title: HIV-1 viral DNA is present in ejaculated abnormal spermatozoa of seropositive subjects.
abstract: 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.
authors.affiliation: Department of Histology and Medical Embryology, Sapienza University of Rome, Rome, Italy.
authors.forename: B
authors.initials: B
authors.lastname: Muciaccia
authors.forename: S
authors.initials: S
authors.lastname: Corallini
authors.forename: E
authors.initials: E
authors.lastname: Vicini
authors.forename: F
authors.initials: F
authors.lastname: Padula
authors.forename: L
authors.initials: L
authors.lastname: Gandini
authors.forename: G
authors.initials: G
authors.lastname: Liuzzi
authors.forename: A
authors.initials: A
authors.lastname: Lenzi
authors.forename: M
authors.initials: M
authors.lastname: Stefanini
mesh_terms: D000328:Adult; D002469:Cell Separation; D053938:DNA Fragmentation; D004279:DNA, Viral / Q000378:metabolism*; D005434:Flow Cytometry; D006679:HIV Seropositivity / Q000378:metabolism* / Q000821:virology*; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008297:Male; D008875:Middle Aged; D009995:Osmosis; D011205:Povidone / Q000494:pharmacology; D012661:Semen / Q000821:virology*; D012822:Silicon Dioxide / Q000494:pharmacology; D013094:Spermatozoa / Q000821:virology*
chemical_list: D004279:DNA, Viral; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone