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Clinical

Protein Antigen in Serotype k Streptococcus mutans Clinical Isolates

K. Nakano1, R. Nomura1, H. Nemoto1, J. Lapirattanakul1, N. Taniguchi1, L. Grönroos2, S. Alaluusua2,3 and T. Ooshima1,*

1 Department of Pediatric Dentistry, Osaka University Graduate School of Dentistry, 1-8 Yamada-oka, Suita, Osaka 565-0871, Japan;
2 Department of Oral and Maxillofacial Diseases, Helsinki University Central Hospital, and
3 Department of Pediatric and Preventive Dentistry, Institute of Dentistry, University of Helsinki, Helsinki, Finland

Correspondence: * corresponding author, ooshima{at}dent.osaka-u.ac.jp


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS & METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Streptococcus mutans, a major pathogen of dental caries and infective endocarditis, is classified into serotypes c, e, f, and k, with serotype k strains recently reported to be frequently detected in persons with infective endocarditis. Thus, we hypothesized that common properties associated with infective endocarditis are present in those strains. Fifty-six oral S. mutans strains, including 11 serotype k strains, were analyzed. Western blotting analysis revealed expression of the 3 types of glucosyltransferases in all strains, while expression of the approximately 190-kDa cell-surface protein (PA) was absent in 12 strains, among which the prevalence of serotype k (7/12) was significantly high. Furthermore, cellular hydrophobicity and phagocytosis susceptibility were lower in the group of serotype k strains. These results indicate that the absence of PA expression, low cellular hydrophobicity, and phagocytosis susceptibility are common bacterial properties associated with serotype k strains, which may be associated with virulence for infective endocarditis.

Key Words: Streptococcus mutans • serotype • expression • protein antigen


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS & METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Streptococcus mutans is a known pathogen of dental caries, and its surface protein antigens have been investigated, with 3 types of glucosyltransferases (GTFB/GTFC/GTFD) shown to be involved in sucrose-dependent adhesion of the bacterium to tooth surfaces (Aoki et al., 1986; Pucci et al., 1987; Hanada and Kuramitsu, 1989). In addition, the approximately 190-kDa cell-surface protein PA (Okahashi et al., 1989), also referred to as SpaP (Lee et al., 1989) and antigen I/II (Russell and Lehner, 1978), has been shown to be correlated with cellular hydrophobicity and sucrose-independent initial adhesion to tooth surfaces (Okahashi et al., 1989). The putative amino acid sequence of PA was reported to be composed of alanine-rich repeat (A-region), proline-rich repeat (P-region), and central variable (V-region) regions (Brady et al., 1991; Seifert et al., 2004).

S. mutans is sometimes isolated from the blood of persons with bacteremia or infective endocarditis, and there have been several studies regarding its virulence factors in blood. Further, PA was reported to be immunodominant toward S. mutans antigens when serum antibody responses to systemic infection by oral streptococci were analyzed in humans (Russell et al., 1992). However, the protein was also shown to be unimportant for the virulence of infective endocarditis in a rat model (Ryd et al., 1996). In contrast, we previously found that PA defects resulted in resistance to phagocytosis by human polymorphonuclear leukocytes in vitro and induction of a longer duration of bacteremia in a rat model (Nakano et al., 2006).

S. mutans is serologically classified into 4 groups (c/e/f/k), based on the serotype-specific rhamnose-glucose polysaccharides, which have a backbone of rhamnose polymers and a side-chain of glucose polymers (Linzer et al., 1986; Nakano et al., 2004a). As for oral S. mutans strains, approximately 70–80% can be classified as serotype c, followed by e (approximately 20%), while the distribution frequencies of serotypes fand k are extremely low (each less than 5%) (Nakano et al., 2004b). Recently, serotype k was reported to be frequently identified in extirpated heart valve specimens from persons with infective endocarditis (Nakano et al., 2007b). In the present study, we analyzed the expression of the major surface proteins GTFs and PA, as well as the biological properties of serotype k strains, and compared them to other serotypes.


