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E Dulioust,
A Le Du,
D Costagliola,
J Guibert,
J-M Kunstmann,
I Heard,
J-C Juillard,
D Salmon,
M Leruez-Ville,
L Mandelbrot,
C Rouzioux,
D Sicard,
J-R Zorn,
P Jouannet, M De Almeida
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ABSTRACT: Couples in whom the man is infected by human immunodeficiency virus (HIV) increasingly request assisted reproductive technology (ART) to allow safe procreation. Semen quality is critical in such situations.
Semen characteristics were evaluated in 189 HIV-infected men requesting ART. At the time of semen analysis all men were healthy and 177 were receiving anti-retroviral therapy. Comparisons were made with HIV-seronegative men, partners of women requiring IVF because of tubal infertility, after matching for age and sexual abstinence delay.
The most significant semen alterations found in the HIV-infected men were reduced percentages of rapidly progressive sperm [median (range), 10% (0-30%) compared with 15% (5-30%) in the controls, P < 0.001], and increased concentrations of non-spermatic cells [3 x 10(6)/ml (0.2-16 x 10(6)/ml) compared with 1.1 x 10(6)/ml (0.1-14 x 10(6)/ml) in the controls, P < 0.001]. HIV-infected men also showed lower ejaculate volumes [2.8 ml (0.6-9.3 ml) compared with 3.6 ml (1.1-11 ml), P < 0.05] and total sperm counts [262.5 x 10(6) (0-1003 x 10(6)) compared with 310.5 x 10(6) (48.3-1679 x 10(6)), P < 0.05].
Semen evaluation in a large population of HIV-infected men requesting ART evidenced several alterations. Some of these anomalies might be related to anti-retroviral treatments.
Human Reproduction 08/2002; 17(8):2112-8. · 4.47 Impact Factor
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ABSTRACT: Antisperm antibodies eluted from the surface of spermatozoa obtained from infertile men recognised several common epitopes. We tested whether these epitopes were relevant to fertility by isolating the immunodominant 37/36 and 19/18 protein zones. These protein zones were cut out of preparative slab gels and electro-eluted. The isolated proteins, P36 and P18, were used for biochemical characterisation and to produce specific antibodies in rabbits. The specific reactivity of P36 and P18 with WGA and AAL lectins, respectively, indicated the presence of lactosaminyl structures with sialic acid termini in P36 and of fucosylated residues in P18. Isoelectric focusing showed that the two proteins consist of several polypeptides. Some of these polypeptides were recognised by both human and rabbit antibodies: the pl of these epitopes was around 5.5 for P36 and 8.3-10.3 for P18. Rabbit antibodies detected the corresponding proteins on the sperm heads of methanol-fixed and of live acrosome-reacted spermatozoa. Anti-P36 antibodies bound mainly to the equatorial segment. They reduced the binding and, consequently, the penetration of zona-free hamster oocytes by human spermatozoa. Anti-P18 antibodies gave more diffuse staining of the acrosomal and post-acrosomal regions and reduced sperm-oocyte penetration without a significant effect on sperm binding. These results suggest that P36 and P18 antigens located in different compartments of the sperm head may participate in the sperm-oolemma interaction. We are currently investigating the physiological role of these antigens by sequencing the proteins isolated from the gels.
Molecular Reproduction and Development 01/2001; 57(4):393-405. · 2.53 Impact Factor
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ABSTRACT: We detected hepatitis C virus (HCV) RNA in the semen of one third of HCV viraemic men. Seminal viral loads were low, but the semen could be infectious and the role of sexual transmission in the spread of HCV infection should not be underestimated.
The Lancet 08/2000; 356(9223):42-3. · 38.28 Impact Factor
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ABSTRACT: To assess HIV burden in both acellular and cellular fractions of semen in men with different levels of blood plasma HIV RNA by a cross-sectional study.
Fifty-two HIV-1-seropositive men (21 receiving antiretroviral therapy) with CD4 cell counts ranging from 1 to 1170 x 10(6)/l.
Semen was separated into seminal plasma and fractions enriched in motile spermatozoa or non-spermatozoal cells. HIV RNA was quantified by the HIV-Monitor technique (Roche) in blood plasma, seminal plasma and spermatozoa fractions. HIV DNA or infectious virions in cellular fractions were detected by either PCR or qualitative viral culture.
