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Paul T. Rose et al Pediatric Dentistry: July/August 1994 Volume 16, Number 4 http://www.aapd.org/upload/articles/rose-16-0...
What is sIgA? <ul><li>Principal immunoglobulin </li></ul><ul><li>Humoral element in secretory immune system </li></ul><ul>...
Goal of Study <ul><li>Compare levels of IgA1 and IgA2 antibodies to saliva  </li></ul><ul><li>Quantify S.mutans in caries-...
Methods <ul><li>Random selection </li></ul><ul><li>Fluoridated communities </li></ul><ul><li>Min of 20 subjects per group ...
Saliva sampling <ul><li>One clinician collected samples to reduce variability b/w 8am-noon for unstimulated parotid and wh...
<ul><li>ELISA sandwich and direct techniques were used to determine concentrations of whole salivary total IgA, IgA1, IgA2...
 
 
Key Points <ul><li>Individual differences in flow rate and IgA antibody content for each subject were considered, there wa...
Main Fxn of IgA <ul><li>IgA functions at three anatomical levels in relation to mucosal   epithelium: 1) luminal SIgA Ab p...
Lawrence A.Tabak, DDS, PhD Pediatric Dentistry – 28:2 2006 http://www.aapd.org/upload/articles/tabak-28-2.pdf In Defense o...
The secret is in the whole <ul><li>Analyzed salivary proteins assigned them to a protein family </li></ul><ul><li>Each ind...
Current view of salivary function
Saliva in Children <ul><li>At Birth, IgA not detectable </li></ul><ul><li>4-6 wks levels rapidly rise </li></ul><ul><li>Ig...
http://www.sciencedaily.com/releases/2005/02/ &quot;The eyes may be the window to the soul, but many scientists would say ...
<ul><li>Saliva and other oral fluids contain many of the same proteins and molecules as blood and urine do. </li></ul><ul>...
Gcamerica.com Saliva testing: good practice, good sense! Saliva-Check Buffer Kit
Patient Education
Patient Education <ul><li>Carbs and acidic foods amplify S.mutans and S.sobrinus in dental plaque which are related to the...
Patient Education <ul><li>Once the salivary problem is identified the practitioner can recommend a course of preventive ca...
10 min of testing: <ul><li>Step 1: </li></ul><ul><li>Hydration </li></ul><ul><li>The rate of production of resting saliva ...
2. Consistency <ul><li>The viscosity of resting saliva as a surrogate for measuring its relative water and mucin content. ...
3. Resting saliva pH <ul><li>The pH of resting saliva </li></ul>
4. Stimulated saliva flow (5 min) <ul><li>The rate of production of stimulated saliva </li></ul>
5. Stimulated saliva pH <ul><li>Higher concentration of bicarbonate ions in parotid saliva  and should be a higher pH than...
6. Buffering capacity <ul><li>Buffering capacity of stimulated saliva in an aqueous env w/ differing amts of lactic acid. ...
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  1. 1. Paul T. Rose et al Pediatric Dentistry: July/August 1994 Volume 16, Number 4 http://www.aapd.org/upload/articles/rose-16-04.pdf IgA Antibodies to Streptococcus mutans in caries-resistant and -susceptible children
  2. 2. What is sIgA? <ul><li>Principal immunoglobulin </li></ul><ul><li>Humoral element in secretory immune system </li></ul><ul><li>Has a neutralizing component to prevent attachment and adherence of oral bacteria </li></ul><ul><li>SIgA adheres selectively to M cells in intestinal Peyer’s patches, thus mediating the transepithelial transport of the Ab molecule from the intestinal lumen to underlying gut-associated organized lymphoid tissue </li></ul>
  3. 3. Goal of Study <ul><li>Compare levels of IgA1 and IgA2 antibodies to saliva </li></ul><ul><li>Quantify S.mutans in caries-resistant (CR) and caries-susceptible (CS) children </li></ul><ul><li>Analyze sIgA and IgA1 and IgA2 subclass levels in whole and parotid using ELISA. </li></ul><ul><li>IgA1made by bone marrow B cells </li></ul><ul><li>IgA2 made by B cells in mucosa </li></ul>
  4. 4. Methods <ul><li>Random selection </li></ul><ul><li>Fluoridated communities </li></ul><ul><li>Min of 20 subjects per group </li></ul><ul><li>21 CR (DMFS less than or equal to 1) </li></ul><ul><li>20 CS (DMFS greater than 5) </li></ul><ul><ul><li>B/w 7-11 yrs of age </li></ul></ul><ul><ul><li>Negative PMH </li></ul></ul><ul><ul><li>No meds </li></ul></ul><ul><ul><li>Mixed dentition w/ 6’s, 1’s, 2’s </li></ul></ul><ul><ul><li>Consent </li></ul></ul>
  5. 5. Saliva sampling <ul><li>One clinician collected samples to reduce variability b/w 8am-noon for unstimulated parotid and whole saliva </li></ul><ul><li>Whole saliva samples </li></ul><ul><li>Parotid saliva samples </li></ul><ul><li>Unstim saliva samples were diluted 1:10 and 1:100 in sterile saline and spiral plated in duplicate on mitis salivarius (MSS) agar supplemented w/15% sucrose and bacitracin for enumeration of total oral streptococci and S.mutans. </li></ul><ul><li>S.mutans was quantified from whole saliva by colonial morphology and incubated for 3 days and the % of S. mutans per total oral streptococci was calculated </li></ul>
