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. 1995 Dec;63(12):4682–4685. doi: 10.1128/iai.63.12.4682-4685.1995

Molecular characterization and T-cell-stimulatory capacity of Mycobacterium leprae antigen T5.

B Wieles 1, E Spierings 1, J van Noort 1, B Naafs 1, R Offringa 1, T Ottenhoff 1
PMCID: PMC173672  PMID: 7591123

Abstract

A Mycobacterium leprae lambda gt11 clone designated T5 has previously been selected with sera from tuberculoid leprosy patients. Sequence analysis of this clone revealed the presence of two overlapping open reading frames (ORFs) present on the two cDNA strands. The first ORF codes for the serologically recognized antigen, which was fused with the lacZ gene in the lambda gt11 clone. The second ORF, present on the complementary strand, displays strong sequence homology with the aspartyl-tRNA synthetase genes of Escherichia coli and Thermus thermophilus. Here we show that the purified T5-derived product, overexpressed in E. coli, is recognized by T cells of the majority of the leprosy patients tested, including lepromatous leprosy patients who do not respond to whole M. leprae bacilli.

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Selected References

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  1. Carothers A. M., Urlaub G., Mucha J., Grunberger D., Chasin L. A. Point mutation analysis in a mammalian gene: rapid preparation of total RNA, PCR amplification of cDNA, and Taq sequencing by a novel method. Biotechniques. 1989 May;7(5):494-6, 498-9. [PubMed] [Google Scholar]
  2. Clark-Curtiss J. E., Jacobs W. R., Docherty M. A., Ritchie L. R., Curtiss R., 3rd Molecular analysis of DNA and construction of genomic libraries of Mycobacterium leprae. J Bacteriol. 1985 Mar;161(3):1093–1102. doi: 10.1128/jb.161.3.1093-1102.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Eriani G., Delarue M., Poch O., Gangloff J., Moras D. Partition of tRNA synthetases into two classes based on mutually exclusive sets of sequence motifs. Nature. 1990 Sep 13;347(6289):203–206. doi: 10.1038/347203a0. [DOI] [PubMed] [Google Scholar]
  4. Eriani G., Dirheimer G., Gangloff J. Aspartyl-tRNA synthetase from Escherichia coli: cloning and characterisation of the gene, homologies of its translated amino acid sequence with asparaginyl- and lysyl-tRNA synthetases. Nucleic Acids Res. 1990 Dec 11;18(23):7109–7118. doi: 10.1093/nar/18.23.7109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Marsh J. L., Erfle M., Wykes E. J. The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene. 1984 Dec;32(3):481–485. doi: 10.1016/0378-1119(84)90022-2. [DOI] [PubMed] [Google Scholar]
  6. Ottenhoff T. H., Converse P. J., Gebre N., Wondimu A., Ehrenberg J. P., Kiessling R. T cell responses to fractionated Mycobacterium leprae antigens in leprosy. The lepromatous nonresponder defect can be overcome in vitro by stimulation with fractionated M. leprae components. Eur J Immunol. 1989 Apr;19(4):707–713. doi: 10.1002/eji.1830190421. [DOI] [PubMed] [Google Scholar]
  7. Poterszman A., Plateau P., Moras D., Blanquet S., Mazauric M. H., Kreutzer R., Kern D. Sequence, overproduction and crystallization of aspartyl-tRNA synthetase from Thermus thermophilus. Implications for the structure of prokaryotic aspartyl-tRNA synthetases. FEBS Lett. 1993 Jul 5;325(3):183–186. doi: 10.1016/0014-5793(93)81069-c. [DOI] [PubMed] [Google Scholar]
  8. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Sathish M., Esser R. E., Thole J. E., Clark-Curtiss J. E. Identification and characterization of antigenic determinants of Mycobacterium leprae that react with antibodies in sera of leprosy patients. Infect Immun. 1990 May;58(5):1327–1336. doi: 10.1128/iai.58.5.1327-1336.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Schimmel P. Aminoacyl tRNA synthetases: general scheme of structure-function relationships in the polypeptides and recognition of transfer RNAs. Annu Rev Biochem. 1987;56:125–158. doi: 10.1146/annurev.bi.56.070187.001013. [DOI] [PubMed] [Google Scholar]
  11. Wieles B., van Agterveld M., Janson A., Clark-Curtiss J., Rinke de Wit T., Harboe M., Thole J. Characterization of a Mycobacterium leprae antigen related to the secreted Mycobacterium tuberculosis protein MPT32. Infect Immun. 1994 Jan;62(1):252–258. doi: 10.1128/iai.62.1.252-258.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  13. Young R. A., Mehra V., Sweetser D., Buchanan T., Clark-Curtiss J., Davis R. W., Bloom B. R. Genes for the major protein antigens of the leprosy parasite Mycobacterium leprae. Nature. 1985 Aug 1;316(6027):450–452. doi: 10.1038/316450a0. [DOI] [PubMed] [Google Scholar]

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