Bagasra Omar, Patel Deepa, Bobroski Lisa, Abbasi Jamil A, Bagasra Alex U, Baidouri Hasna, Harris Twaina, El-Roeiy Albert, Lengvarszky Zsolt, Farzadegan Homayoon, Wood Charles
South Carolina Center for Biotechnology, Department of Biology, Claflin University, 400 Magnolia Street, Orangeburg, SC 29115, USA.
J Mol Histol. 2005 Sep;36(6-7):401-12. doi: 10.1007/s10735-005-9010-9. Epub 2006 Jan 10.
Defining the mechanism of infection with human herpesvirus-8 (HHV-8) or Kaposi's sarcoma associated herpesvirus (KSHV) is an important clinical issue. HHV-8 has been linked to Kaposi's sarcoma (KS) development in HIV-1-infected individuals, and KS develops in 40% of those infected with both viruses. A series of epidemiological data suggest that sexual transmission is one of the routes of transmission for HHV-8. In our studies, we sought to assess the cellular reservoirs of HHV-8 DNA in the semen of HIV-1-infected men and the potential role of HHV-8 infected spermatozoa in horizontal transmission.
A nested polymerase chain reaction (PCR), in situ PCR (ISPCR) and a sodium iodide (NaI) DNA isolation technique that extracts both nuclear and episomal DNA were utilized to amplify specific genes in vitro and within intact cells to evaluate the types of seminal cells infected with HHV-8 in HIV-1-infected and uninfected men.
HHV-8 was present in the spermatozoa and mononuclear cells of the semen in 64 of 73 (88%) HIV-1 infected individuals. Both the sperms as well as the mononuclear cells of the semen specimens of HIV-1 infected men were found to be infected with HHV-8. Multiplex ISPCR revealed that a significantly higher percentage of semen cells were infected with HHV-8 than HIV-1 (p>0.001). Rare (less than one in a 100,000) sperm cells were co-infected with both viruses. A co-culture of HHV-8 infected sperm with uninfected 293 or Sup-T1 cell lines resulted in an abortive infection of these cells with HHV-8. DNA isolation by NaI yielded 73% of the positive sperm, whereas the standard phenol/chloroform method resulted in significantly lower positives (45%) from the same specimens.
Design and methods: Our data strongly suggest a potential sexual/horizontal route of transmission of HHV-8, via the HHV-8 infected sperm and other semen cells, where a large percentage of HIV-1 infected men's sperm and other semen cells are infected with HHV-8. Co-culture studies have further supported the observations that HHV-8 in the sperm cells is infectious and capable of transmission of the virus to uninfected cells.
确定人类疱疹病毒8型（HHV-8）或卡波西肉瘤相关疱疹病毒（KSHV）的感染机制是一个重要的临床问题。HHV-8与HIV-1感染个体的卡波西肉瘤（KS）发展有关，在同时感染这两种病毒的个体中，40%会发展为KS。一系列流行病学数据表明，性传播是HHV-8的传播途径之一。在我们的研究中，我们试图评估HIV-1感染男性精液中HHV-8 DNA的细胞储存库以及HHV-8感染的精子在水平传播中的潜在作用。
采用巢式聚合酶链反应（PCR）、原位PCR（ISPCR）以及一种能提取核DNA和游离型DNA的碘化钠（NaI）DNA分离技术，在体外和完整细胞内扩增特定基因，以评估HIV-1感染和未感染男性精液中感染HHV-8的精细胞类型。
在73名HIV-1感染个体中，有64名（88%）的精液精子和单核细胞中存在HHV-8。发现HIV-1感染男性精液标本中的精子以及单核细胞均感染了HHV-8。多重ISPCR显示，感染HHV-8的精液细胞百分比显著高于感染HIV-1的细胞（p>0.001）。极少（每10万个中少于1个）精子细胞同时感染了这两种病毒。将HHV-8感染的精子与未感染的293或Sup-T1细胞系共培养，导致这些细胞被HHV-8进行了顿挫感染。用NaI进行DNA分离得到73%的阳性精子，而标准的酚/氯仿法从相同标本中得到的阳性率显著较低（45%）。
设计与方法：我们的数据有力地表明，HHV-8存在一种潜在的性/水平传播途径，即通过HHV-8感染的精子和其他精液细胞传播，其中很大比例的HIV-1感染男性的精子和其他精液细胞感染了HHV-8。共培养研究进一步支持了以下观察结果：精子细胞中的HHV-8具有传染性，能够将病毒传播给未感染的细胞。
