Recombinantly produced borrelial BBK32 protein fragments originating from Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii were evaluated as antigens in the serology of Lyme borreliosis (LB). In ELISA, a mid-portion hydrophilic fragment reacted with LB patient sera. Of the 23 patients with culture-or PCR-positive erythema migrans (EM), 43 % at diagnosis and 52 % at convalescence were positive for at least one Borrelia species-specific variant BBK32 fragment antigen. In parallel ELISAs with BBK32 whole proteins from the three borrelial subspecies as antigens, 17 % at diagnosis and 26 % at convalescence were positive. These results suggest that BBK32 protein fragments may improve the early IgG serology of LB compared to the BBK32 whole protein.
Complement C4 is a highly polymorphic protein essential for the activation of the classical complement pathway. Most of the allelic variation of C4 resides in the C4d region. Four polymorphic amino acid residues specify the isotype and an additional four specify the Rodgers and Chido determinants of the protein. Rare C4 allotypes have been postulated to originate from recombination between highly homologous C4 genes through gene conversions. Here we describe the development of a de novo C4 hybrid protein with allotypic and antigenic diversity resulting from nonhomologous intra or interchromosomal recombination of the maternal chromosomes. A conversion was observed between maternal C4A3a and C4B1b genes producing a functional hybrid gene in one of the children. The codons determining the isotype, Asp1054, Leu1101, Ser1102, Ile1105 and His1106, were characteristic of C4B gene, whereas the polymorphic sites in exon and intron 28 were indicative of C4A3a sequence. The protein produced by this hybrid gene was electrophoretically similar to C4B5 allotype. It also possesses reversed antigenicity being Rodgers 1, 2, 3 and Chido-1, -2, -3, 4, -5, and -6. Our case describes the development of a rare bimodular C4B-C4B haplotype containing a functional de novo C4 hybrid gene arisen through gene conversion from C4A to C4B. Overall the data supports the hypothesis of gene conversions as an ongoing process increasing allelic diversity in the C4 locus.
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