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Monoclonal Antibodies Reactive with Immunorecessive Epitopes of Glucuronoxylomannan, the Major Capsular Polysaccharide of Cryptococcus neoformans

机译:与葡糖醛酸甘露聚糖(新隐球菌的主要荚膜多糖)的免疫反应性抗原决定簇反应的单克隆抗体

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Cryptococcus neoformans is surrounded by an antiphagocytic capsule whose primary constituent is glucuronoxylomannan (GXM). An epitope shared by GXM serotypes A, B, C, and D is immunodominant when mice are immunized with serotype A GXM. In contrast, an epitope shared only by serotypes A and D is immunodominant when mice are immunized with serotype D. Hybridomas secreting antibodies reactive with subdominant epitopes were identified through a positive-negative screening procedure in which antibody-secreting colonies were characterized by reactivity with both the immunizing polysaccharide and GXMs from each of the four major serotypes. In this manner, a monoclonal antibody (MAb) that was reactive with an epitope shared only by serotypes A and B was identified and designated F10F5. Such an epitope has not been described previously. Immunization of mice with de-O-acetylated serotype A GXM generated a hybridoma that secreted an antibody, designated F12D2, that was reactive with all four serotypes. Unlike previously described monoclonal and polyclonal panspecific antibodies, the reactivity of MAb F12D2 was not altered by de-O-acetylation of GXM. These results indicate that there are at least two panspecific GXM epitopes; one epitope is dependent on O acetylation for antibody reactivity, and the other is independent of O acetylation. This study identifies strategies for production of MAbs that are reactive with subdominant or cryptic GXM epitopes and provides new information regarding the antigenic makeup and the humoral immune response to GXM, an essential virulence factor that is a target for active and passive immunization.
机译:新型隐球菌被抗吞噬胶囊包围,其主要成分为葡糖醛酸甘露聚糖(GXM)。当用血清型A GXM免疫小鼠时,GXM血清型A,B,C和D共有的表位具有免疫优势。相反,当小鼠用血清型D免疫时,仅由血清型A和D共有的表位具有免疫优势。通过阳性-阴性筛选程序鉴定了分泌与主要表位具有反应性的抗体的杂交瘤,其中抗体分泌菌落的特征是与两种抗体都具有反应性四种主要血清型中的每一种的免疫多糖和GXM。以这种方式,鉴定了与仅由血清型A和血清型B共享的表位反应的单克隆抗体(MAb),并将其命名为F10F5。这样的表位先前没有描述。用去O-乙酰化血清型A免疫小鼠GXM产生了一种杂交瘤,该杂交瘤分泌了一种抗体,称为F12D2,可与所有四种血清型反应。与先前描述的单克隆和多克隆全特异性抗体不同,MAb F12D2的反应性不会因GXM的去O-乙酰化而改变。这些结果表明至少有两个全特异性GXM表位;一个表位的抗体反应性取决于O乙酰化,而另一个表位则与O乙酰化无关。这项研究确定了与主要或隐秘GXM表位反应的单克隆抗体的生产策略,并提供了有关抗原组成和对GXM的体液免疫反应的新信息,GXM是一种必需的毒力因子,是主动和被动免疫的目标。

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