![[Feedback]](/icons/banner/feedbackACT.gif)
![[For Subscribers]](/icons/banner/subscriberACT.gif)
![[Archive]](/icons/banner/archiveACT.gif)
![[Search]](/icons/banner/searchACT.gif)
![[Contents]](/icons/banner/tocACT.gif)
Previous Article | Next Article 
Journal of Clinical Microbiology, March 2001, p. 1114-1117, Vol. 39, No. 3
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.3.1114-1117.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Detection of Treponema denticola in
Atherosclerotic Lesions
Katsuji
Okuda,1,2,*
Kazuyuki
Ishihara,1,2
Taneaki
Nakagawa,1,3
Akihiko
Hirayama,1
Yoshiyuki
Inayama,4 and
Kenji
Okuda5
Oral Health Science
Center1 and Departments of
Microbiology2 and
Periodontics,3 Tokyo Dental College,
Chiba, and Departments of Anatomic and Surgical
Pathology4 and
Bacteriology,5 Yokohama City University
School of Medicine, Yokohama, Japan
Received 19 September 2000/Returned for modification 16 October
2000/Accepted 18 December 2000
 |
ABSTRACT |
We examined 26 atherosclerotic lesions and 14 nondiseased aorta
specimens to detect the periodontopathogenic part of the bacterial 16S
rRNA locus by PCR. Treponema denticola sequence of the 16S rRNA locus was found in 6 out of 26 DNA samples (23.1%) from the formalin-fixed, paraffin-embeded atherosclerotic lesions obtained during surgery but not in any of the 14 nondiseased aorta samples from
deceased persons. Utilizing immunofluorescence microscopy, we observed
aggregated antigenic particles reacting with rabbit antiserum against
T. denticola in thin sections of the PCR-positive samples,
but we could not detect any reacting particles in the PCR-negative thin sections.
| |
TEXT |
Recent epidemiological studies have
established that rheological and hemostatic factors are related to
vascular diseases. These factors are potential biological effectors
which may interact with known risk factors such as hyperlipidemia,
smoking, and infections to promote vascular events. A number of studies
have suggested that infectious organisms may play a role in the
etiology and epidemiology of atherosclerosis and related diseases
(5, 6, 9, 29). For one such organism, Chlamydia
pneumoniae, there is mounting evidence associating infections with
a greater risk of atherosclerosis, myocardial infarction, and chronic
coronary heart disease (3, 20, 26). Persistent infections
by the obligate intracellular gram-negative bacteria are involved in a
wide spectrum of respiratory diseases. It is now recognized that
chronic oral infections, such as adult periodontitis, may have
important long-term sequelae (2, 7, 10, 16, 19, 21-24,
28). An inflammatory response to endothelial cell injury and
dysfunction caused by these infections may lead to atherosclerosis (26). Recently, Haraszthey et al. (11)
reported that periodontal organisms such as Actinobacillus
actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides
forsythus, and Prevotella intermedia were detected in
atheromatous plaque by PCR. In the present study, we attempted to
detect periodontopathic bacterial DNA in atherosclerotic vascular lesions.
To detect A. actinomycetemcomitans, P. gingivalis, B. forsythus,
Campylobacter rectus, and Treponema denticola, we used
the PCR and followed a double-blind protocol. The primers for detecting part of the bacterial 16S rRNA locus by PCR were synthesized in accordance with previously reported procedures (1, 30).
Primer pairs for T. denticola
(5'-TAATACCGAATGTGCTCATTTACAT-3' and
5-TCAAAGAAGCATTCCCTCTTCTTCTTA-3'), B. forsythus
(5'-GCGTATGTAACCTGCCCGCA-3' and
5'-TGCTTCAGTGTCAGTTATACCT-3'), and C. rectus
(5'-TTTCGGAGCGTAAAACTCCTTTTC-3' and
5'-TTTCTGCAAGCAGACACTTTT-3') were designed on the basis of
16S rRNA (1). Primer pairs for P. gingivalis
(5'-ATAATGGAGAACAGCAGGAA-3' and
5'-TCTTGCCAACCAGTTCCATTGC-3') and A. actinomycetemcomitans (5'-CAGCAAGCTGCACAGTTTGCAAA-3'
and 5'-CATTAGTTAATGCCGGGCCGTCT-3') were designed on
the basis of fimbriae and leukotoxin (30). Primer pairs
for C. pneumoniae (5'-TGACAACTGTAGAAATACAGC-3' and 5'-GGTTGAGRTCAACGACTTAAGG-3') were designed based
on the 16S rRNA sequence by Jantos et al. (17). The
preparation of the reaction mixture depended on the individual report.
