Detection of <it>Chlamydia </it>in the peripheral blood cells of normal donors using <it>in vitro </it>culture, immunofluorescence microscopy and flow cytometry techniques

• Main Authors: Andrzejewski Chester, Croteau Nancy L, West Corrie, Webley Wilmore C, Cirino Frances, Stuart Elizabeth S
• Format: Article
• Language: English
• Published: BMC 2006-02-01
• Series: BMC Infectious Diseases
• Online Access: http://www.biomedcentral.com/1471-2334/6/23

<p>Abstract</p> <p>Background</p> <p><it>Chlamydia trachomatis </it>(<it>Ct</it>) and <it>Chlamydia pneumoniae </it>(<it>Cp</it>) are medically significant infectious agents associated with various chronic human pathologies. Nevertheless, specific roles in disease progression or initiation are incompletely defined. Both pathogens infect established cell lines <it>in vitro </it>and polymerase chain reaction (PCR) has detected <it>Chlamydia </it>DNA in various clinical specimens as well as in normal donor peripheral blood monocytes (PBMC). However, <it>Chlamydia </it>infection of other blood cell types, quantification of <it>Chlamydia </it>infected cells in peripheral blood and transmission of this infection <it>in vitro </it>have not been examined.</p> <p>Methods</p> <p><it>Cp </it>specific titers were assessed for sera from 459 normal human donor blood (NBD) samples. Isolated white blood cells (WBC) were assayed by <it>in vitro </it>culture to evaluate infection transmission of blood cell borne chlamydiae. Smears of fresh blood samples (FB) were dual immunostained for microscopic identification of <it>Chlamydia</it>-infected cell types and aliquots also assessed using Flow Cytometry (FC).</p> <p>Results</p> <p>ELISA demonstrated that 219 (47.7%) of the NBD samples exhibit elevated anti-<it>Cp </it>antibody titers. Imunofluorescence microscopy of smears demonstrated 113 (24.6%) of samples contained intracellular <it>Chlamydia </it>and monoclonals to specific CD markers showed that <it>in vivo </it>infection of neutrophil and eosinophil/basophil cells as well as monocytes occurs. <it>In vitro </it>culture established WBCs of 114 (24.8%) of the NBD samples harbored infectious chlamydiae, clinically a potentially source of transmission, FC demonstrated both <it>Chlamydia </it>infected and uninfected cells can be readily identified and quantified.</p> <p>Conclusion</p> <p>NBD can harbor infected neutrophils, eosinophil/basophils and monocytes. The chlamydiae are infectious <it>in vitro</it>, and both total, and cell type specific <it>Chlamydia </it>carriage is quantifiable by FC.</p>