Abstract：
The model flatworm Mesocestoides vogae proliferating stage of infection elicits immunosuppression in the host. It was used to investigate the effects of human leukocyte extract (DLE) alone and in combination with anthelmintic albendazole (ABZ) on the reduction in peritoneal infection, peritoneal exudate cells (PECs), their adherent counterparts, and peritoneal exudates after the termination of therapy. Balb/c mice were infected with the larvae of M. vogae. PECs and adherent macrophages were studied via flow cytometry, mRNA transcript levels, and immunofluorescence. The cytokine levels were measured via ELISA and larvae were counted. ABZ significantly reduced larval counts (581.2 ± 65, p < 0.001), but the highest reduction was observed after combined treatment with ABZ and DLE (389.2 ± 119, p < 0.001) in comparison with the control. Compared to an infected group, the proportions of CD11b+CD19- myeloid cells with suppressive ability decreased after albendazole (ABZ) in combination with DLE, which was the most effective in the elevation of B cells and CD11b+F4/80mid/highMHCIIhigh macrophages/monocytes (22.2 ± 5.4%). Transcripts of the M2 macrophage markers (arginase 1, FIZZ-1, and Ym-1) were downregulated after DLE and combined therapy but not after ABZ, and the opposite trend was seen for iNOS. This contrasts with reduced ex vivo NO production by LPS-stimulated PECs from DLE and ABZ+DLE groups, where adherent macrophages/monocytes had elevated transcripts of the INF-γ receptor and STAT1 and reduced expression of STAT3, STAT6, and IL-10. Each therapy differentially modulated transcription profiles and concentrations of IFN-γ, TNF-α, IL-12p40, IL-6, IL-10, and TGF-β cytokines. DLE strongly ameliorated ABZ-induced suppression of INF-γ and IL-12 and preserved downregulation for IL-4, IL-10, and TGF-β. Epigenetic study on adherent macrophages from infected mice showed that ABZ, ABZ-sulfoxide, and DLE could interact with the mRNA of examined markers in a dose-dependent pattern. Co-administration of DLE with ABZ seemed to augment the drug\'s larvicidal effect via modulation of immunity. In comparison with ABZ, combined therapy was the most effective in alleviating parasite-induced Th2/Treg/STAT3/STA6 directed immunosuppression by stimulating the Th1 cytokines, M1 macrophage polarization, and activation of the IFNγ/STAT1 signaling pathway.
摘要：
模型扁虫感染的中胚层增殖阶段在宿主中引起免疫抑制。它被用来研究人白细胞提取物（DLE）单独和联合驱虫药阿苯达唑（ABZ）对减少腹膜感染的影响。腹膜渗出细胞（PECs），他们的追随者，和腹膜渗出后终止治疗。用M.vogae的幼虫感染Balb/c小鼠。通过流式细胞术研究PECs和粘附巨噬细胞，mRNA转录水平，和免疫荧光。通过ELISA测量细胞因子水平并对幼虫进行计数。ABZ显著减少幼虫计数（581.2±65，p<0.001），但与对照组相比，ABZ和DLE联合治疗后的下降幅度最大（389.2±119，p<0.001）。与受感染的群体相比，阿苯达唑（ABZ）联合DLE后，具有抑制能力的CD11bCD19-髓样细胞的比例降低，在B细胞和CD11bF4/80mid/highMHCIIhigh巨噬细胞/单核细胞升高中最有效（22.2±5.4％）。在DLE和联合治疗后，M2巨噬细胞标记物（精氨酸酶1，FIZZ-1和Ym-1）的转录物下调，但在ABZ后未下调，iNOS的趋势相反。这与来自DLE和ABZ+DLE组的LPS刺激的PEC减少的离体NO产生形成对比。其中粘附巨噬细胞/单核细胞的INF-γ受体和STAT1的转录物升高，而STAT3，STAT6和IL-10的表达降低。每种疗法差异调节IFN-γ的转录谱和浓度，TNF-α,IL-12p40、IL-6、IL-10和TGF-β细胞因子。DLE强烈改善了ABZ诱导的INF-γ和IL-12的抑制，并保留了IL-4，IL-10和TGF-β的下调。对感染小鼠粘附巨噬细胞的表观遗传学研究表明，ABZ，ABZ-亚砜，DLE可以以剂量依赖性模式与所检查标记物的mRNA相互作用。DLE与ABZ的共同给药似乎通过调节免疫力来增强药物的杀幼虫作用。与ABZ相比，联合疗法通过刺激Th1细胞因子来缓解寄生虫诱导的Th2/Treg/STAT3/STA6定向免疫抑制是最有效的，M1巨噬细胞极化，和IFNγ/STAT1信号通路的激活。