PCRs were performed according to these reports. Briefly, reactions for
T. denticola, B. forsythus, and C. rectus
included an initial denaturation step at 95°C for 2 min; 36 cycles of
a denaturation step at 95°C for 30 s, an annealing step at
60°C for 1 min, and a final step at 72°C for 1 min, and a final
step of 72°C for 2 min. Reactions for P. gingivalis and
A. actinomycetemcomitans included 32 cycles of a
denaturation step at 95°C for 30 s, an annealing step at 55°C
for 30 s, and a final step at 72°C for 1 min. Reactions for C. pneumoniae included an initial denaturation step at 95°C for 2 min; 40 cycles of a denaturation step at 94°C for 15 s, an
annealing step at 55°C for 15 s, and a final step at 72°C for
35 s; and a final step at 72°C for 10 min. In all experiments,
negative and positive controls were used to confirm the results of the reactions. DNA samples were extracted from formalin-fixed,
paraffin-embedded blocks of atherosclerotic vascular lesions from 26 autopsy samples from individuals with vascular diseases and 14 samples
from nondiseased aorta specimens from 14 deceased persons without
vascular disease, using a DNA extraction solution (Dexpat; Takara,
Otsu, Japan) according to the manufacturer's protocol. A number of
standard precautions were taken to prevent the occurrence of spurious
results due to contamination; these included the use of dedicated
laboratory space, Dexpat reagents, and pipetting with filters for pre-
and post-PCR analysis and the testing of positive and negative control samples which included 103 T. denticola cells
per µl and 5 µl of autoclaved water, respectively, in parallel with
the test samples. In all examinations in this study, we checked for
contamination of the target bacterial species and confirmed that there
was no contamination in the negative samples.
We detected 316-bp bands of T. denticola DNA in the DNA
samples extracted from 6 of 26 sites (23.1%) in diseased
atherosclerotic aortas by 32 amplified cycles, but we could not detect
such bands in samples from the 14 nondiseased aortas. Chi-square
analysis showed that the relationship between atherosclerosis and
detection of T. denticola is not statistically significant
(P = 0.0512), but the P value is close to a
statistically significant level. The amplified DNA bands are summarized
in Fig. 1. All T. denticola DNA-positive samples were obtained from patients who had received surgical operations. The ages, sexes, and clinical diagnoses of the
patients with the positive samples in which T. denticola
sequence (fragment) of the 16S rRNA locus was detected are summarized
in Table 1. All of the six samples in
Table 1 were obtained from surgically excised tissues. We noted that
patients 1, 2, 4, 5, and 6 had periodontitis lesions. To confirm that
the amplified products were from T. denticola, the amplified
DNA products from these samples were cloned into the pCR-100 vector
using a cloning kit (Invitrogen, San Diego, Calif.) and sequenced by
the dideoxy-chain termination method (27). We confirmed
that the DNA sequences of the six amplified DNA samples were identical
to those of T. denticola ATCC 33520 strain rRNA. Homology
searches of the amplified bands were performed using the BLAST system.
The degree of identity between the amplified fragment and the sequence
deposited in the National Center for Biotechnology Information was
98%. The observed variation is minor, and in every case the sequence
with the highest score in BLAST analysis was that from T. denticola. We examined the six samples in which the T. denticola sequence of the 16S rRNA locus was detected for the
presence of a DNA band of C. pneumoniae using previously
described primers (17), but we could not find any such
band. Unexpectedly, no PCR-amplified DNA bands of A. actinomycetemcomitans, P. gingivalis, B. forsythus, or C. rectus were observed in 26 samples with atherosclerotic vascular
lesions using the primers in this study.
View larger version (66K):
[in this window]
[in a new window]
|
FIG. 1.
