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Chlamydia pneumoniae DNA in non-coronary atherosclerotic plaques and circulating leukocytes
M Berger1, B Schröder, G Daeschlein
1Franz Volhard Clinic and Max Delbrück Center for Molecular Medicine, Medical Faculty of the Charité, Humboldt University, Berlin, Germany.
Abstract:
Earlier studies have associated atherosclerosis with Chlamydia pneumoniae infection. C. pneumoniae may circulate via monocytes and migrate into plaques by leukocyte infiltration; however, detection is difficult. We developed a novel polymerase chain reaction (PCR) method to test the hypothesis that C. pneumoniae DNA in circulating leukocytes is correlated with C. pneumoniae DNA in plaque material and that C. pneumoniae copy number is associated with disease severity. We obtained plaques from 130 patients who underwent surgery for carotid stenosis, aneurysm, or peripheral vascular disease. From 60 patients and 51 normal control subjects we also obtained circulating leukocytes. The C. pneumoniae 16 S rRNA gene was amplified with a highly specific quantitative PCR protocol relying on the TaqMan technology. Immunohistochemistry was performed with antibody against the C. pneumoniae outer membrane protein. C. pneumoniae DNA was present in 25% of atherosclerotic plaques and 20% of circulating leukocytes from patients. The copy number was not correlated with disease severity. C. pneumoniae DNA was more common in younger patients and smokers. C. pneumoniae antibody titers, C-reactive protein, fibrinogen, leukocyte count, cholesterol, and diabetes were not associated with C. pneumoniae DNA. Although immunostaining of plaque and PCR results were highly correlated, we found no relationship between C. pneumoniae DNA in plaques and that in circulating leukocytes. Finally, 13% of normal control subjects had positive leukocytes; however, their copy number was significantly lower than that of the patients. C. pneumoniae DNA is frequent in atherosclerotic plaques and is correlated with positive immunohistochemistry. C. pneumoniae DNA may also be found in circulating leukocytes; however, infected leukocytes and plaques do not coincide. Serology is unreliable in predicting C. pneumoniae DNA. Smoking increases the risk of harboring C. pneumoniae DNA. Our results do not suggest that either test for antibodies or C. pneumoniae DNA from leukocytes in blood is of value in predicting infected plaques.
Insights
Chlamydia pneumoniae DNA is frequently found in atherosclerotic plaques, but not correlated with disease severity. Testing for this bacteria in circulating leukocytes is unreliable for predicting plaque infection.
Area of Science:
- Microbiology
- Cardiovascular Disease Research
Background:
- Atherosclerosis has been linked to Chlamydia pneumoniae infection.
- C. pneumoniae may spread via monocytes and infiltrate plaques, but detection is challenging.
Purpose of the Study:
- To investigate the correlation between C. pneumoniae DNA in circulating leukocytes and plaque material.
- To determine if C. pneumoniae copy number is associated with disease severity in patients with vascular disease.
Main Methods:
- Developed a novel quantitative polymerase chain reaction (PCR) method using TaqMan technology.
- Analyzed atherosclerotic plaques and circulating leukocytes from patients and control subjects.
- Utilized immunohistochemistry to detect C. pneumoniae outer membrane protein.
Main Results:
- C. pneumoniae DNA was detected in 25% of atherosclerotic plaques and 20% of patient leukocytes.
- No correlation was found between C. pneumoniae DNA copy number and disease severity.
- C. pneumoniae DNA was more prevalent in younger patients and smokers.
- No relationship observed between C. pneumoniae DNA in plaques and circulating leukocytes.
- C. pneumoniae DNA copy number in leukocytes was significantly lower in normal controls compared to patients.
Conclusions:
- C. pneumoniae DNA is common in atherosclerotic plaques and correlates with positive immunohistochemistry.
- While C. pneumoniae DNA can be present in circulating leukocytes, infected leukocytes and plaques do not coincide.
- Serological tests and leukocyte DNA testing are unreliable for predicting infected plaques.
- Smoking is a risk factor for harboring C. pneumoniae DNA.