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The patterns of cross-reaction of 30 monoclonal antibodies and three lectins were determined by ELISA with 21 ABH, Ii or Lewis related synthetic oligosaccharides coupled to bovine serum albumin. At least seven main groups of cross-reactive patterns were identified among the antibodies, plus several isolated antibodies which had intermediate patterns between two of the main antibody groups. The three lectins had different cross-reaction patterns, Galactia tenuiflora was different from all the antibodies, Ulex europaeus lectin 1 and Lotus tetragonolobus were similar, but not identical to groups III and V of antibodies respectively. The anti-H antibodies cross-reacting with A type 2 gave similar agglutination scores with all the normal ABO erythrocytes, while the anti-H antibodies not cross-reacting with A type 2 reacted with different scores: O > A2 > A2B > B > A1 > A1B > O(h), suggesting that these antibodies react better with the free H epitopes and do not recognize the H in A or B epitopes. Based on the ELISA and agglutination results and the lowest energy conformations of each oligosaccharide obtained by computer modelling, the most probable oligosaccharide surface areas recognized by each antibody main group are illustrated.

Comparing the reactivities of murine and human monoclonal antibodies directed against the antigens of A, B, and H blood groups by using different techniques (agglutination, inhibition by synthetic oligosaccharides and salivary antigens, tissue immunofluorescence, the use of an anti-idiotype antibody) enabled us to show a high heterogeneity of anti-A, anti-AB antibodies and, to some extend, anti-B and anti-H antibodies. Despite this diversity, the determination of the antibody specificity thanks to synthetic antigens made it possible to distinguish several groups of anti-A, B, AB, and H antibodies and to establish a classification that takes also their reactivity towards salivary and tissue antigens into account. On the contrary, few correlations were found between these tests and the ability of antibodies to agglutinate red blood cells of the different ABO-system variants.

The idiotypy of antibodies (Ab) specific for oligosaccharide determinants of blood groups of the human ABO system was studied through a cascade. Xenogenic antiidiotypic Ab (Ab2) raised in rabbits to the murine monoclonal anti-A61 (Ab1) were screened for reactivity with various anti-ABH Ab. Three anti-A and three anti-A,B monoclonal antibodies (mAb) which were developed in the same mouse strain as that producing Ab1, as well as a human polyclonal anti-A, were found to share cross-reactive idiotopes (CRI) with Ab1. CRI on murine mAb could be due to a Biozzi recurrent Id on anti-A Ab reacting with anti-Id "à la Oudin", while CRI on human anti-A Ab suggested the presence of paratope-induced anti-Id. Inhibition by Ab2 of haemagglutination of A, B or O human red blood cells by many murine anti-ABH mAb, and by polyclonal or monoclonal human anti-A, strongly supported the occurrence of anti-Id mimicking ABH epitopes belonging to type 2 determinants carried by human erythrocytes. Furthermore, a rabbit immunized with Ab2 produced a potent Ab3 response characterized by anti-H-type-2 specificity. Altogether, these results are consistent with the first successful production of anti-Id Ab that mimics the tridimensional shape of a well defined and strictly carbohydrate epitope, eliciting a haemagglutinating Ab3.

ABH antigens are normally present on the surface of the healthy urinary epithelium. In a significant number of patients with tumour of the bladder the epithelium loses these antigens. The loss appears to be unrelated to the grade or stage of the tumour, but the prognosis seems to be brighter when the ABH antigens are retained.

Eight anti-A and seven anti-B monoclonal reagents were tested in parallel, with normal and weak ABH red cell phenotypes. A whole range of different reactivity patterns was found, but by making a comparison with the results obtained using polyclonal standard reagents, two major categories of reagents were distinguished: (a) stronger and more specific reagents, and (b) reagents similar to, or weaker than, the standard polyclonal controls. The analysis of the specificity of the reagents by tissue fluorescence staining and reactivity with synthetic oligosaccharides and purified glycolipids confirmed the existence of broad and restricted specificities. Two kinds of anti-A1 reagents are described. One related to type 3/4 structures, which stains the Golgi apparatus, and another with broad anti-A specificity which cross-reacts with 'A-like' structures. The inhibition of anti-A reagents with salivas and synthetic oligosaccharide antigens gave parallel results for the secretor salivas and the difucosylated A antigens.