1887

Abstract

Urogenital infection is the most common sexually transmitted bacterial infection throughout the world. While progress has been made to better understand how type strains develop and respond to environmental stress , very few studies have examined how clinical isolates behave under similar conditions. Here, we examined the development and persistence phenotypes of several clinical isolates, to determine how similar they are to each other, and the type strain D/UW-3/Cx. The type strain was shown to produce infectious progeny at a higher magnitude than each of the clinical isolates, in each of the six tested cell lines. All chlamydial strains produced the highest number of infectious progeny at 44 h post-infection in the McCoy B murine fibroblast cell line, yet showed higher levels of infectivity in the MCF-7 human epithelial cell line. The clinical isolates were shown to be more susceptible than the type strain to the effects of penicillin and iron deprivation persistence models in the MCF-7 cell line. While subtle differences between clinical isolates were observed throughout the experiments conducted, no significant differences were identified. This study reinforces the importance of examining clinical isolates when trying to relate data to clinical outcomes, as well as the importance of considering the adaptations many type strains have to being cultured .

Funding
This study was supported by the:
  • JaneS Hocking , National Health and Medical Research Centre , (Award APP1023239)
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2021-02-19
2021-03-08
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References

  1. Craig AP, Bavoil PM, Rank RG, Wilson DP. Biomathematical Modeling of Chlamydia Infection and Disease. Intracellular Pathogens I: Chlamydiales American Society of Microbiology; 2012
    [Google Scholar]
  2. Menon S, Timms P, Allan JA, Alexander K, Rombauts L et al. Human and pathogen factors associated with Chlamydia trachomatis-related infertility in women. Clin Microbiol Rev 2015; 28: 969 985 [CrossRef]
    [Google Scholar]
  3. Callan T, Debattista J, Berry B, Brown J, Woodcock S et al. A retrospective cohort study examining STI testing and perinatal records demonstrates reproductive health burden of Chlamydia and gonorrhea. Pathog Dis 2020; 78: [CrossRef]
    [Google Scholar]
  4. Reekie J, Donovan B, Guy R, Hocking JS, Kaldor JM et al. Risk of ectopic pregnancy and tubal infertility following gonorrhoea and Chlamydia infections. Clin Infect Dis. 2019
    [Google Scholar]
  5. Reekie J, Donovan B, Guy R, Hocking JS, Kaldor JM et al. Risk of pelvic inflammatory disease in relation to Chlamydia and gonorrhea testing, repeat testing, and positivity: a population-based cohort study. Clin Infect Dis 2018; 66: 437 443 [CrossRef]
    [Google Scholar]
  6. Geisler WM, Uniyal A, Lee JY, Lensing SY, Johnson S et al. Azithromycin versus doxycycline for urogenital Chlamydia trachomatis infection. N Engl J Med 2015; 373: 2512 2521 [CrossRef] [PubMed]
    [Google Scholar]
  7. Kong FYS, Tabrizi SN, Law M, Vodstrcil LA, Chen M et al. Azithromycin versus doxycycline for the treatment of genital Chlamydia infection: a meta-analysis of randomized controlled trials. Clin Infect Dis 2014; 59: 193 205 [CrossRef]
    [Google Scholar]
  8. Zhanel GG, Dueck M, Hoban DJ, Vercaigne LM, Embil JM et al. Review of macrolides and ketolides: focus on respiratory tract infections. Drugs 2001; 61: 443 498
    [Google Scholar]
  9. Carlier MB, Garcia-Luque I, Montenez JP, Tulkens PM, Piret J. Accumulation, release and subcellular localization of azithromycin in phagocytic and non-phagocytic cells in culture. Int J Tissue React 1994; 16: 211 220 [PubMed]
    [Google Scholar]