   MATERIALS & METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
S. mutans Strains
Eleven serotype k oral strains of S. mutans (NN2011, NN2105, NN2111, NN2193-1 NN2323M-1, OR22P1, LJ23, SA31, SA53, SA72, and SA98) were analyzed. In addition, 45 oral strains composed of serotypes c, e, and f (15 strains each) were selected from our laboratory stock strains, and used in the present study. The NN and LJ series of strains were isolated from Japanese persons in 2002 and 2006, respectively, while the SA series was isolated from Finnish persons in the early 1990s (Nakano et al., 2004a,b, 2007a). Strains MT8148 (c) (Ooshima et al., 1983) and UA159 (c) (Ajdic et al., 2002) were used as reference strains. The study procedure was approved by the Ethics Committee of Osaka University Graduate School of Dentistry, and informed consent was obtained from each person.

GTF and PA Expression in Clinical Isolates
The detection of GTFB, GTFC, GTFD, and PA expression was carried out by Western blotting analysis, as described previously (Nakano et al., 2005). The protein amount was quantified with Bio-Rad Protein Assay (Bio-Rad, Helcules, CA, USA), and an equal amount of each protein (1 µg) was analyzed. Protein expression of each test strain was compared with that of MT8148 or UA159.

Nucleotide Alignment of the Genes Encoding PA
The nucleotide alignment of the genes encoding PA in all of the strains with an absence of PA expression and of those with a reaction similar to MT8148 or UA159 (4 strains per serotype) was determined as follows. The 3 fragments of the genes were amplified with the following sets of primers: PA1F, 5'-TTT GTG CTT TAG AAT TAA TGT TGG-3' and PA1R, 5'-GAC AAT TTT AGA AAT CTT TTT ACC-3'; PA2F, 5'-GGG ATC GGT ACT TTT AGA GCG CGG-3' and PA2R, 5'-CAC TGT TGG ATA AAT CGT TGC CAC-3'; and PA3F, 5'-AAC TAA TAC AGT CAC CTT CAA GGC-3' and PA3R, 5'-GCT CAA TCT GTG ATT TAT CGC TTC-3', based on the pac sequence of MT8148 and UA159 (Appendix Fig. A). The complete sequences of the genes were determined by dye termination reaction with a DNA sequencing system (ABI PRISM 310 Genetic Analyzer; Applied Biosystems, Foster City, CA, USA) and a BigDye terminator cycle sequencing kit. Each complete sequence was registered in the GenBank database, and the accession numbers of each strain are listed in the TableGo. The putative amino acid sequence of each tested strain was compared with that of UA159 in the Oral Pathogen Sequence Databases (http://www.oralgen.lanl.gov/).


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  		Table. Summary of Findings Regarding the Putative Amino Acid Sequences of PA
 
mRNA Expression of the Gene Encoding PA
To confirm transcription of the gene encoding PA in strains with an absence of PA, we used a reverse transcriptase-mediated PCR (RT-PCR) method as described previously, with some modifications (Nomura et al., 2005). In addition, we amplified 16S rRNA to confirm that cDNA was successfully extracted by the primers constructed in a previous study (Matsumoto-Nakano and Kuramitsu, 2006). The assay was carried out 3 times for each strain.

Biological Properties
Cellular hydrophobicity and phagocytosis susceptibility by human polymorphonuclear leukocytes of all of the tested strains were analyzed by methods described by Rosenberg et al.(1980) and Nakano et al.(2004a), respectively.

Statistical Analysis
Statistical analyses were carried out with the computational software packages StatView 5.0 (SAS Institute Inc., Cary, NC, USA) and Prism 4 (GraphPad Software Inc., San Diego, CA, USA). We used Fisher’s protected least-significant-difference test to compare the occurrence of strains with an absence of PA expression within each serotype. Intergroup differences of various factors were estimated by a statistical analysis of variance (ANOVA) for factorial models. The group of strains with PA expression similar to the reference strains and those with an absence of PA expression were compared by Student’s t test. We conducted regression analysis to analyze the relationship between cellular hydrophobicity and phagocytosis rate.


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
GTF and PA Expression
The expression of GTFs and PA in representative strains from each serotype by Western blotting analyses is illustrated in Fig. 1Go. All of the tested strains expressed GTFB/C/D in a manner similar to the reference strains of MT8148 and UA159. In contrast, no visible bands for PA expression were identified in 12 of the 56 strains, of which 7 strains belonged to serotype k. In addition, there were no visible bands in 1 serotype c and 4 serotype f strains. Thus, the prevalence of strains with PA expression similar to the reference strains in the serotype k group (36.4%) was significantly lower than that in the serotype c and e groups (P < 0.01).


