Placenta
Volume 29, Supplement 2 , Pages 160-165, October 2008

Reproductive Assistance for Infected Couples with Bloodborne Viruses

Department of Obstetrics and Gynaecology, Sacco Clinical Sciences Institute, University of Milan Medical School, Via GB Grassi 74, 20157 Milan, Italy

Accepted 7 August 2008.

Article Outline

Abstract 

During the past 15 years Assisted Reproduction has been facing a new demand from patients requiring ART: couples at risk of partner to partner, and mother to child transmission of viral infections, mainly HIV-1, HCV and HIV–HCV co-infected partners. The general conditions and life expectancy of many patients with HIV infection are very good, and three-quarters of these individuals are in their reproductive years. For these reasons, a large number of young couples are expected to make future plans to have children. This desire is not easy to realize for serodiscordant couples, if we consider that, in order to avoid HIV virus transmission, it is necessary to encourage the condom use in vaginal and anal contacts. On the other hand infertile discordant HCV couples need to be included in protocols of controlled assisted reproduction procedures to avoid any risk of HCV transmission to the partner. In this paper we consider assisted reproduction in discordant couples for HIV or HCV-positive men.

Keywords: Couple serodiscordant, ART, HIV virus, HCV virus

 

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1. Introduction 

During the past 15 years, assisted reproduction has been facing a new demand from patients requiring assisted reproductive technique (ART): couples at risk of partner-to-partner viral infection, and mother-to-child transmission of viral infections – mainly HIV-1, HCV and HIV–HCV co-infected partners. The request for reproductive assistance has been mainly for two medical reasons: either to overcome an infertility problem or to decrease the risk of horizontal transmission. This is directly correlated to three different scenarios: both partners being infected, female-only infection and male-only infection.

When it is the woman who is HIV-infected or HCV-infected, irrespective of the partner's infectious status, we should consider not only the reproductive problem but also the risk of, and factors associated to, vertical transmission which is beyond the scope of this paper.

In this paper we describe the available ART options for discordant couples for HIV or HCV male positivity specifically to intrauterine insemination (IUI), in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) according to their fertility need. It is beyond the scope of this paper to define the reproductive choice for each ART technique in the general population, but the primary goal of assisting discordant couples is to protect the uninfected woman from viral transmission and consequently remove the risk of vertical transmission.

Most of the published reports concern gamete banking and donation but also cross-contamination between patients during assisted reproductive techniques. Hepatitis B contamination has been described during artificial insemination [1] as well as HCV cross-contamination between patients during assisted conception [2] and during artificial insemination [3]. HIV infection in artificial insemination with donor semen has been described in Australia [4], Canada, the US [5], [6], [7] and Germany [8]. These accidents demonstrated that sperm alone, independent of any sexual contact, can transmit the virus and this risk can be comparable to that from occasional sexual intercourse (4/8 (50%) in Stewart et al. [4] and 7/199 (3.52%) in Araneta et al. [6]). Nowadays, patients who are virus carriers are considered as higher risk and thus a systematic screening for HIV, HBV and HCV may be performed in assisted reproduction programmes [9], [10]. The need for sperm donors to be screened and for sperm to be frozen and placed in quarantine for a period of 6 months, has been stressed in national [11], [12], [13], [14] and in European [15] recommendations. Although sperm can be used for ART after the quarantine period, this is possible when a new test is carried out on the donor with a negative result.

According to the latest United Nations Joint Programme on HIV (UNAIDS)/World Health Organization (WHO) [16] update (December 2006), the total number of people living with HIV-1 infection had reached 39.5 million. In developed countries, the major risk for HIV-1 transmission is by heterosexual intercourse and the number of cases is increasing [17]. In the past, drug users were considered the most important reservoir for the virus, with the majority of them being men. For these reasons, today, women are considered at higher risk for infection by heterosexual intercourse. In the era of highly active anti-retroviral therapy (HAART), life expectancy for HIV-infected patients has dramatically improved, they are feeling well and they know that the risk of vertical transmission has decreased to less than 1%. Three-quarters of these individuals are in their reproductive years and it is therefore expected that a large number of young couples will make future plans to have children [18]. This desire is not easy to realize for serodiscordant couples if we consider that, in order to avoid HIV virus transmission, it is necessary to encourage condom use in vaginal and anal contacts. Many of them now express the desire for parenthood as a fundamental part of healthy family life and return to normality. Published guidelines from the American Society for Reproductive Medicine (ASRM) (2002) and the American College of Obstetrics and Gynecological Committee on Ethics (2001), recommendations concerning assisted reproduction on people infected by HIV, have been modified to allow assisted reproduction in HIV-serodiscordant couples in order to encourage a policy of non-discrimination for these couples.

Hepatitis C virus (HCV) infection is the most common chronic bloodborne virus in developed countries, with an average of twenty three thousand new infections occurring each year (CDC, unpublished data). The estimated worldwide prevalence of HCV infection is 2.2% [19], and 1.5% in Europe [20], [21], [22]. In assisted reproduction, HCV transmission may pose a risk for the newborn, for gametes or embryos from non-contaminated parents and also for technicians. On the other hand, infertile discordant HCV couples need to be included in protocols of controlled assisted reproduction procedures to avoid any risk of HCV transmission to the partner. International guidelines should be developed through studies on larger populations of HCV chronically infected individuals [23], [24]. For example, until 1999 in France, assisted reproduction in HCV discordant couples has been denied [25] and only in 2001 French legislation has dictated national rules that require the systematic testing of seminal plasma for the presence of HCV RNA in HCV-infected men during ART [26].