  10. Bavoil PM. What’s in a word: the use, misuse, and abuse of the word "persistence" in Chlamydia biology. Front Cell Infect Microbiol 2014; 4: 27 [CrossRef]
    [Google Scholar]
  11. Hocking JS, Vodstrcil LA, Huston WM, Timms P, Chen MY et al. A cohort study of Chlamydia trachomatis treatment failure in women: a study protocol. BMC Infect Dis 2013; 13: 379 [CrossRef]
    [Google Scholar]
  12. Weisblum B. Erythromycin resistance by ribosome modification. Antimicrob Agents Chemother 1995; 39: 577 585 [CrossRef]
    [Google Scholar]
  13. Shkarupeta MM, Lazarev VN, Akopian TA, Afrikanova TS, Govorun VM. Analysis of antibiotic resistance markers in Chlamydia trachomatis clinical isolates obtained after ineffective antibiotic therapy. Bull Exp Biol Med 2007; 143: 713 717 [CrossRef]
    [Google Scholar]
  14. Bhengraj AR, Srivastava P, Mittal A. Lack of mutation in macrolide resistance genes in Chlamydia trachomatis clinical isolates with decreased susceptibility to azithromycin. Int J Antimicrob Agents 2011; 38: 178 179 [CrossRef]
    [Google Scholar]
  15. Hong KC, Schachter J, Moncada J, Zhou Z, House J et al. Lack of macrolide resistance in Chlamydia trachomatis after mass azithromycin distributions for trachoma. Emerg Infect Dis 2009; 15: 1088 1090 [CrossRef]
    [Google Scholar]
  16. Ljubin-Sternak S, Mestrovic T, Vilibic-Cavlek T, Mlinaric-Galinovic G, Sviben M et al. In vitro susceptibility of urogenital Chlamydia trachomatis strains in a country with high azithromycin consumption rate. Folia Microbiol 2013; 58: 361 365 [CrossRef]
    [Google Scholar]
  17. Misyurina OY, Chipitsyna EV, Finashutina YP, Lazarev VN, Akopian TA et al. Mutations in a 23S rRNA gene of Chlamydia trachomatis associated with resistance to macrolides. Antimicrob Agents Chemother 2004; 48: 1347 1349 [CrossRef]
    [Google Scholar]
  18. Zhu H, Wang H-P, Jiang Y, Hou S-P, Liu Y-J et al. Mutations in 23S rRNA and ribosomal protein L4 account for resistance in Chlamydia trachomatis strains selected in vitro by macrolide passage. Andrologia 2010; 42: 274 280 [CrossRef] [PubMed]
    [Google Scholar]
  19. Binet R, Maurelli AT. Frequency of development and associated physiological cost of azithromycin resistance in Chlamydia psittaci 6BC and C. trachomatis L2. Antimicrob Agents Chemother 2007; 51: 4267 4275 [CrossRef]
    [Google Scholar]
  20. Hokynar K, Rantakokko-Jalava K, Hakanen A, Havana M, Mannonen L et al. The finnish new variant of Chlamydia trachomatis with a single nucleotide polymorphism in the 23S rRNA target escapes detection by the APTIMA Combo 2 test. Microorganisms 2019; 7: 227 [CrossRef]
    [Google Scholar]
  21. Bonner CA, Byrne GI, Jensen RA. Chlamydia exploit the mammalian tryptophan-depletion defense strategy as a counter-defensive cue to trigger a survival state of persistence. Front Cell Infect Microbiol 2014; 4: 17 [CrossRef]
    [Google Scholar]
  22. Chacko A, Barker CJ, Beagley KW, Hodson MP, Plan MR et al. Increased sensitivity to tryptophan bioavailability is a positive adaptation by the human strains of C hlamydia pneumoniae . Mol Microbiol 2014; 93: 797 813 [CrossRef]
    [Google Scholar]
  23. Beatty WL, Byrne GI, Morrison RP. Morphologic and antigenic characterization of interferon gamma-mediated persistent Chlamydia trachomatis infection in vitro . Proc Natl Acad Sci U S A 1993; 90: 3998 4002 [CrossRef]
    [Google Scholar]
  24. Harper A, Pogson CI, Jones ML, Pearce JH. Chlamydial development is adversely affected by minor changes in amino acid supply, blood plasma amino acid levels, and glucose deprivation. Infect Immun 2000; 68: 1457 1464 [CrossRef]
    [Google Scholar]
  25. Kokab A, Jennings R, Eley A, Pacey AA, Cross NA. Analysis of modulated gene expression in a model of interferon-gamma-induced persistence of Chlamydia trachomatis in HEp-2 cells. Microb Pathog 2010; 49: 217 225 [CrossRef] [PubMed]
