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Hum. Reprod. Advance Access published online on September 12, 2007
Human Reproduction, doi:10.1093/humrep/dem288
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© The Author 2007. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

HIV-1 viral DNA is present in ejaculated abnormal spermatozoa of seropositive subjects

B. Muciaccia1, S. Corallini1, E. Vicini1, F. Padula1, L. Gandini2, G. Liuzzi3, A. Lenzi2 and M. Stefanini1,4

1 Department of Histology and Medical Embryology, Sapienza University of Rome, Rome, Italy 2 Department of Medical Physiopathology, Sapienza University of Rome, Rome, Italy 3 National Institute for Infectious Disease Spallanzani (INMI), Rome, Italy

4 Correspondence address. Tel: +39(0)6 4976 6570; Fax: +39(0)6 4462 854; E-mail: mario.stefanini{at}uniroma1.it

BACKGROUND: 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.

METHODS: 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.

RESULTS: 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.

CONCLUSIONS: 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.

Key words: osmotic shock/sperm chromatin decondensation/in situ PCR/HIV-1 nested PCR/Alu-LTR PCR

Submitted on January 21, 2007; resubmitted on July 27, 2007; accepted on August 21, 2007.


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