Apoptosis, inflammatory response and parasite load in skin of Leishmania (Leishmania) chagasi naturally infected dogs: A histomorphometric analysis
Apoptosis, inflammatory response and parasite load in skin of Leishmania (Leishmania) chagasi naturally infected dogs: A histomorphometric analysis
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DOI：
10.1016/j.vetpar.2012.04.035
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发表时间：
2012-10-26
影响因子：
2.6
通讯作者：
Vasconcelos, Anilton Cesar
中科院分区：
文献类型：
--
作者：
Araujo Vercosa, Barbara Laurice;Melo, Maria Norma;Vasconcelos, Anilton Cesar
皮肤在恰加斯利什曼原虫感染中起重要作用。细胞凋亡调节炎症反应，对病变的进展或消退起独特作用。寄生虫与多种调节系统相互作用，诱导宿主细胞凋亡，在细胞入侵，稳定和繁殖的病原体。在这种情况下，本研究的目的是评估细胞死亡内的炎症浸润，并将这些结果与寄生虫负荷和临床特征的狗自然感染L恰加斯。皮肤耳廓碎片（8个有症状的+8个无症状的+6个阴性对照）用于表征和测量炎症反应、寄生虫负荷和细胞凋亡。通过IFA和ELISA检测血清中的抗利什曼原虫抗体，直接观察寄生虫和脾、肝、耳廓、骨髓和淋巴结中的培养物，以及PCR（耳廓）来确认犬利什曼病的诊断。在光学显微镜下，用从20个代表性组织学视野获得的图像进行组织形态计量学。将超薄切片固定在300目网格上，与2%乙酸双氧铀和柠檬酸铅对照，并在透射电子显微镜下检查。无鞭毛体仅在有症状动物的皮肤中发现（31.94 +/- 18.81）。有症状的犬中炎性浸润的病灶数量和细胞结构高于其他组，无症状犬中炎性浸润的病灶数量和细胞结构高于对照组（p < 0.05; Tukey）。在有症状的狗中获得的炎性浸润物的平均面积、周长和最大直径高于对照组（p < 0.05; Tukey）。有症状组的凋亡指数高于其他组，无症状组和对照组之间没有差异（p < 0.05; Tukey）。超微结构上，凋亡细胞皱缩，核染色质和胞浆浓缩。凝聚的核经常碎裂。核小体间DNA断裂仅发生在有症状的病例中。在中性粒细胞和巨噬细胞内观察到无鞭毛体。在自然感染的犬中，细胞凋亡与寄生虫负荷、炎症反应强度和临床表现直接相关。(C)2012爱思唯尔有限公司版权所有。
The skin has an important role in infection by Leishmania chagasi. Apoptosis modulates the inflammatory response acting distinctively either on the progression or regression of the lesions. The parasites interact with multiple regulatory systems inducing apoptosis in host cells, during cell invasion, stabilization and multiplication of pathogens. In this context, the aim of this study was to evaluate cell death within the inflammatory infiltrates, and to correlate these results with parasite load and clinical features of dogs naturally infected with L chagasi. Fragments of skin pinnas (8 symptomatic + 8 asymptomatic + 6 negative controls) were used to characterize and measure the inflammatory response, parasite load and apoptosis. Diagnosis of canine leishmaniasis was confirmed by the detection of anti-Leishmania antibodies by IFA and ELISA in serum, direct visualization of the parasite and culture in spleen, liver, pinna, bone marrow and lymph nodes, and PCR (pinna). Histomorphometry was performed with images obtained from 20 representative histological fields in a light microscope. Ultra-thin sections were mounted over a 300 mesh grids, contrasted with 2% uranyl acetate and lead citrate and examined under a Transmission Electronic Microscopy. Amastigotes were only found in the skin of symptomatic animals (31.94 +/- 18.81). The number of foci and cellularity of the inflammatory infiltrates in symptomatic dogs were higher than in other groups and in asymptomatics were higher than in controls (p < 0.05; Tukey). The average area, perimeter and extreme diameters of the inflammatory infiltrates obtained in symptomatic dogs were higher than in controls (p < 0.05; Tukey). The apoptotic index was higher in symptomatic than in other groups and there was no difference between asymptomatics and controls (p < 0.05; Tukey). Ultrastructurally, apoptotic cells were shrunken, with condensed nuclear chromatin and cytoplasm. Condensed nuclei were frequently fragmented. Internucleosomal DNA fragmentation occurred only in symptomatic cases. Amastigotes were observed within neutrophils and macrophages. Apoptosis is directly related to parasite load, intensity of inflammatory response and clinical manifestations in L chagasi naturally infected dogs. (C) 2012 Elsevier B.V. All rights reserved.