The most important point is that the laboratory used for assisted reproductive technology was considered a ‘viral risk’ area, separated from laboratory facilities used for couples negative for HIV, hepatitis B virus (HBV) and hepatitis C virus (HCV). The ART laboratory complied with standard recommended safety precautions [16]. Specific precautions were implemented against the risk of HIV, HCV and HBV contamination as recommended by the French decree of May 10, 2001 [27], and the potentially infected gametes and embryos were handled separately. A special biosafety cabinet workstation was used for all tasks that involved handling sperm, oocytes and embryos.

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2. Virus in the semen 

2.1. HIV in semen 

Araneta et al. [6] and Matz et al. [8] reported that semen used for donor artificial inseminations can transmit HIV-1 infection. In the past, studies on the presence of HIV in sperm had also yielded contradictory results. Using different approaches, Baccetti et al. [28] detected HIV-1 particles and HIV-1 DNA in ejaculated sperm of HIV-seropositive patients; the same group identified an alternative receptor to CD4+, on sperm membrane: this molecule is a galactosyl-alkyl-acylglycerol (GalAAG), a glycolipid structurally related to galactosylceramide, the receptor for HIV identified in CD4+ cells [18], [29]. On the contrary, other authors emphasized the total absence of HIV particles and nucleic acids in the sperm [30], [31], [32], demonstrating that separation of seminal fluid and cellular elements from the sperms, by washing techniques, reduces the viral load of semen detected by PCR and RT-PCR. Semprini et al. [33] were the first to use washed sperm of HIV-1-infected men for intrauterine inseminations (IUI). There are several reports indicating that HIV-1 DNA cannot be found in washed spermatozoa isolated from nonspermatozoa seminal cells and seminal plasma [31], [34], [35]. In contrast to this reassuring epidemiological and laboratory background, there are reports indicating the possibility that HIV-1 virions are found attached to the sperm cells surface and even within its cytoplasm, by transmission electronic microscopy technique. Other papers report that proviral HIV-1 DNA can be detected in spermatozoa of men with AIDS, by extraction PCR, or found in spermatozoa of men infected with HIV-1, by in-situ PCR (IS-PCR) [28], [36], [38], [39], [40], [41], [42], [43], [44], [45], [46], [47], [48], [49], [50].

To investigate these contradictory findings and to assess the role of sperm-washing technique in eliminating both HIV-1 RNA and HIV-1 DNA from semen infected with HIV-1, we tested the ejaculates of men infected with HIV-1 before and after processing semen into the three main seminal fractions – nonspermatozoa cells, cell-free seminal plasma and spermatozoa – by highly sensitive extractive nested PCR and by IS-PCR. All samples of spermatozoa recovered after separation by gradient centrifugation and swim-up (sperm washing) were free of HIV-1 RNA above our threshold of 50copies/ml and of proviral DNA [51].

This confirms findings of previous reports in which nested PCR [35], [52], [53] was used to assess the validity of sperm washing in HIV-infected semen. Although other more recent methodologies of sperm washing [54] confirm the validity of the general principle of removing the cellular component of semen, contradictory reports could be due to the inaccuracy of PCR techniques, such as in older studies [55] or to a too-lower threshold (one viral copy) of the PCR assay used to detect viral copies [53], [56], [57] or to an improper use of the definition of sperm washing [58], without the final swim-up of spermatozoa. In our paper [51], of the seven seminal plasma samples testing positive for HIV-1 RNA, six were from patients on HAART. Four men had elevated blood viral load and three had an undetectable viremia. These findings confirm findings of previous reports of discrepancies between haematic and seminal HIV-1 concentrations [35], [59], either due to subtherapeutic concentrations of anti-retroviral drugs in the male seminal tract, or due to local production of HIV-1 RNA from localized cells that poorly respond to treatment. The false-positive detection of HIV-1 DNA by IS-PCR in semen of HIV-1-non-infected men confirms that this technique is not adequate for studying the presence of provirus in semen fractions. The presence of the virus in spermatozoa pellet samples could be due to the presence of NSCs not completely eliminated during semen separation by discontinuous gradient centrifugation before swim-up. Alternatively, these could be real false-positive results because of nonspecific hybridization of IS-PCR. Bagasra et al. [38], Nuovo et al. [49] and Muciaccia et al. [50] showed the presence of provirus by IS-PCR in spermatozoa and in germ cells at all stages of differentiation, from spermatogonium to round spermatidi. However, in none of these studies proper standards for IS-PCR specificity were assessed on non-infected males. These methodological limitations of IS-PCR probably explain why in recent works this technique had been abandoned. Fiore et al. [60] evaluated the relationship between the seminal HIV-1 viral load and the efficiency of a standardized sperm-washing procedure in removing HIV-1 RNA from semen samples. The results of his paper indicated that the amount of virus present in the original sample affects the efficiency of the procedure and suggested that previous studies on sperm washing were performed on semen sample containing limited amounts of virus generally less than 5×104copies/ml. The bias of his study is that he used eight semen samples from eight healthy HIV-1 seronegative men and he tested semen for HIV-1 RNA after adding different dilutions of the virus. He concluded that the genital tract represents a distinct compartment compared with the blood and that viral loads in semen are variable irrespective of plasma loads and/or anti-retroviral treatment. On the contrary, Vernazza [61] holds as the risk of HIV-1 virus transmission depends exclusively on the viral load of the HIV-infected partner and he takes a strong medical position and in his paper reports that “an HIV-infected person on anti-retroviral therapy with completely suppressed viremia cannot propagate HIV through sexual contact”.

2.2. HCV in semen 

HCV transmission is known to occur essentially by the parenteral route [62], [63]. However, it is estimated that in 40–50% of cases the parenteral risk factor is not identified [64]. Another potential mode of non-parenteral transmission in HCV-positive-infected patients is through body secretions as saliva, ascites, breast milk, urine, faeces [65], [66], [67], [68], [69], [70], [71].

