Reference-strand-mediated conformation analysis of MHC alleles: a new method for high-resolution typing of the Ovar-DQB genes.
Explore the source record for details and available documents.
Alleles↗
Biomedical subjects
M J Stear
Publications and source records attributed to M J Stear.
The use of a gamma-type function to assess the relationship between the number of adult Teladorsagia circumcincta and total egg output.
The relationship between faecal nematode egg count and the number of adult Teladorsagia circumcincta was examined in 508 naturally infected lambs at 6-7 months of age. The relationship was found to be convex and was empirically described by a gamma-type function of the form y = an(b) e(-cn), where n is the number of adult nematodes present and y is the number of nematode eggs per gramme (epg) of faeces. This equation predicted that the peak expected egg count (277 epg) would occur at a worm burden of 2,167 adult worm is consistent with the existence of severe density-dependent constraints on the fecundity of T. circumcincta.
Animals↗
Molecular markers and their use in animal breeding.
The use of DNA markers to define the genetic makeup (genotype) and predict the performance of an animal is a powerful aid to animal breeding. One strategy is known as marker-assisted selection (MAS). MAS facilitates the exploitation of existing genetic diversity in breeding populations and can be used to improve a whole range of desirable traits. DNA markers are, by definition, polymorphic, and the methods used to define DNA markers include restriction fragment length polymorphisms (RFLPs), microsatellites, and single nucleotide polymorphisms (SNPs). Linkage analysis, association analysis and analysis of gene function can be used to determine which polymorphisms are useful markers for desirable traits. Future prospects include the use of high throughput DNA microarray (DNA chip) technology which could revolutionize animal breeding in the next millennium.
Animal Husbandry↗
How lambs control infection with Ostertagia circumcincta.
Two of the most important questions for immunologists studying nematode infections are what effect does the host response have on the parasite and which components of the host response are responsible for these effects. The number of nematodes and the mean length of adult female Ostertagia circumcincta was measured in over 500 6-7 month old lambs of the Scottish Blackface breed. Quantitative genetic analyses indicated that there was substantial genetic variation among lambs in faecal egg counts and in worm length but the analyses provided no evidence for genetic variation in worm numbers. Separate analyses have shown a strong relationship between decreased worm length and decreased worm fecundity. Therefore, genetic resistance in lambs appears to be due to control of worm growth and not to control of worm numbers. The only immune response that is consistently associated with reduced worm length is the IgA response to fourth-stage larvae. The association is remarkably strong. After allowing for the influence of worm number on worm length (density-dependence) essentially all of the variation among deliberately infected lambs in worm length can be accounted for, in a statistical sense, by variation in the strength and specificity of the local IgA response. Therefore, the IgA mediated suppression of worm growth and fecundity appears to be the major mechanism of resistance to O. circumcincta in lambs.
Animals↗
Genetic characterisation of protective vaccine responses in sheep using multi-valent Dichelobacter nodosus vaccines.
Protective vaccine responses to nine distinct serogroups of Dichelobacter nodosus (serogroups A-I) can be readily measured by serogroup-specific K-agglutinating antibody titres. On the basis of a large quantitative genetic experiment (1200 progeny from 129 sire groups), it was shown that variation in antibody responses following vaccination with a multi-valent pilus antigen D. nodosus vaccine (serogroups A-I) is, in part, under genetic control and thus heritable. Based on the genetic relationships between antibody responses to all nine antigens, results suggested that both genes for a broad-based and genes for serogroup-specific response contributed to genetic variation in vaccine response. Furthermore, preliminary data in 389 progeny showed that polymorphism within the ovine major histocompatibility (MHC) based on serological classification accounted for a significant proportion of the variation in vaccine responses. In subsequent experimentation, we examined the importance of genetic polymorphism within the ovine MHC, and the possibility of genes outside the MHC for their involvement in antigen-specific and broad-based vaccine response. Within two large half sib families(131, and 143 progeny), four MHC haplotypes were investigated and found to be associated with differential antibody responses to six out of eight distinct vaccine-antigens presented to the host in a multi-valent vaccine. The model used here shows how well characterised immunogens, quantitative genetic experimentation, and molecular gene mapping tools can be used to unravel genetic differences in host responses to commercial vaccines.
Agglutination Tests↗
Relationships among peripheral eosinophilia, eosinophil peroxidase activity, interleukin-5 concentration and faecal nematode egg count during natural, mixed gastrointestinal nematode infection.
