Antibody Reactivity of Synthetic Peptides Belonging to Proteins Encoded by Genes Located in Mycobacterium tuberculosis-Specific Genomic Regions of Differences
The comparisons of mycobacterial genomes have identified several Mycobacterium tuberculosis-specific genomic regions that are absent in other mycobacteria and are known as regions of differences. Due to M. tuberculosis-specificity, the peptides encoded by these regions could be useful in the specific diagnosis of tuberculosis. To explore this possibility, overlapping synthetic peptides corresponding to 39 proteins predicted to be encoded by genes present in regions of differences were tested for antibody-reactivity with sera from tuberculosis patients and healthy subjects. The results identified four immunodominant peptides corresponding to four different proteins, with three of the peptides showing significantly stronger antibody reactivity and rate of positivity with sera from tuberculosis patients than healthy subjects. The fourth peptide was recognized equally well by the sera of tuberculosis patients as well as healthy subjects. Predication of antibody epitopes by bioinformatics analyses using ABCpred server predicted multiple linear epitopes in each peptide. Furthermore, peptide sequence analysis for sequence identity using BLAST suggested M. tuberculosis-specificity for the three peptides that had preferential reactivity with sera from tuberculosis patients, but the peptide with equal reactivity with sera of TB patients and healthy subjects showed significant identity with sequences present in nob-tuberculous mycobacteria. The three identified M. tuberculosis-specific immunodominant peptides may be useful in the serological diagnosis of tuberculosis.
[1] WHO Report. 2017. Global tuberculosis report. http://www.who.int/tb/publications/global_report/en/.
[2] World Health Organization. 2017. Tuberculosis country profiles. TB burden estimates and country-reported TB data. Kuwait. http://www.who.int/tb/country/data/profiles/en/.
[3] Mustafa AS. 2009. Vaccine potential of Mycobacterium tuberculosis-specific genomic regions: in vitro studies in humans, Expert Rev Vaccines 8: 1309-1312.
[4] Mustafa AS. 2012. What's New in the Development of Tuberculosis Vaccines. Med Princ Pract 21:195-196.
[5] Mustafa AS. 2012. Proteins and peptides encoded by M. tuberculosis-specific genomic regions for immunological diagnosis of tuberculosis. J Mycobac Dis 2:e114.
[6] Mustafa AS. 2013. Diagnostic and vaccine potentials of ESAT-6 family proteins encoded by M. tuberculosis genomic regions absent in M. bovis BCG. J Mycobac Dis 3:129.
[7] Mustafa AS. 2013. In silico analysis and experimental validation of Mycobacterium tuberculosis-specific proteins and peptides of Mycobacterium tuberculosis for immunological diagnosis and vaccine development. Med Princ Pract 22 (Suppl 1): 43-51.
[8] Mustafa AS. 2014. The future of Mycobacterium tuberculosis-specific antigens/peptides in tuberculin skin testing for the diagnosis of tuberculosis. J Mycobac Dis 4:3.
[9] Mustafa AS, Al-Attiyah R. 2004. Mycobacterium tuberculosis antigens and peptides as new vaccine candidates and immunodiagnostic reagents against tuberculosis. Kuwait Med J 36: 171-176.
[10] Al-Attiyah R, Mustafa AS. 2008. Characterization of human cellular immune responses to novel Mycobacterium tuberculosis antigens encoded by genomic regions absent in Mycobacterium bovis BCG. Infect Immun 76, 4190-4198.
[11] Al-Attiyah R, Mustafa AS. 2010. Characterization of human cellular immune responses to Mycobacterium tuberculosis proteins encoded by genes predicted in RD15 genomic region that is absent in Mycobacterium bovis BCG. FEMS Immunol Med Microbiol 59: 177-87.
[12] Mustafa AS, Al-Saidi F, El-Shamy ASM, Al-Attiyah R. 2011. Cytokines in response to proteins predicted in genomic regions of difference of Mycobacterium tuberculosis. Microbiol Immunol 55:267-278.
[13] Mustafa AS. 2014. T-helper 1, T-helper 2, pro-inflammatory and anti-inflammatory cytokines in tuberculosis. IJPMB 3:1-14.
[14] Mustafa AS. 2014. Characterization of a cross-reactive, immunodominant and HLA-promiscuous epitope of Mycobacterium tuberculosis-specific major antigenic protein PPE68. PLoS One 9:e103679.
[15] Hanif SNM, Al-Attiyah R, Mustafa AS. 2010. Molecular cloning, expression, purification and immunological characterization of three low molecular weight proteins encoded by genes in genomic regions of difference of Mycobacterium tuberculosis. Scand J Immunol 71:353-361.
[16] Al-Khodari NY, Al-Attiyah R, and Mustafa AS. 2011. Identification, diagnostic potential and natural expression of immunodominant seroreactive peptides encoded by five Mycobacterium tuberculosis-specific genomic regions. Clin Exp Immunol 18:477-482.
[17] Mustafa AS. 2009. HLA-promiscuous Th1-cell reactivity of MPT64 (Rv1980c), a major secreted antigen of Mycobacterium tuberculosis, in healthy subjects. Med Princ Pract 18:385-392.
