Year : 2010 | Volume
: 28 | Issue : 2 | Page : 91--94
Natural prevalence of antibody titers to Glucostltransferase of Streptococcus mutans in serum in high and low caries active children
BC Kirtaniya1, HS Chawla2, A Tiwari3, NK Ganguly4, V Sachdev5,
1 Professor, Department of Pedodontics, Himachal Dental College, Sunder Nagar, Himachal Pradesh, India
2 Former Professor and Head, Oral health science Centre, PGIMER, Chandigarh, India
3 Former Dean, PGIMER, former Professor and Head, Oral Health Science Centre, PGIMER, Chandigarh, India
4 Director, ICMR, New Delhi, former Professor and Head, Experimental Medicine, PGIMER, Chandigarh, India
5 Principal, Professor and Head, Himachal Dental College, Sunder Nagar, Himachal Pradesh, India
B C Kirtaniya
Department of Pedodontics, Himachal Dental College, Sunder Nagar, Himachal Pradesh
In this investigation, serum immunoglobulin G (IgG) and immunoglobulin A (IgA) titers, as well as total immunoglobulin concentration (IgG + IgA + IgM), were found to be raised with the increase in the number of dental caries. Only the total serum antibody titer in high dental caries (HDC) group was found to be significantly raised as compared to no dental carries (NDC) group. Although the IgG and IgA titers were raised in blood with the increased number of caries, the results were not statistically significant. However, we could not find any correlation between serum antibodies and dental caries except that there was an increased trend of serum antibodies to GTF with the increased number of carious lesions.
|How to cite this article:|
Kirtaniya B C, Chawla H S, Tiwari A, Ganguly N K, Sachdev V. Natural prevalence of antibody titers to Glucostltransferase of Streptococcus mutans in serum in high and low caries active children.J Indian Soc Pedod Prev Dent 2010;28:91-94
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Kirtaniya B C, Chawla H S, Tiwari A, Ganguly N K, Sachdev V. Natural prevalence of antibody titers to Glucostltransferase of Streptococcus mutans in serum in high and low caries active children. J Indian Soc Pedod Prev Dent [serial online] 2010 [cited 2020 May 28 ];28:91-94
Available from: http://www.jisppd.com/text.asp?2010/28/2/91/66744
The first study of successful immunization of monkeys (Macaca fascicularis) against dental caries was done by Bowen in 1969  with whole cell of Streptococcus mutans injected through intravenous route and he found a significant rise in serum antibody titer. There was 80% reduction of dental caries in the vaccinated group. Since then, various researchers throughout the world have worked on various antigenic components of S. mutans administered in various routes to many animal models. In the later years, the focus was on finding out whether natural immunity exists or not, against dental caries in man, keeping in view the fact that among the individuals in a population with almost similar local factors, some do not develop caries whereas others are highly susceptible. In our previous investigation, it was found that because of lack of local immunity, children are more prone to dental caries. This study was conducted to find out the role of systemic antibodies to GTF of S. mutans and dental caries. Specificity to GTF of S. mutans was chosen because of its various advantages (discussed in our earlier article). Antibodies to several other oral cariogenic organisms including S. sanguis, A. visosus and L. acidophilus did not show any relationship with immunity and dental caries. In this investigation, systemic antibodies are chosen because some authors have claimed that the humoral response in caries appears to be similar to that found in other infective microbial diseases. Hence, we try to find out if systemic antibodies play any role in the prevention of dental caries.
Materials and Methods
Forty-six children with an average age of 9 years (6-14 years) were selected according to their caries status. They were divided into four groups as no dental caries (NDC) group consisting of 11 children, low dental caries (LDC) group consisting of 10 children with a DMFT/deft of 1-2, moderate dental caries (MDC) group consisting of 10 children with a DMF/deft of 3-4 and high dental caries (HDC) group consisting of 15 children with a DMFT/deft of 5-15. Care was taken to select the healthy children without any disease for about 6 months or not suffering from hypogammaglobulinemia or not tonsillectomized.
The blood samples were taken from the median cubital vein of the left arm using disposable syringe and needles in the morning hours between 9.00 AM to 12 noon. The blood was transferred to a 10-ml disposable plastic vial and was kept stationary for 1 hour at 37°C for clotting. Then, the serum was collected and it was centrifuged at 2000 r.p.m. for 15 minutes. The supernatant was transferred to a 2-ml sterile plastic vial and stored at −20°C till enzyme-linked immunosorbent assay (ELISA) was done.
