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Vol. 41, No. 6, 2007
Issue release date: November 2007
Free Access
Caries Res 2007;41:445–450
(DOI:10.1159/000107930)

Salivary Glucosyltransferase B as a Possible Marker for Caries Activity

Vacca Smith A.M.a · Scott-Anne K.M.b · Whelehan M.T.c · Berkowitz R.J.c · Feng C.d · Bowen W.H.b
aDepartment of Microbiology and Immunology, bCenter for Oral Biology, cEastman Department of Dentistry and dDepartment of Biostatistics and Computational Biology, University of Rochester, Rochester, N.Y., USA
email Corresponding Author

Abstract

Bacteria-derived glucosyltransferases (Gtf) (EC 2.4.1.5), through synthesizing glucan polymers from sucrose and starch hydrolysates, play an essential role in the etiology and pathogenesis of caries. We attempted to correlate the levels of Gtf in whole saliva with the prevalence of carious lesions in young children. We examined saliva from children who were either free of overt carious lesions, or had severe early childhood caries (mean dmfs = 18.72 ± 9.0 SD), for Gtf by direct enzyme assay. The levels of GtfB, GtfC and GtfD from Streptococcus mutans in the saliva using monoclonal/specific antibodies in an enzyme-linked immunosorbent assay were determined. Multiple logistic regression analyses with model selection showed that GtfB levels correlated with dmfs values of the subjects (p = 0.006). There was no correlation between total Gtf activity as measured by direct enzyme assay and dmfs values. There was a strong correlation between mutans streptococci populations in saliva and caries activity. Collectively, these data show that GtfB levels in saliva correlate strongly with presence of clinical caries and with number of carious lesions in young children. It is also possible to measure different Gtfs, separately, in whole saliva. These observations may have important clinical implications, may lead to development of a chair side caries activity test and support the importance of GtfB in the pathogenesis of dental caries.


 goto top of outline Key Words

  • Dental caries
  • Glucosyltransferase B
  • Saliva
  • Streptococcus mutans

 goto top of outline Abstract

Bacteria-derived glucosyltransferases (Gtf) (EC 2.4.1.5), through synthesizing glucan polymers from sucrose and starch hydrolysates, play an essential role in the etiology and pathogenesis of caries. We attempted to correlate the levels of Gtf in whole saliva with the prevalence of carious lesions in young children. We examined saliva from children who were either free of overt carious lesions, or had severe early childhood caries (mean dmfs = 18.72 ± 9.0 SD), for Gtf by direct enzyme assay. The levels of GtfB, GtfC and GtfD from Streptococcus mutans in the saliva using monoclonal/specific antibodies in an enzyme-linked immunosorbent assay were determined. Multiple logistic regression analyses with model selection showed that GtfB levels correlated with dmfs values of the subjects (p = 0.006). There was no correlation between total Gtf activity as measured by direct enzyme assay and dmfs values. There was a strong correlation between mutans streptococci populations in saliva and caries activity. Collectively, these data show that GtfB levels in saliva correlate strongly with presence of clinical caries and with number of carious lesions in young children. It is also possible to measure different Gtfs, separately, in whole saliva. These observations may have important clinical implications, may lead to development of a chair side caries activity test and support the importance of GtfB in the pathogenesis of dental caries.

