|Year : 2013 | Volume
| Issue : 4 | Page : 270-274
Correlation of total salivary secretory immunoglobulin A (SIgA) and mutans specific SIgA in children having different caries status
Soumen Pal1, Malay Mitra2, Jiban Mishra3, Subrata Saha2, Bhaswar Bhattacharya2
1 Department of Pedodontics and Preventive Dentistry, Awadh Dental College and Hospital, Jamshedpur, Jharkhand, India
2 Department of Pedodontics and Preventive Dentistry, Dr. R. Ahmed Dental College and Hospital, Kolkata, West Bengal, India
3 Department of Pedodontics and Preventive Dentistry, Burdwan Dental College and Hospital, Kolkata, West Bengal, India
|Date of Web Publication||21-Nov-2013|
Departments of Pedodontics and Preventive Dentistry, Jamshedpur, Jharkhand
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background: Relation between secretory immunoglobulin A (SigA) and dental caries still imprecise. Studies have been conducted either for total SIgA or mutans specific SIgA with bizarre results. Aim: The aim of this study is to find out the relationship between mutans antigen specific SIgA and total salivary SIgA and its influence in caries status in children. Materials and Methods: A total of 45 children aged between 6-14 years were divided based on their caries index (decayed, missing, filled, extracted tooth) score in high moderate and no caries group and their saliva were analyzed with enzyme linked immunosorbent assay for total SIgA and mutans specific SIgA. Results: High caries group showed high mutans specific SIgA, but less total SIgA, whereas low caries group showed the reverse. Conclusion: The mutans specific SIgA and total SIgA has a weak, but negative correlation in children potentiating caries inhibitory action of SIgA.
Keywords: Antigen, caries, saliva, secretory immunoglobulin A, Streptococcus mutans
|How to cite this article:|
Pal S, Mitra M, Mishra J, Saha S, Bhattacharya B. Correlation of total salivary secretory immunoglobulin A (SIgA) and mutans specific SIgA in children having different caries status. J Indian Soc Pedod Prev Dent 2013;31:270-4
|How to cite this URL:|
Pal S, Mitra M, Mishra J, Saha S, Bhattacharya B. Correlation of total salivary secretory immunoglobulin A (SIgA) and mutans specific SIgA in children having different caries status. J Indian Soc Pedod Prev Dent [serial online] 2013 [cited 2020 Feb 17];31:270-4. Available from: http://www.jisppd.com/text.asp?2013/31/4/270/121831
| Introduction|| |
Salivary secretory immunoglobulin A (SIgA) has an immunological control over dental caries and persuambly prevents the adherence of cariogenic microorganisms to hard surfaces and may also inhibit the activity of glucosyltransferases. , Studies, which evaluated the role of total salivary SIgA and caries prevalence or mutans antigen specific SIgA and prevalence of caries in children showed variant relationship between them. However,this leads to some confusion. With total salivary SigA, there have been results of positive, negative even no correlation with dental caries status of the children.  The same thing can be noticed with mutans specific SIgA also. Since total SIgA in saliva can be formed against other common oral pathogens,  so it can not be conclusive to evaluate its role against specifically in caries protection. On the other hand, it is also true that the amount of total salivary SIgA will determine the immunity status of the patient.  Again the amount of mutans specific SIgA in the same patient will give evidence of protection of salivary SIgA against that particular pathogen. However only a few studies have evaluated the interrelation between the total SIgA of saliva and mutans specific SIgA of saliva in the same patient having different caries status. Hence,aim of this study is to evaluate the interrelation of total salivary SIgA and mutans specific SIgA in children having different caries status in mixed dentition by enzyme linked immunosorbent assay (ELISA).
| Materials and Methods|| |
In this cross-sectional analytical study, 45 children in the age group of 6-14 years were selected with caries index ranging from 0 to 15 (decayed, missing, filled, extracted tooth) based on the criteria that they should not be immune compromised and having a good quality to expectorate.
Before the commencement of the study, the parents or legal guardian were adequately informed in a clear concise terms in a language familiar to them about the procedure and a written consent of the parents or legal guardians was taken.
Children selected for the study were called on a prefixed day and made to sit comfortably in the dental chair. The children were accompanied by their parents or legal guardian during the procedure. The procedure included thorough history taking along with intraoral, extra oral and physical examination of the child. Intraoral examinations were made using dental mirror and explorer under proper illumination. Examination for dental caries was performed according to the World Health Organization criteria and method (1997). 