Many studies have analysed semen for the possible presence of HCV RNA with controversial results. The majority of these studies failed to document the presence of HCV RNA in seminal plasma [23], [72], [73], [74], [75], [76] while other publications have indicated its presence [3], [23], [53], [57], [65], [68], [69], [70], [71], [72], [73], [74], [75], [76], [77], [78], [79], [80], [81], [82], [83], [84], [85], [86], [87], [88], [89], [90], [91], [92]. The discrepancy in results on the presence of HCV RNA in seminal plasma of men chronically infected by this agent is due to various factors, such as the molecular techniques used, the sensitivity of the assays designed to detect HCV RNA, to the wide range of protocols for RNA extraction [83], [84], and eventually the presence in semen of inhibitors lactoferrin, peroxides, and mostly zinc residues that might interfere with the action of Taq polymerases [63], [93]. According to Messeguer [57], nested PCR appears to be most appropriate technique among these various methodologies for HCV virus relevance. The detection limit of this methodology is as low as one copy of viral RNA [93]. In 2002, the same author [57] demonstrated that samples tested negative by use of commercial methods for HIV/HCV detection, proved positive at nested PCR examination.

In addition to this, Levy et al. [82] observed that the viral concentration in seminal plasma changes rapidly over time. This variability of HCV concentration in seminal plasma is a critical point, one of its main consequences being that each sample of seminal plasma should be tested individually for the presence of HCV RNA prior to use in ART.

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3. Assisted reproduction in HIV-1 patients 

An assisted reproduction technology (ART) program was offered to serodiscordant couples for male HIV infection seeking medical assistance. Inclusion criteria were adopted to protect not only the couple, but also the possible child as well; partners were to engage only in protected sexual relations. HIV status had to be either monitored or treated or both and a long-term compliance had to be assessed by the infectious disease physician. Standard laboratory criteria were adopted: CD4+ lymphocytes of more than 200/mm3 at least twice in the 4 months prior to treatment; stable viral load, with no increase of more than 0.5log in two successive samples during the 4 months prior to treatment; infection by quantifiable amplifiable strain of HIV-1. Each couple was interviewed by a psychologist at inclusion and thereafter whenever necessary. Female fertility was assessed by standard procedures. In the clinical practice it is important to screen the HIV discordant couples in order to evidence the infertility factors, because of the high prevalence of sub-fertility factors. One of the most important factors is the genital tract infection in both males and females [93], [94]. The exact mechanisms involved in male-to-female transmission of HIV-1 are as yet undefined, but circumstantial evidence indicates that genital tract infections may act as facilitating factors. In sub-Saharan and Latin-American countries, where heterosexual transfer of the virus is the leading cause of infection, there is also a high prevalence of carriers of genital infections. The presence of a sexually transmitted pathogen recruits inflammatory cells in both the male and female genital tract. It is speculative to think that the biological mechanism behind this might be an increase in the number of HIV-1-infected cells in the semen or vaginal fluid of the seropositive subject, leading to a higher risk of infection for the seronegative partner. Conversely, when the infection is present in the seronegative partner, the uninfected inflammatory cells may become a specific target for the virus. As genital tract infections may facilitate male-to-female transmission of HIV and because HIV-infected subjects often ignore the recommendation for condom-protected intercourse, their detection and eradication may have an impact on the reduction of male-to-female sexual transfer of the virus.

All the semen samples may be treated with “sperm washing”. Semen analyses were performed and samples were processed using a 40–80% density gradient (Pureception kit, Sage) to separate motile spermatozoa from non-sperm cells, immotile spermatozoa and seminal plasma. The ejaculate was layered over the gradient and centrifuged at 400g for 30min. After centrifugation, the supernatant was removed and the sperm pellet recovered and resuspended in 3ml of fresh medium (sperm-washing medium, Sage). A centrifugation at 400g for 10min was performed and the supernatant was discarded. One millilitre of medium was subsequently gently layered on the pellet and the tube was incubated at 37°C for 1h. After swim-up, a supernatant volume of about 500μl was recovered [51].

The technique of choice in serodiscordant couples for HIV-positive male partner, when no other infertility problem is AIH after sperm washing, that is efficiency, safe, relatively low cost [95]. The efficiency of the sperm-washing technique [95], [96] was proved by the large number of AIH (3000) with safe pregnancies published in 2007. A previous review [96] reported 3221 cycle worldwide, most of them in Europe. In September 2007 issue of AIDS [97] a paper was presented where eight European centres offering assisted reproduction with sperm washing to HIV-1-serodiscordant couples. All the centres presented their results in these papers. A total of 1036 couples underwent 3390 assisted reproduction cycles resulting in 580 pregnancies. No transmission of HIV to the female partner was observed after 3272 cycles with complete follow-up information. The upper level of the 95% CI of the transmission risk was thus 0.09%. The study presented by AIDS is multicentric, but retrospective and each centre has a different clinical approach with consequent possible bias.

When the female partner was suffering from infertility factors, or the male partner had less than 1×106 total motile cells in the final fraction after sperm washing, or both partners had a combination of sub-fertility conditions in vitro fertilization (IVF)/ICSI was performed. The pregnancy rate per embryo transfer was in agreement both with similar smaller HIV series [52] and larger non-HIV series [98]. Savasi recently reported their experience on a large group of patients (500 FIVET/ICSI cycles) without HIV-1 transmission to the female partners with an adequate follow-up. The problem with ICSI to all serodiscordant couples is the high multiple pregnancy rate and the possible obstetrical and neonatal complications associated to these pregnancies (14% in Garrido et al. [53], 57.1% in Pena et al. [99], 10% in 95). We should also consider the possible additional costs determined by prenatal and neonatal care in multiple pregnancies [100], [101].

