Intranasal immunization of guinea pig with trivalent influenza antigen adjuvanted by Cyclamen europaeum tubers extract

Abstract

Cyclamen europaeum tubers extract (CTE) with concentration commonly used for human rhinosinusitis treatment was tested as mucosal adjuvant in experimental intranasal immunization of guinea pigs with concentrated commercially available influenza trivalent vaccine and subsequent infection with influenza strain A/California/04/2009 H1N1pdm. Dual intranasal immunization with vaccine compound consisting of 7.5 µg of each hemagglutinin and 500 µg of CTE in 50 µl induced reciprocal GMT on day 21 after immunization 40 (5-640) against H1N1pdm; 43.20 (5-1280) against H3N2; 10.80 (5-80) against influenza B. Animals with HI titers 1/80 against cell-derived antigen were completely protected against challenge with A/California/04/2009 H1N1pdm09.

Saponins extracted from Polemonium caeruleum have adjuvant activity in guinea pig intranasal immunization with trivalent influenza antigens

“…Performed study shows that saponins from pharmacopeia plant Polemonium caeruleum has immunogenic activities in guinea pig model showed by pharmacopeia vaccine control method as hemagglutinin inhibition reaction. Saponins extracted from different plants (Polemonium caeruleum, Cyclamen europaeum) standardized by haemolitical reaction have almost the same immunogenic effects after intranasal administration performed identically [13]. Effectiveness of proposed immunization scheme, supposing dual intranasal injection on day 1 and 14, effective immunogenic saponin dose, was confirmed.…”

“…Biological activity and dose was standardized in hemolytic reaction. Reaction was performed in polystyrol V-shape microplates (Greiner Bio One) as described earlier (13). Dilution with conversion titer 1/64 (hemolytic) -1/128 (non hemolytic) per 50 µl was taken for immunization experiment.…”

“…For statistical calculations reciprocal titers were used. Analysis was performed with Excel (Microsoft) as described previously [13].…”

Development and laboratory production of virus-like immune-stimulating complexes based on saponins and evaluation of their adjuvant potential using mice immunisation with influenza antigens

“…Гемолитическую реакцию проводили в микроварианте в 96-луночных полистироловых планшетах [16]. На одном планшете готовили двукратные разведения исследуемых растворов ИСКОМ (Матрикс-BP и Матрикс-BQ) и стандартного раствора детергента в повторах, после чего добавляли по 50 мкл 0,5% суспензии эритроцитов петуха.…”

“…РТГА и гемолитическую реакцию проводили согласно методике, описанной V. Evseenko c соавт. [16]. Для вычисления значений средних геометрических титров (СГТ) и погрешностей измерения считали, что титр ниже предела детекции равен 1:10.…”