Comparative Assessment of Growth and Gene Regulation between 4 Serotypes of <i>Streptococcus pneumoniae</i> in Broth and Cell Culture — Oak Academic Publishing
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Comparative Assessment of Growth and Gene Regulation between 4 Serotypes of <i>Streptococcus pneumoniae</i> in Broth and Cell Culture
Department of Biological Sciences, Tarleton State University, Stephenville, TX, USA
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Texas A&M AgriLife Research and Extension Center, Stephenville, TX, USA
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Department of Biological Sciences, Tarleton State University, Stephenville, TX, USA
1 Department of Biological Sciences, Tarleton State University, Stephenville, TX, USA
2 Texas A&M AgriLife Research and Extension Center, Stephenville, TX, USA
3 Department of Biological Sciences, Tarleton State University, Stephenville, TX, USA
Streptococcus pneumoniae is a medically important pathogen capable of causing human infections of pneumonia, bacteremia, otitis media, and meningitis. Although there are vaccinations available, infections with S. pneumoniae still remains a global problem. S. pneumoniae is a highly adaptable bacterial species with numerous serotypes based on capsular polysaccharides. The different serotypes vary in their ability to colonize and causing pathology. Here we compared the regulation of five different virulence factors from four common serotypes of S. pneumoniae that vary in their carriage, morbidity, and mortality rates in the human population using two different in vitro methods, broth and cell culture. We determine that there is variation of virulence factor gene regulation within a serotype using two different culture methods, and variation between the serotypes in the same culture condition. The regulation of genes appeared to have a correlation with the ability of the various serotypes to grow in broth culture, adhere to cultured lung cells, and invade the cultured lung cells, as serotypes that shared similar regulation of virulence factors tended to behave similarly in culture. Many studies with S. pneumoniae rely on the use of one selected serotype, but since there is a wide variation in the growth and regulatory mechanisms of these bacteria. As demonstrated here, future studies should utilize more strains in models before concluding mechanisms of pathobiology.
Frankel, R., Virata, M., Hardalo, C., Altice, F. and Friedland, G. (1996) Invasive Pneumococcal Disease: Clinical Features, Serotypes, and Antimicrobial Resistance Patterns in Cases Involving Patients with and without Human Immunodeficiency Virus Infection. Clinical Infectious Diseases, 23, 577-584. https://doi.org/10.1093/clinids/23.3.577
Alanee, S., McGee, L., Jackson, D., Chiou, C., Feldman, C., et al. (2007) Association of Serotypes of Streptococcus pneumoniae with Disease Severity and Outcome in Adults: An International Study. Clinical Infectious Diseases, 45, 46-51. https://doi.org/10.1086/518538
Hamborsky, J., Kroger, A. and Wolfe, S. (2015) Epidemiology and Prevention of Vaccine-Preventable Diseases. 13rd Edition, Centers for Disease Control and Prevention, Public Health Foundation, Washington DC.
Shiri, T., Datta, S., Madan, J., Tsertsvadze, A., Royle, P., et al. (2017) Indirect Effects of Childhood Pneumococcal Conjugate Vaccination in Invasive Pneumococcal Disease: A Systematic Review and Meta-Analysis. The Lancet, 5, e51-e59. https://doi.org/10.1016/S2214-109X(16)30306-0
O’Brien, K., Wolfson, L., Watt, J., Henkle, E., Deloria-Knoll, M., et al. (2009) Burden of Disease Caused by Streptococcus pneumoniae in Children Younger than 5 Years: Global Estimates. The Lancet, 374, 893-902. https://doi.org/10.1016/S0140-6736(09)61204-6
Henrichsen, J. (1995) Six Newly Recognized Types of Streptococcus pneumoniae. Journal of Clinical Microbiology, 33, 2759-2762.
Sjöström, K., Spindler, C., Ortqvist, A., Kalin, M., Sandgren, A., et al. (2006) Clonal and Capsular Types Decide Whether Pneumococci Will Act as a Primary or Opportunistic Pathogen. Clinical Infectious Diseases, 42, 451-459. https://doi.org/10.1086/499242
Morona, J., Morona, R. and Paton, J. (2006) Attachment of Capsular Polysaccharide to the Cell Wall of Streptococcus pneumoniae Type 2 Is Required for Invasive Disease. PNAS, 103, 8505-8510. https://doi.org/10.1073/pnas.0602148103
Rensheng, L., Mann, B., Lewis, W., Rowe, A., Heath, R., et al. (2005) Solution Structure of Choline Binding Protein A, the Major Adhesin of Streptococcus pneumoniae. The EMBO Journal, 24, 34-43. https://doi.org/10.1038/sj.emboj.7600490
Howard, L. and Gooder, H. (1974) Specificity of the Autolysin of Streptococcus (Diplococcus) pneumoniae. Journal of Bacteriology, 117, 796-804.
