Grange Philippe A, Marcelin Anne-Geneviève, Calvez Vincent, Chauvel Caroline, Escande Jean-Paul, Dupin Nicolas
Laboratoire de Microbiologie Cutanée, Pavillon Tarnier, Groupe Hospitalier Cochin-Port Royal, Université Paris V, Paris, France.
J Invest Dermatol. 2005 Jun;124(6):1249-58. doi: 10.1111/j.0022-202X.2005.23756.x.
Human herpesvirus-8 (HHV-8) is commonly detected in all epidemiologic forms of Kaposi's sarcoma. Despite the broad cellular tropism of HHV-8, studies on mucosal shedding of HHV-8 have shown that infectious particles are restricted to saliva isolated from the oropharynx. We used biotinylated purified HHV-8 particles in a direct binding assay to whole clarified human salivary samples isolated from HHV-8-infected and uninfected individuals. We found that the major binding activity was carried out by a protein of 78-kDa size, which was further characterized as human lactoferrin (hLf) using 2-D electrophoresis and MALDI-ToF analysis. Preliminary comparison of HHV-8 binding activity of 76 salivary samples from HHV-8-infected and uninfected individuals showed that 7.8% of the uninfected population exhibited a form of Lf not recognized by HHV-8. Deglycosylation of hLf by PNGase F did not reduce HHV-8 binding activity, whereas endoproteinase cleavage of native hLf generated a non-glycosylated 8-kDa peptide recognized by HHV-8 particles and was located at the position Ala606-Tyr679 in the native hLf amino acid sequence, corresponding to the C-terminal region of the glycoprotein. This work identify the lactoferrin in saliva as a ligand for HHV-8 and suggests that this glycoprotein could be used as a carrier for the viral particles.
人类疱疹病毒8型（HHV - 8）在卡波西肉瘤的所有流行病学形式中均普遍被检测到。尽管HHV - 8具有广泛的细胞嗜性，但关于HHV - 8黏膜脱落的研究表明，感染性颗粒仅限于从口咽部分离的唾液。我们在直接结合试验中使用生物素化的纯化HHV - 8颗粒，与从HHV - 8感染和未感染个体中分离的全澄清人类唾液样本进行结合。我们发现主要的结合活性由一种78 kDa大小的蛋白质执行，使用二维电泳和基质辅助激光解吸电离飞行时间分析（MALDI - ToF）进一步鉴定为人类乳铁蛋白（hLf）。对来自HHV - 8感染和未感染个体的76份唾液样本的HHV - 8结合活性进行初步比较，结果显示7.8%的未感染人群表现出一种HHV - 8无法识别的Lf形式。用PNGase F对hLf进行去糖基化处理并未降低HHV - 8的结合活性，而天然hLf经内蛋白酶切割产生了一种能被HHV - 8颗粒识别的非糖基化8 kDa肽段，该肽段位于天然hLf氨基酸序列中的Ala606 - Tyr679位置，对应糖蛋白的C末端区域。这项工作确定唾液中的乳铁蛋白是HHV - 8的一种配体，并表明这种糖蛋白可作为病毒颗粒的载体。
