Gizzarelli Manuela, Fiorentino Eleonora, Ben Fayala Nour El Houda, Montagnaro Serena, Torras Raquel, Gradoni Luigi, Oliva Gaetano, Foglia Manzillo Valentina
Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Naples, Italy.
Unit of Vector-Borne Diseases, Department of Infectious Diseases, Istituto Superiore di Sanità, Rome, Italy.
Front Vet Sci. 2020 May 19;7:273. doi: 10.3389/fvets.2020.00273. eCollection 2020.
Canine leishmaniasis (CanL) is a disease characterized by a large variety of clinical alterations, the majority of which being due to immune mediated mechanisms. Sick dogs usually produce high levels of -specific immunoglobulins which may give rise to circulating immune complexes (CICs) whose defective clearance by scavenging macrophages induces vasculitis and their deposition in specific organs. The aim of this study was to assess the serum level of CICs in dogs exposed to natural and experimental infection. Fifty-two sera were examined, belonging to untreated groups of naïve beagles previously studied to assess the performance of anti-leishmanial vaccines under natural (no. 22 dogs) or experimental (no. 30 dogs) transmission. Sera were classified in five groups according to the dog's health condition, IFAT titer, and the bone marrow (BM) nested (n)-PCR result. A: no.10 healthy dogs before the experimental infection; B: no.10 clinically healthy dogs infected experimentally, IFAT negative (= reciprocal titer <160) and n-PCR positive; C: no.10 clinically healthy dogs naturally infected, IFAT positive at titers 160-320 and n-PCR negative; D: no.10 sick dogs experimentally infected, IFAT positive at titer >320 and n-PCR positive; E: no.12 sick dogs naturally infected, IFAT positive at titer >320 and n-PCR positive. CICs levels were assessed by ELISA method (canine CIC assay-Cloude-Clone Corporation, USA). The two groups characterized by negative IFAT (A and B) had the lowest median level of CICs (16.09 and 12.78 μg/ml, respectively). CICs value increased progressively in the group C and reached the highest levels in the groups D and E, both characterized by high antibodies titer and severe disease, independently from the mode of infection. Significant differences in CICs concentration ( < 0.0001) were demonstrated between A, B, and C groups when compared with D or E groups of dogs. No differences were found inside the first three groups, while differences were recorded between the last two groups of sick dogs. CICs serum concentration increased with the progress of leishmaniasis, being significantly correlated with the increase of specific antibodies over time. High CICs levels detectable by commercial ELISA proved specific to an established infection in dogs in the absence of other concomitant infections, as demonstrated by the similar trend assessed in experimentally and naturally infected dogs.
犬利什曼病(CanL)是一种具有多种临床改变特征的疾病,其中大多数是由免疫介导机制引起的。患病犬通常会产生高水平的特异性免疫球蛋白,这可能导致循环免疫复合物(CICs)的产生,清除巨噬细胞对其清除功能的缺陷会诱导血管炎,并使其沉积在特定器官中。本研究的目的是评估自然感染和实验感染犬的血清CICs水平。对52份血清进行了检测,这些血清来自之前未经治疗的初免比格犬组,该组犬之前被用于评估抗利什曼原虫疫苗在自然(22只犬)或实验(30只犬)传播情况下的性能。根据犬的健康状况、间接荧光抗体试验(IFAT)滴度和骨髓(BM)巢式(n)-聚合酶链反应(PCR)结果,将血清分为五组。A组:实验感染前10只健康犬;B组:10只经实验感染的临床健康犬,IFAT阴性(=效价倒数<160)且n-PCR阳性;C组:10只自然感染的临床健康犬,IFAT滴度为160 - 320且n-PCR阴性;D组:10只经实验感染的患病犬,IFAT滴度>320且n-PCR阳性;E组:12只自然感染的患病犬,IFAT滴度>320且n-PCR阳性。通过酶联免疫吸附测定法(ELISA)(犬CIC检测试剂盒 - Cloude - Clone Corporation,美国)评估CICs水平。IFAT阴性的两组(A组和B组)CICs的中位数水平最低(分别为16.09和12.78μg/ml)。C组的CICs值逐渐升高,在D组和E组达到最高水平,这两组均具有高抗体滴度和严重疾病的特征,且与感染方式无关。与D组或E组犬相比,A、B和C组之间CICs浓度存在显著差异(<0.0001)。前三组内部未发现差异,而两组患病犬之间存在差异。CICs血清浓度随着利什曼病的进展而升高,与特异性抗体随时间的增加显著相关。如在实验感染和自然感染犬中评估的相似趋势所示,通过商业ELISA可检测到的高CICs水平证明在无其他伴随感染的情况下对犬已确诊的感染具有特异性。