Another interesting point is the HIV-1 testing after sperm washing: the best prudent advice is to make testing for HIV-1 by PCR after sperm washing to IUI IVF and to also ICSI both for fresh and cryopreserved semen. So far, we agree with Garrido et al. [53] and Gilling-Smith [97] that in order to protect patient from technical errors during semen washing viral detection sampling prior to ART is the method of choice.

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4. Assisted reproduction in HCV patients 

HCV viral loads in seminal plasma showed dramatic and rapid variations through time. This variability of HCV concentration in seminal plasma is a critical point, one of its main consequences being that each sample of seminal plasma should be tested individually for the presence of HCV RNA prior to use in ART [82]. Another interesting finding is the rapid decrease in the HCV viral load in seminal plasma after introduction of an antiviral treatment combining alpha interferon and ribavirin. Unfortunately, it is not possible to recommend the use of such a treatment before starting ART, in order to reduce the risks of HCV transmission, since ribavirin has been shown to induce serious damage to spermatogenesis [102].

The opportunity for HCV-serodiscordant couples, who need artificial insemination or IVF/ICSI, would be to use donor sperm, but in Italy in and Egypt it is illegal, even in the case of the possible transmission of infectious diseases. A second opportunity would be to postpone IUI, IVF and ICSI trials in HCV-positive males until further laboratory testing has occurred. Semen would be submitted to selection by centrifugation in a discontinuous gradient, washing in BM1, followed by the swim-up technique [65]. The motile sperm would be divided into two parts: one half the used to detect HCV RNA, other half, if negative, could be used for assisted reproduction with minimal risk of viral contamination.

In Europe, several ART clinics have had extensive experience in assisting HCV-positive patients with reproduction. Most of these patients were HIV–HCV co-infected, since HCV infection has been found in 25–30% of HIV-positive persons. Most published papers reported ART in HIV–HCV co-infected patients. Chu et al. [103] have reported series of successful pregnancies within vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), without HCV seroconversion of mother or child, in these patients.

Garrido et al. [53] published a paper on 91 discordant couples for HIV, HCV and HIV–HCV male co-infection. The study concluded that sperm wash, nested PCR and ICSI are the safest and most reasonable method to treat HIV and HCV-serodiscordant couples seeking progeny.

Pasquier et al. [83] said that the use of density gradient plus ‘swim-up’ to purify motile spermatozoa reduces HIV-1 and HCV genomes in spermatozoa of doubly-infected men to undetectable concentrations. This method associated with standardized virus assay could be useful for serodiscordant couples wishing to have children. Also, Mencaglia et al. [89] suggest that sperm wash and ICSI could be useful for reducing the risk of HIV and/or HCV transmission in serodiscordant couples with infected males wishing to have a child, irrespective of their fertility status. Halfon et al. [92] demonstrate that the finding of HCV RNA in fractions of semen and in spermatozoa calls for reinforced precaution for men whose blood is chronically infected with HCV and who are candidates for ART. We suggest that seminal plasma has to be systematically evaluated for the absence of viral RNA in order to ensure that ART is performed with samples free of HCV RNA.

Before using sperm from a patient exhibiting HCV RNA in seminal plasma for ART, the following recommendations could be given: (i) semen and corresponding motile sperm selected after gradient and swim-up migration must be cryopreserved in highly secure straws in liquid nitrogen until ART; (ii) the corresponding seminal plasma and motile sperm must be stored at −80°C for HCV RNA testing; (iii) only specimens corresponding to sperm tested negative for HCV RNA must be considered for ART [82].

Viruses have been reported to have the ability to survive, and retain their virulence, in liquid nitrogen [104]. Tedder et al. [105] have previously reported on HBV contamination of negative bone-marrow samples cryopreserved in the same liquid-nitrogen tank. Therefore the threat of HCV transmission via cryopreserved bioproducts is both possible and real. For these reasons, it is necessary to assess the real safety of straws used for cryopreservation of human gametes and embryos, especially for routine use in assisted reproduction in conjunction with transmissible viral diseases.

Regrettably, there are limited data available in the literature on the safety of assisted reproductive treatment performed with testicular sperm. In such cases, blood contamination increases the viral load of the sample and the risk of HCV transmission in comparison with procedures done with ejaculated sperm purified by gradient selection and swim-up [106]. In a report, Manno et al. [106] reported on a case of TESE/ICSI in an HCV chronically infected serodiscordant couple with HCV RNA-positive seminal plasma. In this case, the female partner also did not seroconvert following the assisted reproductive treatment trial.

Since HCV seropositivity does not seem to decrease the success rates of IVF, and the rate of vertical and horizontal transmission is limited, there seems to be no evident reason to discourage HCV seropositive patients from attempting IVF. However, care must be taken on the part of the IVF centre to limit any transmission risk.

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5. Conflict of interest 

The authors have no conflict of interest.