A longitudinal study of sheep, naturally infected with gastrointestinal helminths, was carried out to quantify the relationships among eosinophilia, eosinophil peroxidase activity, interleukin-5 level and faecal egg counts. Faecal egg counts, peripheral eosinophilia and peroxidase activity per eosinophil were moderately repeatable but interleukin-5 concentrations had a remarkably high repeatability. Animals with higher concentrations of interleukin-5 had greater peripheral eosinophilia and those animals with higher numbers of eosinophils had lower faecal egg counts. These associations were statistically significant but quite weak. Variation in interleukin-5 levels does not appear to be responsible for most of the variation in eosinophil responses in outbred sheep.
Animals↗
The distribution of pepsinogen within the abomasa of cattle and sheep infected with Ostertagia spp. and sheep infected with Haemonchus contortus.
The effect of nematode infections on the production of pepsinogen by ruminants was investigated immunohistochemically and biochemically. Abomasal tissues were collected from parasite-naive cattle and sheep, from sheep infected with predominantly Ostertagia circumcincta, sheep infected experimentally with Haemonchus contortus and cattle infected with Ostertagia ostertagi. Pepsinogen was also assayed biochemically in homogenates of fundic mucosae from sheep infected with predominantly O. circumcincta. Infection with Ostertagia spp. parasites was associated mainly with nodular hyperplasia, resulting in increased numbers of cells that produce both pepsinogen and mucus. Measured biochemically, nodules contained more pepsinogen than adjacent more normal mucosa (p < 0.05), and this effect was largely attributable to the greater mass of nodules. Infection of sheep with H. contortus was associated with generalised hyperplasia, characterised by increased numbers of mucopeptic cells and in at least one animal with reductions in parietal cell numbers. At the same time, the zymogen granule content of chief cells was reduced. Similar changes were occasionally seen in sheep infected predominantly with O. circumcincta. Generalised hyperplasia is likely to be indicative of the presence of ambulatory parasitic stages as opposed to those confined to nodules. The potential for the enhanced production of pepsinogen by increased numbers of cells with a joint mucous cell and zymogenic cell phenotype may offset decreases in the numbers of chief cells or reductions in chief cell activity.
Abomasum↗
Variation in the number of expressed MHC genes in different cattle class I haplotypes.
Analysis of cattle major histocompatibility complex (MHC) (BoLA) class I gene expression using serological and biochemical methods has demonstrated a high level of polymorphism. However, analysis of class I cDNA sequences has failed to produce conclusive evidence concerning the number and nature of expressed genes. Such information is essential for detailed studies of cattle immune responses, and to increase our understanding of the mechanisms of MHC evolution. In this study a selective breeding programme has been used to generate a number of MHC homozygous cattle expressing common serologically defined class I specificities. Detailed analysis of five class I haplotypes was carried out, with transcribed class I genes identified and characterized by cDNA cloning, sequence analysis, and transfection/expression studies. Surface expression of the gene products (on lymphocytes) was confirmed using monoclonal antibodies of defined BoLA specificity. Phylogenetic analysis of available transcribed cattle MHC class I sequences revealed complex evolutionary relationships including possible evidence for recombination. The study of individual haplotypes suggests that certain groupings of related sequences may correlate with loci, but overall it was not possible to define the origin of individual alleles using this approach. The most striking finding of this study is that none of the cattle class I genes is consistently expressed, and that in contrast to human, haplotypes differ from one another in both the number and composition of expressed classical class I genes.
Amino Acid Sequence↗
Mechanisms underlying resistance to nematode infection.
Lambs show considerable genetic variation in faecal egg count following natural, predominantly Ostertagia circumcinta infection. This genetic variation is acquired and not innate. Worm length is positively associated with worm fecundity. The genetic variation in faecal egg count is a consequence of genetic variation in worm length and hence worm fecundity, and not of genetic variation in worm burdens. In contrast to lambs, mature sheep may be able to regulate both fecundity and worm numbers. In lambs, three factors account for the majority of the variation in worm length: the strength of the local IgA response against fourth-stage larvae, the specificity of this response against four molecules in particular, and the density-dependent influence of worm number.
Animals↗
The curvilinear relationship between worm length and fecundity of Teladorsagia circumcincta.
