Development and validation of monoclonal antibodies specific for Candida albicans Als2, Als9-1, and Als9-2.
Development and validation of monoclonal antibodies specific for Candida albicans Als2, Als9-1, and Als9-2.
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DOI：
10.1371/journal.pone.0269681
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发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
真菌凝集素样序列（Als）细胞表面糖蛋白，最好的特点是在白色念珠菌，介导粘附和聚集与宿主细胞，其他微生物和非生物表面的相互作用。对每种C. Als蛋白是研究Als蛋白定位和功能的重要试剂。本文描述了C. albicans Als 2和C.白色念珠菌Als 9 -1和Als 9 -2，由ALS 9基因座产生的两种蛋白变体。印第安人角白色念珠菌ALS 9表达水平不足以在野生型细胞表面上产生可检测的Als 9蛋白，因此MAb验证需要产生过表达菌株，每个菌株具有两个ALS 9等位基因之一。针对N-糖基化形式的蛋白质免疫原以及内切糖苷酶H处理的免疫原产生抗Als 2 MAb。抗N-糖基化免疫原的单克隆抗体证明具有上级和免疫标记的C.白色念珠菌酵母细胞和芽管，以及都柏林念珠菌和热带念珠菌酵母的表面。Als 2在C.从口腔念珠菌病的鼠模型中回收的白色念珠菌酵母细胞，证明了体内和体外的Als 2产生。这些新的单克隆抗体增加了抗Als单克隆抗体的收集，这些单克隆抗体是更好地理解Als蛋白在C.白色念珠菌的生物学和发病机制。
Fungal agglutinin-like sequence (Als) cell-surface glycoproteins, best characterized in Candida albicans, mediate adhesive and aggregative interactions with host cells, other microbes, and abiotic surfaces. Monoclonal antibodies (MAbs) specific for each C. albicans Als protein are valuable reagents for gaining insight into Als protein localization and function. This manuscript describes development and validation of MAbs specific for C. albicans Als2, as well as for C. albicans Als9-1 and Als9-2, two protein variants produced from the ALS9 locus. Native C. albicans ALS9 expression levels were not sufficiently high to produce detectable Als9 protein on the wild-type cell surface so MAb validation required production of overexpression strains, each featuring one of the two ALS9 alleles. An anti-Als2 MAb was raised against an N-glycosylated form of the protein immunogen, as well as an Endoglycosidase H-treated immunogen. The MAb raised against the N-glycosylated immunogen proved superior and immunolabeled C. albicans yeast cells and germ tubes, and the surface of Candida dubliniensis and Candida tropicalis yeasts. Als2 was visible on C. albicans yeast cells recovered from a murine model of oral candidiasis, demonstrating Als2 production both in vivo and in vitro. These new MAbs add to the collection of anti-Als MAbs that are powerful tools to better understand the role of Als proteins in C. albicans biology and pathogenesis.
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影响因子：
5.2
作者：
Hoyer LL;Cota E
通讯作者：
Cota E
通过催化蛋白激酶A亚基TPK1对白色念珠菌细胞壁的功能控制。
影响因子：
3.6
作者：
Fanning S;Xu W;Beaurepaire C;Suhan JP;Nantel A;Mitchell AP
通讯作者：
Mitchell AP
影响因子：
5.7
作者：
Oh SH;Schliep K;Isenhower A;Rodriguez-Bobadilla R;Vuong VM;Fields CJ;Hernandez AG;Hoyer LL
通讯作者：
Hoyer LL
影响因子：
7
作者：
Jackson, Andrew P.;Gamble, John A.;Berriman, Matthew
通讯作者：
Berriman, Matthew
影响因子：
10.7
作者：
Maguire SL;ÓhÉigeartaigh SS;Byrne KP;Schröder MS;O'Gaora P;Wolfe KH;Butler G
通讯作者：
Butler G