Share This Article:

Identification of Growth Promoting Effect of rBCG/BCG Culture Supernatant and Its Potential Applications

Abstract Full-Text HTML XML Download Download as PDF (Size:472KB) PP. 32-38
DOI: 10.4236/wjv.2013.32006    3,689 Downloads   5,823 Views   Citations

ABSTRACT

Background: Although BCG is the most widely administered vaccine in the world, there have never been as many cases of TB as there are now. Globally, more than 8.8 million people developed active TB and 1.4 millionmany of themdied in 2010. It is estimated that half of pulmonary TB cases arise from latent Mtb infection, making the study of latency and reactivation of utmost importance. Methods: Widely administered BCG vaccines and a gene modified recombinant BCG (rBCG) strain, AERAS-422, were used as models to investigate the growth promoting function of resuscitation-promoting factors (Rpfs) in different bacilli culture phases. Different supernatant fractions were prepared by ultrafiltration, and the promoting function of each fraction containing secreted Rpf(s) was evaluated by growth curve monitoring and colony counting on 7H10 agar plates. Results: The promoting effect of culture supernatants was mainly associated with the high molecular weight fraction (>30 kDa), which stimulated bacterial growth, but did not extend the exponential phase of stimulated culture. Anti-RpfB antibody showed significant growth restriction of the tested cultures. When comparing rBCG cultures containing 7H9 medium, the 10 - 30 kDa fraction, or the >30 kDa fraction, only the >30 kDa fraction was displayed with down-regulation of the secretion of RpfC, D and E. In colony counting tests, the plates containing the >30 kDa fraction had total countable colony numbers 2 to 3 fold higher than the plates with the 10 - 30 kDa fraction, and colonies appeared one to two weeks earlier than on the regular plates. The potential applications of the prepared supernatant fractions containing RpfA and RpfB are discussed, which may include accelerating diagnosis of Mtb infection and future TB vaccine development.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

T. Jin, T. Qu, A. Raina and E. Tsao, "Identification of Growth Promoting Effect of rBCG/BCG Culture Supernatant and Its Potential Applications," World Journal of Vaccines, Vol. 3 No. 2, 2013, pp. 32-38. doi: 10.4236/wjv.2013.32006.