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References 

  1. Berry W, Gottesfeld R, Alter H, Vierling J. Transmission of hepatitis B virus by artificial insemination. J Am Med Assoc. 1987;257:1079–1081
  2. Lesourd F, Izopet J, Mervan C, Payen JL, Sandres K, Monrozies X, et al. Transmissions of hepatitis C virus during the ancillary procedures for assisted conception. Hum Reprod. 2000;15:1083–1085
  3. McKee T, Avery S, Majid A, Brinsden P. Risks for transmission of hepatitis C during artificial insemination. Fertil Steril. 1996;66:161–163
  4. Stewart GJ, Tyler JP, Cunningham AL, Barr JA, Driscoll GL, Gold J, et al. Transmission of human T-cell lymphotropic virus type III (HTLV-III) by arti®cial insemination by donor. Lancet. 1985;2:581–585
  5. Chiasson M, Stoneburner R, Joseph S. Human immunodeficiency virus transmission through artificial insemination. J Acquir Immune Defic Syndr. 1990;3:69–72
  6. Araneta MR, Mascola L, Eller A, O'Neil L, Ginsberg MM, Bursaw M, et al. HIV transmission through donor artificial insemination. J Am Med Assoc. 1995;273:854–858
  7. Wortley P, Hammett T, Fleming P. Donor insemination and human immunodeficiency virus transmission. Obstet Gynecol. 1998;91:515–518
  8. Matz B, Kupfer B, Ko Y, Walger P, Vetter H, Eberle J, et al. HIV-1 infection by artificial insemination. Lancet. 1998;351:728
  9. Edelstein M, Kruithoff C, Brothman L, Veeck L, Alexander N, Muasher S. The prevalence of human immunodeficiency virus in patients and their spouse entering a large in vitro fertilization program. J In Vitro Fert Embryo Transf. 1990;7:254–256
  10. Abusheikha N, Akagbosu F, Marcus S, Lass A, Cousins C, Brinsden P. Viral screening and assisted conception treatment-the bourn hall experience. J Assist Reprod Genet. 1999;16:337–339
  11. American Fertility Society . Revised new guidelines for the use of semen donor insemination. Fertil Steril. 1988;49:211
  12. Centers for Disease Control and Prevention . Perspectives in disease prevention and health promotion update: universal precautions for prevention of transmission of human immunode®ciency virus, hepatitis B virus and other bloodborne pathogens in health-care settings. MMWR Morb Mortal Wkly Rep. 1988;37:377–388
  13. Human Fertilisation and Embryology Authority. Code of practice: HIV screening for gamete donors; 1991.
  14. Barratt C, Matson P, Holt W. British andrology society guidelines for the screening of semen donors for donor insemination. Hum Reprod. 1993;8:1521–1523
  15. Barratt C, Englert Y, Gottlieb C, Jouannet P. Gamete donation guidelines. The corsendonk consensus statements for the European Union. Hum Reprod. 1998;13:500–501
  16. World Health Organization . WHO laboratory manual for the examination of human semen and semen±cervical mucus interaction. 4th ed.. Cambridge: Cambridge University Press; 1999;
  17. Ethics Committee of the American Society for Reproductive Medicine . Human immunodeficiency virus and infertility treatment. Fertil Steril. 2002;77:218–222
  18. Englert Y, Van Vooren JP, Place I, Liesnard C, Laruelle C, Delbaere A. ART in HIV-infected couples. Hum Reprod. 2001;16:1309–1315
  19. Alter MJ. Epidemiology of viral hepatitis and HIV co-infection. J Hepatol. 2006;44:S6–S9
  20. Botta-Fridlund D. Risk of hepatitis virus B and C transmission during medically assisted reproduction. Contracept Fertil Sex. 1994;22:298–301
  21. Zeuzem S, Teuber G, Lee JH, Rüster B, Roth WK. Risk factors for the transmission of hepatitis C. J Hepatol. 1996;24:3–10
  22. Pawlotsky JM. Chronic hepatitis C virus infection with repeatedly normal aminotransferase levels: disease or not disease?. Am J Gastroenterol. 1997;92:1775–1777
  23. Caldwell SH, Sue M, Bowden JH, Dickson RC, Driscoll CJ, Yeaton P, et al. Hepatitis C virus in body fluids after liver transplantation. Liver Transpl Surg. 1996;2:124–129
  24. Englert Y, Lesage B, Van Vooren JP, Liesnard C, Place I, Vannin AS, et al. Medically assisted reproduction in the presence of chronic viral diseases. Hum Reprod Update. 2004;10:149–162
  25. Arrêté du 12-1-1999. Journal official du 28-2-1999 relatif aux rêgles de Bonnes Pratiques cliniques et biologiques en Assistance Médicale à la Procrèation.
  26. Arrêté du 10 Mai modifiant l'arrêté du 12 Janvier 1999 relatif aux rěgles de bonnes pratiques cliniques et biologiques en assistance médicale à la procréation. J Off Répub Fr. 2001;112:7735–7736
  27. Decree of May 10 Prise en charge en assistance médicale à la procréation des patients à risque viral. J Off Répub Fr. mai 2001;15:7735;Arrêté du 10 mai 2001
  28. Baccetti B, Benedetto A, Burrini AG, Collodel G, Ceccarini EC, Crisa N, et al. HIV-particles in spermatozoa of patients with AIDS and their transfer into the oocyte. J Cell Biol. 1994;127:903–914
  29. Brogi A, Presentini R, Moretti E, Strazza M, Piomboni P, Costantino-Ceccarini EE. New insights into the interaction between the gp120 and the HIV receptor in human sperm. J Reprod Immunol. 1998;41:213–231
  30. Lasheeb AS, King J, Ball JK, Curran R, Barratt CL, Afnan M, et al. Semen characteristics in HIV-1 positive men and the effect of semen washing. Genitourin Med. 1997;73:303–305