Copyright © 2007 S. Karger AG, Basel


 goto top of outline References
  1. Alaluusua S, Renkonen OV: Streptococcus mutans establishment and dental caries in children from 2 to 4 years old. Scand J Dent Res 1983;91:453–457.
  2. Beighton D: The value of salivary bacterial counts in the prediction of caries activity; in Johnson NW (ed): Risk Markers for Oral Diseases. Dental Caries. Cambridge, Cambridge University Press, 1991, pp 313–326.
  3. Berkowitz R: Etiology of nursing caries: a microbiologic perspective. J Public Health Dent 1996;56:51–54.
  4. Bowden GH: Does assessment of microbial composition of plaque/saliva allow for diagnosis of disease activity of individuals? Community Dent Oral Epidemiol 1997;25:76–81.
  5. Clark BR, Engval E: Enzyme Immunoassay. Boca Raton, CRC Press Inc, 1980.
  6. Duchin S, van Houte J: Relationship of Streptococcus mutans and lactobacilli to incipient smooth surface dental caries in man. Arch Oral Biol 1978;23:779.
  7. Eisenberg AD, Mundorff SA, Featherstone JD, Leverett DH, Adair SM, Billings RJ, Proskin HM: Associations of microbiological factors and plaque index with caries prevalence and water fluoridation status. Oral Microbiol Immunol 1991;6:139–145.
  8. Gold OG, Jordan HV, van Houte J: A selective medium for Streptococcus mutans. Arch Oral Biol 1973;18:1357–1364.
  9. Grainger RM, Jarrett M, Honey SL: Swab test for dental caries activity: an epidemiological study. J Can Dent Assoc 1965:31:515.
  10. Grainger RM, Nikiforuk G: Determination of relative caries experience. J Can Dent Assoc 1960;26:531.
  11. Grindefjord M, Dahllof G, Nilsson B, Modeer T: Prediction of dental caries development in 1-year-old children. Caries Res 1995;29:343–348.
  12. Hamada S, Slade HD: Biology, immunology, and cariogenicity of Streptococcus mutans. Microbiol Rev 1980;44:331–384.
  13. Hanada N, Kuramitsu HK: Isolation and characterization of the gtfC gene, coding for synthesis of both soluble and insoluble glucans. Infect Immun 1988;56:1999–2005.
  14. Hanada N, Kuramitsu HK: Isolation and characterization of the Streptococcus mutans gtfD gene, coding for primer-dependent soluble glucan synthesis. Infect Immun 1989;57:2079–2085.
  15. Hausen H: Caries prediction – state of the art. Community Dent Oral Epidemiol 1997;25:87–96.
  16. Holbrook WP, de Soet JJ, de Graaff J: Prediction of dental caries in pre-school children. Caries Res 1993;27:424–430.
  17. Ivanyi J, Davies P: Monoclonal antibodies against human growth hormone. Mol Immunol 1980;17:287–290.
  18. Köhler B, Andreèn I, Jonsson B: The earlier the colonization by mutans streptococci, the higher the caries prevalence at 4 years of age. Oral Microbiol Immunol 1988;3:14–17.
  19. Köhler B, Bratthall D: Practical method to facilitate estimation of Streptococcus mutans levels in saliva. J Clin Microbiol 1979;9:584–588.
  20. Leverett DH, Featherstone JD, Proskin HM, Adair SM, Eisenberg AD, Mundorff-Shrestha SA, Shields CP, Shaffer CL, Billings RJ: Caries risk assessment by a cross-sectional discrimination model. J Dent Res 1993a;72:529–537.
  21. Leverett DH, Proskin HM, Featherstone JD, Adair SM, Eisenberg AD, Mundorff-Shrestha SA, Shields CP, Shaffer CL, Billings RJ: Caries risk assessment in a longitudinal discrimination study. J Dent Res 1993b;72:538–543.
  22. Loesche WJ: Role of Streptococcus mutans in human dental decay. Microbiol Rev 1986;50:353–380.
  23. Mattos-Graner RO, Smith DJ, King WF, Mayer MPA: Water-insoluble glucan synthesis by mutans streptococcal strains correlates with caries incidence in 12- to 30-month-old children. J Dent Res 2000;79:1371–1377.
  24. Messer LB: Assessing caries risk in children. Aust Dent J 2000;45:6–10.

    External Resources

  25. Milnes AR: Description and epidemiology of nursing caries. J Public Health Dent 1996;56:38–50.
  26. Moore S, Stein WH: Photometric ninhydrin method for use in the chromatography of amino acids. Biol Chem 1948;176:367–368.
  27. Petti S, Hausen HW: Caries prediction by multiple salivary mutans streptococcal counts in caries-free children with different levels of fluoride exposure, oral hygiene and sucrose intake. Caries Res 2000;34:380–387.
  28. Powell LV: Caries prediction: a review of the literature. Community Dent Oral Epidemiol 1998;26:361–371.
  29. Radike AW: Criteria for diagnosing dental caries; in Proceedings of the Conference on Clinical Testing of Cariostatic Agents, 1968. Chicago, American Dental Association, 1972, pp 87–88.
  30. Rogosa M, Mitchell JA, Wiseman RE: Selective medium for the isolation and enumeration of oral lactobacilli. J Dent Res 1951;30:682–684.
  31. Schilling KM, Blitzer MH, Bowen WH: Adherence of Streptococcus mutans to glucans formed in situ in salivary pellicle. J Dent Res 1989;68(spec iss):678–680.
  32. Schilling KM, Bowen WH: The activity of glucosyltransferases adsorbed onto saliva-coated hydroxyapatite. J Dent Res 1988;67:2–8.
  33. Schilling KM, Bowen WH: Glucan synthesized in situ in experimental pellicle functions as specific binding sites for Streptococcus mutans. Infect Immun 1992;60:284–295.
  34. Snyder ML: Laboratory methods in the clinical evaluation of caries activity. J Am Dent Assoc 1951;41:400.
  35. Stamm JW, Disney JA, Beck JD, Weintraub JA, Stewart PW: The University of North Carolina caries risk assessment study: final results and some alternative modeling approaches; in Bowen WH, Tabak LA (eds): Cariology for the Nineties. Rochester, University of Rochester Press, 1993, pp 209–234.
  36. Tinanoff N: Introduction to the early childhood caries conference: initial description and current understanding. Community Dent Oral Epidemiol 1998;23(suppl 1):5–7.
  37. Vacca-Smith AM, Bowen WH: In situ studies of pellicle formation on hydroxyapatite discs. Arch Oral Biol 2000;45:277–291.
  38. Vacca-Smith AM, Venkitaraman AR, Quivey RG, Bowen WH: Interactions of streptococcal glucosyltransferases with alpha-amylase and starch on the surface of saliva-coated hydroxyapatite. Arch Oral Biol 1996a;30:291–298.

