Muciaccia B, Corallini S, Vicini E, Padula F, Gandini L, Liuzzi G, Lenzi A, Stefanini M
Department of Histology and Medical Embryology, Sapienza University of Rome, Rome, Italy.
Hum Reprod. 2007 Nov;22(11):2868-78. doi: 10.1093/humrep/dem288. Epub 2007 Sep 12.
Semen is the major vehicle for HIV-1 infection as it contains free and cell-associated virions and infected cells. However, the presence of HIV-1 in spermatozoa has been a matter of debate, since the sperm cell fraction may contain somatic infected cells that jeopardize the attribution of the detected virus to the spermatozoa.
Spermatozoa from 12 HIV-1 seropositive subjects were purified by multilayered Percoll gradient followed by osmotic shock. Residual presence of non-seminal cells (NCS) in purified spermatozoa, was then evaluated by cytometric and molecular analysis. HIV-1 DNA was revealed by nested PCR and in situ PCR after sperm chromatin decondensation. DNA-fragmented ejaculated spermatozoa in semen of infected subjects were detected by terminal deoxynucleotidyl transferase-mediated dUDP nick-end labeling (TUNEL) analysis.
Purification procedure adopted allowed complete removal of NCS. On purified sperm cells, HIV-1 DNA was detected in 5 out of 12 subjects by nested-PCR. On crude semen of 10 out of 12 subjects, HIV-1 DNA was in situ detected in a small percentage of abnormal spermatozoa with a wide range of structural alterations. TUNEL analysis revealed an increased percentage of DNA-fragmented ejaculated spermatozoa in semen of infected subjects.
We report molecular evidence demonstrating that HIV-1 infected subjects can ejaculate small amounts of HIV-1 DNA-positive abnormal spermatozoa. Their possible role in HIV-1 sexual transmission remains to be clarified.
精液是HIV-1感染的主要载体,因为它含有游离的和与细胞相关的病毒粒子以及被感染的细胞。然而,HIV-1在精子中的存在一直存在争议,因为精子细胞部分可能包含体细胞感染细胞,这会影响将检测到的病毒归因于精子。
通过多层Percoll梯度离心随后进行渗透休克,对12名HIV-1血清阳性受试者的精子进行纯化。然后通过细胞计数和分子分析评估纯化精子中非生殖细胞(NCS)的残留情况。在精子染色质解聚后,通过巢式PCR和原位PCR检测HIV-1 DNA。通过末端脱氧核苷酸转移酶介导的dUDP缺口末端标记(TUNEL)分析检测感染受试者精液中DNA片段化的射精精子。
采用的纯化程序能够完全去除NCS。通过巢式PCR在12名受试者中的5名的纯化精子细胞上检测到HIV-1 DNA。在12名受试者中的10名的粗精液中,在一小部分具有广泛结构改变的异常精子中原位检测到HIV-1 DNA。TUNEL分析显示感染受试者精液中DNA片段化的射精精子百分比增加。
我们报告了分子证据,证明HIV-1感染的受试者可以射出少量HIV-1 DNA阳性的异常精子。它们在HIV-1性传播中的可能作用仍有待阐明。