  31. Quayle AJ, Xu C, Mayer KH, Anderson Dj. T lymphocytes and macrophages, but not motile spermatozoa are a significant source of human immunodeficiency virus in semen. J Infect Dis. 1997;176:960–968
  32. Pudney J, Nguyen H, Xu C, Anderson DJ. Microscopic evidence against HIV-1 infection of germ cells or attachment to sperm. J Reprod Immunol. 1998;4:105–125
  33. Semprini AE, Levi-Setti P, Bozzo M, Ravizza M, Taglioretti A, Sulpizio P, et al. Insemination of HIV-negative women with processed semen of HIV-positive partners. Lancet. 1992;28:1317–1319
  34. Borzy MS, Connell RS, Kiessling AA. Detection of human immunodeficiency virus in cell-free seminal fluid. J Acquir Immune Defic Syndr. 1988;1:419–424
  35. Bujan L, Pasquier C, Labeyrie E, Lanusse-Crousse P, Morucci M, Daudin M. Insemination with isolated and virologically tested spermatozoa is a safe way for human immunodeficiency type 1 virus-serodiscordant couples with an infected male partner to have a child. Fertil Steril. 2004;82:857–862
  36. Bagasra O, Freund M, Weidmann J, Harley . Interaction of human immunodeficiency virus with human sperm in vitro. J Acquir Immune Defic Syndr. 1988;1:431–435
  37. Bagasra O, Farzadegan H, Seshamma T, Oakes JW, Saah A, Pomerantz RJ. Detection of HIV-1 proviral DNA in sperm from HIV-1 infected men. AIDS. 1994;8:1669–1674
  38. Anderson DJ, et al. Presence of HIV-1 in semen. In:  Alexander NJ,  Gabelnick HL,  Spieler JM editor. Heterosexual transmission of AIDS. Proceedings of the Second Contraceptive Research and Development (CONRAD) Program International Workshop, Norfolk, Virginia, February 1–3, 1989. New York: Wiley-Liss; 1990;p. 167–180
  39. Baccetti B, Benedetto A, Burrini AG, Collodel G, Elia C, Piomboni P, et al. HIV particles detected in spermatozoa of patients with AIDS. J Submicrosc Cytol Pathol. 1991;23:339–345
  40. Baccetti B, Benedetto A, Collodel G, Di Caro A, Garbuglia AR, Piomboni P. The debate on the presence of HIV-1 in human gametes. J Reprod Immunol. 1998;41:41–67
  41. Gobert B, Amiel C, Tang JQ, Barbarino P, Bene MC, Faure G. CD4-like molecules in human sperm. FEBS Lett. 1990;261:339–342
  42. Miller VE, Scofield VL. Transfer of HIV-1 by semen: role of sperm. In:  Alexander NJ,  Gabelnick HL,  Spieler JM editor. Heterosexual transmission of AIDS. Proceedings of the Second Contraceptive Research and Development (CONRAD) Program International Workshop, held in Norfolk, Virginia, February 1–3, 1989. New York: Wiley-Liss; 1990;p. 147–154
  43. Pudney J. Caveats associated with identifying HIV-1 using transmission electron microscopy. In:  Alexander NJ,  Gabelnick HL,  Spieler JM editor. Heterosexual transmission of AIDS. Proceedings of the Second Contraceptive Research and Development (CONRAD) Program International Workshopheld in Norfolk, Virginia, February 1–3, 1989. New York: Wiley-Liss; 1990;p. 197–204
  44. Mermin JH, Holodniy M, Katzenstein DA, Merigan TC. Detection of human immunodeficiency virus DNA and RNA in semen by the polymerase chain reaction. J Infect Dis. 1991;164:769–772
  45. Van Voorhis BJ, Martinez A, Mayer K, Anderson DJ. Detection of HIV-1 in semen from seropositive men using culture and polymerase chain detection deoxyribonucleic acid amplification techniques. Fertil Steril. 1991;55:588–594
  46. Scofield VL, Rao B, Broder S, Kennedy C, Wallace M, Graham B, et al. HIV interaction with sperm. AIDS. 1994;8:1733–1736
  47. Dussaix E, Guetard D, Dauget C, D'Almeida M, Auer J, Ellrodt A, et al. Spermatozoa as potential carriers of HIV-1. Res Virol. 1993;144:487–495
  48. Nuovo GJ, Becker J, Simsir A, Margiotta M, Khalife G, Shevchuk M. HIV-1 nucleic acids localize to the spermatogonia and their progeny. A study by polymerase chain reaction in situ hybridization. Am J Pathol. 1994;144:1142–1148
  49. Muciaccia B, Uccini S, Filippini A, Ziparo E, Paraire F, Baroni CD, et al. Presence and cellular distribution of HIV-1 in the testes of seropositive subjects: an evaluation by in situ PCR hybrydization. FASEB J. 1998;12:151–163
  50. Persico T, Savasi V, Ferrazzi E, Oneta M, Semprini AE, Simoni G. Detection of human immunodeficiency virus-1 RNA and DNA by extractive and in situ PCR in unprocessed semen and seminal fractions isolated by semen-washing procedure. Hum Reprod. 2006;21:1525–1530
  51. Ohl J, Partisani M, Wittemer C, Schmitt MP, Cranz C, Stoll-Keller F, et al. Assisted reproduction techniques for HIV serodiscordant couples: 18 months of experience. Hum Reprod. 2003;18:1244–1249
  52. Garrido N, Meseguer M, Bellver J, Remohi J, Simon C, Pellicer AA. Report of the results of a 2 year programme of sperm wash and ICSI treatment human immunodeficiency virus and hepatitis C virus serodiscordant couples. Hum Reprod. 2004;19:2581–2586
  53. Politch JA, Xu C, Tucker L. Anderson DJ Separation of human immunodeficiency virus type 1 from motile sperm by the double tube gradient method versus other methods. Fertil Steril. 2004;81:440–447
  54. Marina S, Marina F, Alcolea R, Exposito R, Huguet J, Nadal J, et al. Human immunodeficiency virus type 1-serodiscordant couples can bear healthy children after undergoing intrauterine insemination. Fertil Steril. 1998;70:35–39