  6. 6. <ul><li>ELISA sandwich and direct techniques were used to determine concentrations of whole salivary total IgA, IgA1, IgA2 and parotid salivary total IgA. </li></ul><ul><li>The direct detection method originated in the 1940s when Coons and colleagues labeled antibodies with a fluorescent tag to mark tissue antigens. In this technique, a labeled primary antibody reacts directly with the antigen. </li></ul><ul><li>Results analyzed by student’s t-test </li></ul><ul><li>P values greater than 0.05 were significant </li></ul>
  7. 9. Key Points <ul><li>Individual differences in flow rate and IgA antibody content for each subject were considered, there was a significant difference b/w CR and CS individuals for IgA antibodies to S. mutans in whole saliva. </li></ul><ul><li># of salivary S.mutans higher in CS than CR children </li></ul><ul><li>IgA antibodies levels to S.mutans in whole saliva was higher in CR than CS children. </li></ul>
  8. 10. Main Fxn of IgA <ul><li>IgA functions at three anatomical levels in relation to mucosal epithelium: 1) luminal SIgA Ab prevents adhesion and entry of Ag into the epithelium; 2) IgA Ab in the lamina propria binds and excretes Ag to the lumen; and 3) IgA Ab in transit through the epithelium can inhibit virus production or neutralize proinflammatory Ags . An additional property of IgA is its inability to trigger the release of inflammatory mediators through receptors specific for its Fc domain (http://www.jimmunol.org/cgi/content/full/178/1/27) </li></ul>
  9. 11. Lawrence A.Tabak, DDS, PhD Pediatric Dentistry – 28:2 2006 http://www.aapd.org/upload/articles/tabak-28-2.pdf In Defense of the Oral Cavity: The Protective Role of the Salivary Secretions
  10. 12. The secret is in the whole <ul><li>Analyzed salivary proteins assigned them to a protein family </li></ul><ul><li>Each individualized molecule plays a role </li></ul><ul><li>Protein roles: </li></ul><ul><ul><li>Adhesins </li></ul></ul><ul><ul><li>Agglutinins </li></ul></ul><ul><ul><li>Antimicrobial </li></ul></ul><ul><ul><li>pH modulating </li></ul></ul>
  11. 13. Current view of salivary function
  12. 14. Saliva in Children <ul><li>At Birth, IgA not detectable </li></ul><ul><li>4-6 wks levels rapidly rise </li></ul><ul><li>IgA1 dominates, by 20 wks IgA2 increases to adult levels. </li></ul><ul><li>Early childhood: lysozyme and salivary peroxidase are at adult levels </li></ul><ul><li>MUC5B dominates over MUC7 from 1mo-1yr </li></ul><ul><li>Antimicrobial peptides: </li></ul><ul><ul><li>Beta-defensin-3 </li></ul></ul><ul><ul><li>Cathelicidin LL37 </li></ul></ul><ul><ul><li>Alpha-defensins 1,2,3 </li></ul></ul><ul><ul><li>HNP1-3 were greater in cavity free children </li></ul></ul>
  13. 15. http://www.sciencedaily.com/releases/2005/02/ &quot;The eyes may be the window to the soul, but many scientists would say the mouth is the window to the body&quot; –Science Daily
  14. 16. <ul><li>Saliva and other oral fluids contain many of the same proteins and molecules as blood and urine do. </li></ul><ul><li>Saliva can reveal disease, i.e. cancer </li></ul><ul><li>Developing technology for identifying molecules “biomarkers” for disease </li></ul><ul><li>Determines who vulnerable pt is to caries </li></ul><ul><li>“ -human’s sugar chain makeup (oligosaccharides) is 100% genetically determined and cannot be changed. The USC researchers found that the sugar chain makeup in saliva can predict a child’s future cavity history to plus or minus one cavity with greater than 98% confidence”. </li></ul>
  15. 17. Gcamerica.com Saliva testing: good practice, good sense! Saliva-Check Buffer Kit
  16. 18. Patient Education
  17. 19. Patient Education <ul><li>Carbs and acidic foods amplify S.mutans and S.sobrinus in dental plaque which are related to the initiation and progression of dental caries. </li></ul>
  18. 20. Patient Education <ul><li>Once the salivary problem is identified the practitioner can recommend a course of preventive care (i.e. fluoride, chlorhexidine), lifestyle changes, altering oral hygiene, increasing saliva production. </li></ul>
  19. 21. 10 min of testing: <ul><li>Step 1: </li></ul><ul><li>Hydration </li></ul><ul><li>The rate of production of resting saliva as a surrogate measure of hydration. </li></ul>
  20. 22. 2. Consistency <ul><li>The viscosity of resting saliva as a surrogate for measuring its relative water and mucin content. </li></ul>
  21. 23. 3. Resting saliva pH <ul><li>The pH of resting saliva </li></ul>
  22. 24. 4. Stimulated saliva flow (5 min) <ul><li>The rate of production of stimulated saliva </li></ul>
  23. 25. 5. Stimulated saliva pH <ul><li>Higher concentration of bicarbonate ions in parotid saliva and should be a higher pH than the resting pH. </li></ul>
  24. 26. 6. Buffering capacity <ul><li>Buffering capacity of stimulated saliva in an aqueous env w/ differing amts of lactic acid. A small pipette is used to withdraw a small quantity of saliva, which is placed on test pads. Low buffer capacity – bright red (0pts) </li></ul><ul><li>Normal buffer – green (4pts) </li></ul><ul><li>Intermediate buffer – blue (2pts) </li></ul><ul><li>12 pt scale is created to keep track of any changes in buffer capacity over time. </li></ul><ul><li>Score b/w 10-12 is normal capacity </li></ul><ul><li>The remaining sample can be used for immuno-assay for S. mutans levels. </li></ul>
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