Morphologic and molecular evaluation of Chlamydia trachomatis growth in human endocervix reveals distinct growth patterns.

Maria E Lewis,Luis Marrero,Alison J Quayle,Sheila J Greene,David J Tate,Wandy L Beatty,Lyndsey R Buckner,Robert J Belland,David H Martin,Pamela A Kozlowski,Michelle O'Brien,Arnold H Zea,Chris L Mcgowin,Rebecca A Lillis,Ashok A Aiyar,Yasser M Abdelrahman

doi:10.3389/fcimb.2014.00071

Abstract

In vitro models of Chlamydia trachomatis growth have long been studied to predict growth in vivo. Alternative or persistent growth modes in vitro have been shown to occur under the influence of numerous stressors but have not been studied in vivo. Here, we report the development of methods for sampling human infections from the endocervix in a manner that permits a multifaceted analysis of the bacteria, host and the endocervical environment. Our approach permits evaluating total bacterial load, transcriptional patterns, morphology by immunofluorescence and electron microscopy, and levels of cytokines and nutrients in the infection microenvironment. By applying this approach to two pilot patients with disparate infections, we have determined that their contrasting growth patterns correlate with strikingly distinct transcriptional biomarkers, and are associated with differences in local levels of IFNγ. Our multifaceted approach will be useful to dissect infections in the human host and be useful in identifying patients at risk for chronic disease. Importantly, the molecular and morphological analyses described here indicate that persistent growth forms can be isolated from the human endocervix when the infection microenvironment resembles the in vitro model of IFNγ-induced persistence.

Highlights

Chlamydia trachomatis is an obligate intracellular bacterium and serovars D-K are tropic for the columnar and transitional epithelial cells of the genital tract
Women aged 18–28 years and attending the Delgado STD Clinic were asked to participate and enrolled in this study if they had a high probability of chlamydial infection based on the following criteria: a recent positive NAAT screening test for C. trachomatis; recent sexual contact with a male suspected of chlamydial infection; or clinical evidence of cervicitis
IFNγ AND INDOLES IN SECRETIONS IDENTIFY POTENTIAL CORRELATES FOR DIFFERING CHLAMYDIAL GROWTH PATTERNS BETWEEN PATIENTS we examined the infecting ompA genotype, clinical history, and key immune and environmental elements that might explain the contrasting growth patterns seen in Patient 1 and Patient 2

Summary

Introduction

Chlamydia trachomatis is an obligate intracellular bacterium and serovars D-K are tropic for the columnar and transitional epithelial cells of the genital tract. Chlamydial infections in women are generally asymptomatic and often go undetected and untreated (Brunham and Rey-Ladino, 2005). Natural history studies indicate untreated infections can be asymptomatic for substantial periods of time, can spontaneously resolve or can progress to cause complications (Parks et al, 1997; Golden et al, 2000; Joyner et al, 2002; Morre et al, 2002; Molano et al, 2005; Geisler et al, 2008). If bacteria ascend into the endometrium and Fallopian tubes chronic infection can lead to pelvic inflammatory disease (PID). 11% of women with PID will subsequently develop tubal factor infertility, but, as many of these infections are clinically silent, they remain undiscovered until reproductive consequences ensue (Cohen and Brunham, 1999)

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