Amplified bands of the T. denticola 16S
rRNA. Lanes 1 to 6, DNA samples extracted from the atherosclerotic
aortas obtained from surgically excised tissue of patients with
vascular disease (Table 1); Lane P, positive control; lane M, positive
base pair markers. Strong bands are present in lanes 1 to 4, and faint
bands are present in lanes 5 and 6 at positions, corresponding to the
316-bp T. denticola 16S rRNA fragment.
|
|
View this table:
[in this window]
[in a new window]
|
TABLE 1.
Age, sex, and clinical diagnosis of patients with
atherosclerosis in whom T. denticola 16S rRNA was detected
|
|
We examined the PCR-positive samples by an immunofluorescence technique
for the presence of antigens of T. denticola. Rabbit antiserum against T. denticola was obtained by repeated
inoculation of the cells into male New Zealand White rabbits as
described in our previous papers (14, 15). Thin sections
of the paraffin block samples were incubated with rabbit antiserum
diluted 1:250 or 1:500 with phosphate-buffered saline (PBS) (pH 7.2) at
room temperature for 60 min and then washed thoroughly with PBS. The samples were then incubated with fluorescence-conjugated goat anti-rabbit immunoglobulin G (Cappel, ICN Pharmaceuticals, Inc., Aurora, Ohio) diluted 1:1,000 with PBS. The samples were observed by
immunofluorescence microscopy (Axiophot 2; Carl Zeiss, Jena, Germany).
Panels 1A, 2A, 3A, and 4A in Fig. 2 are
thin sections of atherosclerotic lesions which were stained with
hematoxylin-eosin and in which we detected T. denticola DNA,
as shown in Table 1 and Fig. 1. Panels 1B, 2B, 3B, and 4B in Fig. 2 are
the immunofluorescence photographs corresponding to panels 1A, 2A, 3A,
and 4A. The stained particles were observed in these sections. We
detected some fluorescent particles in the foam cells within the
fibrolipid lesions. The locations of the immunostained antigenic
particles were similar to those reported for C. pneumoniae
by Kuo et al. (20). However, we could not detect any
spiral cells in the thin sections stained with the sera examined. It is
possible that aggregated antigens of T. denticola were
stained with the rabbit antiserum. No distinct fluorescent particle was
observed when the sections were stained with rabbit serum absorbed with
whole T. denticola cells. No fluorescent particle was
observed in sections in which we could not detect the DNA band of 316 bp.
View larger version (96K):
[in this window]
[in a new window]
|
FIG. 2.
(1A, 2A, 3A, and 4A) Thin sections of atherosclerotic
lesions, stained with hematoxylin and eosin, in which strong T. denticola DNA bands were detected (Table 1 and Fig. 1). (1B, 2B,
3B, and 4B) Thin sections corresponding to panels 1A through 4A,
respectively, stained with rabbit antiserum against T. denticola. Clear immunofluorescent particles can be observed in
localized areas of foam cells or between small muscles in panels 1B
through 4B. Bars, 20 µm.
|
|
Oral gram-positive and gram-negative bacteria have frequently been
identified in bacteremia and may play a role in vascular diseases
(8, 12, 13). In addition, phagocytes in periodontal lesions may engulf various bacterial cells and their antigens (19). The bacterial cells and phagocytes may then
penetrate the gingival tissues, be transported via the circulation to
the heart, and adhere to the endothelium of an artery. These deposited bacteria can then stimulate the release of inflammatory cytokines and
initiate the formation of the characteristic foam cells associated with
atherosclerosis. Recently, Chiu (4) reported that oral bacteria, including Streptococcus sanguis and P. gingivalis, could be detected in vascular lesions. We demonstrated
earlier that a protease, dentilisin, of T. denticola can
degrade the intracellular matrix (14, 15). In addition to
the motility of T. denticola cells, it is possible that the
protease activity may also play a role in their penetration of
periodontal tissues by infiltrating between the gingival cells and into
the bloodstream. Recently, Peters et al. (25) showed that
oral treponemes penetrated endothelial cells. The affinity of
Treponema pallidum for vascular structures is well known
(18), but the molecular basis for this property is
unknown. We found that five of the six T. denticola-positive aorta samples were from patients with periodontitis. In order to
confirm that T. denticola can penetrate into the gingival
tissues and that the spirochetes carried by phagocytes can play a role in atherosclerosis, additional studies with humans and studies with
animal models will be needed. Moreover, the presence of oral spirochetes in some aorta lesions does not necessarily imply an etiological role for the spirochetes in atherosclerosis.
| |
ACKNOWLEDGMENTS |
We gratefully acknowledge H. Kuramitsu for his thoughtful review of
the manuscript. We thank C. Reynolds for assistance in preparing the manuscript.