[18] Mustafa AS, Al-Attiyah R 2009 Identification of Mycobacterium tuberculosis-specific genomic regions encoding antigens those induce qualitatively opposing cellular immune responses. Ind J Exp Biol 47:498-504.
[19] Mustafa, AS, Al-Attiyah R, Hanif SNM, Shaban FA. 2008. Efficient testing of large pools of Mycobacterium tuberculosis RD1 peptides and identification of major antigens and immunodominant peptides recognized by human Th1 cells. Clin Vaccine Immunol 15:916-924
[20] Mustafa AS. 2009. Th1-cell reactivity and HLA-DR binding prediction for promiscuous recognition of MPT63 (Rv1926c), a major secreted protein of Mycobacterium tuberculosis. Scand J Immunol 69:213-222.
[21] Al‐Attiyah R, Mustafa AS. 2004. Computer-assisted prediction of HLA-DR binding and experimental analysis for human promiscuous Th1-cell peptides in the 24 kDa secreted lipoprotein (LppX) of Mycobacterium tuberculosis. Scand J Immunol 59:16-24.
[22] El-Shazly S, Mustafa AS, Ahmad S, Al-Attiyah R. 2007. Utility of three mammalian cell entry proteins of Mycobacterium tuberculosis in serodiagnosis of tuberculosis. Inter J Tuber Lung Dis 11:676-682.
[23] Saha S, Raghava GPS. 2006. Prediction of continuous B-Cell epitopes in an antigen using recurrent neural network. Proteins 65:40–48.
[24] IEDB. June 2010, updating date. Epitope prediction and analysis tools. http://tools.immuneepitope.org.
[25] Protein BLAST: search protein databases using a protein query - NIH. https://blast.ncbi.nlm.nih.gov/Blast.cgi?PAGE=Proteins (accessed on December 10, 2017).
[26] TubercuList - GenoList - Institut Pasteur. http://genolist.pasteur.fr/TubercuList/ (accessed on December 17, 2017).
[27] Okkels LM, Brock I, Follmann F, Agger EM, SArend SM, Ottenhoff THM, Oftung F, Rosenkrands I, Andersen P. 2003. PPE protein (Rv3873) from DNA segment RD1 of Mycobacterium tuberculosis: strong recognition of both specific T-cell epitopes and epitopes conserved within the PPE family. Infect Immun 71:6116-6123.
[1] WHO Report. 2017. Global tuberculosis report. http://www.who.int/tb/publications/global_report/en/.
[2] World Health Organization. 2017. Tuberculosis country profiles. TB burden estimates and country-reported TB data. Kuwait. http://www.who.int/tb/country/data/profiles/en/.
[3] Mustafa AS. 2009. Vaccine potential of Mycobacterium tuberculosis-specific genomic regions: in vitro studies in humans, Expert Rev Vaccines 8: 1309-1312.
[4] Mustafa AS. 2012. What's New in the Development of Tuberculosis Vaccines. Med Princ Pract 21:195-196.
[5] Mustafa AS. 2012. Proteins and peptides encoded by M. tuberculosis-specific genomic regions for immunological diagnosis of tuberculosis. J Mycobac Dis 2:e114.
[6] Mustafa AS. 2013. Diagnostic and vaccine potentials of ESAT-6 family proteins encoded by M. tuberculosis genomic regions absent in M. bovis BCG. J Mycobac Dis 3:129.
[7] Mustafa AS. 2013. In silico analysis and experimental validation of Mycobacterium tuberculosis-specific proteins and peptides of Mycobacterium tuberculosis for immunological diagnosis and vaccine development. Med Princ Pract 22 (Suppl 1): 43-51.
[8] Mustafa AS. 2014. The future of Mycobacterium tuberculosis-specific antigens/peptides in tuberculin skin testing for the diagnosis of tuberculosis. J Mycobac Dis 4:3.
[9] Mustafa AS, Al-Attiyah R. 2004. Mycobacterium tuberculosis antigens and peptides as new vaccine candidates and immunodiagnostic reagents against tuberculosis. Kuwait Med J 36: 171-176.
[10] Al-Attiyah R, Mustafa AS. 2008. Characterization of human cellular immune responses to novel Mycobacterium tuberculosis antigens encoded by genomic regions absent in Mycobacterium bovis BCG. Infect Immun 76, 4190-4198.
[11] Al-Attiyah R, Mustafa AS. 2010. Characterization of human cellular immune responses to Mycobacterium tuberculosis proteins encoded by genes predicted in RD15 genomic region that is absent in Mycobacterium bovis BCG. FEMS Immunol Med Microbiol 59: 177-87.
[12] Mustafa AS, Al-Saidi F, El-Shamy ASM, Al-Attiyah R. 2011. Cytokines in response to proteins predicted in genomic regions of difference of Mycobacterium tuberculosis. Microbiol Immunol 55:267-278.
[13] Mustafa AS. 2014. T-helper 1, T-helper 2, pro-inflammatory and anti-inflammatory cytokines in tuberculosis. IJPMB 3:1-14.
