PENGEKSPRESIAN DAN KAJIAN IMUNOLOGI KE ATAS GEN mtp40 DARIPADA Mycobacterium tuberculosis

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Date
2001-03
Authors
AZIAH, ISMAIL
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Publisher
Pusat Pengajian Sains Perubatan Universiti Sains Malaysia
Abstract
The development of improved vaccines is considered a high priority in the effort to control tuberculosis worldwide. The focus of this project is the MTP40 protein of Mycobacterium tuberculosis which have been previously reported to contain both B and T cell epitopes. In this study, immune response to 7 peptides (P1-P7) derived from the MTP40 protein of M. tuberculosis were tested by enzyme linked immunosorbent assay (ELISA) and lymphoblastic transformation test (L TT) using specimens from tuberculosis patients (TB), healthy contacts (HC) and normal controls (NC). Antibody response to MTP40 peptides were generally found to be higher in the TB group as compared to the HC group but only in peptides P1 (p<O.005) and P2 (p<O.01) were the difference between the two groups found to be statistically significant. On the other hand, lymphocyte responses to these peptides were generally found to be higher in the He group as compared to the TB group with P1, P2, P5 and P6 showing statistically significant differences. This results suggest that MTP40 may be useful as a vaccine candidate. Further work was done to explore this possibility. The mtp40 gene was cloned into two vectors. Cloning of the gene into pJW4303 created the recombinant clone pJWmtp40-24 which was suitable for use as a DNA vaccine. Cloning of the gene into the second vector, pGEX-2T, an expression vector, resulted in the recombinant clone pGEXmtp40-1 which allowed for high level expression of MTP40 as a fused protein to the glutathione s-transferase (GST) enzyme. xiii pJWmtp40-24 was used to immunize C57BL/6J mice. Serum from the immunized mice were shown by Western blotting to be capable of recognizing the MTP40 protein exposed by pGEXmtp40-1. Through ELISA, it was shown that pJWmtp40-24 was able to stimulate antibody response to all 7 peptides described above although responses to each peptide differed from mice to mice. In L TT test, specimens from the immunized mice showed statistically significant responses to peptides P4 and P6. The results described above showed that the recombinant clone pJWmtp40-24 has a good potential to be developed into a DNA vaccine candidate against tuberculosis.
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Keywords
Mycobacterium tuberculosis
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