Abstract:Streptococcus mutans is a common principal causative agent of dental caries. In this communication, we describe that the antibodies raised against purified dextransucrase effectively inhibited the growth of S. mutans. The purified enzyme showed 58-fold enrichment, 17.5% yield and a specific activity of 3.96 units/mg protein. Purified IgG fraction of the antibody showed significant affinity with the antigenic protein. Immunotritation of the enzyme with dextransucrase antibody showed a gradual increase in inhibi… Show more
“…Although showing promising anticaries effects, but their use was restricted of having cross reactivity with human heart tissues and skeleton muscles 52 . However western blot analysis revealed that antibodies against dextransucrase did not show any cross reactivity with human heart, liver, gall bladder and other mammalian tissues tested as described previously 16 . In the present study various human samples were tested for cross reactivity with dextransucrase antibody by immunohistochemistry which also showed negative results con rming no cross reactivity of dextransucrase antibodies with the human tissues.…”
Section: Discussionsupporting
confidence: 53%
“…Dextransucrase was puri ed to homogeneity from the culture supernatant of S. mutans MTCC-890 grown at 37 °C for 36 h by 55% ammonium sulphate precipitation followed by gel ltration chromatography using Sephadex G-200 and treatment with PEG-400 (polyethylene glycol 400) as reported earlier 16 .…”
Section: Bacterial Strain and Growth Conditionsmentioning
confidence: 99%
“…Polyclonal antisera against dextransucrase puri ed from S. mutans was raised in New Zealand White rabbits immunised subcutaneously with puri ed dextransucrase (500μg) mixed with complete Freund's Complete Adjuvant (F5881, Sigma, USA). Serum was collected after two weeks of last booster and analysed for dextransucrase speci c antibody by Dot blot assay, confocal microscopy, ELISA and western blot analysis as described previously 16 . Serum IgG was puri ed by a nity chromatography using Protein A Sepharose Column (Bio Vision, USA).…”
Section: Bacterial Strain and Growth Conditionsmentioning
confidence: 99%
“…However antibodies against these molecules are reported to have cross reactivity against skeleton muscles and heart tissues 15 , but antibodies against dextransucrase when tested for cross reactivity with human heart tissues and various mammalian tissues revealed negative results 16 .…”
Dextransucrase produced by Streptococcus mutans play an essential role in the formation of dental caries by synthesizing exopolysaccharides from sucrose, an important metabolite of the organism. In this study we report the location of dextransucrase in Streptococcus mutans cells and describe that antibodies raised against dextransucrase inhibited biofilm formation and reduced the adherence and hydrophobic properties of Streptococcus mutans. Western blot analysis and immunoelectron microscopy revealed that dextransucrase is located abundantly in the membrane fraction in S. mutans cells. Scanning electron microscopy and fluorescence microscopy revealed reduced cell density, impaired bioflim (plaque) formation in presence of dextransucrase antibodies. Genes associated with bioflim formation in S. mutans such as GtfB, GtfC, BrpA, relA, Smu630, vicK were down regulated (50–97%) in presence of the enzyme antibody. Presence of enzyme antibodies reduced adherence of S. mutans cells to glass surfaces by 58% and hydrophobicity by 55.2%. However dextransucrase antibodies did not affect acid production by S. mutans, under the experimental conditions. Immunohistochemistry studies with certain human samples displayed no cross reactivity with dextransucrase antibody. These findings suggest that antibodies against dextransucrase exhibit a profound inhibitory effect on the vital cariogenic factors of S. mutans and have no cross reactivity with human tissues tested, thus implying that dextransucrase could be a promising antigen to study its anticariogenic potential.
