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Biomedical subjects

R A Oliver

Publications and source records attributed to R A Oliver.

At least 37 records · Page 2Linked to original sources

Bovine T cells recognize antigen in association with MHC class II haplotypes defined by one-dimensional isoelectric focusing.

A recently established, one-dimensional isoelectric focusing (IEF) method for distinguishing major histocompatibility complex (MHC) class II polymorphisms in an outbred species, cattle, has allowed us to analyse the involvement of the MHC in the recognition of antigen by bovine T cells. Bovine T-cell lines of Th cell phenotype (BoCD4+) specific for ovalbumin were generated from six individual high responder animals. These animals were bovine MHC (BoLA) class II typed using the IEF technique which detects bovine DR-like products. Four of the animals were shown to be heterozygous and two were homozygous for the IEF specificities. Six out of the 13 IEF specificities (EDF types) detected so far were represented by this group of animals. The cell lines were tested against a panel of IEF-typed antigen-presenting cells (APC) from unrelated donors. The lines only responded to antigen in proliferation assays when the APC shared at least one MHC class II EDF specificity with the BoCD4+ cell line. The responses did not correlate with BoLA class I specificities. However, lines from one of the animals were consistently generated to one of the two haplotypes only. This suggests that there are non-responder alleles to a multi-epitope antigen, present in the cattle population. The results demonstrate that IEF of bovine MHC class II products defines haplotypes of functional relevance, and may indeed be identifying the actual restriction elements involved in presentation of ovalbumin. These results have important implications for future vaccine design in an outbred species, particularly in terms of immune response gene effects and disease associations.

Animals↗

Associations between the bovine major histocompatibility complex (BoLA) and milk production in Icelandic dairy cattle.

Associations between BoLA type and milk production were investigated using information from 80 Icelandic cattle from four dairy farms in Iceland. The specificity w6 reduced milk, fat and protein yield. Although w6 has been shown to be associated with increased susceptibility to mastitis, there is no evidence to suggest that the reduced yields here were due to mastitis. The association may be due to increased disease susceptibility or due to a direct effect of BoLA Class I antigens.

Alleles↗

Variation in T cell responses to ovalbumin in cattle: evidence for Ir gene control.

Genetic control of immune responsiveness in cattle was investigated using an antigen-dependent T cell proliferation assay in vitro. Bovine T cell proliferative responses to ovalbumin were dependent upon major histocompatibility complex (MHC) class II molecules. Responses of an unrelated panel of animals to a limiting concentration of ovalbumin after a single immunization were compared. Two discrete patterns of response were observed. One group of animals had low or non-responses which were not significantly different from the preimmune levels. Another group of animals showed significant responses. After a second immunization the majority of low responders remained low responders. There was no significant correlation between bovine MHC class I BoLA haplotype and magnitude of response within this group of unrelated animals. However, the magnitude of the T cell responses by two half-sib family groups segregated with BoLA haplotypes inherited from the sire. In contrast no significant correlation with antibody responses in vivo could be demonstrated. We suggest that the observed variation in T cell response is linked to bovine MHC class II immune response (Ir) genes.

Animals↗

The analyses of antigen and DNA polymorphism within the bovine major histocompatibility complex: 1. The class I antigens.

Serology, isoelectric focusing (IEF) of expressed antigens, and restriction fragment length polymorphisms (RFLP) were compared for the identification of BoLA class I haplotypes. Expressed antigens identified as bands by IEF correlated well with serological definition confirming and extending our earlier findings (Joosten et al. 1988). Comparison of serology and isoelectric focusing bands with restriction fragments was more complicated; fragments were found which correlated both with broadly reacting and antigen specific sera. We also found correlation of fragments with two or more sera which showed no cross-reactivity. Fragments unique to particular haplotypes were also observed. Serology remains the simplest method of typing BoLA class I antigens. Isoelectric focusing generally agrees with serological definition of antigens and detects antigens not yet defined by serology. It may also be useful in defining the products of other expressed BoLA class I loci. In order to identify RFLPs which could be used for typing, comparison with serology or IEF is essential. Haplotype specific RFLPs could be useful in identifying genes linked to the MHC.

Animals↗

The effect of MHC compatibility between parasite-infected cell line and recipient in immunization against tropical theileriosis.

Lymphoblastoid cell lines, infected and transformed in vitro by a Moroccan stock of Theileria annulata, infected and immunized susceptible taurine cattle, at cell doses of 10(8), 10(6), 10(4) and 10(2), regardless of whether the recipients were BoLA matched or mismatched to the donor cell line. The MHC relationship between the cell line and recipient did affect the severity of the clinical response to cell line immunization which may reflect differences in the specific priming of the immune response. At the highest cell doses the BoLA-mismatched recipients reacted more severely than the BoLA-matched. This study shows that, unlike the closely related parasite T. parva, there is no histocompatibility barrier to immunization using T. annulata-infected cell lines which could be achieved with as few as 10(2) allogeneic infected cells. The role of MHC compatibility between cell line and recipient in the priming of a protective immune response is discussed.