[14] Mustafa AS. 2014. Characterization of a cross-reactive, immunodominant and HLA-promiscuous epitope of Mycobacterium tuberculosis-specific major antigenic protein PPE68. PLoS One 9:e103679.
[15] Hanif SNM, Al-Attiyah R, Mustafa AS. 2010. Molecular cloning, expression, purification and immunological characterization of three low molecular weight proteins encoded by genes in genomic regions of difference of Mycobacterium tuberculosis. Scand J Immunol 71:353-361.
[16] Al-Khodari NY, Al-Attiyah R, and Mustafa AS. 2011. Identification, diagnostic potential and natural expression of immunodominant seroreactive peptides encoded by five Mycobacterium tuberculosis-specific genomic regions. Clin Exp Immunol 18:477-482.
[17] Mustafa AS. 2009. HLA-promiscuous Th1-cell reactivity of MPT64 (Rv1980c), a major secreted antigen of Mycobacterium tuberculosis, in healthy subjects. Med Princ Pract 18:385-392.
[18] Mustafa AS, Al-Attiyah R 2009 Identification of Mycobacterium tuberculosis-specific genomic regions encoding antigens those induce qualitatively opposing cellular immune responses. Ind J Exp Biol 47:498-504.
[19] Mustafa, AS, Al-Attiyah R, Hanif SNM, Shaban FA. 2008. Efficient testing of large pools of Mycobacterium tuberculosis RD1 peptides and identification of major antigens and immunodominant peptides recognized by human Th1 cells. Clin Vaccine Immunol 15:916-924
[20] Mustafa AS. 2009. Th1-cell reactivity and HLA-DR binding prediction for promiscuous recognition of MPT63 (Rv1926c), a major secreted protein of Mycobacterium tuberculosis. Scand J Immunol 69:213-222.
[21] Al‐Attiyah R, Mustafa AS. 2004. Computer-assisted prediction of HLA-DR binding and experimental analysis for human promiscuous Th1-cell peptides in the 24 kDa secreted lipoprotein (LppX) of Mycobacterium tuberculosis. Scand J Immunol 59:16-24.
[22] El-Shazly S, Mustafa AS, Ahmad S, Al-Attiyah R. 2007. Utility of three mammalian cell entry proteins of Mycobacterium tuberculosis in serodiagnosis of tuberculosis. Inter J Tuber Lung Dis 11:676-682.
[23] Saha S, Raghava GPS. 2006. Prediction of continuous B-Cell epitopes in an antigen using recurrent neural network. Proteins 65:40–48.
[24] IEDB. June 2010, updating date. Epitope prediction and analysis tools. http://tools.immuneepitope.org.
[25] Protein BLAST: search protein databases using a protein query - NIH. https://blast.ncbi.nlm.nih.gov/Blast.cgi?PAGE=Proteins (accessed on December 10, 2017).
[26] TubercuList - GenoList - Institut Pasteur. http://genolist.pasteur.fr/TubercuList/ (accessed on December 17, 2017).
[27] Okkels LM, Brock I, Follmann F, Agger EM, SArend SM, Ottenhoff THM, Oftung F, Rosenkrands I, Andersen P. 2003. PPE protein (Rv3873) from DNA segment RD1 of Mycobacterium tuberculosis: strong recognition of both specific T-cell epitopes and epitopes conserved within the PPE family. Infect Immun 71:6116-6123.
@article{"International Journal of Medical, Medicine and Health Sciences:77237", author = "Abu Salim Mustafa", title = "Antibody Reactivity of Synthetic Peptides Belonging to Proteins Encoded by Genes Located in Mycobacterium tuberculosis-Specific Genomic Regions of Differences", abstract = "The comparisons of mycobacterial genomes have identified several Mycobacterium tuberculosis-specific genomic regions that are absent in other mycobacteria and are known as regions of differences. Due to M. tuberculosis-specificity, the peptides encoded by these regions could be useful in the specific diagnosis of tuberculosis. To explore this possibility, overlapping synthetic peptides corresponding to 39 proteins predicted to be encoded by genes present in regions of differences were tested for antibody-reactivity with sera from tuberculosis patients and healthy subjects. The results identified four immunodominant peptides corresponding to four different proteins, with three of the peptides showing significantly stronger antibody reactivity and rate of positivity with sera from tuberculosis patients than healthy subjects. The fourth peptide was recognized equally well by the sera of tuberculosis patients as well as healthy subjects. Predication of antibody epitopes by bioinformatics analyses using ABCpred server predicted multiple linear epitopes in each peptide. Furthermore, peptide sequence analysis for sequence identity using BLAST suggested M. tuberculosis-specificity for the three peptides that had preferential reactivity with sera from tuberculosis patients, but the peptide with equal reactivity with sera of TB patients and healthy subjects showed significant identity with sequences present in nob-tuberculous mycobacteria. The three identified M. tuberculosis-specific immunodominant peptides may be useful in the serological diagnosis of tuberculosis.
", keywords = "Genomic regions of differences, Mycobacterium tuberculosis, peptides, serodiagnosis.", volume = "12", number = "4", pages = "132-6", }