  55. Garrido N, Meseguer M, Bellver J. In vitro fertilization with intracytoplasmic sperm injection for human immunodeficiency virus-1 serodiscordant couples. Am J Obstet Gynecol. 2002;187:1121
  56. Meseguer M, Garrido N, Gimeno C, Remohı J, Simòn C, Pellicer C. Comparison of polymerase chain reaction-dependent methods for determining the presence of human immunodeficiency virus and hepatitis C virus in washed sperm. Fertil Steril. 2002;78:1199–1202
  57. Leruez-Ville M, de Almeida M, Tachet A, Dulioust E, Guibert J, Mandelbrot L, et al. Assisted reproduction in HIV-1-serodifferent couples: the need for viral validation of processed semen. AIDS. 2002;16:2267–2273
  58. Xu C, Politch JA, Mayer KH, Anderson DJ. Human immunodeficiency virus type-1 episomal cDNA in semen. AIDS Res Ther. 2005;2:9
  59. Fiore RJ, Lorusso F, Vacca M, Ladisa N, Greco P, De Palo R. The efficiency of sperm washing in removing human immunodeficienncy type varies according to the seminal viral load. Fertil Steril. 2005;84:232–234
  60. Vernazza P, Hirsch B, Bernasconi E. HIV-infected persons on effective anti-retroviral therapy are sexually non-infectious. <http://www.nadironlus.org/modules>; Last access July 2008.
  61. Choo QL, Kuo G, Weiner AJ, Overby LR, Bradley DW, Houghton M. Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science. 1989;244:359–362
  62. Moyer LA, Mast EE, Alter MJ. Hepatitis C: part II. Prevention counseling and medical evaluation. Am Fam Physician. 1999;59:349–354
  63. Everhart JE, Di Bisceglie AM, Murray LM, Alter HJ, Melpolder JJ, Kuo G, et al. Risk for non-A, non-B (type C) hepatitis through sexual or household contact with chronic carriers. Ann Intern Med. 1990;112:544–545
  64. Levy R, Tardy JC, Bourlet T, Cordonier H, Mion F, Lornage J, et al. Transmission risk of hepatitis C virus in assisted reproductive techniques. Hum Reprod. 2000;15:810–816
  65. Belec L, Legoff J, Si-Mohamed A, Bloch F, Matta M, Mbopi-Keou FX, et al. Cell-associated, non-replicating strand(þ) hepatitis C virus-RNA shedding in cervicovaginal secretions from chronically HCV infected women. J Clin Virol. 2003;27:247–251
  66. Beld M, Sentjens R, Rebers S, Weel J, Wertheim-van Dillen P, Sol C, et al. Detection and quantitation of hepatitis C virus RNA in feces of chronically infected individuals. J Clin Microbiol. 2000;38:3442–3444
  67. Liou TC, Chang TT, Young KC, Lin XZ, Lin CY, Wu HL. Detection of HCV RNA in saliva, urine, seminal fluid, and ascites. J Med Virol. 1992;37:197–202
  68. Numata N, Ohori H, Hayakawa Y, Saitoh Y, Tsunoda A, Kanno A. Demonstration of hepatitis C virus genome in saliva and urine of patients with type C hepatitis: usefulness of the single round polymerase chain reaction method for detection of the HCV genome. J Med Virol. 1993;41:120–128
  69. Young KC, Chang TT, Liou TC, Wu HL. Detection of hepatitis C virus RNA in peripheral blood mononuclear cells and in saliva. J Med Virol. 1993;41:55–60
  70. Kumar RM, Shahul S. Role of breast-feeding in transmission of hepatitis C virus to infants of HCV-infected mothers. J Hepatol. 1998;29:191–197
  71. Hsu HH, Wright TL, Luba D, Martin M, Feinstone SM, Garcia G, et al. Failure to detect hepatitis C virus genome in human secretions with the polymerase chain reaction. Hepatology. 1991;14:763–767
  72. Fried MW, Shindo M, Fong TL, Fox PC, Hoofnagle JH, Di Bisceglie AM. Absence of hepatitis C viral RNA from saliva and semen of patients with chronic hepatitis C. Gastroenterology. 1992;102:1306–1308
  73. Terada S, Kawanishi K, Katayama K. Minimal hepatitis C infectivity in semen. Ann Intern Med. 1992;117:171–172
  74. Semprini AE, Persico T, Thiers V, Oneta M, Tuveri R, Serafini P, et al. Absence of hepatitis C virus and detection of hepatitis G virus/GB virus C RNA sequences in the semen of infected men. J Infect Dis. 1998;177:848–854
  75. Debono E, Halfon P, Bourliere M, Gerolami-Santandrea V, Gastaldi M, Castellani P, et al. Absence of hepatitis C genome in semen of infected men by polymerase chain reaction, branched DNA and in situ hybridization. Liver. 2000;20:257–261
  76. Kotwal GJ. Routine laboratory diagnosis of hepatitis C virus infection. J Hepatol. 1993;17:S83–S89
  77. Kotwal GJ, Baroudy BM, Kuramoto IK, McDonald FF, Schiff GM, Holland PV, et al. Detection of acute hepatitis C virus infection by ELISA using a synthetic peptide comprising a structural epitope. Proc Natl Acad Sci U S A. 1992;89:4486–4489
  78. Liu FH, Tian GS, Fu XX. Detection of plus and minus strand hepatitis C virus RNA in peripheral blood mononuclear cells and spermatid. Zhonghua Yi Xue Za Zhi. 1994;74:284–286
  79. Fiore RJ, Potenza D, Monno L, Appice A, Di Stefano M, Giannelli A, et al. Detection of HCV RNA in serum and seminal fluid from HIV-1 co-infected intravenous drug addicts. J Med Virol. 1995;46:364–367
  80. Leruez-Ville M, Kunstmann JM, De Almeida M, Rouzioux C, Chaix ML. Detection of hepatitis C virus in the semen of infected men. Lancet. 2000;356:42–43
  81. Levy R, Bourlet T, Maertens A, Salle B, Lornage J, Laurent JL, et al. Pregnancy after safe IVF with hepatitis C virus RNA-positive sperm. Hum Reprod. 2002;17:2650–2653