The GTF was prepared from S. mutans serotype c and purified following the method of Chludzinski et al. The estimation of antibody was done using ELISA as per the method of Engvall and Permann, 1972 .
The serum immunoglobulin concentrations estimated by ELISA was expressed in optical density (OD) and are shown in [Table 1],[Table 2],[Table 3].
The mean immunoglobulin G (IgG) concentrations in serum in NDC group was 0.260 ± 0.071, in LDC group it was 0.251 ± 0.073, in the MDC group it was 0.267 ± 0.082 and in the HDC group it was 0.305 ± 0.079. The mean IgG titer to GTF of S. mutans seemed to be raised with the increase in number of carious lesions. However, this raised titer was not statistically significant between NDC verses LDC, NDC verses MDC and NDC verses HDc.
The mean immunoglobulin A (IgA) concentration to GTF in NDC group was 0.182 ± 0.061, in the LDC group it was 0.251 ± 0.135, in the MDC group it was 0.262 ± 0.102 and in the HDC group it was 0.269 ± 0.134. There was an increase in serum IgA level with the increase in the number of dental caries. However, neither of the above raised titers was statistically significant among various groups, i.e., NDC verses LDC, NDC verses MDC and NDC verses HDC groups.
Total immunoglobulin concentrations
The mean total immunoglobulin concentration in the NDC group was 1.787 ± 0.381, in the LDC group it was 1.990 ± 0.275, in the MDC group it was 2.086 ± 0.356 and in the HDC group it was 2.133 ± 0.317. The mean total serum immunoglobulin concentration was observed to be raised with the increase in the number of caries. The mean total immunoglobulin concentration in serum in the HDC group was significantly higher than that in the NDC group (P < 0.05). The differences between NDC verses LDC and NDC verses MDC were not statistically significant.
The mean IgA, IgG and total immunoglobulin concentrations in serum are shown in the [Figure 1].
In this investigation, serum IgG titer was found to be raised with increase in the number of dental caries. The differences among NDC verses LDC, NDC verses MDC and NDC verses HDC were not statistically significant. These findings are in agreement with those of Challacomb and Lehner  who found a positive correlation between serum antibodies and active dental caries and S. mutans count. They explained that development of carious lesions was associated with a rise in serum antibody titer to S. mutans and treatment of caries led to a decrease of antibody titer in serum. On the contrary, Challacombe  had reported a significant increase in serum IgG titer in low caries group. Challacomb concluded that low caries was associated with significantly greater serum IgG antibody titer. Bhatia et al0. found that serum antibodies increased in active caries children than caries free children. The significant results of serum IgG of various authors may be due to the fact that S. mutans and/or GTF were properly absorbed in adults through the GALT. Therefore, the subjects in the above mentioned studies were adults except in that of Bhatia et al. In this investigation, all the subjects were children. Since the antigen was not in direct contact with blood, the IgG level was not significantly raised in serum which was evident in this study. Moreover, dental caries is a localized disease which is not in direct contact with blood (unless the pulp is exposed); so, the role of systemic antibodies might not be that significant. The presence of IgG antibody in saliva might be attributed to an exaggerated role of IgG antibody against dental caries, which reaches the mouth through gingival crevicular epithelium by passive diffusion. Therefore, IgG concentration may be raised in patients suffering from gingival disease or plaque accumulation in and around gingiva rather than dental caries.
IgA was found to be raised with the increase in number of carious lesions. The raised IgA titer was not statistically significant between NDC verses LDC, NDC verses MDC and NDC verses HDC. This result was in agreement with that of Challacomb and Lehner  and Challocomb,  wherein the authors could not find any significant role of serum IgA antibody in the immunological response to dental caries. It has been explained that since serum antibodies are primarily of IgG type, IgA has little or no role against dental caries. This may be the reason why the results of this investigation did not show a significant increase in serum IgA titer. The slight rise of IgA in serum could be the local antigenic challenge, thereby, a weak systemic response.  On the other hand Lehner et al, found that serum IgA levels were significantly higher in adult subjects with a high caries experience. But in our present study, all the subjects were children of 6-14 years of age with immature immune system and with open carious lesions for a long period of time. This rise of antibody A in serum in our view may be the result of local antigenic stimulation for dimerization of serum IgA to s-lgA for secretion into saliva or other body secretions.