This study was supported in part by a research grant from the Fund for
Comprehensive Research on Aging and Health from the Ministry of Health
and Welfare in Japan and by a grant from the Oral Health Science Center
of Tokyo Dental College.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Department of
Microbiology and Oral Health Science Center, Tokyo Dental College,
1-2-2, Masago, Mihama-ku, Chiba, Japan. Phone: 81-43-270-3741. Fax:
81-43-270-3744. E-mail: kokuda{at}tdc.ac.jp.
| |
REFERENCES |
| 1.
|
Ashimoto, A.,
C. Chen, and J. Slots.
1996.
Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesions.
Oral Microbiol. Immunol.
11:266-273[Medline].
|
| 2.
|
Beck, J.,
R. Garcia,
G. Heiss,
P. S. Vokomas, and S. Offenbacher.
1996.
Periodontal disease and cardiovascular disease.
J. Periodontol.
67:1123-1137[Medline].
|
| 3.
|
Blasi, F.,
F. Denti,
M. R. Cosentini,
R. Raccanelli,
A. Rinaldi,
L. Fagetti,
G. Esposito,
U. Ruberti, and L. Allegra.
1996.
Detection of Chlamydia pneumoniae but not Helicobacter pylori in atherosclerotic plaques of aortic aneurysms.
J. Clin. Microbiol.
34:2766-2769[Abstract].
|
| 4.
|
Chiu, B.
1999.
Multiple infection in carotid atherosclerotic plaques.
Am. Heart J.
138:S534-S536[CrossRef][Medline].
|
| 5.
|
Coles, K. A.,
A. J. Plant,
T. V. Riley, and D. W. Smith.
1998.
Cardiovascular disease: an infectious aetiology.
Med. Microbiol.
9:17-27.
|
| 6.
|
Danesh, J.,
R. Collins, and R. Peto.
1997.
Chronic infections and coronary heart disease: is there a link?
Lancet
350:430-436[CrossRef][Medline].
|
| 7.
|
DeStefano, F.,
R. F. Anda,
H. S. Kahn,
D. F. Williamson, and C. M. Russell.
1993.
Dental disease and risk of coronary heart disease and mortality.
Br. Med. J.
306:688-691.
|
| 8.
|
Dorn, B. R.,
W. A. Dunn, Jr., and A. Progulske-Fox.
1999.
Invasion of human coronary artery cells by periodontal pathogens.
Infect. Immun.
67:5792-5798[Abstract/Free Full Text].
|
| 9.
|
Ellis, R. W.
1997.
Infection and coronary heart disease.
J. Med. Microbiol.
46:535-539[Abstract].
|
| 10.
|
Genco, R. J.,
S. Chadda,
S. Grossi,
R. Dunford,
G. Taylor,
W. Knowler, and D. Pettitt.
1997.
Periodontal disease is a predictor of cardiovascular disease in a native American population.
J. Dent. Res.
76:409.
|
| 11.
|
Haraszthey, V. I.,
J. J. Zambon,
M. Trevism,
R. Shah,
M. Zeid, and R. J. Genco.
1998.
Identification of pathogens in atheromatous plaque.
J. Dent. Res.
77:666.
|
| 12.
|
Herzberg, M. C.,
K. L. Brintzenhofe, and C. C. Clawson.
1983.
Aggregation of human platelets and adhesion of Streptococcus sanguis.
Infect. Immun.
39:145-1469.
|
| 13.
|
Herzberg, M. C.,
K. Gong,
G. D. MacFarlane,
P. R. Erikson,
A. H. Sobery,
P. K. Krebsbach,
G. Manjula,
K. Schilling, and F. A. Hoff.
1990.
Phenotypic characterization of Streptococcus sanguis virulence factors associated with bacterial endocarditis.