The 45 children were divided into three groups according to the severity of dental caries as: 
Dental examination and saliva sampling were carried out in different days. On the day of saliva collection, the children were asked to perform regular oral hygiene procedure after breakfast (1.5 h before collection) and during this period children were not allowed to eat or drink. Children were seated in dental chairs and 4 cc of unstimulated saliva were collected in special tubes using the method described by Collins and Dawes.  All samples were collected between 10 and 11 a.m., and the time spent on each procedure didn't exceed 30 min.
- Group A: High dental caries with caries index of 5-15 consisting of 15 children.
- Group B: Moderate dental caries consisting of 15 children with caries index between 1- 4.
- Group C: No dental caries consisting of 15 children with caries index of 0.
Saliva samples mixed with 5.0 mM phenyl methylsulfonyl fluoride (Sigma, St. Louis, USA) and 0.002% sodium azide (Merck, Darmstadt, Germany) were stored at -20°C until antibody analysis.
Saliva samples that stored at -20°C were taken from the deep fridge and brought to room temperature. The samples were centrifuge at 3000 rpm for 10 min and sample supernatant was divided in two parts in equal volume for measuring Streptococcus mutans specific SIgA and total SIgA of each sample.The antigen were prepared first from the strain of Streptococcus mutans (ATCC 25175) by the procedure proposed by Burges and Edwards  , and the extracted antigen were stored at -20°C for further use. The amount of protein present in the antigen was measured by Bradford reagent. Salivary total SIgA level was quantified by ELISA method  using a commercial SIgA ELISA kit (DRG Diagnostica, Germany). And the Streptococcous mutans specific antigen was calculated by procedures described by Koga-Ito et al.  , by the reagent from the same aforesaid commercial ELISA kit (DRG Diagnostica, Germany). Salivary SIgA level was calculated using standard samples with known level of SIgA provided by the manufacturer and was expressed in milligrams.
Data was analyzed using SPSS version 11 (SPSS Inc, Chicago, IL, USA) software. A univariate descriptive analysis was made expressing the results as percentages and frequencies. Statistical significance was considered for P values < 0.05 in all cases. Pearson correlation and analysis of variance (ANOVA) test were used to compare the variables at different time intervals.
| Results and Observations|| |
These groups were used to analyze the amount of SIgA present in saliva sample specifically against Streptococcus mutans whole cell antigen along with total SIgA present in each samples were also measured.
In Group A (high caries group) mean SIgA level calculated against S. mutans was 33.71 ± 7.20 μg/ml, with mean age of the samples being 9.667 ± 2.469 years and mean caries index 6.600 ± 2.098. Whereas total SIgA calculated was 145.80 ± 20.85 μg/ml [Table 1].
Group B (moderate caries group) showed a mean SIgA level against S. mutans of 27.86 ± 4.49 μg/ml, and total SIgA was 189.13 ± 26.74 μg/ml ,mean age was 9.333 ± 2.498 years and mean caries index being 2.200 ± 0.941, presented in [Table 1].
Group C (low caries group) presented with S. mutans specific mean SIgA level 24.660 ± 1.715 μg/ml, with total SIgA was 215.40 ± 26.71 μg/ml caries index recorded 0, and mean age being 10.000 ± 2.591 years [Table 1].
The mean total salivary SIgA recorded in Group A, Group B and Group C were 145.80 ± 19.94 μg/ml, 189.47 ± 25.99 μg/ml, and 215.40 ± 26.71 μg/ml respectively.
To compare the average mutans specific SIgA and total SIgA level, the mean and standard deviation for the parameter of mutans specific SIgA level and total SIgA level of each group were tabulated. ANOVA of data were performed for the Fisher's 'F' test and 'P' value were calculated for both variables as shown in [Table 2] and [Table 3]. The differences of variances of the respective parameters in the three categories were computed to find whether the difference was significant at probability value equal to 0.05.
|Table 2: ANOVA test performed for S. Mutans specific SIgA level in different groups|
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From ANOVA [Table 2] values it can be seen that there is statistical differences present between mean values of mutans specific SIgA level between three groups (P < 0.05). After performing Fisher's individual family error rate at 0.05% value between the three groups and comparing the mean values from [Table 1] and Graph 1 [Additional file 1] it can be concluded that highest amount of mutans specific SIgA level was found in Group A and lowest amount was found in Group C with the Group B showing intermediate values.