HIV RNA was detected in 86.5% of seminal plasma specimens and in 14.6% of spermatozoa fractions; HIV DNA was detected in 57.1% of non-spermatozoal cell fractions. HIV RNA levels in blood plasma and seminal plasma were correlated (r5 = 0.56, P < 0.0001, Spearman's rank test). A majority of men had lower levels in seminal plasma than in blood plasma: one-third had HIV-positive seminal cell fractions. However, 20 men (38.5%) with HIV RNA levels in seminal plasma (median: 4.65 log10 copies/ml) comparable to or higher than those in blood plasma had all HIV-positive non-spermatozoal cells or spermatozoa fractions with a high frequency of positive cultures.
A high frequency of men had detectable HIV in semen. We identified a subpopulation demonstrating high levels of HIV RNA in seminal plasma, comparable to or higher than those in blood plasma, frequently associated with a substantial viral shedding in seminal cells, raising the possibility of viral production within the genital tract and suggesting heterogeneity in the potential of HIV sexual transmission among infected men.
AIDS 05/1999; 13(7):823-31. · 6.24 Impact Factor
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ABSTRACT: It is of critical importance to precisely understand the modalities of HIV presence in semen, especially with regard to procreation. In this study, paired blood and semen samples from 31 human immunodeficiency virus type 1 (HIV-1)-positive men were assessed for cell-free HIV-RNA load in blood plasma (BP) and seminal plasma (SP), and for detection of HIV by culture, PCR and RT-PCR in semen cellular fractions separated by centrifugation on Percoll gradient. HIV-RNA was detected in 94% of BP and 84% of SP samples. For 11 men (35%), HIV-RNA load in SP was equal or superior to that observed in blood. HIV-DNA presence was demonstrated (either by PCR or culture positivity) in 39% of the non-spermatozoal cells (NSC)-enriched fractions, and in one Percoll-selected sperm pellet. HIV-RNA was detected in 17% (4/23) of the sperm pellets. This positivity was associated with an HIV-RNA load in SP equal or superior to the HIV-RNA load in blood, a high rate of HIV-DNA detection in the NSC fraction, and a low CD4+ cell count. In such conditions, a significant viral production inside the genital tract is more likely to be present, and a close association between HIV and gametes might occur. Assisted procreation with selected spermatozoa should be preceded by accurate assessments of viral presence in blood, SP and semen cellular fractions.
Journal of Reproductive Immunology 01/1999; 41(1-2):27-40. · 2.97 Impact Factor
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ABSTRACT: IgA and IgG antibodies eluted from the surface of spermatozoa obtained from 11 infertile men were used to analyse the antigens defined by each class of sperm-associated antibodies. An enhanced chemiluminescent Western blot technique was developed to detect the low concentrations of immunoglobulins present in the eluted samples. The same analysis was performed with the sperm membrane-specific antibodies isolated from the sera of 8 of the patients included in the study. Sperm-eluted antibodies reacted with a total of 18 protein bands migrating with molecular masses of between 110 and 18 kDa. Individual antibody-binding patterns differed. Furthermore, IgA and IgG antibodies from any one patient recognised different sets of antigens. In spite of the apparent heterogeneity of the antigens defined by sperm-associated antibodies, the majority of these antibodies reacted with three protein zones of 68/64, 37/36 and 20/18 kDa. The antigens defined by the sperm surface-specific antibodies obtained from the sera of eight infertile patients differed from one patient to another and, in the majority of the patients, differed from those defined by the corresponding sperm-associated antibodies. Nevertheless, two protein zones of 68/64 and 20/18 kDa were recognised by both local and systemic antibodies in 6 and 4 patients, respectively.
Journal of Reproductive Immunology 10/1997; 34(2):121-36. · 2.97 Impact Factor
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ABSTRACT: Subzonal insemination was the first micromanipulation technique used in the treatment of male factor sterility. Analysis of the sperm pathology prior to patient inclusion in the program allowed us to study gamete interaction in the human and the early embryogenesis. A series of 523 cycles and 3,027 micro-injected oocytes is reported.
Contraception, fertilité, sexualité (1992) 02/1997; 25(1):69-78.
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ABSTRACT: In order to identify the surface antigens of human spermatozoa recognized by the sera of immune infertile men, sperm membrane-specific antibodies were obtained from three serum samples exhibiting high titres of antibodies with different sperm-binding patterns. Serum antibodies were adsorbed onto normal motile spermatozoa and subsequently eluted from the sperm membrane. Using the immunoblotting technique under renaturating conditions, the sperm-eluted antibodies were tested against an electrophoretically fractionated sperm membrane preparation. A total of 15 protein bands ranging from 110 to 16 kDa were defined, but the immunoblot profiles differed quantitatively and qualitatively from one serum to another. Only two polypeptides were recognized by the three sperm membrane-specific antibody preparations; one of 90 kDa and another of 110 kDa. Blots were also used for the affinodetection of specific oligosaccharide side chains. Three lectins were tested (concanavalin A, Pisum sativum, and wheat germ agglutinin). Of the 15 protein zones recognized by the antibodies, 11 bound at least one lectin and should contain glycopeptides with oligosaccharides of four different types: N-linked biantennary complex type, N-linked fucosylated complex type, N-linked lactosaminyl complex type with terminal sialic acid and polysialyl type oligosaccharides. Further analysis of these glycoproteins will be pursued with sperm-associated antibodies eluted from the ejaculates of infertile men in order to define those with a potential role in the fertilization process.
Human Reproduction 04/1995; 10(3):551-7. · 4.47 Impact Factor
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ABSTRACT: To investigate the impact of sperm-associated antibodies in the fertilization process beyond the zona pellucida in human oocytes.
Subzonal insemination (SUZI) was performed with antibody-coated spermatozoa from patients with autoimmune infertility. Sperm parameters and antibody binding were analyzed after Percoll selection and were compared with SUZI outcome.
Patients (n = 29) with an immunobead test higher than 60% for immunoglobulin (Ig)G, IgA, or both classes were included in the study (38 attempts). Twenty-six patients had previous IVF failures, and the semen characteristics for three were unsuitable for IVF.
Diploid fertilization occurred in 52.6% of cycles and 16.0% of oocytes (n = 263). The fertilization rate was inversely correlated to the proportion of spermatozoa coated with IgG in the Percoll-selected sperm suspension. Three of the 16 attempts in which > 90% of the spermatozoa were coated with IgG resulted in a low diploid rate of 4.1%. Fertilization was obtained in 18 of the 22 attempts in which < 90% of spermatozoa were IgG coated, with a diploid fertilization rate of 22.8%. Association of both classes of antibodies (IgG and IgA) further impaired the fertilization rate. Twenty ETs gave four pregnancies: one biochemical, one ongoing, and two giving birth to healty babies (1 singleton and 2 twins).
Subzonal insemination is a potential solution for achieving fertilization in cases of IVF failure because of sperm autoimmunity. However, IgG inhibits in a dose-dependent manner the fusion of gametes' membranes. Though the level of IgA antibodies does not appear as critical as that of IgG antibodies, association of both classes of Igs impairs the probability of fertilization.
Fertility and Sterility 04/1995; 63(3):584-90. · 3.56 Impact Factor
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ABSTRACT: Antibodies directed against sperm membrane antigens located mainly over the sperm heads were eluted from the sperm cell fraction of autoimmune ejaculates and transferred to antibody-negative spermatozoa of fertile donors. The ability of these antibody-coated spermatozoa to bind to the human zona pellucida and to penetrate zona-free hamster oocytes was evaluated in vitro. The majority of the sperm-eluted samples contained both IgA and IgG antibodies. In order to evaluate the effect of each class of antibody on the analyzed sperm functions, each isotype was specifically absorbed before transfer. Sperm-binding to salt-stored zona pellucida, as assayed by FITC and TRITC labeling of antibody-free and antibody-coated spermatozoa incubated with the same zona, was consistently reduced by 60-85% by the five eluted samples tested. Removal of either IgA or IgG antibodies from the eluted samples did not change the overall effect. Sperm penetration of zona-free hamster eggs was variously affected by sperm-associated antibodies. Of the 8 samples of sperm-eluted antibodies tested, only 4 had a significant effect on sperm penetration. Three of them decreased the penetration by 67-78%, while the fourth exhibited a modest increasing effect of 39%. These four samples contained antibodies of the two isotypes. In the samples with a decreasing effect, the elimination of one or another of the two isotypes restored the ability of the sperm to penetrate the hamster oocytes. These results suggest that sperm-associated antibodies may have different effects on zona-binding and gametic fusion events that lead to fertilization. Whereas IgA and IgG antibodies taken together or separately decreased sperm binding to the human zona pellucida, the two classes of antibodies must be associated in order to impair sperm penetration into zona-free hamster oocytes.
