title: Coordinator's report: an assessment of the functional activity of human Rh monoclonal antibodies after their evaluation by nine laboratories.
abstract: The functional activity of 55 anti-D and 26 MAbs of other Rh specificities was determined as part of the Third International Workshop and Symposium on Monoclonal Antibodies Against Human Red Blood Cell and Related Antigens. Most MAbs were IgG1 (45 anti-D, 16 Rh). There was a large range of anti-D and IgG concentrations of the anti-D MAbs (0.4-1680 IU/ml, 1.5-400 micrograms/ml). Correlation between quantitative data from different laboratories was good. Six laboratories tested the anti-Ds in monocyte ADCC assays. Methods varied greatly, especially the effector cells used, the methods of red cell sensitisation, the effector to target cell ratio and the assay incubation times. There was some correlation of results between most laboratories, but results were better when similar assays were performed. The extent of monocyte-mediated haemolysis was related to the number of molecules of IgG anti-D bound to the red cells. Monocyte phagocytosis assays resulted in a greater variability in results between the four evaluating laboratories, though IgG3 MAbs were found more active than IgG1, and mostly promoted red cell adherence rather than phagocytosis. Two monocyte chemiluminescence assays found comparatively low activity of the IgG3 MAbs; the most active IgG1 anti-Ds were MAbs 93 and 95. Correlation between different monocyte-mediated assays was generally poor unless the assays were performed by the same laboratory. Results of a macrophage ADCC assay showed that the IgG3 anti-D's promoted greater haemolysis than most IgG1 MAbs. Only two IgG1 anti-D MAbs (70 and 104) mediated high haemolysis in this assay. Lymphocyte ADCC assays were utilised in four laboratories. The IgG3 antibodies exhibited low activity but one IgG1 anti-D (104) consistently mediated high haemolysis. There was no relationship between quantitation and haemolysis in this assay. A few MAbs exhibited low or no activity in most assays, mainly due to low quantitation. Most of the MAbs of non-D specificity mediated low functional activity which in most cases was not due to low quantitation. However two MAbs, 24 (anti-CcEe) and 25 (anti-CD), consistently showed good activity. This study highlighted the great variability in assay methodology between the nine participating laboratories, which resulted in some apparent differences in functional activity of some of the MAbs. The use of standardised methods as well as fewer MAbs tested and quantitated at different concentrations may help to determine the relevant factors contributing to IgG function.
authors.affiliation: International Blood Group Reference Laboratory, Bristol, UK.
authors.forename: B M
authors.initials: BM
authors.lastname: Kumpel
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000920:Antibody-Dependent Cell Cytotoxicity; D006801:Humans; D008214:Lymphocytes / Q000276:immunology; D008264:Macrophages / Q000276:immunology; D009000:Monocytes / Q000276:immunology; D010587:Phagocytosis; D012204:Rh-Hr Blood-Group System / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D012204:Rh-Hr Blood-Group System