    External Resources

  39. Vacca-Smith AM, Venkitaraman AR, Schilling KM, Bowen WH: Characterization of glucosyltransferase of human saliva adsorbed onto hydroxyapatite surfaces. Caries Res 1996b;30:354–360.
  40. Van Palenstein Helderman WH, Mikx FH, Van’t Hof MA, Truin G, Kalsbeek H: The value of salivary bacterial counts as a supplement to past caries experience as caries predictor in children. Eur J Oral Sci 2001;109:312–315.
  41. Venkitaraman AR, Vacca-Smith AM, Kopec LK, Bowen WH: Characterization of glucosyltransferaseB, gtfC, and gtfD in solution and on the surface of hydroxyapatite. J Dent Res 1995;74:1695–1701.
  42. Voller A, Bidwell DE, Bartlett A: The Enzyme-Linked Immunosorbent Assay (ELISA). A Guide with Abstracts of Microplate Applications. Alexandria, Dynatech Laboratories Inc, 1979.
  43. Wood JM, Critchley P: The extracellular polysaccharide produced from sucrose by cariogenic streptococcus. Arch Oral Biol 1966;11:1039–1042.
  44. Wunder D, Bowen WH: Effects of antibodies to glucosyltransferase on soluble and insolubilized enzymes. Oral Dis 2000;6:289–296.
  45. Yamashita Y, Bowen WH, Burne RA, Kuramitsu HK: Role of the Streptococcus mutans gtf genes in caries induction on the specific-pathogen-free rat model. Infect Immun 1993;61:3811–3817.

 goto top of outline Author Contacts

William H. Bowen, BDS, PhD
University of Rochester, Center for Oral Biology
601 Elmwood Avenue, Box 611
Rochester, NY 14642 (USA)
Tel. +1 585 275 0772, Fax +1 585 276 0190, E-Mail William_Bowen@urmc.rochester.edu


 goto top of outline Article Information

Received: December 8, 2006
Accepted after revision: April 11, 2007
Published online: September 7, 2007
Number of Print Pages : 6
Number of Figures : 0, Number of Tables : 2, Number of References : 45


 goto top of outline Publication Details

Caries Research

Vol. 41, No. 6, Year 2007 (Cover Date: November 2007)

Journal Editor: Shellis, R.P.(Bristol)
ISSN: 0008–6568 (print), 1421–976X (Online)

For additional information: http://www.karger.com/CRE


Copyright / Drug Dosage / Disclaimer

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

Abstract

Bacteria-derived glucosyltransferases (Gtf) (EC 2.4.1.5), through synthesizing glucan polymers from sucrose and starch hydrolysates, play an essential role in the etiology and pathogenesis of caries. We attempted to correlate the levels of Gtf in whole saliva with the prevalence of carious lesions in young children. We examined saliva from children who were either free of overt carious lesions, or had severe early childhood caries (mean dmfs = 18.72 ± 9.0 SD), for Gtf by direct enzyme assay. The levels of GtfB, GtfC and GtfD from Streptococcus mutans in the saliva using monoclonal/specific antibodies in an enzyme-linked immunosorbent assay were determined. Multiple logistic regression analyses with model selection showed that GtfB levels correlated with dmfs values of the subjects (p = 0.006). There was no correlation between total Gtf activity as measured by direct enzyme assay and dmfs values. There was a strong correlation between mutans streptococci populations in saliva and caries activity. Collectively, these data show that GtfB levels in saliva correlate strongly with presence of clinical caries and with number of carious lesions in young children. It is also possible to measure different Gtfs, separately, in whole saliva. These observations may have important clinical implications, may lead to development of a chair side caries activity test and support the importance of GtfB in the pathogenesis of dental caries.



