Except immunoglobulins and components of complement system, the important part of congenital humoral immunity are antimicrobial (poly) peptides (AMP). Main antimicrobial mechanism of AMP action is the destruction of cytoplasmic membrane. In host organism number of AMP act simultaneously, therefore method for the measuring of overall AMP activity would be helpful. The method was recently applied and modified for estimation of AMP activity in serum: aliquots of low-molecular fraction of serum added to test-culture of Candida albicans, and the part of killed cells, i.e. total activity of AMP, calculated by spectroscopy. Aim of present study is the estimation of AMP activity in mice sera after immunization with cells of clinical yeasts compare to the immunoglobulins G. Yeasts strains-Candida albicans, Cryptococcus neoformans, Rhodotorula mucilaginosa, Trichosporon cutaneum, Geotrichum candidum, and Saccharomyces cerevisiae-were cultivated in liquid defined media up to the end of exponential growth phase. Obtained yeasts cells used for immunization of mice, but yeast antigens-for the estimation of IgG-antibodies levels in mice sera by ELISA method. AMP activity were measured as described above. Immunization of mice with yeasts cells was accompanied not only with increase of specific IgG levels, but with elevation of total activity of AMP: for example specific IgG in sera of mice immunized by C. albicans cells was 16-fold higher than in non-immunized (control), and AMP activity-1,5 fold higher than in control. Correlation between specific humoral response at the immunoglobulins level and humoral response at the AMP-activity level (Pirson`s coefficient r=0,735; p ≤ 0,05) as a result of immunization of mice by fungal cells was demonstrated for the first time. The method may be used for study of humoral immunity associated with action of AMP in various animal and human models.
Vera Arzumanian*, Olga Shmeleva and Natalia Michailova
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