Figure 1
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  		Figure 1. GTF and PA expression in S. mutans oral isolates. Test strains were grown overnight, then washed and adjusted to OD550 = 1.0 with phosphate-buffered saline. Whole bacterial cells or the culture supernatants concentrated by ammonium sulfate precipitation (50%) were then dissolved with SDS gel loading buffer. An equal amount of each protein (1 µg) was separated by 10% SDS polyacrylamide gel electrophoresis and then transferred onto a polyvinylidene difluoride membrane. The transferred protein bands were reacted with rabbit antibodies against cell-associated GTF for detection of GTFB and GTFC, those against cell-free GTF for GTFD detection, and those against PA for PA detection. Finally, the bands were visualized with an alkaline-phosphatase-conjugated antirabbit immunoglobulin G antibody and 5-bromo-4chloro-3-indolylphosphate-nitroblue tetrazolium substrate.

 
Molecular Characterization of Genes Encoding PA
We amplified the genes encoding PA in all 12 strains with an absence of PA expression and compared the results with those of 16 strains (4 strains of each serotype) that had similar reactions to the reference strains in the Western blotting assays for PA. PCR amplification of the genes encoding PA using the primers designed for this study successfully produced the 3 fragments, and the complete sequence of all tested strains was determined. Next, the amino acid sequence was estimated based on the nucleotide alignment of each strain. The total number of putative amino acids of the strains with PA expression similar to the reference strains ranged from 1562 to 1566, which indicated a high homology (95–99% identity and 97–99% similarity) to UA159 (TableGo).

As for the strains with an absence of PA expression, the 5-nucleotide deletion identified in SA31 (k ) and SA72 (k) (position 737–741 of UA159) resulted in a frameshift and appeared at the stop codon (codon 252) in the A-region (Appendix Fig. B). In addition, the stop codon was identified in the V-region (codon 654) in the putative PA of NN2193-1 (k), in which a deletion of 82 amino acids was identified in the A-region (Appendix Fig. C). The total number of putative amino acid sequences of the other 9 strains with an absence of PA expression ranged from 1560 to 1566, which showed a high homology (97–99% identity and 98–99% similarity) to UA159 (TableGo). The differences in putative amino acid sequences were mostly identified in the V-region, while there were no strains with an alteration of the anchor region (LPNTG motif). As for mRNA expression, representative results of RT-PCR analyses are shown in Fig. 2Go. mRNA expression of the gene encoding PA in all of the strains with an absence of PA expression (1 serotype c, 4 serotype f , and 7 serotype k strains) was shown to be extremely weak, while all of the strains with expression similar to that of MT8148 or UA159 showed a strong expression of the gene encoding PA.


Figure 2
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  		Figure 2. Detection of transcription of genes encoding PA by RT-PCR. Total RNA was prepared from the tested strains, then treated for 30 min at 37°C with 0.1 U of RQ1 RNase-Free DNase per µL, to remove contaminating DNA, after which amplification of cDNA synthesized from mRNA was carried out. Successive PCR assays were performed under the following conditions: 30 cycles at 94°C for 30 sec, 50°C for 30 sec, and 72°C for 30 sec, using the following primers: PAtF, 5'-TAG TAA AAC ACT GTG TGG TGC TGT-3', and PAtR, 5'-CCA GCT TGG TTT GAC TTT GTT CAG-3' (Appendix Fig. A). Amplification of 16S rRNA was performed using the following sets of primers: 5'-GTG GGA CGC AAG GAA ACA CAC TGT GC-3', and 5'-CGT CGC CTT GGT AAG CTC TTA CCT TAC C-3' to confirm that cDNA was successfully extracted.

 
Correlation to Biological Properties
The serotype k group showed the lowest hydrophobicity rate, which was significantly lower than that of the serotype c group (P < 0.001) (Fig. 3AGo), while the hydrophobicity rates for the serotype e and f groups were also lower than that for the serotype c group (P < 0.05 and P < 0.001, respectively). In addition, the group of strains with an absence of PA expression was shown to be significantly less hydrophobic than those with expression similar to the reference strains (P < 0.001) (Fig. 3BGo).


