desferrioxamine prevents islet allograft destruction in NOD mice.

Nonobese diabetic (NOD) mice get spontaneous diabetes with clinical and pathological manifestations similar to those seen in human type I diabetes. NOD mice will destroy transplants of treated allogeneic islet tissue by a recurrence of the disease process that destroyed the original islet tissue. This may be prevented by treatment of the animals with combined desferrioxamine and nicotinamide. Transplanted animals become normoglycemic and remain so for the duration of the treatment. This suggests that oxygen-derived free radicals may be involved in islet damage in spontaneous diabetes.

Animals↗

PubMed1986

Reversal of diabetes in outbred mice by islet allotransplantation.

The combination of donor pretreatment with cyclophosphamide, organ culture in 95% O2:5% CO2 for 7-10 days, and short-term immunosuppression of recipients with cyclosporin A (CsA) were necessary to obtain 100% survival of single-cluster BALB/c islet allografts in outbred mice. In vivo and in vitro pretreatment of the donor tissue alone resulted in the acceptance of 45% of the islet allografts in nonimmunosuppressed outbred mice. CsA treatment of recipients alone yielded 40% survival of the untreated allografts. CsA treatment played an important role in maintaining the capacity of islet allografts to function in outbred mice. During CsA treatment, 88% of streptozocin-treated mice showed graft-dependent reversal of diabetes; the remainder showed no evidence of graft function, and CsA treatment failed to prevent acute graft rejection. After withdrawal of CsA immunosuppression, 38% of this total group remained normoglycemic. These findings suggest that modulation of both donor-tissue immunogenicity and recipient responsiveness will be required for successful pancreatic islet transplantation in diabetic humans.

Animals↗