Hrčková Gabriela, Mačak Kubašková Terézia, Mudroňová Dagmar, Jurčacková Zuzana, Ciglanová Denisa
Institute of Parasitology, The Slovak Academy of Sciences, Hlinkova 3, 040 01 Košice, Slovakia.
Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy, Komenského 68/73, 041 81 Košice, Slovakia.
Pharmaceutics. 2023 Feb 6;15(2):541. doi: 10.3390/pharmaceutics15020541.
The model flatworm 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 . 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, < 0.001), but the highest reduction was observed after combined treatment with ABZ and DLE (389.2 ± 119, < 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+F4/80MHCII 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.
模型扁虫感染的增殖阶段会引发宿主的免疫抑制。本研究利用该模型来探究人白细胞提取物（DLE）单独使用以及与驱虫药阿苯达唑（ABZ）联合使用，对治疗结束后腹膜感染、腹膜渗出细胞（PEC）、其黏附对应物以及腹膜渗出物减少的影响。将Balb/c小鼠感染 的幼虫。通过流式细胞术、mRNA转录水平和免疫荧光研究PEC和黏附巨噬细胞。通过酶联免疫吸附测定法测量细胞因子水平并对幼虫进行计数。与对照组相比，阿苯达唑（ABZ）显著减少了幼虫数量（581.2±65，<0.001），但阿苯达唑与DLE联合治疗后幼虫数量减少最多（389.2±119，<0.001）。与感染组相比，阿苯达唑（ABZ）联合DLE后具有抑制能力的CD11b + CD19 - 髓样细胞比例降低，这对B细胞以及CD11b + F4/80MHCII巨噬细胞/单核细胞的升高最为有效（22.2±5.4%）。DLE及联合治疗后M2巨噬细胞标志物（精氨酸酶1、FIZZ - 1和Ym - 1）的转录本下调，但阿苯达唑治疗后未下调，而诱导型一氧化氮合酶（iNOS）则呈现相反趋势。这与DLE组和ABZ + DLE组中LPS刺激的PEC体外一氧化氮（NO）产生减少形成对比，在这些组中，黏附巨噬细胞/单核细胞的INF - γ受体和信号转导和转录激活因子1（STAT1）转录本升高，而信号转导和转录激活因子3（STAT3）、信号转导和转录激活因子6（STAT6）和白细胞介素10（IL - 10）的表达降低。每种治疗方法对干扰素 - γ（IFN - γ）、肿瘤坏死因子 - α（TNF - α）、白细胞介素12p40（IL - 12p40）、白细胞介素6（IL - 6）、白细胞介素10（IL - 10）和转化生长因子 - β（TGF - β）细胞因子的转录谱和浓度有不同的调节作用。DLE强烈改善了ABZ诱导的IFN - γ和IL - 12抑制，并维持了IL - 4、IL - 10和TGF - β的下调。对感染小鼠黏附巨噬细胞的表观遗传学研究表明，ABZ、ABZ - 亚砜和DLE可以剂量依赖性方式与检测标志物的mRNA相互作用。DLE与ABZ联合给药似乎通过调节免疫增强了药物的杀幼虫作用。与ABZ相比，联合治疗在通过刺激Th1细胞因子、M1巨噬细胞极化和IFNγ/STAT1信号通路激活来减轻寄生虫诱导的Th2/Treg/STAT3/STA6定向免疫抑制方面最为有效。
