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We repeatedly detected DNA of Borrelia garinii or B. afzelii and Borrelia-like structures in the blood, joint fluid or in the synovium of 10 patients with Lyme arthritis by means of the polymerase chain reaction and immunoelectron microscopy at 2-4-month intervals in the course of two years. All samples were analyzed using primers which amplified the 16S rRNA gene sequence of Borrelia burgdorferi sensu lato and nucleotide sequences for the OspA gene. No cross hybridization occurred with DNA from human cells and with DNA from other bacteria. Capture and labelling with monoclonal antibodies of aggregated antigens, membranes and flagellae were evident in the blood of 7 patients, in 4 synovial membranes and 2 synovial fluids. Borreliae were found in blood capillaries, in collagen and in clusters surrounding inflammatory cells in the synovium of patients with recurrent infections who carried IgM and IgG antibodies to OspA and to 83 kDa core protein. After significant improvement for several weeks after treatment, arthritis recurred in six patients. Synoviocyte hyperplasia, inflammatory infiltration and concentric adventitial fibroplasia were seen in the synovium of the patients with persisting borreliae. Only two patients were infected with B. afzelii, the others with B. garinii.

Summary Borrelia burgdorferi glycoconjugates with different oligosaccharide structures were characterized by a blotting technique with peroxidase-labelled lectins. The localization of surface carbohydrates was studied using electron microscopy with lectin-gold complexes. A high-mannose glycan structure was detected in 83 kDa glycoprotein (major extracellular protein); at least four carbohydrates (glucose or mannose, galactose, N-acetylgalactosamine and N-acetylglucosamine) were present in other Borrelia glycoconjugates. N-acetylneuraminic (sialic) acid was detected on the Borrelia surface. Two sialidases with different specificities were used in an attempt to cleave off the Borrelia N-acetylneuraminic acid. The attempt was successful by using Vibrio cholerae sialidase which has a broad substrate specificity, while the mumps-virus sialidase with restricted substrate specificity had no effect. Endogenous activity of N-acetylneuraminidase was not demonstrated in B. burgdorferi K5 and B31 strains. Zusammenfassung Glycokonjugate von Borrelia burgdorferi mit verschiedenen Oligosaccharidstrukturen wurden durch ein Blotting-Verfahren mit Peroxidase-markierten Lektinen charakterisiert. Die Lokalisierung der Oberflachen-Kohlenhydrate wurde unter Verwendung von Lektin-Gold-Komplexen elektronenmikroskopisch untersucht. In dem wichtigsten extrazellularen Protein, einem 83 kDa Glycoprotein, wurde eine mannosereiche Glycanstruktur entdeckt. Mindestens vier Kohlenhydrate (Glucose oder Mannose, Galactose, N-Acetylgalactosamin und N-Acetylglucosamin) waren in anderen Borrelia -Glycokonjugaten vorhanden. N-Acetylneuramin (Sialin)-Saure wurde auf der Oberflache der Borrelien entdeckt. Zwei Sialidasen unterschiedlicher Spezifitat wurden verwendet, um die N-Acetylneuraminsaure der Borrelien abzuspalten. Dies gelang unter Verwendung der Sialidase von Vibrio cholerae , die eine breite Substratspezifitat aufweist, wogegen die Sialidase des Mumpsvirus mit eingeschrankter Substratspezifitat wirkungslos war. In den B. burgdorferi -Stammen K5 and B31 konnte eine endogene Aktivitat der N-Acetylneuraminidase nicht nachgewiesen wurden.