ABSTRACT
After the preliminary clustering of the whole workshop panel using flowcytometry data, we selected those clusters that were composed of mAbs that bound a broad range of cells. To obtain evidence that mAbs in each of these preliminary clusters detected the same antigen, they were tested for their capacity to compete with each other for binding to a target cell and the molecular weights of their antigens were estimated after immunoprecipitation. Most preliminary clusters in the non-lineage panel contained control mAbs that had been characterised in one of the previous workshops, and this study therefore increased the number of mAbs available to each of these non-lineage antigens. One new interesting cluster, BoWC14, was described which defined an antigen on myeloid cells and a subpopulation of CD4, CD8 and WC1 T lymphocytes (BT3/8.12 and IL-A155). An additional cluster, which did not fit into any other panel, contained mAbs specific for an antigen restricted to the erythroid lineage and received the nomenclature BoWC15 (ANA8, IL-A135, IL-A137, IL-A138 and IL-A160).
Subject(s)
Antibodies, Monoclonal/chemistry , Granulocytes/immunology , Leukocytes, Mononuclear/immunology , Animals , Antibody Specificity/immunology , Binding Sites, Antibody/immunology , Binding, Competitive/immunology , Cattle , Flow Cytometry , PhenotypeABSTRACT
A set of monoclonal antibodies (MoAbs) to leucocyte antigens is an essential tool to identify different cell types and functional membrane molecules involved in immune responses. Since no MoAbs existed to bovine integrins, except against the beta 2 subfamily, we generated MoAbs to beta 3 integrin after the immunization of mice with bovine platelets. Two MoAbs, IL-A164 (IgG2a) and IL-A166 (IgG1), were selected that reacted specifically with bovine platelets and detected the same membrane molecule. The antigen was a heterodimer of two polypeptide chains of 122 kDa and 95 kDa as resolved by SDS-PAGE under reducing conditions. Although the Mr of the smaller subunit is identical to that of beta 2 integrin, preabsorption with an antibody to beta 2 (or CD18) did not remove the bovine antigen. Comparing the molecular masses of the two subunits in reduced and non-reduced forms showed a pattern that was similar to that of human GPIIb/IIIa (also called alpha IIb beta 3 or CD41a). Reduction of the bovine molecule increased the apparent Mr of the light chain from 76 kDa to 95 kDa, while the heavy subunit changed from 136 kDa to 122 kDa. As with human GPIIb, the decrease in Mr of the alpha-subunit is probably a result of a small disulphide-linked polypeptide, although no additional evidence for this was detected for the bovine integrin. Sequencing of the N-terminal amino acids of both bovine polypeptides showed identity of the bovine integrin with human GPIIb/IIIa.
Subject(s)
Antibodies, Monoclonal/immunology , Platelet Glycoprotein GPIIb-IIIa Complex/immunology , Platelet Membrane Glycoproteins/immunology , Amino Acid Sequence , Animals , Cattle , Female , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Molecular Weight , Precipitin TestsABSTRACT
Three monoclonal antibodies (mAbs), P13 (#90), IL-A78 (#76) and IL-A79 (#77), defined a new cluster, called BoWC8. The antigen defined by these mAbs is a molecule of 160 kDa under reducing conditions, and is expressed on lymphocytes late (6 days) after activation. The molecule can be expressed on multiplying T cells, but not on B cells.
Subject(s)
Antibodies, Monoclonal/immunology , Cattle/immunology , Receptors, Very Late Antigen/immunology , T-Lymphocytes/immunology , Animals , Antibodies, Protozoan/immunology , Antibody Specificity/immunology , B-Lymphocytes/immunology , Cell Line , Cells, Cultured , Epitopes/immunology , Flow Cytometry/veterinary , Lymphocyte Activation , Lymphoid Tissue/immunology , Mice , Precipitin Tests/veterinary , Theileria parva/immunology , Trypanosoma brucei brucei/immunology , Trypanosomiasis, Bovine/immunologyABSTRACT
PIP: Masturbatory semen specimen from 49 fertile Black African males in Kenya whose wives were pregnant was obtained after at least 3 days of abstinence for the analysis of parameters which included volume motility, vitality, sperm concentration, pH, fructose and acid phosphatase levels. About 1/2 the spermatozoa was actively progressive in motility, while 40% was nonmotile. Vitality in the 1st hour revealed that 81.4% of the sperm was alive. About 90% of the semen specimens had more than 40% idea forms of spermatozoa. Spermatozoal abnormalities were a frequent feature. There was no correlation between age and the testicular volume, but seminal fluid volume and sperm density tended to decrease with age. Serum levels of Follicle Stimulating Hormone, Luteinizing Hormone, Prolactin and Testosterone were determined in the subjects' sera, enabling the establishment of reference values for these parameters in African Kenyan males.^ieng