References

[1] World Health Organization, “Global Tuberculosis Control Report 2011,” 2011.
[2] B. R. Bloom and C. J. Murray, “Tuberculosis: Commentary on a Reemergent Killer,” Science, Vol. 257, No. 5073, 1992, pp. 1055-1064. doi:10.1126/science.257.5073.1055
[3] S. M. Arend and J. T. van Dissel, “Evidence of Endogenous Reactivation of Tuberculosis after a Long Period of Latency,” The Journal of Infectious Diseases, Vol. 186, No. 6, 2002, pp. 876-877. doi:10.1086/342604
[4] J. Chan and J. Flynn, “The Immunological Aspects of Latency in Tuberculosis,” Clinical Immunology, Vol. 110, No. 1, 2004, pp. 2-12. doi:10.1016/S1521-6616(03)00210-9
[5] G. V. Mukamolova, A. S. Kaprelyants, D. I. Young, M. Young, D. B. Kell, “A Bacterial Cytokine,” Proceeding National Academy Science, Vol. 95, No. 15, 1998, pp. 8916-8921. doi:10.1073/pnas.95.15.8916
[6] G. V. Mukamolova, S. S. Kormer, D. B. Kell and A. S. Kaprelyants, “Stimulation of the Multiplication of Micrococcus Luteus by an Autocrine Growth Factor,” Archives of Microbiology, Vol. 172, No. 1, 1999, pp. 9-14. doi:10.1007/s002030050733
[7] G. V. Mukamolova, A. G. Murzin, E. G. Salina, G. R. Demina, D. B. Kell, A. S. Kaprelyants and M. Young, “Muralytic Activity of Micrococcus Luteus Rpf and Its Relationship to Physiological Activity in Promoting Bacterial Growth and Resuscitation,” Molecular Microbiology, Vol. 59, No. 1, 2006, pp. 84-98. doi:10.1111/j.1365-2958.2005.04930.x
[8] G. V. Mukamolova, O. A. Turapov, K. Kazarian, M. Telkov, A. S. Kaprelyants, D. B. Kell and M. Young, “The Rpf Gene of Micrococcus Luteus Encodes an Essential Secreted Growth Factor,” Molecular Microbiology, Vol. 46, No. 3, 2002, pp. 611-621. doi:10.1046/j.1365-2958.2002.03183.x
[9] G. V. Mukamolova, N. D. Yanopolskaya, D. B. Kell and A. S. Kaprelyants, “On Resuscitation from the Dormant State of Micrococcus Luteus,” Antonie Van Leeuwenhoek, Vol. 73, No. 3, 1998, pp. 237-243. doi:10.1023/A:1000881918216
[10] S. Biketov, G. V. Mukamolova, V. Potapov, E. Gilenkov, G. Vostroknutova, D. B. Kell, M. Young and A. S. Kaprelyants, “Culturability of Mycobacterium tuberculosis Cells Isolated from Murine Macrophages: A Bacterial Growth Factor Promotes Recovery,” FEMS Immunology Medical Microbiology, Vol. 29, No. 4, 2000, pp. 233-240. doi:10.1111/j.1574-695X.2000.tb01528.x
[11] G. V. Mukamolova, O. A. Turapov, D. I. Young, A. S. Kaprelyants, D. B. Kell and M. Young, “A Family of Autocrine Growth Factors in Mycobacterium Tuberculosis,” Molecular Microbiology, Vol. 46, No. 3, 2002, pp. 623-635. doi:10.1046/j.1365-2958.2002.03184.x
[12] V. V. Yeremeev, T. K. Kondratieva, E. I. Rubakova, S. N. Petrovskaya, K. A. Kazarian, M. V. Telkov, S. F. Biketov, A. S. Kaprelyants and A. S. Apt, “Proteins of the Rpf Family: Immune Cell Reactivity and Vaccination Efficacy against Tuberculosis in Mice,” Infection and Immunity, Vol. 71, No. 8, 2003, pp. 4789-4794. doi:10.1128/IAI.71.8.4789-4794.2003
[13] R. K. Gupta, B. S. Srivastava and R. Srivastava, “Comparative Expression Analysis of Rpf-Like Genes of Mycobacterium tuberculosis H37Rv Under Different Physiological Stress and Growth Conditions,” Microbiology, Vol. 156, No. Pt9, 2010, pp. 2714-2722. doi:10.1099/mic.0.037622-0
[14] J. M. Tufariello, W. R. Jacobs Jr. and J. Chan, “Individual Mycobacterium tuberculosis Resuscitation-Promoting Factor Homologues are Dispensable for Growth in Vitro and in Vivo,” Infection and Immunity, Vol. 72, No. 1, 2004, pp. 515-526. doi:10.1128/IAI.72.1.515-526.2004
[15] J. M. Tufariello, K. Mi, J. Xu, Y. C. Manabe, A. K. Kesavan, J. Drumm, K. Tanaka, W. R. Jacobs Jr. and J. Chan, “Deletion of the Mycobacterium tuberculosis Resuscitation-Promoting Factor Rv1009 Gene Results in Delayed Reactivation from Chronic Tuberculosis,” Infection and Immunity, Vol. 74, No. 5, 2006, pp. 2985-2995.