 goto top of outline Author Contacts

William H. Bowen, BDS, PhD
University of Rochester, Center for Oral Biology
601 Elmwood Avenue, Box 611
Rochester, NY 14642 (USA)
Tel. +1 585 275 0772, Fax +1 585 276 0190, E-Mail William_Bowen@urmc.rochester.edu


 goto top of outline Article Information

Received: December 8, 2006
Accepted after revision: April 11, 2007
Published online: September 7, 2007
Number of Print Pages : 6
Number of Figures : 0, Number of Tables : 2, Number of References : 45


 goto top of outline Publication Details

Caries Research

Vol. 41, No. 6, Year 2007 (Cover Date: November 2007)

Journal Editor: Shellis, R.P.(Bristol)
ISSN: 0008–6568 (print), 1421–976X (Online)

For additional information: http://www.karger.com/CRE


Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

References

  1. Alaluusua S, Renkonen OV: Streptococcus mutans establishment and dental caries in children from 2 to 4 years old. Scand J Dent Res 1983;91:453–457.
  2. Beighton D: The value of salivary bacterial counts in the prediction of caries activity; in Johnson NW (ed): Risk Markers for Oral Diseases. Dental Caries. Cambridge, Cambridge University Press, 1991, pp 313–326.
  3. Berkowitz R: Etiology of nursing caries: a microbiologic perspective. J Public Health Dent 1996;56:51–54.
  4. Bowden GH: Does assessment of microbial composition of plaque/saliva allow for diagnosis of disease activity of individuals? Community Dent Oral Epidemiol 1997;25:76–81.
  5. Clark BR, Engval E: Enzyme Immunoassay. Boca Raton, CRC Press Inc, 1980.
  6. Duchin S, van Houte J: Relationship of Streptococcus mutans and lactobacilli to incipient smooth surface dental caries in man. Arch Oral Biol 1978;23:779.
  7. Eisenberg AD, Mundorff SA, Featherstone JD, Leverett DH, Adair SM, Billings RJ, Proskin HM: Associations of microbiological factors and plaque index with caries prevalence and water fluoridation status. Oral Microbiol Immunol 1991;6:139–145.
  8. Gold OG, Jordan HV, van Houte J: A selective medium for Streptococcus mutans. Arch Oral Biol 1973;18:1357–1364.
  9. Grainger RM, Jarrett M, Honey SL: Swab test for dental caries activity: an epidemiological study. J Can Dent Assoc 1965:31:515.
  10. Grainger RM, Nikiforuk G: Determination of relative caries experience. J Can Dent Assoc 1960;26:531.
  11. Grindefjord M, Dahllof G, Nilsson B, Modeer T: Prediction of dental caries development in 1-year-old children. Caries Res 1995;29:343–348.
  12. Hamada S, Slade HD: Biology, immunology, and cariogenicity of Streptococcus mutans. Microbiol Rev 1980;44:331–384.
  13. Hanada N, Kuramitsu HK: Isolation and characterization of the gtfC gene, coding for synthesis of both soluble and insoluble glucans. Infect Immun 1988;56:1999–2005.
  14. Hanada N, Kuramitsu HK: Isolation and characterization of the Streptococcus mutans gtfD gene, coding for primer-dependent soluble glucan synthesis. Infect Immun 1989;57:2079–2085.
  15. Hausen H: Caries prediction – state of the art. Community Dent Oral Epidemiol 1997;25:87–96.
  16. Holbrook WP, de Soet JJ, de Graaff J: Prediction of dental caries in pre-school children. Caries Res 1993;27:424–430.
  17. Ivanyi J, Davies P: Monoclonal antibodies against human growth hormone. Mol Immunol 1980;17:287–290.
  18. Köhler B, Andreèn I, Jonsson B: The earlier the colonization by mutans streptococci, the higher the caries prevalence at 4 years of age. Oral Microbiol Immunol 1988;3:14–17.
  19. Köhler B, Bratthall D: Practical method to facilitate estimation of Streptococcus mutans levels in saliva. J Clin Microbiol 1979;9:584–588.
  20. Leverett DH, Featherstone JD, Proskin HM, Adair SM, Eisenberg AD, Mundorff-Shrestha SA, Shields CP, Shaffer CL, Billings RJ: Caries risk assessment by a cross-sectional discrimination model. J Dent Res 1993a;72:529–537.
  21. Leverett DH, Proskin HM, Featherstone JD, Adair SM, Eisenberg AD, Mundorff-Shrestha SA, Shields CP, Shaffer CL, Billings RJ: Caries risk assessment in a longitudinal discrimination study. J Dent Res 1993b;72:538–543.
  22. Loesche WJ: Role of Streptococcus mutans in human dental decay. Microbiol Rev 1986;50:353–380.
  23. Mattos-Graner RO, Smith DJ, King WF, Mayer MPA: Water-insoluble glucan synthesis by mutans streptococcal strains correlates with caries incidence in 12- to 30-month-old children. J Dent Res 2000;79:1371–1377.
  24. Messer LB: Assessing caries risk in children. Aust Dent J 2000;45:6–10.