    [Google Scholar]
  26. Matsumoto A, Manire GP. Electron microscopic observations on the effects of penicillin on the morphology of Chlamydia psittaci . J Bacteriol 1970; 101: 278 285 [CrossRef]
    [Google Scholar]
  27. Belland RJ, Nelson DE, Virok D, Crane DD, Hogan D et al. Transcriptome analysis of chlamydial growth during IFN-gamma-mediated persistence and reactivation. Proc Natl Acad Sci U S A 2003; 100: 15971 15976 [CrossRef] [PubMed]
    [Google Scholar]
  28. Wyrick PB. Chlamydia trachomatis Persistence In Vitro: An Overview. J Infect Dis 2010; 201: 88 95 [CrossRef]
    [Google Scholar]
  29. Witkin SS, Minis E, Athanasiou A, Leizer J, Linhares IM. Chlamydia trachomatis: the Persistent Pathogen. Clin Vaccine Immunol 2017; 24: e00203 00217 [CrossRef] [PubMed]
    [Google Scholar]
  30. Raulston JE. Response of Chlamydia trachomatis serovar E to iron restriction in vitro and evidence for iron-regulated chlamydial proteins. Infect Immun 1997; 65: 4539 4547 [CrossRef]
    [Google Scholar]
  31. Dill BD, Dessus-Babus S, Raulston JE. Identification of iron-responsive proteins expressed by Chlamydia trachomatis reticulate bodies during intracellular growth. Microbiology 2009; 155: 210 219 [CrossRef]
    [Google Scholar]
  32. Nairz M, Fritsche G, Brunner P, Talasz H, Hantke K et al. Interferon‐γ limits the availability of iron for intramacrophage Salmonella typhimurium . Eur J Immunol 2008; 38: 1923 1936 [CrossRef]
    [Google Scholar]
  33. Shima K, Klinger M, Solbach W, Rupp J. Activities of first-choice antimicrobials against gamma interferon-treated Chlamydia trachomatis differ in hypoxia. Antimicrob Agents Chemother 2013; 57: 2828 2830 [CrossRef]
    [Google Scholar]
  34. Reveneau N, Crane DD, Fischer E, Caldwell HD. Bactericidal activity of first-choice antibiotics against gamma interferon-induced persistent infection of human epithelial cells by Chlamydia trachomatis . Antimicrob Agents Chemother 2005; 49: 1787 1793 [CrossRef]
    [Google Scholar]
  35. Perry LL, Su H, Feilzer K, Messer R, Hughes S et al. Differential sensitivity of distinct Chlamydia trachomatis isolates to IFN-gamma-mediated inhibition. Journal of immunology 1999; 162: 3541 3548
    [Google Scholar]
  36. Kintner J, Lajoie D, Hall J, Whittimore J, Schoborg RV. Commonly prescribed β-lactam antibiotics induce C. trachomatis persistence/stress in culture at physiologically relevant concentrations. Front Cell Infect Microbiol 2014; 4: 44 [CrossRef]
    [Google Scholar]
  37. Stevens MP, Twin J, Fairley CK, Donovan B, Tan SE et al. Development and evaluation of an ompA quantitative real-time PCR assay for Chlamydia trachomatis serovar determination. J Clin Microbiol 2010; 48: 2060 2065 [CrossRef]
    [Google Scholar]
  38. Thompson CC, Carabeo RA. An optimal method of iron starvation of the obligate intracellular pathogen, Chlamydia trachomatis . Front Microbiol 2011; 2: 20 [CrossRef]
    [Google Scholar]
  39. Suchland RJ, Geisler WM, Stamm WE. Methodologies and cell lines used for antimicrobial susceptibility testing of Chlamydia spp. Antimicrob Agents Chemother 2003; 47: 636 642 [CrossRef]
    [Google Scholar]
  40. Huston WM, Swedberg JE, Harris JM, Walsh TP, Mathews SA et al. The temperature activated HtrA protease from pathogen Chlamydia trachomatis acts as both a chaperone and protease at 37 °C. FEBS Lett 2007; 581: 3382 3386 [CrossRef]
    [Google Scholar]
  41. Huston WM, Theodoropoulos C, Mathews SA, Timms P. Chlamydia trachomatis responds to heat shock, penicillin induced persistence, and IFN-gamma persistence by altering levels of the extracytoplasmic stress response protease HtrA. BMC Microbiol 2008; 8: 190 [CrossRef]
    [Google Scholar]
  42. Fudyk T, Olinger L, Stephens RS. Selection of mutant cell lines resistant to infection by Chlamydia trachomatis and Chlamydia pneumoniae . Infect Immun 2002; 70: 6444 6447 [CrossRef]
    [Google Scholar]
  43. Kägebein D, Gutjahr M, Große C, Vogel AB, Rödel J et al. Chlamydia trachomatis-infected epithelial cells and fibroblasts retain the ability to express Surface-Presented major histocompatibility complex class I molecules. Infect Immun 2014; 82: 993 1006 [CrossRef]
    [Google Scholar]
  44. Rota TR. Chlamydia trachomatis in cell culture. II. susceptibility of seven established mammalian cell types in vitro. adaptation of trachoma organisms to McCoy and BHK-21 cells. In vitro. 1977; 13: 280 292
    [Google Scholar]
  45. Croy TR, Kuo CC, Wang SP. Comparative susceptibility of eleven mammalian cell lines to infection with trachoma organisms. J Clin Microbiol 1975; 1: 434 439 [CrossRef]
    [Google Scholar]
  46. Chen JC-R, Stephens RS. Trachoma and LGV biovars of Chlamydia trachomatis share the same glycosaminoglycan-dependent mechanism for infection of eukaryotic cells. Mol Microbiol 1994; 11: 501 507 [CrossRef]
    [Google Scholar]
  47. Sompolinsky D, Richmond S. Growth of Chlamydia trachomatis in McCoy cells treated with cytochalasin B. Appl Microbiol 1974; 28: 912 914 [CrossRef]
    [Google Scholar]
  48. Ripa KT, Mardh PA. Cultivation of Chlamydia trachomatis in cycloheximide-treated mccoy cells. J Clin Microbiol 1977; 6: 328 331
    [Google Scholar]
  49. Sabet SF, Simmons J, Caldwell HD. Enhancement of Chlamydia trachomatis infectious progeny by cultivation of HeLa 229 cells treated with DEAE-dextran and cycloheximide. J Clin Microbiol 1984; 20: 217 222 [CrossRef]
    [Google Scholar]
  50. Borges V, Ferreira R, Nunes A, Sousa-Uva M, Abreu M et al. Effect of long-term laboratory propagation on Chlamydia trachomatis genome dynamics. Infection, Gen Evol 2013; 17: 23 32 [CrossRef]
    [Google Scholar]
  51. Morrison RP. Differential sensitivities of Chlamydia trachomatis strains to inhibitory effects of gamma interferon. Infect Immun 2000; 68: 6038 6040 [CrossRef]
    [Google Scholar]
  52. Wyrick PB, Knight ST. Pre-Exposure of infected human endometrial epithelial cells to penicillin in vitro renders Chlamydia trachomatis refractory to azithromycin. J Antimicrob Chemother 2004; 54: 79 85 [CrossRef]
    [Google Scholar]
  53. Lambden PR, Pickett MA, Clarke IN. The effect of penicillin on Chlamydia trachomatis DNA replication. Microbiology 2006; 152: 2573 2578 [CrossRef]
    [Google Scholar]
  54. Carrasco JA, Tan C, Rank RG, Hsia R-ching, Bavoil PM. Altered developmental expression of polymorphic membrane proteins in penicillin-stressed Chlamydia trachomatis . Cell Microbiol 2011; 13: 1014 1025 [CrossRef]
    [Google Scholar]
  55. LaRue RW, Dill BD, Giles DK, Whittimore JD, Raulston JE. Chlamydial Hsp60-2 is iron responsive in Chlamydia trachomatis serovar E-Infected human endometrial epithelial cells in vitro. Infect Immun 2007; 75: 2374 2380 [CrossRef]
    [Google Scholar]
  56. Thompson CC, Carabeo RA. An optimal method of iron starvation of the obligate intracellular pathogen, Chlamydia Trachomatis . Front Microbiol 2011; 2: 20 [CrossRef]
    [Google Scholar]
  57. Al-Younes HM, Rudel T, Brinkmann V, Szczepek AJ, Meyer TF. Low iron availability modulates the course of Chlamydia pneumoniae infection. Cell Microbiol 2001; 3: 427 437 [CrossRef]
    [Google Scholar]
  58. Pokorzynski ND, Thompson CC, Carabeo RA. Ironing out the unconventional mechanisms of iron acquisition and gene regulation in Chlamydia. Front Cell Infect Microbiol 2017; 7: 394 [CrossRef]
    [Google Scholar]
  59. Mpiga P, Ravaoarinoro M. Chlamydia trachomatis persistence: An update. Microbiol Res 2006; 161: 9 19 [CrossRef]
    [Google Scholar]
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