Identification of Recombinant Antigens of Mycobacterium Leprae Which React With Antibodies from Lepromatous and Borderline Leprosy Patients
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Date
1991-06
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Addis Ababa University
Abstract
Recombinant antigens of Mycobacterium leprae were identified by
screening a lambda gt-11 M. leprae library with absorbed
lepromatous (LL), borderline tuberculoid (BT) leprosy and a healthy
contact sera in separate experiments. Six, three, and one clones
were respectively isolated by successive screenings of the plaques
by immunodetection of antigens on nitrocellulose filters. Of
these, a total of six lysogens were generated. Four were made from
the clones isolated using LL serum and two were made fromclo~es
isolated using BT serum. Analysis of the apparent molecular mass
of the recombinant proteins by either SDS-PAGE or Western blotting,
showed the fusion proteins to be in the range of 116 kD to 1.44 kD.
The reactivity of recombinant proteins with patients sera
throughout the spectrum of the disease was found to be heterogenous
regardless of the molecular sizes of the proteins. The
characterization of the clones with regard to DNA insert mass was
made by amplification with Taq polymerase chain reaction. The
analysis on 1.5% agarose gel electrophoresis showed the range to be
from 0.3 Kb to 2.0 Kb. The proliferation of T-cell as a result of
stimulation by the recombinant proteins (antigens) looks promising
and further study for confirmation is in progress.
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Biology