The variation among sheep in fecundity of Teladorsagia (Ostertagia) circumcincta was estimated by dividing the faecal egg count by the worm number following deliberate infection of mature Scottish Blackface lambs. Fecundity was skewed and ranged from 0 to 350 eggs per worm per day. Most animals had relatively low worm fecundities, but a small number of individuals had relatively high worm fecundities. However, as fecundity is a ratio of two imprecise estimates, extreme values may be statistical artefacts. Following both deliberate and natural infection, differences in worm fecundity were associated with differences in adult female worm length. In both infections, fecundity varied with worm length to the power 0.4. This relationship should assist the measurement of fecundity in studies of host immunity, in epidemiological modelling and in estimating the influence of density-dependent relationships.
Animals↗
The recognition of molecules from fourth-stage larvae of Ostertagia circumcincta by IgA from infected sheep.
The recognition of antigens from fourth-stage Ostertagia circumcincta by plasma IgA was studied in a group of 30, ten-month-old Scottish Blackface sheep which had been naturally, then deliberately infected. Western blotting revealed 49 bands that were recognized by antibody from one or more of the 30 sheep. There was a heterogeneous pattern of antigen recognition amongst the sheep. No individual recognized all the 49 bands and only one of the 49 bands was recognized by all sheep. Two antigens with approximate molecular weights 87,000 Da and 129,000 Da were significantly associated with a reduction in mean adult worm lengths. The observed variation in recognition of these two antigens on fourth-stage larval preparations accounted, in a statistical sense, for nearly 40% of the total variation in worm length. In addition to the variation in antibody mediated recognition of these two parasite molecules, three other components have been implicated in regulating worm length. They are a 37,000 Da band from adult worms, the amount of fourth-stage larval specific IgA in the abomasal mucosa and the density-dependent influence of adult worm burden. Together, these components and their interactions accounted for over 90% of the observed variation in worm length. These results indicate that the parasite-specific IgA response, or something extremely closely associated with it, is the major immunological mechanism controlling worm length.
Animals↗
The relationship between the number and size of nematodes in the abomasum and the concentration of pepsinogen in ovine plasma.
Six-month-old lambs that had been naturally infected with predominantly Teladorsagia (Ostertagia) circumcincta were tested for plasma pepsinogen concentrations because pepsinogen concentrations may reflect the extent of damage to the abomasum. The distribution of pepsinogen concentrations among these lambs was positively skewed with most individuals having relatively low concentrations. Pepsinogen concentrations were more strongly associated with variation in the mean length of the adult female worms than with variation in the number of nematodes present. Previous trials have suggested that genetic variation in the growth of lambs is strongly influenced by genetic variation in worm length. Together these results imply that variation among lambs in the pathogenic effects of T. circumcincta depends upon the mean size of the worms as well as the number of worms present.
Abomasum↗
Studies on host resistance to tick infestations among trypanotolerant Bos indicus cattle breeds in east Africa.
Recent epidemiological studies carried out in East Africa have indicated that some Bos indicus cattle breeds such as the Orma Boran and Maasai Zebu have a degree of trypanotolerance worth exploitation by their introduction into trypanosomosis endemic areas where other cattle breeds cannot survive. However, in most areas of East Africa, trypanosomosis, ticks, and tick-borne diseases occur together. It is therefore important to obtain information on the susceptibility of these breeds to tick infestation and tick-borne diseases. This study was therefore designed to determine the susceptibility of these cattle breeds to tick infestations. They were compared with the Galana Boran (trypanosusceptible) and the Friesian (susceptible to tick infestations, tick-borne diseases, and trypanosomosis). The four breeds of cattle were exposed to natural tick challenge for a period of seven months and whole body weekly tick counts were done on each animal. Significant differences to tick infestations among the four breeds were observed. For both Rhipicephalus appendiculatus and Boophilus decoloratus, susceptibility to infestation increased in the order, Maasai Zebu, Orma Boran, Galana Boran and Friesian. The results generated by this pilot study so far suggest that variation in susceptibility to tick infestations exists among the four breeds. The Orma Boran and Maasai Zebu showed greater resistance to tick-infestations than the Galana Boran and Friesian. This suggests that utilization of these trypanotolerant cattle breeds could be feasible even in the face of tick challenge and should therefore be considered when planning integrated trypanosomosis and tick control strategies.
Africa, Eastern↗
The influence of dietary supplementation with urea on resilience and resistance to infection with Haemonchus contortus.
