Chaves Luciana Botelho, Achkar Samira Maria, Silva Andréa de Cássia Rodrigues da, Caporale Graciane Maria Medeiros, Cruz Paula Sônia, Batista Alexandre Mendes, Scheffer Karin Corrêa, Fernades Irene, Carrieri Maria Luiza, De Gaspari Elizabeth N
Instituto Pasteur, São Paulo, SP, Brasil.
J Infect Dev Ctries. 2015 Nov 30;9(11):1238-49. doi: 10.3855/jidc.6959.
In Brazil, various isolates of rabies virus (RABV) show antigenic profiles distinct from those established by the reduced panel of eight monoclonal antibodies (MAbs) determined by the Centers for Disease Control and Prevention (CDC), utilized for the antigenic characterization of RABV in the Americas. The objective of this study was to produce MAbs from RABV isolates from insectivorous bats with an antigenic profile incompatible with the pre-established one.
An isolate of RABV from the species Eptesicus furinalis that showed an antigenic profile incompatible with the panel utilized was selected. Hybridomas were produced utilizing the popliteal lymph nodes of mice immunized with ribonucleoproteins purified from the isolate.
Two MAbs-producing clones were obtained, BR/IP1-3A7 and BR/IP2-4E10. Fifty-seven isolates of RABV from different species of animals and different regions of Brazil were analyzed utilizing the MAbs obtained. In the analysis of 23 RABV isolates from non-hematophagous bats, the MAbs cross-reacted with ten isolates, of which four were of the species Nyctinomops laticaudatus, one of the species Eptesicus furinalis, and five of the genus Artibeus. Of the nine isolates of non-hematophagous isolates that displayed an incompatible profile analyzed, characteristic of insectivorous bats, BR/IP1-3A7 reacted with five (55.55%) and BR/IP2-4E10 with four (44.44%).
The MAbs obtained were able to recognize epitopes common between the three genera, Artibeus, Eptesicus, and Nyctinomops, thereby allowing the antigenic characterization of RABV isolates in Brazil.
在巴西,狂犬病病毒(RABV)的各种分离株显示出与美国疾病控制与预防中心(CDC)确定的用于美洲RABV抗原特性鉴定的八种单克隆抗体(MAb)简化组合所确定的抗原谱不同。本研究的目的是从食虫蝙蝠的RABV分离株中制备抗原谱与预先确定的抗原谱不兼容的MAb。
选择一株来自棕蝠(Eptesicus furinalis)的RABV分离株,其抗原谱与所用组合不兼容。利用从该分离株中纯化的核糖核蛋白免疫小鼠的腘窝淋巴结制备杂交瘤。
获得了两个产生MAb的克隆,即BR/IP1-3A7和BR/IP2-4E10。利用获得的MAb对来自巴西不同动物物种和不同地区的57株RABV分离株进行了分析。在对23株非吸血蝙蝠的RABV分离株的分析中,这些MAb与10株分离株发生交叉反应,其中4株属于大耳蝠属(Nyctinomops laticaudatus),1株属于棕蝠属(Eptesicus furinalis),5株属于叶口蝠属(Artibeus)。在所分析的9株显示出不兼容谱的非吸血分离株中,具有食虫蝙蝠特征的,BR/IP1-3A7与5株(55.55%)反应,BR/IP2-4E10与4株(44.44%)反应。
获得的MAb能够识别叶口蝠属、棕蝠属和大耳蝠属这三个属之间共有的表位,从而能够对巴西的RABV分离株进行抗原特性鉴定。