Chlamydia pneumoniae : A link between Periodontitis and Cardiovascular Disease?

1
Chlamydia pneumoniae A link between
Periodontitis and Cardiovascular Disease?
Debra Holtham
Department of Biology, York College of
Pennsylvania
Photo taken from (www.cardiologytoday.com)

• Determine if nested Polymerase Chain Reaction
  system (nPCR) is as reliable for testing
  Chlamydia pneumonie in tissue samples as for
  dental plaque
• 2. Verify that the genotype of Chlamydia
  pneumoniae in dental plaque and atherosclerotic
  lesion are the same

Project Summary Chlamydia pnuemoniae is a
bacteria found in patients with periodontal and
cardiovascular disease. Periodontal disease
exists from bacterial infection within gums and
cardiovascular diseases range from hardened
arterial walls to build up of plaque within
heart walls. The objective is to determine
whether these two regions, oral and
cardiovascular, in fact contain the same
genotypic Chlamydia pneumoniae that is found
since we already know it exists in each region.
Methods involved are polymerase chain reaction
(PCR) to amplify the sample from each designated
area followed by electrophoresis to determine if
any common base pair regions exist between the
samples and the Chlamydia pneumoniae control.
Gene typing will determine if the genes are in
fact identical. I suspect to find both regions
to contain Chlamydia pneumoniae.
Objectives
Research Design Part 2
Hypothesis (1) confirmed
nPCR samples
DNA Sequencing
diff gene sequence
same gene sequence
Review of Literature

• Chlamydia pneumoniae has been identified in
  aortic, coronary, carotid, iliac and femoral
  atherosclerotic lesions by various methods
  including PCR, electron microscopy, microbial
  culture and immunohistochemistry.
• Antigen traces have been found specifically in
  atherosclerotic lesions using various
  immunological assays such as Enzyme-Linked
  Immunosorbent Assay (ELISA) and Direct
  Fluorescent Antibody Test (DFA) and DNA traces
  have been detected by means of Polymerase Chain
  Reaction (PCR).
• Seroepidemiological association of C. pneumoniae
  with atherosclerosis demonstrated that patients
  with coronary heart disease(CHD) and acute
  myocardial infarction(AMI) had C. pneumoniae
  antibodies more often than the control
  population. Since then, other seroepidemiological
  studies have confirmed these findings, with IgA
  antibody being used as the marker.
• Chlamydia pneumoniae, periodontal pathogens,
  cause arthrosclerosis in experimental animals and
  have been found in human atherosclerotic lesions.
  C. pneumoniae is capable of producing
  inflammatory atherosclerosis-like changing in the
  aortas of infected rabbits.
• The prevalence of cardiovascular disease (CVD)
  seems to be highest in those individuals in whom
  periodontitis coexists.
• Patients with periodontal disease are, on
  average, at twice the risk for coronary vascular
  disease including myocardial infarction.
• Indicates that periodontitis is a CVD risk
  factor for individuals who react to an infection
  with a systemic inflammatory and immune response.
• Nested PCR (nPCR) amplification with 2 different
  primers increases both sensitivity and
  specificity which is fundamental for detection of
  unique Chlamydia pneumoniae base pairs.

Confirms Hypothesis (2)
Hypothesis (2) Incorrect
Haraszthy VI., Zambon, J.J., Trevisan, M., Zeid,
M., Genco, R.J. Identification of Periodontal
Pathogens in Atheromaotus Plaques. Journal of
Periodontal. 2000 October. 1554-1560.
Expected Results

• nPCR is successful with tissue samples and
  Chlamydia pneumoniae is detected
• 2. Chlamydia pneumoniae has the same genotype in
  the oral and cardiovascular samples

Introduction
This study involves genetic methods to
discover if one possible linking factor between
Chlamydia pneumoniae infection and cardiovascular
disease is colonization from oral infection.
Chlamydia pneumoniae is a gram negative, obligate
intracellular bacterium. It lacks a peptidoglycan
layer and was recognized to cause infections in
humans as early as 1965. Interestingly, this
bacterium has been found in gingival infections
of patients with gum disease as well as in
arterial walls of patients with hardened and
clogged arteries in the heart. Periodontal
diseases (gum disease), including gingivitis and
periodontitis, are serious conditions that, if
left untreated, can lead to tooth loss, blood,
and bone infections. Periodontitis is a chronic
infection that affects the teeth by creating
pockets between the tooth, supporting bone, and
soft tissue. Cardiovascular disease (CVD) refers
to the class of diseases that involve malfunction
of the heart and/or blood vessels (arteries and
veins) which supply oxygen to all areas of the
body. While the term technically refers to any
disease that affects the cardiovascular system,,
it is usually used to refer to those related to
atherosclerosis (hardening and clogging of the
arteries), heart attack, and stroke. From this
study of Chlamydia pneumoniae from bacterial film
samples in the oral cavity and hardened and
clogged arterial walls of the arteries, a
discovery can be made whether or not these two
areas contain the same genotypic Chlamydia
pneumoniae.
Literature Cited Haraszthy VI., Zambon, J.J.,
Trevisan, M., Zeid, M., Genco, R.J.
Identification of Periodontal Pathogens in
Atheromaotus Plaques. Journal of Periodontal.
2000 October. 1554-1560. Mattila, Kimmo J.,
Pussinen, Pirkko J., Paju, Susanna. Dental
Infections and Cardiovascular Diseases A
review. Journal of Periodontal. 2005 November.
2085-2088. N. SINADA, J.K. SPITZNAGEL, and P.
BAVOIL. Detection of Chlamydia pneumoniae in
Human Dental Plaque. Abstract 2133, J. Dent.
Res. Annual Meeting Abstract Issue. March, 2006
IADR, Orlando, FL Reed, S. G., D. E. Lopatin, B.
Foxman, and B. A. Burt. 2000. Oral Chlamydia
trachomatis in patients with established
periodontitis. Clin. Oral Investig. 4226-232.
Haraszthy VI., Zambon, J.J., Trevisan, M., Zeid,
M., Genco, R.J. Identification of Periodontal
Pathogens in Atheromaotus Plaques. Journal of
Periodontal. 2000 October. 1554-1560.

• Results from this study specifically state that
  80 of the 50 endarterectomy specimens were in
  one or more PCR assays with 18 for
  C.pneumoniae

Research Design Part 1
preparation of control and experimental samples
Nested PCR of Samples
Electrophoresis
Analyze Result
diff size/ no products
same size products
Confirms Hypothesis (1)
Hypothesis (1) Incorrect
Acknowledgements A short and sweet Thank you to
Dr. Spitznagel for the stepping stone and Dr.
Thompson for the helpful hints during Senior
Thesis