Ecology, Environment and Conservation Paper

STUDY EXPRESSION OF FOX3, GATA3, T-BET MARKERS USING IMMUNOHISTOCHEMICAL TECHNIQUE IN NORMAL AND IMMUNOCOMPROMISED MICE AFTER EXPOSED TO PENICILLIUM MARNEFFEI AN EXPERIMENTAL STUDY

Milad A. Mezher

Abstract

Penicillium marneffei can cause a lethal systemic infection (Penicilliosis) with fever and anaemia similar to disseminated cryptococcosis. The incidence of P. marneffei is increasing as HIV spreads throughout Asia. The major function of Fox3, Gata3, T-bet markers is probably related to its ability to mediate substrate adhesion to mammalian cells, a process that involves the binding of specific cell surface receptors to discrete domains in the fibronectin molecule. Fox3, Gata3, T-bet markers were investigated in mice infected intravenously with 5×106 virulent P. marneffei conidia by immunohistochemistry technique to study the immune response using detection kit (The Novolink TM Polymer Detection Systems) using commercial kit (Novocastra, UK) in the King’s College Medical School at King’s College Hospital. Three groups of animals were studied; including control group (mice infected with only with P. marneffei) and the groups only with cortisone and the third groups the fungus with cortisone these groups were used to study the expression of immune markers. Expression of Fox3, Gata3, T-bet markers were in the studied organs were shown in the cytoplasm of the tissue cells and detected by IHC technique. Depending on the scoring system used for the Anti Fox3, Gata3, T-bet markers, the intensity of the staining of the cytoplasim used as a parameter dependent. The intensity of the cytoplasim of the stained cells was negative if there is no expression. Expression was found positive in most of the studied organs, but by different intensity, in lung over expression represented by strong staining in the cortisone group the expression rate was Fox3+, Gata3 +++, T-bet + , lymphatic site, at the group infected with fungus and immune suppressed with cortisone the expression were Fox3+++, +++ Gata3, Fox3-, while in the control group the expression scored as Fox3+++,Gata3++, T-bet-. Results showed significant difference (P value >0.01) in the expression between the studied organs. In conclusion, Fox3, Gata3, T-bet markers expression levels were time-dependent i.e. increased and related with the progress of infection that can significantly enhance resistance to P. marneffei.