Manente Francine Alessandra, Quinello Camila, Ferreira Lucas Souza, de Andrade Cleverton Roberto, Jellmayer Juliana Aparecida, Portuondo Deivys Leandro, Batista-Duharte Alexander, Carlos Iracilda Zeppone
School of Pharmaceutical Sciences, São Paulo State University - UNESP, Araraquara, SP, Brazil.
Med Mycol. 2018 Aug 1;56(6):711-722. doi: 10.1093/mmy/myx098.
This report describes a model of host resistance for Sporothrix schenckii, an opportunistic fungi in immunosuppressed mice with cyclophosphamide (CY) to be used in studies of immunotoxicology and immunopharmacology. Two doses of CY were administered intraperitoneally: 200 mg/kg and a booster of 150 mg/kg at 9-day intervals. Three days after the first dose of CY the animals were infected subcutaneously with 1.8 × 108 yeast/ml (S. schenckii ATCC 16345). At 7 and 14 days post-infection, the animals were euthanized and analyzed the fungal load by unit forming colony count in the spleen and popliteal lymph nodes. The relative weight of thymus and spleen, splenic index, the frequency of T and B cells in spleen by flow cytometry, the hind paw inflammation index and cytokine (interleukin [IL]-17, IL-10, and interferon [IFN]-γ) profile were measured. Histopathological studies of the spleen and the hind paw were also assessed. The immunosuppression status was confirmed at the evaluated days by reduction of relative weight of thymus, reduction of the splenic white pulp, the population of B and T lymphocytes, and the cytokine profile in the treated mice with CY in comparison with nontreated groups, associated to higher fungal load in hind paw and spleen in the infected mice. The described model reveals an increasing in susceptibility to infection and severity when associated with immunosuppression. This model can serve as a reference for studies of S. schenckii host resistance in pharmaceutical and toxicological studies.
本报告描述了一种用于申克孢子丝菌的宿主抗性模型，该菌是一种机会性真菌，用于在免疫抑制小鼠中使用环磷酰胺（CY）进行免疫毒理学和免疫药理学研究。腹腔注射两剂CY：200mg/kg，9天后再注射150mg/kg的加强剂量。在首次注射CY后3天，将动物皮下感染1.8×108酵母/ml（申克孢子丝菌ATCC 16345）。在感染后7天和14天，对动物实施安乐死，并通过脾脏和腘窝淋巴结中的单位形成菌落计数分析真菌载量。测量胸腺和脾脏的相对重量、脾脏指数、通过流式细胞术检测脾脏中T细胞和B细胞的频率、后爪炎症指数以及细胞因子（白细胞介素[IL]-17、IL-10和干扰素[IFN]-γ）谱。还评估了脾脏和后爪的组织病理学研究。与未处理组相比，通过CY处理的小鼠胸腺相对重量降低、脾脏白髓减少、B和T淋巴细胞数量减少以及细胞因子谱变化，证实了在评估日的免疫抑制状态，这与感染小鼠后爪和脾脏中较高的真菌载量相关。所描述的模型揭示了与免疫抑制相关时对感染的易感性和严重程度增加。该模型可作为药物和毒理学研究中申克孢子丝菌宿主抗性研究的参考。