Previous research has indicated that supplementing an apparently adequate diet with additional protein improves both host resistance and resilience in lambs infected with Haemonchus contortus. The present study tested the influence of supplementation with non-protein nitrogen (urea). Helminth-naive Hampshire Down lambs were given an apparently adequate basal diet or a diet supplemented with urea. The lambs were then infected with Haemonchus contortus for 10 weeks. Supplementation with urea had no discernible effect on resistance to infection; faecal egg counts, worm burdens, worm lengths and mean number of eggs per adult female worm did not differ between the 2 groups. However, lambs on the supplemented diet showed better resilience; they had greater packed red cell volumes, higher plasma albumin concentrations and increased liveweight gain compared to lambs on the basal diet. The loss of appetite following infection was less in lambs fed the urea-supplemented diet. The observed effect of urea supplementation was seemingly due to greater food consumption as well as the better diet.
Animal Nutritional Physiological Phenomena↗
Changes in the zymogenic cell populations of the abomasa of sheep infected with Haemonchus contortus.
The effects of dietary urea supplementation and of a 10-week trickle infection regime, simulating chronic exposure to Haemonchus contortus, on the zymogenic population of the abomasa of Hampshire Down lambs was examined. At necropsy a variety of parameters including plasma pepsinogen concentration, the wet weights of abomasal fundic mucosal pieces and the amounts of pepsinogen contained in them, were assessed. Tissue pepsinogen concentration was measured as the total, acid-stable proteolytic activity present in mucosal homogenates, as well as immunohistochemically. The immunohistochemical findings were quantified using computer-aided image analysis. Elevation of plasma pepsinogen concentrations in infected animals was of borderline significance (P = 0.06). The fundic mucosae of infected animals were heavier (P < 0.02) than those of control animals, but there was no overall change in the pepsinogen content of tissues. Immunohistochemistry revealed that infected animals had increased numbers of zymogenic cells, due to mucous cell hyperplasia and the adaptation of cells to produce both mucins and pepsinogen. The pepsinogen content of chief cells, the major source of pepsinogen in uninfected animals, was reduced in infected lambs. Image analysis confirmed that at a mid-point of the mucosa of infected animals there was increased pepsinogen-specific immunoreactivity that corresponded with areas of mucosal hyperplasia. Mucous cell hyperplasia might therefore allow the maintenance of pepsinogen secretion in infected animals even if chief cell output is reduced.
Abomasum↗
The processes influencing the distribution of parasitic nematodes among naturally infected lambs.
The impact of mixed, nematode infection upon a group of animals will depend upon the number of nematodes present, how they are distributed among hosts and whether individuals that are heavily parasitized with one species are more likely to be heavily parasitized with other species. A survey of over 500 six-month-old, Scottish Blackface lambs from a single farm in Southwest Strathclyde identified 7 different categories of nematodes in the abomasum and small intestine. There were considerable differences among years and among nematodes in the prevalence and mean intensity of infection. Ostertagia circumcincta was present in nearly all lambs and judged by prevalence and intensity is one of the most successful of all parasitic nematodes. Each category of nematodes had a skewed distribution; most animals had relatively few worms but a small proportion had many worms. The variance of the number of nematodes in each category were approximately equal to the square of the mean. The counts of adult O. circumcincta followed a negative binomial distribution, but the negative binomial distribution did not provide a good description of the observed values for the other species. These other species had a lower prevalence and possibly some sheep were not exposed to infection. There was no significant genetic variation among lambs in the number of nematodes present and therefore the differences among these lambs were unlikely to be a consequence of genetic differences in host susceptibility. Lambs with increased numbers of one species were more likely to be have increased numbers of the other species, but the correlations were weak and may reflect covariation in exposure to different parasites.
Abomasum↗
DNA typing for BoLA class I using sequence-specific primers (PCR-SSP).
The analysis of cattle MHC (BoLA) class I gene expression is an essential component of studies on immune responses and susceptibility to disease. International BoLA workshops have generated data and reagents that allow discrimination of class I molecules at the haplotype level, but progress has been limited by difficulties encountered in defining single alleles. Our aim in this study was to develop a DNA-based system for improved identification of expressed class I alleles, utilizing available cDNA sequences derived from cattle carrying a series of serologically defined class I specificities. This method has allowed more accurate typing of animals for expression of the class I genes present within a small number of haplotypes. The method has also reliably differentiated between allelic variants (identified by prior sequence analysis) and has split existing serological specificities. The data show that MHC class I genes in cattle are more polymorphic than demonstrated by serology and biochemical analysis.
Alleles↗