Total immunoglobulin concentration of serum was raised quite significantly in HDC group than in the NDC, LDC and MCD groups. When total immunoglobulin concentration of HDC group was compared with NDC group, it was found to be statistically significant at 5% level (P < 0.05). The rise of immunoglobulins in serum was observed by Hayashi et al. in rats and they found a reduction of dental caries upto 54.4% in the experimental group immunized with GTF through intraperitoneal injections. In another experimental study on M. fascicularis immunized with highly purified GTF of S. mutans serotype c through subcutaneous route, Russel and Colman,  found higher level of antibodies to GTF. In in vitro conditions, the serum of these animals inhibited the synthesis of dextran and mutan. Our findings in serum are in partial agreement with those of the above mentioned authors but we found higher total immunoglobulin concentration only in high caries active children than in caries free children. When the total immunoglobulin concentration of NDC group was compared with LDC and MDC groups, the results were not statistically significant. These findings are in agreement with those of various authors as we mentioned earlier. The rise of total immunoglobulin concentration in serum in HDC group as compared with NDC group in this investigation should not be pinpointed since the IgG and IgA antibody titers, which are the predominant antibodies in serum, were not raised to a significant level in HDC group.
To conclude we can say that the increase in serum IgG, IgA and total antibody titer showed a direct correlation with the increase in number of carious lesions. The insignificant rise of serum IgG and IgA as well as total antibody titer could be due to these reasons: (i) GTF was not directly stimulating the hemopoetic system as it was not in direct contact with blood and (ii) GTF ingested naturally from oral route through GALT might be insufficient or of a low dose to produce a significant immune response.
|1||Bowen WH, Cohen B, Cole MF, Colman G. Immunisation against dental caries. Br Dent J 1975;39:45-58.|
|2||Chludzinski AM, Germaine GR, Schachtele CF. Streptococcus mutans dextransuucrase, purification, properties and requirements for primer dextran. J Dent Res 1976;55:75-86.|
|3||Russell MW, Challacombe SJ, Lehner T. Serum glucosyltranferase inhibiting antibodies and dental caries in rhesus monkeys immunized against S. mutans. Immunology 1976;30:619-27.|
|4||Challacombe SJ. Serumand salivary antibodies to streptococcusmutans in relationto the development and treatment of human dental caries. Arch Oral Biol 1980;25:495-502.|
|5||Bhatia S, Chawla HS, Tewari A, Ganguli NK. Status of serum and salivary antibodies specific to LTA derived from S. mutans in relation to dental caries activity. M.D.S. Thesis in Pedodontics and Preventive dentistry, Punjab University, 1986.|
|6||McGhee JR, Michalek SM. Immunobiology of dental caries; microbiological aspects and local immunity. Annu Rev Microbiol 1981;35:595-638.|
|7||Lehner T, Wilton JM, Ward RG. Serum antibodies in dental caries in man. Arch Oral Biol 1970;15:481-90.|
|8||Hayashi JA, Shklair IL, Bahn AN. Immunization with dextransucrases and glucosidichydrolases. J Dent Res 1972;51:436-42.|
|9||Russell RR, Colman G. Immunization of monkeys (Macaca fascicularis) with purified S. mutans glucosyltranferase. Arch Oral Biol 1981;26:23-8.|
|10||Everhart DL, Grigsby WR, Carter WH Jr. Evaluation of dental caries experience and salivary immunoglobulins in whole saliva. J Dent Res 1972;51:1487-92.|
|11||Van de Rijn I, Bleiweis AS, Zabriskie JB. Antigens in Streptococcus mutans cross reactive with human heart muscle. J Dent Res 1976;55:C59-64.|
|12||Ferretti JJ, Shea C, Humphrey MW. Cross-reactivity of S. mutans antigens and humanheart tissue. Infect Immun 1980;30:69-73.|
|13||Bolton RW, Hlava GL. Evaluation of salivary IgA antibody to cariogenic microorganisms in children correlation with dental caries activity. J Dent Res 1982;61:1225-8.|
|14||Challacombe SJ, Bergmeier LA, Rees AS. Natural antibodies in man to a protein antigen from the bacterium S.mutans related to dental caries experience. Arch Oral Biol 1984;29:179-84.|
|15||Cole MF, Emilson CG, Hsu SD, Li SH, Bowen WH. Effect of pre-oral immunization of humans with streptococcus mutans on induction of salivary and serum antibodies and inhibition of experimental infection. Infect Immun 1984;46:703-9.|