From ANOVA [Table 3] values it can be seen that there is statistical differences present between mean values of total SIgA level between three groups (P < 0.05). After performing Fisher's individual family error rate at 0.05% value between the three groups and comparing the mean values and Graph 2, [Additional file 2] it can be concluded that highest amount of total SIgA level was found in Group C and lowest amount was found in Group A with Group B showing intermediate values.
Comparing the levels of mutans specific and total SIgA level among three groups [Graph 3] [Additional file 3] it can be seen that although mutans specific SIgA level is more in high caries group, but the total amount of SIgA is less than the other two groups. Whereas no caries group shows highest amount of total SIgA than other two groups, but mutans specific SIgA level is least in this.
Now considering the Pearson correlation value [Table 4] for total SIgA level and caries index is -0.631 suggesting a weak, but negative correlation, i.e. with increase of caries total SIgA level is decreasing.
|Table 4.: Pearson correlation values of age and S. Mutans specifi c and total SIgA level|
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In case of S. mutans specific SIgA level and caries index, the Pearson correlation value [Table 4] found was also weak, but positive suggesting that with the increase of caries in patient mouth there will be increase in the mutans specific SIgA, which is statistically significant also (P < 0.05).
Whereas comparing the values of S. mutans specific SIgA level and total SIgA level in Pearson correlation value [Table 4] a little strong negative correlation was seen, which is statistically significant also.
| Discussion|| |
In this study, it has been found that the mutans specific SIgA is present is more in amount in the high caries group (33.71 μg/ml) and less amount in no caries group (24.660 μg/ml) and the differences are statistically significant among the three groups. Again having Pearson correlation values we can say that the mutans specific SIgA and caries index has a weak, but positive correlation [Table 4]. According to Challacombe  salivary SIgA would not be related to a direct protection against dental caries, but would reflect a past exposure of the host to the cariogenic microorganisms. In the absence of a continuous stimulation, which does not happen on the natural progress of dental caries, the immunological titers would tend to decline, which seems to be a characteristic of the immunological system. One such study showed that when the antigenic load was removed with the successful treatment of carious cavity in patients mouth the postoperative amount of mutans specific SIgA was also decreased.  So from the present study result it can be concluded that the higher amount of mutans specific SIgA in the saliva of the high caries group was due to the need of reducing the high antigenic load. 
Although in the present study mutans specific SIgA was found more in high caries group than the no or moderate caries group, but comparing the total SIgA a statistically significant difference is found in high, moderate and no caries group with a weak negative correlation with caries index. Highest amount of total SIgA was found in no caries group, followed by moderate caries group and lowest amount was found in high caries group. The levels of SIgA in subjects without any systemic or immunological disease ranges from 40 to 300 μg/ml.  In our study, the total SIgA level correlated with this value (range 125-238 μg/ml). Studies conducted by Gregory et al.,  Everhart et al.,  Ranadheer et al.,  in their studies found similar results. They reported an increase in the level of salivary SIgA with decrease in caries activity. Whereas, studies conducted by de Farias and Bezerra  and Thaweboon et al.  found an increase in SIgA level with increase in caries activity. Like de Farias and Bezerra found a high total SIgA level in patients having early childhood caries, but their result may be due to the system used in determining the level of SIgA. However in their study de Farias and Bezerra potentiate the anti caries role of salivary SIgA by saying that this increased amount of SIgA in Early Childhood Caries group may be due to high antigenic load of S. mutans present in the saliva of those children. From the different studies it can be seen that the total salivary SIgA and mutans specific SIgA although may share positive or negative correlation with caries status of patient, but all the authors tried to promote the caries protective role of the SIgA. In our study, the high mutans specific antibody found in the high caries group may be due to the need of neutralization of high antigenic load present in those cases. Again comparing the correlation between total salivary SIgA and mutans antigen specific SIgA we can say that it has a negative correlation in our study. That proves that if patient is having higher total SIgA then its immunity against caries will be more on the other hand patient with low amount of total SIgA will have more mutans infection and in those cases mutans specific SIgA will be more in high caries group to neutralize the high antigen present in those cases.