Figure 3
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  		Figure 3. Biological properties of clinical isolates of S. mutans. Cellular hydrophobicity rates are shown for each serotype (A) and based on the presence or absence of PA expression (B). Phagocytosis rates for each serotype are shown (C) and based on the presence or absence of PA expression (D). The phagocytosis rate was calculated as follows. Tested strains were grown overnight, then washed and adjusted with PBS to 1.0 x 108 CFU/mL. Next, a 500-µL quantity of each suspension was mixed with human peripheral blood (500 µL) and incubated for 10 min at 37°C. Interactions between polymorphonuclear leukocytes (PMNs) and the bacteria were observed under a microscope following Giemsa staining. The phagocytosis rate is expressed as mean ratio of PMNs with bacterial phagocytosis of per 100 PMNs, with 500 PMNs examined. PA+ indicates strains with PA expression similar to the reference strains, and PA- indicates those with an absence of PA expression as compared with the reference strains. Statistical analysis was carried out by ANOVA (A,C) or a t test (B,D) (*P < 0.05, ***P < 0.001).

 
As for phagocytosis susceptibility, the serotype f and k strains exhibited significantly lower phagocytosis rates than the serotype c and e strains (P < 0.001) (Fig. 3CGo). In addition, the group of strains with an absence of PA expression was shown to be less susceptible to phagocytosis than those with expression similar to the reference strains (P < 0.001) (Fig. 3DGo). Regression analysis also revealed that phagocytosis rate and cellular hydrophobicity were positively correlated (P < 0.0001, R = 0.53) (data not shown).

There were 1 serotype c and 4 serotype f strains with an absence of PA expression, all of which were shown to have lower cellular hydrophobicity values and phagocytosis rates than the average for each serotype group. In contrast, 4 strains with expression similar to the reference strains had higher values of cellular hydrophobicity than the average for the serotype k strains. As for phagocytosis rate, 2 of those strains were higher than the average for the serotype k strains, while strain NN2011 was similar to the average and strain LJ23 was lower.

As for the 3 strains with non-functional PA (SA31, SA72, and NN2193-1), their values of cellular hydrophobicity were lowest among all of the strains analyzed in the present study. In contrast, their phagocytosis rates were shown to be higher than the average for the serotype k strains, though significantly lower than the strains with PA expression similar to the reference strains.


   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
It is well-known that PA is one of the most important surface proteins for the virulence of S. mutans. However, there are no known reports describing the clinical prevalence of strains with an absence of PA expression. In the present study, none of the tested strains demonstrated an attenuated expression of GTFs, but PA expression could not be identified in 12 strains (21.4%), among which serotype k strains were the most prevalent. We considered that the actual rates of strains present in the oral cavity with an absence of PA should be standardized by calculating the frequency in combination with the prevalence of each serotype. Since the prevalence of c, e, f, and k strains in the oral cavity has been reported to be 78%, 17%, 3% and 2%, respectively (Nakano et al., 2004a), the estimated clinical rate of strains with an absence of PA in the oral cavity was calculated to be 7.2%.

Twelve strains with an absence of PA expression were investigated in this study. The putative amino acid sequences of 9 of those strains were shown to have a high homology with the reference strains. However, RT-PCR analysis revealed that the absence of PA expression in these strains was derived from a lack of mRNA expression of the gene encoding PA. In contrast, those of the other 3 strains represented non-functional proteins, and all belonged to the serotype k. It has been shown that a frameshift mutation of PA in strain GS-5 resulted in premature termination and loss of anchoring (Murakami et al., 1997). The reason why PA expression was found to be absent in these 3 strains is because of the occurrence of premature termination and loss of PA anchoring.

Cellular hydrophobicity is considered to be mainly influenced by cell-surface PA expression (Okahashi et al., 1989). In this study, cellular hydrophobicity of the 12 strains with an absence of PA expression was significantly lower than that of strains with positive PA expression (P < 0.001). Bacterial hydrophobicity is known to be associated with the interaction between bacteria and phagocytotic cells, and subsequent phagocytosis (Tsuda et al., 2000). Our results clearly showed that the strains lacking PA expression were less susceptible to phagocytosis (P < 0.001). In addition, regression analysis revealed that the rates of cellular hydrophobicity and phagocytosis were positively correlated (P < 0.0001). Thus, strains with an absence of PA could survive in blood for a longer duration, which might be a crucial factor contributing to their virulence in blood.