Animals↗

Qualitative variation in the immune response to ovarian follicular fluid proteins in cattle.

In two experiments 44 heifers were immunised with inhibin preparations of partially purified ovarian follicular fluid (PPFF) of ovine, porcine and equine origin in non-ulcerative Freund's adjuvant. In the first experiment 20 cattle were immunised with ovine PPFF1. In the second, a further 24 were immunised with either a different sheep PPFF (ovine PPFF2) or PPFF from pigs or horses. Large molecules of over 30 kDa initiated a strong immune response in some animals, but none in others, although there was no obvious relationship with the MHC Class I (BoLA) phenotype. Immunisation with ovine partly purified follicular fluid affected ovulation rate in some cattle, and only these cows produced detectable antibodies to an approximately 40 kDa protein. These data show that the immunological response of cattle to a mixture of antigens is highly variable between individuals and may explain the variation in the ovarian response to immunological manipulations of ovulation rate seen in this species.

Animals↗

Characterization of class I bovine lymphocyte antigens (BoLA) by one-dimensional isoelectrofocusing.

BoLA class I antigens were characterized in a group of British and Dutch Friesian cattle by one-dimensional isoelectric focusing (1D-IEF) and the results compared with serology using alloantisera and microcytotoxicity. For IEF analysis, non-stimulated peripheral blood mononuclear cells (PBM) were metabolically labelled with 35S methionine, detergent lysates were prepared and MHC molecules precipitated with the monoclonal antibodies (mAbs) W6/32 or B1.1G6. Staphylococcus protein A precipitated antigens were separated on a vertical slab gel under denaturing conditions. The banding patterns seen for the W6/32 precipitated molecules obtained by 1D-IEF were compared with the serological specificities. Characteristic banding patterns were observed for most serological specificities as well as workshop undefined haplotypes. These patterns were seen both in families and the outbred population. In families IEF haplotypes segregated with serotypes. Additional MHC class I products were suggested by variable banding patterns for different w10 haplotypes and when using the different mAbs. A pulse chase experiment with a w12 animal also suggested more than one expressed product. The w2 and w5 specificities were not precipitated by either W6/32 or B1.1G6 and w6.2 and w6.4 were precipitated by W6/32 but not by B1.1G6. These results show that 1D-IEF is useful for BoLA typing. For the characterization of class I antigens, however, much depends on the mAbs used.

Animals↗

Use of bovine lymphocyte antigens in diagnosis of parentage.

Lymphocyte antigens controlled by alleles at the BoLA-A locus were used to indicate which of two bulls could be eliminated as the sire in 18 cases of disputed parentage. The use of bovine lymphocyte antigens (BoLA) significantly increased the exclusion probability over that of the standard red cell and electrophoretic techniques.

Animals↗

A method for freezing bovine lymphocytes.

A simple two-stage technique for preserving bovine lymphocytes is described. Lymphocytes from animals chosen at random were used. The experiments indicate that the optimum temperature for freezing and the optimum concentration of dimethylsulphoxide (DMSO) as cryoprotectant were in the range -29 degrees C to -31 degrees C and 17.5% to 20% respectively. These concentrations of DMSO are much greater than those reported in most other studies.

Animals↗

A genetic study of bovine lymphocyte antigens (BoLA) and their frequency in several breeds.

Using sera which defined the BoLA specificities at the two International BoLA workshops (Edinburgh, 1978 and Wageningen, 1980) and the European Regional workshop (Paris, 1979), 142 informative matings from 15 bulls have been studied. On the basis of this data, 11 of the 15 internationally agreed specificities and one of the regionally defined specificities behave as if controlled by alleles at a single autosomal locus. Data has not as yet been obtained for the other four internationally agreed specificities which are also believed to be at this locus. - The frequencies of 13 of the internationally agreed specificities and one of the regionally defined specificities have been studied for both sexes in one breed and for a single sex in another five breeds. The other two internationally agreed specificities are very recent and the populations have not been tested for them. The frequencies between sexes within a breed and within sexes between breeds differ significantly.

Alleles↗

Electrophoretic mobility of N- and C-terminal monoferric fragments of bovine transferrin phenotypes AA, D1D1, D2D2, and EE, and N-terminal amino acid sequences.