    External Resources

  25. Milnes AR: Description and epidemiology of nursing caries. J Public Health Dent 1996;56:38–50.
  26. Moore S, Stein WH: Photometric ninhydrin method for use in the chromatography of amino acids. Biol Chem 1948;176:367–368.
  27. Petti S, Hausen HW: Caries prediction by multiple salivary mutans streptococcal counts in caries-free children with different levels of fluoride exposure, oral hygiene and sucrose intake. Caries Res 2000;34:380–387.
  28. Powell LV: Caries prediction: a review of the literature. Community Dent Oral Epidemiol 1998;26:361–371.
  29. Radike AW: Criteria for diagnosing dental caries; in Proceedings of the Conference on Clinical Testing of Cariostatic Agents, 1968. Chicago, American Dental Association, 1972, pp 87–88.
  30. Rogosa M, Mitchell JA, Wiseman RE: Selective medium for the isolation and enumeration of oral lactobacilli. J Dent Res 1951;30:682–684.
  31. Schilling KM, Blitzer MH, Bowen WH: Adherence of Streptococcus mutans to glucans formed in situ in salivary pellicle. J Dent Res 1989;68(spec iss):678–680.
  32. Schilling KM, Bowen WH: The activity of glucosyltransferases adsorbed onto saliva-coated hydroxyapatite. J Dent Res 1988;67:2–8.
  33. Schilling KM, Bowen WH: Glucan synthesized in situ in experimental pellicle functions as specific binding sites for Streptococcus mutans. Infect Immun 1992;60:284–295.
  34. Snyder ML: Laboratory methods in the clinical evaluation of caries activity. J Am Dent Assoc 1951;41:400.
  35. Stamm JW, Disney JA, Beck JD, Weintraub JA, Stewart PW: The University of North Carolina caries risk assessment study: final results and some alternative modeling approaches; in Bowen WH, Tabak LA (eds): Cariology for the Nineties. Rochester, University of Rochester Press, 1993, pp 209–234.
  36. Tinanoff N: Introduction to the early childhood caries conference: initial description and current understanding. Community Dent Oral Epidemiol 1998;23(suppl 1):5–7.
  37. Vacca-Smith AM, Bowen WH: In situ studies of pellicle formation on hydroxyapatite discs. Arch Oral Biol 2000;45:277–291.
  38. Vacca-Smith AM, Venkitaraman AR, Quivey RG, Bowen WH: Interactions of streptococcal glucosyltransferases with alpha-amylase and starch on the surface of saliva-coated hydroxyapatite. Arch Oral Biol 1996a;30:291–298.

    External Resources

  39. Vacca-Smith AM, Venkitaraman AR, Schilling KM, Bowen WH: Characterization of glucosyltransferase of human saliva adsorbed onto hydroxyapatite surfaces. Caries Res 1996b;30:354–360.
  40. Van Palenstein Helderman WH, Mikx FH, Van’t Hof MA, Truin G, Kalsbeek H: The value of salivary bacterial counts as a supplement to past caries experience as caries predictor in children. Eur J Oral Sci 2001;109:312–315.
  41. Venkitaraman AR, Vacca-Smith AM, Kopec LK, Bowen WH: Characterization of glucosyltransferaseB, gtfC, and gtfD in solution and on the surface of hydroxyapatite. J Dent Res 1995;74:1695–1701.
  42. Voller A, Bidwell DE, Bartlett A: The Enzyme-Linked Immunosorbent Assay (ELISA). A Guide with Abstracts of Microplate Applications. Alexandria, Dynatech Laboratories Inc, 1979.
  43. Wood JM, Critchley P: The extracellular polysaccharide produced from sucrose by cariogenic streptococcus. Arch Oral Biol 1966;11:1039–1042.
  44. Wunder D, Bowen WH: Effects of antibodies to glucosyltransferase on soluble and insolubilized enzymes. Oral Dis 2000;6:289–296.
  45. Yamashita Y, Bowen WH, Burne RA, Kuramitsu HK: Role of the Streptococcus mutans gtf genes in caries induction on the specific-pathogen-free rat model. Infect Immun 1993;61:3811–3817.