Inhibition of Candida albicans adhesion by recombinant human antibody single-chain variable fragment specific for Als3p.
Inhibition of Candida albicans adhesion by recombinant human antibody single-chain variable fragment specific for Als3p.
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DOI：
10.1111/j.1574-695x.2008.00465.x
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发表时间：
2008-11
影响因子：
--
通讯作者：
Bliss, Joseph M.
中科院分区：
文献类型：
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作者：
Laforce-Nesbitt, Sonia S.;Sullivan, Mark A.;Hoyer, Lois L.;Bliss, Joseph M.
白念珠菌粘附素Als 3 p被鉴定为先前描述的人抗体片段（scFv）的潜在同源抗原，这是基于scFv的结合模式与该蛋白在菌丝表面上的分布的相似性。尽管所有的scFv都与野生型强烈结合，但通过免疫荧光测定（IFA），scFv 3显示与含有纯合缺失的ALS 3的菌株1843没有可检测的结合。保留了与ALS 3重组菌株2322的结合，并且scFv 3也与S.酿酒酵母表达ALS 3。其他scFv保留与1843的结合，但具有显著改变的模式。为了确定scFv 3是否可以干扰Als 3 p功能，使用人上皮或内皮细胞作为靶标进行粘附测定。野生型C.具有scFv 3的白色念珠菌将真菌对两种细胞类型的粘附降低至与als 3 Δ/als 3 Δ突变体相当的水平。这些实验证实噬菌体展示是分离对涉及C.白念珠菌毒力，并且scFv干扰对人细胞的粘附。保留与als 3 Δ/als 3 Δ突变体结合的其他scFv的免疫染色模式改变表明Als 3 p也可能在C.白色念珠菌细胞表面。
The Candida albicans adhesin, Als3p, was identified as a potential cognate antigen for previously described human antibody fragments (scFv) based on similarity of the binding pattern of the scFv to the distribution of this protein on the hyphal surface. Although all scFv bound avidly to wild-type, scFv3 showed no detectable binding via immunofluorescence assay (IFA) to strain 1843, containing a homozygous deletion of ALS3. Binding to the ALS3 reintegrant strain, 2322, was preserved, and scFv3 also bound to S. cerevisiae expressing ALS3. Other scFv retained binding to 1843, but with a markedly altered pattern. To determine if scFv3 could interfere with Als3p function, adhesion assays were conducted using human epithelial or endothelial cells as target. Treatment of wild-type C. albicans with scFv3 reduced adhesion of the fungus to both cell types to levels comparable to the als3Δ/als3Δ mutant. These experiments confirm that phage display is a viable method to isolate human scFv specific to an antigen implicated in C. albicans virulence, and that the scFv interfere with adhesion to human cells. The altered pattern of immunostaining with other scFv that retain binding to the als3Δ/als3Δ mutant suggest that Als3p may also have a role in structural organization of the C. albicans cell surface.