In the present study, serotype k strains had the highest rate of an absence of PA expression as compared with the other serotypes. Furthermore, based on our comparison of cellular hydrophobicity and phagocytosis susceptibility in each serotype group, the serotype k strains were less hydrophobic and not as susceptible to phagocytosis as compared with serotype c (P < 0.001). Considering the prevalence of serotype k strains in the oral cavity, only a limited number of strains present in the oral cavity possess these properties. Nevertheless, it is important to note that detection of serotype k strains of S. mutans is a possible marker for the presence of virulent strains in blood.


   ACKNOWLEDGMENTS
 
This study was supported by the 21st Century COE program entitled "Origination of Frontier BioDentistry" at Osaka University Graduate School of Dentistry, supported by the Ministry of Education, Culture, Sports, Science and Technology of Japan, by Grants-in-Aid for Scientific Research (A) 19209063 and (B) 16390605 from the Japan Society for Promotion of Science, and by Grants-in-Aid for Young Scientists (A) 18689050 and (B) 19791572 from the Ministry of Education, Culture, Sports, Science and Technology of Japan.


   FOOTNOTES
 
A supplemental appendix to this article is published electronically only at http://jdr.iadrjournals.org/cgi/content/full/87/10/964/DC1.

Received for publication November 1, 2007. Revision received May 30, 2008. Accepted for publication July 5, 2008.