“…Although showing promising anticaries effects, but their use was restricted of having cross reactivity with human heart tissues and skeleton muscles 52 . However western blot analysis revealed that antibodies against dextransucrase did not show any cross reactivity with human heart, liver, gall bladder and other mammalian tissues tested as described previously 16 . In the present study various human samples were tested for cross reactivity with dextransucrase antibody by immunohistochemistry which also showed negative results con rming no cross reactivity of dextransucrase antibodies with the human tissues.…”
Section: Discussionsupporting
confidence: 53%
“…Dextransucrase was puri ed to homogeneity from the culture supernatant of S. mutans MTCC-890 grown at 37 °C for 36 h by 55% ammonium sulphate precipitation followed by gel ltration chromatography using Sephadex G-200 and treatment with PEG-400 (polyethylene glycol 400) as reported earlier 16 .…”
Section: Bacterial Strain and Growth Conditionsmentioning
confidence: 99%
“…Polyclonal antisera against dextransucrase puri ed from S. mutans was raised in New Zealand White rabbits immunised subcutaneously with puri ed dextransucrase (500μg) mixed with complete Freund's Complete Adjuvant (F5881, Sigma, USA). Serum was collected after two weeks of last booster and analysed for dextransucrase speci c antibody by Dot blot assay, confocal microscopy, ELISA and western blot analysis as described previously 16 . Serum IgG was puri ed by a nity chromatography using Protein A Sepharose Column (Bio Vision, USA).…”
Section: Bacterial Strain and Growth Conditionsmentioning
confidence: 99%
“…However antibodies against these molecules are reported to have cross reactivity against skeleton muscles and heart tissues 15 , but antibodies against dextransucrase when tested for cross reactivity with human heart tissues and various mammalian tissues revealed negative results 16 .…”
Dextransucrase produced by Streptococcus mutans play an essential role in the formation of dental caries by synthesizing exopolysaccharides from sucrose, an important metabolite of the organism. In this study we report the location of dextransucrase in Streptococcus mutans cells and describe that antibodies raised against dextransucrase inhibited biofilm formation and reduced the adherence and hydrophobic properties of Streptococcus mutans. Western blot analysis and immunoelectron microscopy revealed that dextransucrase is located abundantly in the membrane fraction in S. mutans cells. Scanning electron microscopy and fluorescence microscopy revealed reduced cell density, impaired bioflim (plaque) formation in presence of dextransucrase antibodies. Genes associated with bioflim formation in S. mutans such as GtfB, GtfC, BrpA, relA, Smu630, vicK were down regulated (50–97%) in presence of the enzyme antibody. Presence of enzyme antibodies reduced adherence of S. mutans cells to glass surfaces by 58% and hydrophobicity by 55.2%. However dextransucrase antibodies did not affect acid production by S. mutans, under the experimental conditions. Immunohistochemistry studies with certain human samples displayed no cross reactivity with dextransucrase antibody. These findings suggest that antibodies against dextransucrase exhibit a profound inhibitory effect on the vital cariogenic factors of S. mutans and have no cross reactivity with human tissues tested, thus implying that dextransucrase could be a promising antigen to study its anticariogenic potential.
“…indicated that dextransucrase antibodies inhibited acid production and reduced hydrophobicity of S. mutans , indicating the further enhancement of anticariogenic potential of dextransucrase antibodies. [ 51 ] Moreover, no cross-reactivity with dextransucrase antibody was seen indicating no harmful physiological effects in the humans. This has given the hope that effective, safer, and acceptable vaccines against human dental caries can be achieved in the near future.…”
Despite advances in the 21
st
century, dental caries still remains to be one of the most common infectious diseases. Its prevalence was confirmed by the World Health Organization and affirms dental caries as a major health problem in all over the world. Even though the process of tooth decay is multifactorial, the oral bacteria, mutans streptococci, such as
Streptococcus mutans
and Streptococcus sobrinus, are considered to be causative agents of dental caries in human. Numerous studies carried out on animals and various categories of vaccines were developed such as whole cell vaccine, subunit vaccine, and synthetic peptides. Irrespective of success from active and passive immunization based on animal trials, it is the phenomenon of human heart reactivity that limited the applicability of these trials in humans. Continuous efforts are being made to overcome these limitations and for further success in human trials. With the advent of various antibodies against antigens of mutans streptococci, local passive immunization has become the safer approach in humans against the colonization of bacteria and caries induction. This review provided insight into epidemiology, active and passive immunization in both animal and human trials, as well as the prospects of caries vaccination.
Introduction. Dextransucrase produced by
Streptococcus mutans
plays a vital role in the formation of dental caries by synthesizing exopolysaccharides from sucrose, which helps in the attachment of microbes to the tooth surface, causing caries. Exploring antibody production against
S. mutans
antigens could be an effective method to protect against dental caries.
Hypothesis. Dextransucrase antibodies may help in the prevention of caries formation by inhibiting essential cariogenic factors.
Aims. The aim of this study was to investigate the effects of dextransucrase antibodies on biofilm formation and certain associated cariogenic factors of
S. mutans
.
Methodology. Dextransucrase was purified from culture of
S. mutans
. The antisera against the enzyme were raised in rabbits. The effect of dextransucrase antibodies on biofilm formation was studied using scanning electron microscopy, fluorescence microscopy and quantitative real-time polymerase chain reaction. The effects of the antibodies on associated cariogenic factors were examined using established methods. The cross-reactivity of antibodies with human lung, liver, heart, thyroid and kidney tissues was evaluated by immunohistochemistry.
Results. Our findings showed impaired biofilm formation in
S. mutans
in the presence of dextransucrase antibodies. Genes associated with biofilm formation such as gtfB, gtfC, brpA, relA, Smu.630 and vicK were downregulated (50–97 %) by dextransucrase antibodies in
S. mutans
. The adherence of
S. mutans
to glass surface was reduced by 58 % and hydrophobicity was reduced by 55.2 % in the presence of the antibodies compared to the controls. Immunohistochemistry studies revealed no cross-reactivity of human tissues with dextransucrase antibodies.
Conclusions. These findings suggest that antibodies raised against dextransucrase exhibit a profound inhibitory effect on biofilm formation and vital cariogenic factors of
S. mutans
, which supports the contention that dextransucrase could be a promising antigen to study for its anticariogenic potential.
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