  82. Pasquier C, Daudin M, Righi L, Berges L, Thauvin L, Berrebi A, et al. Sperm washing and virus nucleic acid detection to reduce HIV and hepatitis C virus transmission in serodiscordant couples wishing to have children. AIDS. 2000;14:2093–2099
  83. Bourlet T, Levy R, Maertens A, Tardy JC, Grattard F, Cordonier H, et al. Detection and characterization of hepatitis C virus RNA in seminal plasma and spermatozoon fractions of semen from patients attempting medically assisted conception. J Clin Microbiol. 2002;40:3252–3255
  84. Cassuto NG, Sifer C, Feldmann G, Bouret D, Moret F, Benifla JL, et al. A modified RT-PCR technique to screen for viral RNA in the semen of hepatitis C viruspositive men. Hum Reprod. 2002;17:3153–3156
  85. Nyamathi A, Robbins WA, Fahey JL, Wiley D, Pekler VA, Longshore D, et al. Presence and predictors of hepatitis C virus RNA in the semen of homeless men. Biol Res Nurs. 2002;4:22–30
  86. Pasquier C, Bujan L, Daudin M, Righi L, Berges L, Thauvin L, et al. Intermittent detection of hepatitis C virus (HCV) in semen from men with human immunodeficiency virus type 1 (HIV-1) and HCV. J Med Virol. 2003;69:344–349
  87. Pekler VA, Robbins WA, Nyamathi A, Yashina TL, Leak B, Robins TA. Use of versant TMA and bDNA 3.0 assays to detect and quantify hepatitis C virus in semen. J Clin Lab Anal. 2003;17:264–270
  88. Mencaglia L, Falcone P, Lentini GM, Consigli S, Pisoni M, Lofiego V, et al. ICSI for treatment of human immunodeficiency virus and hepatitis C virus-serodiscordant couples with infected male partner. Hum Reprod. 2005;20:2242–2246
  89. Loskutoff NM, Huyser C, Singh R, Walker DL, Thornhill AR, Morris L, et al. Use of a novel washing method combining multiple density gradients and trypsin for removing human immunodeficiency virus-1 and hepatitis C virus from semen. Fertil Steril. 2005;84:1001–1010
  90. Briat A, Dulioust E, Galimand J, Fontaine H, Chaix ML, Letur-Konirsch H, et al. Hepatitis C virus in the semen of men coinfected with HIV-1: prevalence and origin. AIDS. 2005;19:1827–1835
  91. Halfon P, Giorgetti C, Bourliere M, Chabert-Orsoni V, Khiri H, Penaranda G, et al. Medically assisted procreation and transmission of hepatitis C virus: absence of HCV RNA in purified sperm fraction in HIV co-infected patients. AIDS. 2006;20:241–246
  92. Cohen MS, Hoffman IF, Royce RA, Kasembe P, Dyer JR, Daly CC, et al. Reduction of concentration of HIV-1 in semen after treatment of urethritis: implications for prevention of sexual transmission of HIV-1. AIDSCAP Malawi Research Group. Lancet. 1998;349:1868–1873
  93. Rotchford K, Strum A, Wilkson D. Effect of coinfection with STDs and of STD treatment on HIV sheddind in genital tract secretions: systematic review and data syntesis. Sex Trans Dis. 2000;27:243–248
  94. Savasi V, Ferrazzi E, Lanzani C, Oneta M, Parrilla B, Persico T. Safety of sperm washing and ART outcome in 741 HIV-1-serodiscordant couples. Hum Reprod. 2007;22:772–777
  95. Sauer MV. Sperm washing techniques address the fertility needs of HIV-seropositive men: a clinical review. Reprod Biomed Online. 2005;1:135–140
  96. Bujan L, Hollander L, Coudert M, Gilling-Smith C, Vucetich A, Guibert J, et al. Safety and efficacy of sperm washing in HIV-1-serodiscordant couples where the male is infected: results from the European CREAThE network. AIDS. 2007;12:1909–1914
  97. ESHRE . Assisted reproductive technology in Europe, 2002. Results generated from European registers by ESHRE. Hum Reprod. 2006;21:1680–1697
  98. Pena JE, Thornton MH, Sauer MV. Assessing the clinical utility of in vitro fertilization with intracytoplasmic sperm injection in human immunodeficiency virus type 1 serodiscordant couples: report of 113 consecutive cycles. Fertil Steril. 2003;80:356–362
  99. Nakhuda GS, Sauer MV. Addressing the growing problem of multiple gestations created by assisted reproductive therapies. Semin Perinatol. 2005;29:355–362
  100. Oliviennes F. Double trouble: yes a twin pregnancy is an adverse outcome. Hum Reprod. 2000;15:1663–1665
  101. Narayana K, D'Souza UJ, Rao KP. Effect of ribavirin on epididymal sperm count in rat. Ind J Physiol Pharmacol. 2002;46:97–101
  102. Chu MC, Pena JE, Nakhuda GS, Thornton MH, Sauer MV. Assessing the reproductive performance of men co-infected with HIV-1 and hepatitis C undergoing assisted reproduction. Arch Gynecol Obstet. 2006;274:155–159
  103. Letur-KonõÈrsch H, Collin G, Sifer C, Devaux A, Kuttenn F, Madelenat P, et al. Safety of cryopreservation straws for human gametes or embryos: a study with human immunodeficiency virus-1 under cryopreservation conditions. Hum Reprod. 2003;18:140–144
  104. Tedder RS, Zuckerman MA, Goldstone AH, Hawkins AE, Fielding A, Briggs EM, et al. Hepatitis B transmission from contaminated cryopreservation tank. Lancet. 1995;346:137–140
  105. Manno M, Marchesan E, Crovatto M, Martelli P, Tomei F, Adamo V. Preliminary evidence on the safety of ICSI with testicular spermatozoa in HCV-infected male: a case report. Hum Reprod. 2003;18:1666–1668

PII: S0143-4004(08)00243-9

doi:10.1016/j.placenta.2008.08.006

Placenta
Volume 29, Supplement 2 , Pages 160-165, October 2008

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