Comprehensive detection of immunorelevant Borrelia garinii antigens by two-dimensional electrophoresis

Lyme disease is caused by spirochetes of the Borrelia burgdorferi complex a nd has been reported in many temperate parts of the Northern Hemisphere. Th e B. burgdorferi complex consists of at least five different species and fi ve genotypes with different pathogenicity. Serodiagnosis was achieved by de tection of antigens on one-dimensional (1-D) immunoblots. A systematic and comprehensive approach to elucidate antigens has been started here by the c ombination of two-dimensional electrophoresis (2-DE) immunoblotting with ma trix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Ant igens in the proteome of B. garinii BITS were analyzed for their reactivity with sera from patients in early stage (erythema migrans) and late manifes tations (neuroborreliosis late, arthritis and acrodermatitis chronica athro phicans) of borreliosis. A strategy to handle the enormous amount of data w as developed and 65 antigens were detected, of which 20 were identified. Th ese comprise the known antigens from 1-D immunoblots used routinely in sero diagnosis and additionally the two new antigens, GAPDH and the ABC transpor ter oligopeptide permease. Several disease-stage unique proteins were detec ted and some of them identified. The genetic variability between B. garinii strains BITS and 20047, B. afzelii, and B. burgdorferi, sensu stricto, see n on the 2-DE patterns underlines the necessity of the search for additiona l antigens to improve the serodiagnosis and development of vaccines to be u sed outside of Northern America. A 2-DE database of B. garinii was built up and is available on the World Wide Web (http://www.mpiib-berlin.mpg.de/2D- PAGE).