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Affinity and avidity of polyclonal antibodies towards the major antigenic determinant of slime-producing Staphylococcus epidermidis and their protective effect in experimental keratitis

, : Affinity and avidity of polyclonal antibodies towards the major antigenic determinant of slime-producing Staphylococcus epidermidis and their protective effect in experimental keratitis. Abstracts of the General Meeting of the American Society for Microbiology 102: 11-12

Staphylococcus epidermidis is an important cause of bacterial keratitis. Certain S. epidermidis strains produce an extracellular slime layer rich in a 20 kDa acidic polysaccharide (20-kDa PS). The purpose of this study was to produce rabbit polyclonal antibodies against 20-kDa PS, examine the specificity, affinity and avidity of these antibodies and examine the protective and therapeutic effects of their administration in a rabbit keratitis model. Rabbits were actively immunized with 20-kDa PS (0.9 mg) emulsified with Freund's complete adjuvant for the first injection and incomplete adjuvant for the next two challenges. The specificity of produced sera for the homologous antigen was studied by competitive ELISA. Obtained results suggest that 20-kDa PS reacts specifically with produced antibodies. The avidity of the polyclonal sera against 20-kDa PS was studied by use of disruptive means (formamide) in the ELISA system. Results show that immune sera exhibit significantly higher avidity (P<0.001) for the antigen than pre-immune sera. Estimation of the affinity of polyclonal antibodies for the 20-kDa PS was based on the assumption that antibody-antigen interactions are similar to the enzyme-substrate interactions and therefore their kinetics can be well described by the Michaelis-Menten model. Results show that polyclonal antibodies exhibit high affinity for the homologous antigen. Thus, specific polyclonal antibodies (about 70 mg) were administered to twenty rabbits one day before induction of slime-producing S. epidermidis keratitis. Clinical observations were made weekly for one month and levels of anti-20-kDa PS IgG in serum and aqueous humor were determined on days 0, 3, 14 and 30. The levels of anti-20-kDa PS IgG in immunized rabbits were significantly higher than those of the control group (P<0.001). A significant decrease in serum was observed on day 30 due to physiological catabolism. Passively immunized rabbits were significantly better protected than control. Obtained results suggest that administration of specific polyclonal antibodies significantly protects against corneal S. epidermidis pathology and damage.


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