Iron-saturated bovine transferrins A, D1, D2 and E were cleaved by trypsin yielding monoferric fragments. The N-terminal fragments (F) of transferrins A and D2 had identical mobility in cellulose acetate electrophoresis, that of transferrin D1 a slower mobility, and that of E a still slower mobility. The C-terminal fragments (S) gave multiple bands which were essentially identical in the case of transferrins A, D1, and E, but of slower mobility in the case of transferrin D2. All four variants had identical N-terminal amino acid sequences. The electrophoretic mobility of the C-terminal fragments was reduced by neuraminidase treatment, but the N-terminal fragments were unaffected. The four transferrin variants therefore appear to be made up from three electrophoretically distinguishable N-terminal halves and two C-terminal halves. The feature responsible for the electrophoretic double banding of homozygous bovine asialotransferrins is consistently associated with the C-terminal half of the molecule.

Amino Acid Sequence↗

Absorption analysis of boLA sera.

BoLA specificities w1 w2 w5 w8 w9 w10 and w11 have been re-examined by absorption. All of the clusters except w9 showed evidence of sub-group specificities, but only in some cases (w1, w5, w8 and w11) is it suggested that the definition should be revised. The results show a very high degree of reproducibility between sera from different laboratories with clusters of narrow and wide sera for most specificities. These behave in all the cases reported here as linear sub groups or inclusion groups.

Absorption↗

Effect of cold submersion on intramuscular temperature of the gastrocnemius muscle.

This study investigated the effect of a 30-minute, 10 degrees C water bath on the intramuscular temperature of a lower leg and the contralateral lower leg. Intramuscular temperature was measured in 10 subjects using hypodermic thermistor probes inserted 25.3 mm into the lateral head of the gastrocnemius muscles of both legs. One lower leg was submersed in a 10 degrees C cold bath with the water level maintained 5 cm above the patella and with the subject in a nonweight-bearing position. Intramuscular temperature significantly decreased in both lower legs during treatment, although the intramuscular temperature of the treatment lower leg was significantly lower than that of the contralateral lower leg. A temperature difference continued for four hours after treatment; however, the temperature of both lower legs was significantly lower after four hours than it was before the cold bath treatment.

Adipose Tissue↗

Analysis of alloantisera against bovine lymphocytes. Joint report of the 1st International Bovine Lymphocyte Antigen (BoLA) workshop.

The results and agreements of the 1 international BoLA workshop, held in Edinburgh, Scotland in August 1978, are reported. Most of these concern the results from a comparison test of 249 alloantisera to bovine lymphocytes, the antisera being contributed by 9 laboratories. These sera were compared directly in Edinburgh on a panel of lymphocytes from 130 cattle of 21 breeds. In the microlymphocytotoxicity test used 75% of the sera reacted. Sixty eight of these sera were grouped into clusters according to their reaction patterns against the lymphocyte panel. Eleven of these clusters were clearly defined and were given workshop BoLA designations. In addition 22 sera were assigned to subgroups of the agreed clusters. There was no evidence that the method of production of the sera had any effect on their specificity. Although genetic data was not available, the phenotypes of the test panel of lymphocytes are consistent with the clusters detecting antigens controlled by multiple alleles at a single autosomal locus. It was agreed to name the genetic region where this putative locus is located BoLA (bovine lymphocyte antigen).

Animals↗

The production and analysis of antilymphocyte sera following pregnancy and skin grafting of cattle.

This paper presents evidence to show that high titre antilymphocyte sera can be prepared more easily by skin grafting than from parous sera. It also shows that it is possible to analyse complex antilymphocyte sera by absorption with lymphocytes to produce working reagents. Following the 1 International BoLA Workshop the genotype of some of the animals used for skin grafting was ascertained and it was found that in each case where the resulting serum had been studied the main antiserum specificity that had been produced reacted with one of the BoLA specificities present in the skin donor.

Animals↗

Isometric muscle contraction response during recovery from reduced intramuscular temperature.

This study was done to determine the effects of a 30-minute cold water bath on intramuscular temperature and plantar flexion strength, immediately after treatment and during a 3-hour posttreatment recovery period. Twenty persons were tested twice, receiving treatment once and serving as controls once. Measurements were taken prior to the treatment period, immediately after treatment and then every 30 minutes for 3 hours. The dominant leg was submerged in water at 10 C for 30 minutes. Plantar flexion strength was measured with a cable tensiometer and intramuscular temperature was measured with a hypodermic thermistor probe. Intramuscular temperature significantly decreased immediately posttreatment on the experimental days and then increased significantly during the recovery period. Significant increases in strength were noted during the recovery period. A definite relationship exists between intramuscular temperature and plantar flexion strength.

Adult↗