Effect Of Penicillium Marneffei ANDP.Marneffei Cytoplasmic Yeast Antigen On The Polarization Of Alveolar Macrophages Of BALB/c Mice

Objective To explore the effect of Penicillium marneffei(PM) and P.marneffei cytoplasmic yeast antigen(CYA) on the polarization of alveolar macrophages(AMs)ofBALB/c mice.Methods 150 numbers of Five/six-weeks-oldmale BALB/c mice were randomly in divided into normal group(N group) with 30 numbers of mice,2nd week post-infection group(PM-2W group) with 60 numbers of mice and 4th weekpost-infection group(PM-4W group) with 60 numbers of mice.50μl suspension of PM at the concentration of 5×107CFU(colony-forming-units, CFU)/ml was slowly dripped into the nose of mouse for infection for 2 or 4 weeks. The mice for negative control conducted without any treatment. AMswere harvested from mice which are sacrificed after anesthetized and then 1×106 macrophages were added to each well of a 12-well tissue culture plate and incubated for 24h to collect the suspernatant and cells. To detect the mRNA expression of macrophage polarization key enzyme inducible nitric oxide synthase(iNOS) and arginase1(Arg1)was measured by qRT-PCR; the iNOS and Argl activity was analyzed by Griess assay and arginase assay,respectively; IL-12, TNF-a, IL-10 and TGF-β1evels were quantified by ELISA assay. The AMs of PM infected mice were divided as follow:PM-control group, the PM-IFN-y+LPS group(M1 positive control), the PM-IL-4 group (M2 positive control) and the PM-CYA group. After incubated for 24h, the supernatant and cells were collected for detect the above indicators on the same way.Results1.It showed that 2nd week post-infection group were higher expression of M1-phenotypic markers(iNOSmRNA,NO,IL-12,TNF-a)and M2-phenotypic markers(ArglmRNA,urea) than normal group (P<0.05), but could not promote levels of M2-related cytokine IL-10 increase (P>0.05). The above indicators in 4th weekpost-infection group were decreased significantly when compare with 2nd week post-infection group(P<0.05). All groups failedto produce any detectableM2-related cytokine TGF-β.2. PM-CYA group induced higher expression of iNOS mRNA, NO, IL-12 when compared with PM-control and M1 positive group in 2nd week post-infection(P<0.05). PM-CYA group also induced higher expression of Argl mRNA and urea when compared with PM-control(P<0.05), but the expression lower than the M2 positive group(P<0.05). PM-CYA group could not promote levels of IL-10 increase(P>0.05).CYA induce higher expression of M1 subtype macrophages more than M2 subtype macrophages, and 2nd week post-infection higher expression than 4th week post-infection(P<0.05). All groups failedto produce any detectableTGF-β.Conclusions 1. AMs of PM infected mice easier switched toward M1 and M2 subtype macrophages in 2nd week post-infection and dropped significantly in 4th week post-infection, but M2 subtype macrophages switched were not so complete.2.The polarization tendency of AMs induce by CYA was consistent with PM induce polarition and had a tendency toward increased; CYA more lean to induce M1 subtype macrophages.3. CYA could be responsible for P. marneffei infection induced host immune response.