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

  • Ajdic D, McShan WM, McLaughlin RE, Savic G, Chang J, Carson MB, et al. (2002). Genome sequence of Streptococcus mutans UA159, a cariogenic dental pathogen. Proc Natl Acad Sci USA 99:14434–14439.[Abstract/Free Full Text]
  • Aoki H, Shiroza T, Hayakawa M, Sato S, Kuramitsu HK (1986). Cloning of a Streptococcus mutans glucosyltransferase gene coding for insoluble glucan synthesis. Infect Immun 53:587–594.[Abstract/Free Full Text]
  • Brady LJ, Crowley PJ, Ma JK, Kelly C, Lee SF, Lehner T, et al. (1991). Restriction fragment length polymorphisms and sequence variation within the spaP gene of Streptococcus mutans serotype c isolates. Infect Immun 59:1803–1810.[Abstract/Free Full Text]
  • Hanada N, Kuramitsu HK (1989). Isolation and characterization of the Streptococcus mutans gtfD gene, coding for primer-dependent soluble glucan synthesis. Infect Immun 57:2079–2085.[Abstract/Free Full Text]
  • Lee SF, Progulske-Fox A, Erdos GW, Piacentini DA, Ayakawa GY. Crowley PJ, et al. (1989). Construction and characterization of isogenic mutants of Streptococcus mutans deficient in major surface protein antigen P1 (I/II). Infect Immun 57:3306–3313.[Abstract/Free Full Text]
  • Linzer R, Reddy MS, Levine MJ (1986). Immunochemical aspects of serotype carbohydrate antigens of Streptococcus mutans, In: Molecular microbiology and immunology of Streptococcus mutans. Hamada S, Michalek SM, Kiyono H, Manaker L, McGhee JR, editors. Amsterdam: Elsevier, pp. 29–38.
  • Matsumoto-Nakano M, Kuramitsu HK (2006). Role of bacteriocin immunity proteins in the antimicrobial sensitivity of Streptococcus mutans. J Bacteriol 188:8095–8102.[Abstract/Free Full Text]
  • Murakami Y, Nakano Y, Yamashita Y, Koga T (1997). Identification of a frameshift mutation resulting in premature termination and loss of cell wall anchoring of the PAc antigen of Streptococcus mutans GS-5. Infect Immun 65:794–797.[Abstract]
  • Nakano K, Nomura R, Nakagawa I, Hamada S, Ooshima T (2004a). Demonstration of Streptococcus mutans with a cell wall polysaccharide specific to a new serotype, k, in the human oral cavity. J Clin Microbiol 42:198–202.[Abstract/Free Full Text]
  • Nakano K, Nomura R, Shimizu N, Nakagawa I, Hamada S, Ooshima T (2004b). Development of a PCR method for rapid identification of new Streptococcus mutans serotype k strains. J Clin Microbiol 42:4925–4930.[Abstract/Free Full Text]
  • Nakano K, Nomura R, Nakagawa I, Hamada S, Ooshima T (2005). Role of glucose side chains with serotype-specific polysaccharide in the cariogenicity of Streptococcus mutans. Caries Res 39:262–268.[Medline] [Order article via Infotrieve]
  • Nakano K, Tsuji M, Nishimura K, Nomura R, Ooshima T (2006). Contribution of cell surface protein antigen PAc of Streptococcus mutans to bacteremia. Microbes Infect 8:114–121.[CrossRef][Medline] [Order article via Infotrieve]
  • Nakano K, Lapirattanakul J, Nomura R, Nemoto H, Alaluusua S, Grönroos L, et al. (2007a). Streptococcus mutans clonal variation revealed by multilocus sequence typing. J Clin Microbiol 45:2616–2625.[Abstract/Free Full Text]
  • Nakano K, Nomura R, Nemoto H, Mukai T, Yoshioka H, Shudo Y, et al. (2007b). Detection of novel serotype k Streptococcus mutans in infective endocarditis patients. J Med Microbiol 56:1413–1415[Free Full Text]
  • Nomura R, Nakano K, Ooshima T (2005). Molecular analysis of the genes involved in the biosynthesis of serotype specific polysaccharide in the novel serotype k strains of Streptococcus mutans. Oral Microbiol Immunol 20:303–309.[CrossRef][Medline] [Order article via Infotrieve]
  • Okahashi N, Sasakawa C, Yoshikawa M, Hamada S, Koga T (1989). Cloning of a surface protein antigen gene from serotype c Streptococcus mutans. Mol Microbiol 3:221–228.[CrossRef][Medline] [Order article via Infotrieve]
  • Ooshima T, Izumitani A, Sobue S, Hamada S (1983). Cariostatic effects of palatinose on experimental dental caries in rats. Jpn J Med Sci Biol 36:219–223.[Medline] [Order article via Infotrieve]
  • Pucci MJ, Jones KR, Kuramitsu HK, Macrina FL (1987). Molecular cloning and characterization of the glucosyltransferase C gene (gtfC ) from Streptococcus mutans LM7. Infect Immun 55:2176–2182.[Abstract/Free Full Text]
  • Rosenberg M, Gutnick D, Rosenberg E (1980). Adherence of bacteria to hydrocarbon: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiol Lett 9:29–33.[CrossRef]
  • Russell MW, Lehner T (1978). Characterization of antigens extracted from cells and culture fluids of Streptococcus mutans serotype c. Arch Oral Biol 23:7–15.[CrossRef][Medline] [Order article via Infotrieve]
  • Russell MW, Wu H, White PL, Kilian M, Henrichsen J (1992). Serum antibody responses to Streptococcus mutans antigens in humans systemically infected with oral streptococci. Oral Microbiol Immunol 7:321–325.[Medline] [Order article via Infotrieve]
  • Ryd M, Schennings T, Flock M, Heimdahl A, Flock JI (1996). Streptococcus mutans major adhesion surface protein, P1 (I/II), does not contribute to attachment to valvular vegetations or to the development of endocarditis in a rat model. Arch Oral Biol 41:999–1002.[Medline] [Order article via Infotrieve]
  • Seifert TB, Bleiweis AS, Brady LJ (2004). Contribution of the alanine-rich region of Streptococcus mutans P1 to antigenicity, surface expression, and interaction with the proline-rich repeat domain. Infect Immun 72:4699–4706.[Abstract/Free Full Text]
  • Tsuda H, Yamashita Y, Toyoshima K, Yamaguchi N, Oho T, Nakano Y, et al. (2000). Role of serotype-specific polysaccharide in the resistance of Streptococcus mutans to phagocytosis by human polymorphonuclear leukocytes. Infect Immun 68:644–650.[Abstract/Free Full Text]

Journal of Dental Research, Vol. 87, No. 10, 964-968 (2008)
DOI: 10.1177/154405910808701001
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