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K. Shimazaki, A.M. Chan, J. Braun, K.A. Kelly, L.K. Gordon; Expression of Epithelial Membrane Protein 2 (EMP–2) Controls Chlamydia Infectivity . Invest. Ophthalmol. Vis. Sci. 2006;47(13):289.
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Epithelial membrane protein–2 (EMP2), a 4–transmembrane protein, has recently been shown to be important in chlamydial infectivity in in vitro studies using a human endometrial cell line. EMP2 is highly expressed in the eye, reproductive tract, and lung, sites that are major infectious targets of Chlamydia. In this study we investigate expression of EMP2 in a human corneal limbal cell line (HCLE) and the possible role for EMP2 in chlamydial infectivity in these cells.
The HCLE cell line (IOVS 2003; 44:2496) was used for these studies. EMP2 expression was evaluated through real time RT–PCR using EMP2–specific PCR primers. Protein expression of EMP2 was analyzed by Western blot using a rabbit anti–human EMP2 antibody. The cell line was used in 32 hour infection studies with the C.muridarum. Chlamydia inclusion bodies were identified using an antibody against Chlamydia LPS and detected by fluorescein–conjugated secondary antibody. Evans Blue was used as a counterstain to identify the total number of cells. Inclusion bodies were quantified using an Olympus Provis AX70 confocal fluorescent microscope. A total of 500–1000 cells were counted by both fluorescence and light microscopy in random fields and the percent infection was calculated. An antibody against the second extracellular loop of EMP2 or a control rabbit serum was used to determine if anti–EMP2 antibodies block infection.
EMP2 is highly expressed in the HCLE cell line by both protein and mRNA evaluation. The cells were highly susceptible to Chlamydia infection and blockade of EMP2 using a specific antibody against the 2nd extracellular loop caused both a significant reduction in the numbers and the size of the inclusions.
Similar to previous studies using a human endometrial cell line, the corneal cell line HCLE is susceptible to chlamydial infection and this susceptibility is decreased by blockade of the protein EMP2. This data supports a role for EMP2 in the pathogenesis of chlamydial infections.
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