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T Horsburgh

Publications and source records attributed to T Horsburgh.

At least 55 records · Page 3Linked to original sources

The use of paramagnetic beads for the detection of major histocompatibility complex class I and class II antigens.

Specific immunoadsorbents were prepared using paramagnetic particles (Dynabeads), and their ability to immunoprecipitate major histocompatibility complex (MHC) Class I and Class II antigens compared with conventional protein A Sepharose immunoadsorbents. Lysates of lymphoblastoid cells provided the antigen source which were visualized by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Dynabeads were found to be as effective as protein A Sepharose immunoadsorbents at immunoprecipitating MHC Class I and Class II antigens, but had a much lower nonspecific binding capacity resulting in fewer interference bands and lower backgrounds.

Cells, Cultured↗

Age-matching improves the results of renal transplantation with older donors.

Whilst HLA matching is routine for renal transplantation, the possible benefits of matching donor and recipient age have not been previously examined. In this study we examined the simultaneous effect of donor to recipient age difference on the graft survival of 141 consecutive first cadaver transplant recipients treated by cyclosporin immunosuppression. Multivariate regression analysis, taking into account other variables of moderately matched recipients (i.e. dialysis time and type, donor/recipient sex, local/imported kidneys, recent sensitivity, total ischaemic time, preoperative transfusions), indicated that age-difference was the single most important variable (P less than 0.05). Individually there was no significant effect of recipient age, whilst older donors (aged greater than 50 years) were associated with significantly worse graft survival than those younger (P less than 0.01). When dealing with donors aged greater than or equal to 50 years the corresponding recipient 1-year graft was improved when the donor was no more than 5 years older than the transplant recipient. Donor age to recipient age difference is a potentially important selection criterion in renal transplantation.

Adolescent↗

Receptor-mediated endocytosis of low-density lipoprotein by cultured human glomerular cells.

Lipoproteins might be involved in the pathogenesis of glomerular damage. Uptake of low-density lipoprotein (LDL) by cultured human glomerular cells has been studied using LDL, labelled with the fluorescent probe 1,1'-dioctadecyl-3,3,3'3'-tetramethyl-indocarbocyanine perchlorate (diI). Cells have been characterised using phase-contrast microscopy, monoclonal antibodies and lectins. Differentiated glomerular epithelial cells, epithelial-like cells and mesangial cells all took up diI-LDL. Uptake was specific for LDL, of high affinity and inhibited by excess unlabelled LDL, heparin and preloading the cells with cholesterol. Binding of diI-LDL to the cell surface was restricted to discrete areas which were arranged in linear arrays on mesangial cells. Endocytosis of surface-bound diI-LDL occurred within 3 min and breakdown of internalised diI-LDL within 30 min. These results indicate that cultured human glomerular cells take up LDL by receptor-mediated endocytosis.

Cells, Cultured↗

A simple, rapid, reliable reverse haemolytic plaque assay and positive quality control test for routine use.

A simple, rapid, reliable protein A reverse haemolytic plaque assay is described. Monolayers of protein A-coupled sheep red blood cells, in liquid medium, are formed in shallow 15 mm diameter chambers of the type commercially available for leucocyte migration inhibition assays. No agarose is necessary and the chambers are quick and easy to use, economical of reagents, and of a constant size and volume. Results compare favourably with those obtained using Cunningham chambers. The assay is ideal for clinical studies in which large numbers of samples are assayed daily. A positive quality control test for the protein A reverse hemolytic plaque assays is described. Spleen cells are stimulated with pokeweed mitogen to give high numbers of secreting cells. The cells are harvested on day 6 of culture and stored in aliquots at -70 degrees C. When thawed and tested in the protein A assay, these secreting cells form a sensitive and reproducible monitor of the day-to-day performance of the assay. Variation between operators and between batches of reagents may also be checked if desired, with little additional time, effort or expense.

Animals↗