Infect. Immun.
58:515-522[Abstract/Free Full Text].
|
| 14.
|
Ishihara, K.,
T. Miura,
H. K. Kuramitsu, and K. Okuda.
1996.
Characterization of the Treponema denticola prtP gene encoding a prolyl-phenylalanine-specific protease (dentilisin).
Infect. Immun.
64:5178-5186[Abstract].
|
| 15.
|
Ishihara, K.,
H. K. Kuramitsu,
T. Miura, and K. Okuda.
1998.
Dentilisin activity affects the organization of the outer sheath of Treponema denticola.
J. Bacteriol.
180:3837-3844[Abstract/Free Full Text].
|
| 16.
|
Isoluoma, M.,
K. Hietaniemi, and M. Jokinen.
1989.
Association between dental health and acute myocardial infarction.
Br. Med. J.
298:779-781.
|
| 17.
|
Jantos, C. A.,
R. Roggendorf,
F. N. Wuppermann, and J. H. Hegemann.
1998.
Rapid detection of Chlamydia pneumoniae by PCR-enzyme immunoassay.
J. Clin. Microbiol.
36:1890-1894[Abstract/Free Full Text].
|
| 18.
|
Jordaan, H. F.
1986.
Widespread superficial thrombophlebitis as a manifestation of secondary syphilis a new sign. A report of 2 cases.
S. Afr. Med. J.
70:493-494[Medline].
|
| 19.
|
Kinane, D. F.
1998.
Periodontal diseases' contributions to cardiovascular disease.
Ann. Periodontol.
3:142-150[Medline].
|
| 20.
|
Kuo, C. C.,
A. Shor,
L. A. Campbell,
H. Tsukushi,
D. L. Patton, and J. T. Grayton.
1993.
Demonstration of Chlamydia pneumoniae in atherosclerotic lesions of coronary arteries.
J. Infect. Dis.
167:841-849[Medline].
|
| 21.
|
Kweider, M.,
G. D. O. Lowe,
G. D. Murray,
D. F. Kinane, and D. A. Gowan.
1993.
Dental disease, fibrinogen and white cell count; links with myocardial infection?
Scott. Med. J.
38:73-74[Medline].
|
| 22.
|
Mattila, K. J.,
M. S. Nieminen,
V. V. Valtonen,
V. P. Rasi,
Y. A. Kesaniemi,
S. L. Syrjala,
P. S. Jungell,
M. Isoluoma,
K. Hietaniemi,
M. A. Jokienen, and J. K. Huttunen.
1989.
Association between dental health and acute myocardial infarction.
Br. Med. J.
298:779-782.
|
| 23.
|
Mattila, K.,
M. S. Valle,
M. S. Nieminen,
V. V. Valtonen, and K. L. Hietaniemi.
1993.
Dental infections and coronary atherosclerosis.
Atherosclerosis
103:205-211[CrossRef][Medline].
|
| 24.
|
Mattila, K. J.,
V. V. Valtonen,
V. P. Rasi,
Y. A. Kesaniemi,
S. L. Syrjala,
P. S. Jungell,
A. Progulske-Fox,
E. Kozarov,
B. Dorn,
W. Dunn, Jr.,
J. Burks, and Y. Wu.
1999.
Porphyromonas gingivalis virulence factors and invasion cells of the cardiovascular system.
J. Periodontal Res.
34:393-399[CrossRef][Medline].
|
| 25.
|
Peters, S. R.,
M. Valdez,
G. R. Riviere, and D. D. Thomas.
1999.
Adherence to and penetration through endothelial cells by oral treponemes.
Oral Microbiol. Immunol.
14:379-383[CrossRef][Medline].
|
| 26.
|
Ross, R.
1999.
Atherosclerosisan inflammatory disease.
N. Engl. J. Med.
340:115-126[Free Full Text].
|
| 27.
|
Sanger, F.,
S. Nicklen, and A. R. Coulson.
1977.
DNA sequencing with chain-terminating inhibitors.
Proc. Natl. Sci. USA
74:5463-5467[Abstract/Free Full Text].
|
| 28.
|
Syrjanen, J.,
J. Peltola,
V. V. Valtonen,
M. Iivanainen,
M. Kaste, and J. K. Huttunen.
1989.
