02989nas a2200469   4500000000100000008004100001260001300042653002400055653002400079653001800103653002300121653001800144653002200162653002100184653001400205653001900219653002100238653004000259653002000299653001100319653002600330653002800356653002500384653002700409653002700436653001800463100001400481700001700495700001500512700001200527700001900539700002200558700001300580700001800593700001400611700001600625245011600641300001100757490000600768520173100774022001402505       1992                            d  c1992 Jan10aAmino Acid Sequence10aAntigens, Bacterial10aB-Lymphocytes10aBacterial Proteins10aBase Sequence10aBlotting, Western10aCarrier Proteins10aCell Line10aDNA, Bacterial10aDNA, Recombinant10aElectrophoresis, Polyacrylamide Gel10aGene Expression10aHumans10aLymphocyte Activation10aMolecular Sequence Data10aMycobacterium leprae10aReceptors, Fibronectin10aReceptors, Immunologic10aT-Lymphocytes1 aThole J E1 aSchöningh R1 aJanson A A1 aGarbe T1 aCornelisse Y E1 aClark-Curtiss J E1 aKolk A H1 aOttenhoff T H1 aVries R R1 aAbou-Zeid C00aMolecular and immunological analysis of a fibronectin-binding protein antigen secreted by Mycobacterium leprae.  a153-630 v63 a<p>By screening a Mycobacterium leprae lambda gt11 genomic DNA library with leprosy-patient sera we have previously identified 50 recombinant clones that expressed novel M. leprae antigens (Sathish et al., 1990). In this study, we show by DNA sequencing and immunoblot analysis that three of these clones express a M. leprae homologue of the fibronectin-binding antigen 85 complex of mycobacteria. The complete gene was characterized and it encodes a 327-amino-acid polypeptide, consisting of a consensus signal sequence of 38 amino acids followed by a mature protein of 289 amino acids. This is the first sequence of a member of the M. leprae antigen 85 complex, and Southern blotting analysis indicated the presence of multiple genes of the 85 complex in the genome of M. leprae. The amino acid sequence displays 75-85% sequence identity with components of the antigen 85 complex from M. tuberculosis, M. bovis BCG and M. kansasii. Furthermore, antibodies to the antigen 85 complex of M. tuberculosis and M. bovis BCG reacted with two fusion proteins containing the amino acid regions 55-266 and 266-327 of the M. leprae protein. The M. leprae 30/31 kDa protein induces strong humoral and cellular responses, as judged by Western blot analysis with patient sera and proliferation of T cells derived from healthy individuals and leprosy patients. Amino acid regions 55-266 and 265-327 both were shown to bind to fibronectin, indicating the presence of at least two fibronectin-binding sites on the M. leprae protein. These data indicate that this 30/31 kDa protein is not only important in the immune response against M. leprae, but may also have a biological role in the interaction of this bacillus with the human host.</p>  a0950-382X