[16] H. J. Jin, L. Nguyen, T. Qu and E. Tsao, “Improved Formulation and Lyophilization Cycle for rBCG Vaccine,” Vaccine, 2011, Vol. 29, No. 29-30, pp. 4848-4852. doi:10.1016/j.vaccine.2011.04.056
[17] R. Sun, Y. A. Skeiky, A. Izzo, V. Dheenadhayalan, Z. Imam, E. Penn, et al., “Novel Recombinant BCG Expressing Perfringolysin O and the Over-expression of Key Immunodominant Antigen; Pre-Clinical Characterization, Safety and Protection against Challenge with Mycobacterium tuberculosis,” Vaccine, Vol. 27, No. 33, 2009, pp. 4412-4423. doi:10.1016/j.vaccine.2009.05.048
[18] M. O. Shleeva, K. Bagramyan, M. V. Telkov, G. V. Mukamolova, M. Young, D. B. Kell and A. S. Kaprelyants, “Formation and Resuscitation of ‘Non-Culturable’ Cells of Rhodococcus Rhodochrous and Mycobacterium tuberculosis in Prolonged Stationary Phase,” Microbiology, Vol. 148, No. Pt5, 2002, pp. 1581-1591. doi:10.1016/j.tube.2010.12.006
[19] M. O. Shleeva, G. V. Mukamolova, M. Young, H. D. Williams and A. S. Kaprelyants, “Formation of ‘Non-Culturable’ Cells of Mycobacterium Smegmatis in Stationary Phase in Response to Growth under Suboptimal Conditions and Their Rpf-mediated Resuscitation,” Microbiology, Vol. 150, No. Pt6, pp. 1687-1697. doi:10.1099/mic.0.26893-0
[20] M. Cohen-Gonsaud, P. Barthe, C. Bagnéris, B. Henderson, J. Ward, C. Roumestand and N. H. Keep, “The Structure of a Resuscitation-promoting Factor Domain from Mycobacterium tuberculosis Shows Homology to Lysozymes,” Nature Structural & Molecular Biology, Vol. 12, No. 3, 2005, pp. 270-273. doi:10.1038/nsmb905
[21] E. C. Hett, M. C. Chao, L. L. Deng and E. J. Rubin, “A Mycobacterial Enzyme Essential for Cell Division Synergizes with Resuscitation-Promoting Factor,” PLoS Pathogens, Vol. 4, No. 2, 2008, pp. 1-8. doi:10.1371/journal.ppat.1000001
[22] E. C. Hett, M. C. Chao, A. J. Steyn, S. M. Fortune, L. L. Deng and E. J. Rubin, “A Partner for the Resuscitation-Promoting Factors of Mycobacterium tuberculosis,” Molecular Microbiology, Vol. 66, No. 3, 2007, pp. 658-668. doi:10.1111/j.1365-2958.2007.05945.x
[23] A. Ruggiero, D. Marasco, F. Squeglia, S. Soldini, E. Pedone and C. Pedone, “Structure and Functional Regulation of RipA, a Mycobacterial Enzyme Essential for Daughter Cell Separation,” Structure, Vol. 18, No. 9, 2010, pp. 1184-1190. doi:10.1016/j.str.2010.06.007
[24] A. Ruggiero, F. Squeglia, C. Esposito, D. Marasco, E. Pedone, C. Pedone and R. Berisio, “Expression, Purification, Crystallization and Preliminary X-Ray Crystallographic Analysis of the Resuscitation Promoting Factor Interacting Protein RipA from M. tuberculosis,” Protein & Peptide Letters, Vol. 17, No. 1, 2010, pp. 70-73. doi:10.2174/092986610789909557
[25] A. Ruggiero, B. Tizzano, E. Pedone, C. Pedone, M. Wilmanns and R. Berisio, “Crystal Structure of the Resuscitation-Promoting Factor (DeltaDUF) RpfB from M. tuberculosis,” Journal of Molecular Biology, Vol. 385, No. 1, 2009, pp. 153-162. doi:10.1016/j.jmb.2008.10.042
[26] X. Wu, Y. Yang, Y. Han, J. Zhang, Y. Liang, H. Li, B. Li and L. Wang, “Effect of Recombinant Rv1009 Protein on Promoting the Growth of Mycobacterium tuberculosis,” Journal of Applied Microbiology, Vol. 205, No. 4, 2008, pp. 1121-1127. doi:10.1111/j.1365-2672.2008.03850.x
[27] E. Russell-Goldman, J. Xu, X. Wang, J. Chan and J. M. Tufariello, “A Mycobacterium tuberculosis Rpf Double-Knockout Strain Exhibits Profound Defects in Reactivation from Chronic Tuberculosis and Innate Immunity Phenotypes,” Infection and Immunity, Vol. 76, No. 9, 2008, pp. 4269-4281. doi:10.1128/IAI.01735-07
[28] K. J. Downing, J. C. Betts, D. I. Young, R. A. McAdam, F. Kelly, M. Young and V. Mizrahi, “Global Expression Profiling of Strains Harbouring Null Mutations Reveals That the Five Rpf-Like Genes of Mycobacterium tuberculosis Show Functional Redundancy,” Tuberculosis, Vol. 84, No. 3-4, 2004, pp. 167-179. doi:10.1016/j.tube.2003.12.004

  
comments powered by Disqus

Copyright © 2019 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.