02620nas a2200409   4500000000100000008004100001260001300042653002400055653001200079653002400091653002300115653002300138653001600161653001100177653002500188653002600213653000900239653002400248653002800272653002100300653002500321653003200346653002400378653003400402653002500436653001800461100001200479700001500491700001600506700001400522700001200536245013200548300001200680490001600692520148800708022001402196       1996                            d  c1996 Jun10aAmino Acid Sequence10aAnimals10aAntigens, Bacterial10aBacterial Proteins10aBacterial Vaccines10aGuinea Pigs10aHumans10aLeprosy, Tuberculoid10aLymphocyte Activation10aMice10aMice, Inbred BALB C10aMolecular Sequence Data10aMolecular Weight10aMycobacterium leprae10aNontuberculous Mycobacteria10aPhytohemagglutinins10aSequence Homology, Amino Acid10aSuperoxide Dismutase10aT-Lymphocytes1 aBisht D1 aMehrotra J1 aDhindsa M S1 aSingh N B1 aSinha S00aA major T-cell-inducing cytosolic 23 kDa protein antigen of the vaccine candidate Mycobacterium habana is superoxide dismutase.  a1375-830 v142 ( Pt 6)3 a<p>This study describes the purification and immunochemical characterization of a major 23 kDa cytosolic protein antigen of the vaccine candidate Mycobacterium habana (TMC 5135). The 23 kDa protein alone was salted out from the cytosol at an ammonium sulfate saturation of 80-95%. It represented about 1.5% of the total cytosolic protein, appeared glycosylated by staining with periodic acid/Schiff's reagent, and showed a pl of approximately 5.3. Its native molecular mass was determined as approximately 48 kDa, suggesting a homodimeric configuration. Immunoblotting with the WHO-IMMLEP/IMMTUB mAbs mc5041 and IT61 and activity staining after native PAGE established its identity as a mycobacterial superoxide dismutase (SOD) of the Fe/Mn type. The sequence of the 18 N-terminal amino acids, which also contained the binding site for mc5041, showed a close resemblance, not only with the reported deduced sequences of Mycobacterium leprae and Mycobacterium tuberculosis Fe/MnSODs, but also with human MnSOD. In order to study its immunopathological relevance, the protein was subjected to in vivo and in vitro assays for T cell activation. It induced, in a dose-related manner, skin delayed hypersensitivity in guinea-pigs and lymphocyte proliferation in BALB/c mice primed with M. habana. Most significantly, it also induced lymphocyte proliferative responses, in a manner analogous to M. Ieprae, in human subjects comprising tuberculoid leprosy patients and healthy contacts.</p>  a1350-0872