Dental infections in association with cerebral infection in young middle aged men.
J. Intern. Med.
225:179-184[Medline].
|
| 29.
|
Valtonen, V. V.
1991.
Infection as a risk factor for infarction and atherosclerosis.
Ann. Med.
23:539-543[Medline].
|
| 30.
|
Watanabe, K., and T. O. Frommel.
1996.
Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Treponema denticola detection in oral plaque samples using the polymerase chain reaction.
J. Clin. Periodontol.
23:212-219[CrossRef][Medline].
|
Journal of Clinical Microbiology, March 2001, p. 1114-1117, Vol. 39, No. 3
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.3.1114-1117.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Hashimoto, M., Kadowaki, T., Tsukuba, T., Yamamoto, K.
(2006). Selective Proteolysis of Apolipoprotein B-100 by Arg-Gingipain Mediates Atherosclerosis Progression Accelerated by Bacterial Exposure. J Biochem
140: 713-723
[Abstract]
[Full Text]
-
Nakano, K., Inaba, H., Nomura, R., Nemoto, H., Takeda, M., Yoshioka, H., Matsue, H., Takahashi, T., Taniguchi, K., Amano, A., Ooshima, T.
(2006). Detection of Cariogenic Streptococcus mutans in Extirpated Heart Valve and Atheromatous Plaque Specimens.. J. Clin. Microbiol.
44: 3313-3317
[Abstract]
[Full Text]
-
Beck, J. D., Eke, P., Heiss, G., Madianos, P., Couper, D., Lin, D., Moss, K., Elter, J., Offenbacher, S.
(2005). Periodontal Disease and Coronary Heart Disease: A Reappraisal of the Exposure. Circulation
112: 19-24
[Abstract]
[Full Text]
-
Yoshida, A., Nagashima, S., Ansai, T., Tachibana, M., Kato, H., Watari, H., Notomi, T., Takehara, T.
(2005). Loop-Mediated Isothermal Amplification Method for Rapid Detection of the Periodontopathic Bacteria Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. J. Clin. Microbiol.
43: 2418-2424
[Abstract]
[Full Text]
-
Cavrini, F., Sambri, V., Moter, A., Servidio, D., Marangoni, A., Montebugnoli, L., Foschi, F., Prati, C., Di Bartolomeo, R., Cevenini, R.
(2005). Molecular detection of Treponema denticola and Porphyromonas gingivalis in carotid and aortic atheromatous plaques by FISH: report of two cases. J Med Microbiol
54: 93-96
[Abstract]
[Full Text]
-
Choi, J.-I., Chung, S.-W., Kang, H.-S., Rhim, B.Y., Park, Y.-M., Kim, U.-S., Kim, S.-J.
(2004). Epitope Mapping of Porphyromonas gingivalis Heat-shock Protein and Human Heat-shock Protein in Human Atherosclerosis. J. Dent. Res.
83: 936-940
[Abstract]
[Full Text]
-
Sambri, V., Marangoni, A., Cavrini, F., Leone, O., Magnani, G., Montebugnoli, L., Prati, C., Cevenini, R., Ishihara, K., Akihiro, N., Ito, R., Miyachi, K., Kuramitsu, H. K., Okuda, K.
(2004). Need for Procedural Details in Detection of Periodontopathic Bacterial DNA in the Atheromatous Plaque by PCR. J. Clin. Microbiol.
42: 4914-4915
[Full Text]
-
Ishihara, K., Nabuchi, A., Ito, R., Miyachi, K., Kuramitsu, H. K., Okuda, K.
(2004). Correlation between Detection Rates of Periodontopathic Bacterial DNA in Carotid Coronary Stenotic Artery Plaque and in Dental Plaque Samples. J. Clin. Microbiol.
42: 1313-1315
[Abstract]
[Full Text]
-
Choi, J.-I., Chung, S.-W., Kang, H.-S., Rhim, B.Y., Kim, S.-J., Kim, S.-J.
(2002). Establishment of Porphyromonas gingivalis Heat-shock-protein-specific T-cell Lines from Atherosclerosis Patients. J. Dent. Res.
81: 344-348
[Abstract]
[Full Text]
Top
Abstract
Text