Hirst, R., Kadioglu, A., O’Callaghan, C. and Andrew, P. (2004) The Role of Pneumolysin in Pneumococcal Pneumonia and Meningitis. Clinical & Experimental Immunology, 138, 195-201. https://doi.org/10.1111/j.1365-2249.2004.02611.x
García-Suárez, M., Flórez, N., Astudillo, A., Vázquez, F., Villaverde, R., Fabrizio, K., et al. (2007) The Role of Pnuemolysin in Mediating Lung Damage in a Lethal Pneumococcal Pneumonia Murine Model. Respiratory Research, 8, 3.
Echlin, H., Frank, M., Iverson, A., Chang, T., Johnson, M., et al. (2016) Pyruvate Oxidase as a Critical Link between Metabolism and Capsule Biosynthesis in Streptococcus pneumoniae. PLoS Pathology, 12, e1005951. https://doi.org/10.1371/journal.ppat.1005951
Carvalho, S., Andisi, V., Gradstedt, H., Neef, J., Kuipers, O., et al. (2013) Pyruvate Oxidase Influences the Sugar Utilization Pattern and Capsule Production in Streptococcus pneumoniae. PLoS ONE, 8, e68277. https://doi.org/10.1371/journal.pone.0068277
Livak, K.J. and Schmittgen, T.D. (2001) Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25, 402-408.
Matsumura, Y., Sakai, H., Ban, N. and Inagaki, N. (2007) ABECA3-Mediated Choline Phospholipids Uptake into Intracellular Vesicles in A549 Cells. FEBS Letters, 581, 3139-3144. https://doi.org/10.1016/j.febslet.2007.05.078
Binsker, U., Kohler, T.P., Krauel, K., Kohler, S., Habermeyer, J., et al. (2017) Serotype 3 Pneumococci Sequester Platelet-Derived Human Thrombospondin-1 via the Adhesin and Immune Evasion Protein Hic. The Journal of Biological Chemistry, 292, 5770-5783. https://doi.org/10.1074/jbc.M116.760504
Gen, K.A., Gilbert, G.L., Song, J.Y., Skovsted, I.C., Klungman, K.P., et al. (2015) Pneumococcal Capsules and Their Types: Past, Present, and Future. Clinical Microbiology Reviews, 28, 871-899. https://doi.org/10.1128/CMR.00024-15
Weingerger, D.M., Trzcinski, K., Lu, Y.-J., Bogaert, D., Brandes, A., et al. (2009) Pneumococcal Capsular Polysaccharide Structure Predicts Serotype Prevalence. PLoS Pathogens, 5, e1000479.
Peterson, S., Cline, R.T., Tettelin, H., Sharov, V. and Morrison, D.A. (2000) Gene Expression Analysis of the Streptococcus pneumoniae Competence Regulons by Use of DNA Microarrays. Journal of Bacteriology, 182, 6192-6202. https://doi.org/10.1128/JB.182.21.6192-6202.2000
Hoover, S.E., Perez, A.J., Tsui, H.-C., Sinha, D., Smiley, D.L., et al. (2015) A New Quorum-Sensing System (TprA/PhrA) for Streptococcus pneumoniae D39 That Regulates a Lantibiotic Biosynthesis Gene Cluster. Molecular Microbiology, 97, 229-243. https://doi.org/10.1111/mmi.13029
Sorg, R.A., Kuipers, O.P. and Veening, J.W. (2014) Gene Expression Platform for Synthetic Biology in the Human Pathogen Streptococcus pneumoniae. ACS Synthetic Biology, 4, 228-239. https://doi.org/10.1021/sb500229s
Lanie, J.A., Ng, W.L., Kazmierczak, K.M., Andrzejewski, T.M., Davidsen, T.M., et al. (2007) Genome Sequence of Avery’s Virulent Serotype 2 Strain D39 of Streptococcus pneumoniae and Comparison with That of Unencapsulated Laboratory Strain R6. Journal of Bacteriology, 189, 38-51. https://doi.org/10.1128/JB.01148-06
Kalin, M. (1998) Pneumococcal Serotypes and Their Clinical Relevance. Thorax, 53, 159-162. https://doi.org/10.1136/thx.53.3.159
Song, J.Y., Nahm, M.H. and Moseley, M.A. (2013) Clinical Implications of Pneumococcal Serotypes: Invasive Disease Potential, Clinical Presentations, and Antibiotic Resistance. Journal of Korean Medical Science, 28, 4-15. https://doi.org/10.3346/jkms.2013.28.1.4
Hausdorff, W.P., Bryant, J., Paradiso, P.R. and Siber, G.R. (2000) Which Pneumococcal Serotypes Cause the Most Invasive Disease: Implications for Conjugate Vaccine Formulation and Use? Clinical Infectious Diseases, 30, 100-121. https://doi.org/10.1086/313608