As a whole present result is almost analogous to the study conducted by Everhart et al., who while estimating total SIgA found a negative correlation with caries status of the children but after they estimated the mutans specific SIgA level of those children they found a statistically significant negative correlation with caries index. 
| Conclusion|| |
From the study, it can be concluded that the SIgA present in the saliva confers a specific protection to patient against dental caries. Presence higher amount of the total SIgA will diminish the caries incidence in patients, whereas high caries status present with high amount of antigenic challenge will produce more amount of mutans specific SIgA for neutralization.
| References|| |
|1.||Petersen PE. Challenges to improvement of oral health in the 21st century - The approach of the WHO global oral health programme. Int Dent J 2004;54:329-43. |
|2.||Koga-Ito CY, Martins CA, Balducci I, Jorge AO. Correlation among mutans Streptococci counts, dental caries, and IgA to Streptococcus mutans in saliva. Braz Oral Res 2004;18:350-5. |
|3.||Watanabe Y, Mizoguchi H, Masamura K, Nagaya T. No relationship of salivary flow rate or secretory immunoglobulin A to dental caries in children. Environ Health Prev Med 1997;2:122-5. |
|4.||Amornchat C, Kraivaphan P, Dhanabhumi C, Tandhachoon K, Trirattana T, Choonhareongdej S. Effect of Cha-em Thai mouthwash on salivary levels of mutans Streptococci and total IgA. Southeast Asian J Trop Med Public Health 2006;37:528-31. |
|5.||Orstavik D, Brandtzaeg P. Secretion of parotid IgA in relation to gingival inflammation and dental caries experience in man. Arch Oral Biol 1975;20:701-4. |
|6.||Marcotte H, Lavoie MC. Oral microbial ecology and the role of salivary immunoglobulin A. Microbiol Mol Biol Rev 1998;62:71-109. |
|7.||World Health Organization. Oral Health Surveys. Geneva: World Health Organization; 1997. |
|8.||Kirtaniya BC, Chawla HS, Tiwari A, Ganguly NK, Sachdev V. Natural prevalence of antibody titres to GTF of S. mutans in saliva in high and low caries active children. J Indian Soc Pedod Prev Dent 2009;27:135-8. |
|9.||Collins LM, Dawes C. The surface area of the adult human mouth and thickness of the salivary film covering the teeth and oral mucosa. J Dent Res 1987;66:1300-2. |
|10.||Burgess TE , Edwards JR.Chemical Characterization of a Cell Wall Antigen from Streptococcus mutans FAl. Infect Immun 1973;8:491-93. |
|11.||Carpenter AB. Enzyme-linked immunoassays. In: Rose NR, De Macario EC, Folds JD, Lane HC, Nakamura RM. Manual of Clinical Laboratory Immunology.5 th ed. Washington: ASM Press; 1997; 20-9. |
|12.||Challacombe SJ. Salivary IgA antibodies to antigens from Streptococcus mutans in human dental caries. Adv Exp Med Biol 1978;107:355-67. |
|13.||Everhart DL, Bamgboye PO, Schwartz MS. Salivary anti-Streptococcus mutans changes over a six-month period in children ages two-five years. J Dent Res 1982;61:386-90, |
|14.||de Farias DG, Bezerra AC. Salivary antibodies, amylase and protein from children with early childhood caries. Clin Oral Investig 2003;7:154-7. |
|15.||Sivapathasundaram B, Raghu AR. Dental caries. In: Rajendran R, Sivapathasundaram B, editors. Shafer's Textbook of Oral Pathology. 6 th ed. Philadelphia: Elsevier; 2009. p. 515-48. |
|16.||Gregory RL, Filler SJ, Michalek SM, McGhee JR. Salivary immunoglobulin A and serum antibodies to Streptococcus mutans ribosomal preparations in dental caries-free and caries-susceptible human subjects. Infect Immun 1986;51:348-51. |
|17.||Everhart DL, Klapper B, Carter WH Jr, Moss S. Evaluation of dental caries experiences and salivary IgA in children ages 3-7. Caries Res 1977;11:211-5. |
|18.||Ranadheer E, Nayak UA, Reddy NV, Rao VA. The relationship between salivary IgA levels and dental caries in children. J Indian Soc Pedod Prev Dent 2011;29:106-12. |
|19.||Thaweboon S, Thaweboon B, Nakornchai S, Jitmaitree S. Salivary secretory IgA, pH, flow rates, mutans Streptococci and Candida in children with rampant caries. Southeast Asian J Trop Med Public Health 2008;39:893-9. |
[Table 1], [Table 2], [Table 3], [Table 4]