Characterization of Polyclonal Active Components from Actinomyces Viscosus
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Actinomyces viscosus (AV) is an oral bacterium which has an established relationship with periodontal diseases. Exactly how AV exerts its effects are unknown although its ability to elicit a potentially detrimental immune response probably is involved. Because the components of the oral flora interact in a complex way to establish a microenvironment to which the body's immune system is exposed, it is difficult to ascertain directly the causes of periodontal disease. It has been recommended that a study into the biochemical nature of AV's interaction with human lymphocytes be undertaken as a step towards this understanding. This SIP involves the beginning of that biochemical study. AV was treated in various ways, and its activity (as measured by its ability to stimulate lymphocyte antibody production) was measured. The object of the treatment is to find modes by which AV can be broken down, in a step-by-step fashion, leaving the active component(s) in one fraction. Treatment of AV with enzymes, temperature changes, pH changes, periodate oxidation, and ammonium sulfate precipitation were undertaken. After treatment, AV was incubated with human lymphocytes, and antibody production was measured. If antibody production was high, it was concluded that treatment did not remove, or denature, the active component of AV. It was concluded that pH changes wipe out activity. Temperature changes do not. RNase and DNase enhance activity of AV, possibly by removing material surrounding active components. Lipase destroys activity, suggesting a lipid component in the active moiety. Ammonium sulfate precipitation revealed two primarily stimulatory fractions. This method could be used to eliminate unwanted material. Periodate oxidation did not yield conclusive results. Antibody production was measured by the ELISA method, which is described in detail in MATERIALS AND METHODS.