Journal of Reproductive Immunology 09/1993; 24(3):175-86. · 2.97 Impact Factor
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ABSTRACT: To define sperm factors related to in vitro fertilization (IVF) failure in cases of antisperm autoimmunity.
A detailed analysis of sperm morphology, movement characteristics, acrosomal function, and antibody binding was performed on the sperm population selected on a discontinuous two-layer Percoll gradient and used for IVF. The results were compared retrospectively between fertilizing (n = 13) and nonfertilizing (n = 11) sperm populations.
Twenty-one infertile couples undergoing 24 cycles of IVF treatment because of antisperm autoimmunity were included in this study.
Fertilizing and nonfertilizing sperm populations were not different with respect to the percentage of motility, the normal morphology, the multiple anomalies index, the acrosome abnormalities, and the spontaneous or induced acrosome reaction. If the proportion of spermatozoa coated with either immunoglobulin (Ig)A or IgG antibodies was similar in the two groups, their localization was often different: antibodies were mainly on the sperm heads in the cases of fertilization failure. There were significant differences between fertilizing and nonfertilizing sperm samples in several movement parameters. Among them, the amplitude of lateral head displacement (ALH) was the most significantly correlated with fertilization success. Finally, spermatozoa were poorly bound to the zona pellucida (ZP) when fertilization failed, whereas high numbers of spermatozoa were attached to the ZP when fertilization occurred.
Fertilization failure in patients with antisperm antibodies may be the result of several factors in which the impact of the antibodies on the membrane function play a critical role. Movement parameters, particularly the ALH and the localization of antibodies on migrated spermatozoa could predict the IVF failure or success more accurately than the proportion of antibody-coated spermatozoa in the inseminated populations. The fertilization failure was associated with an incapacity of spermatozoa to bind to ZP.
Fertility and Sterility 04/1993; 59(3):606-12. · 3.56 Impact Factor
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ABSTRACT: Anti-sperm antibodies were eluted from the sperm cell fraction of autoimmune human ejaculates and transferred onto normal motile spermatozoa. The movement and the acrosomal status of these antibody-coated spermatozoa were evaluated after incubation in a capacitating medium. The amplitude of lateral head displacement (ALH) and the straight line velocity (VSL) were analyzed using an HTM automated motility analyser. Acrosomal loss was monitored by an FITC-conjugated lectin binding technique. During the 6-h incubation in BWW-BSA medium, antibody-free and antibody-coated spermatozoa exhibited significant changes of ALH and VSL distribution that evolved differently in the two populations. The dynamics of sperm movement in control spermatozoa were apparently modified by the presence of antibodies on the sperm membrane. The low percentage of spontaneous acrosomal loss obtained in control populations, even after 20 h of incubation, was not modified by the presence of antibodies on spermatozoa. However, the same antibodies decreased the acrosomal loss induced by a calcium ionophore after 3 h of incubation in capacitating conditions. These results suggest that sperm capacitation and acrosome reaction, considered as essential for successful fertilization, can be altered by antisperm antibodies present on human ejaculated spermatozoa.
Journal of Reproductive Immunology 07/1992; 22(1):59-72. · 2.97 Impact Factor
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ABSTRACT: The present study was designed to investigate whether autoantibodies to external domains of the sperm plasma membrane affect the movement of normal motile spermatozoa. Eight sera and 20 seminal plasma samples containing high levels of anti-sperm antibodies as well as antibodies eluted from the sperm fraction of 19 autoimmune ejaculates were incubated with donor's motile spermatozoa, obtained by swim-up migration in Tyrode's solution. Sperm movement was analysed using 1 s exposure microphotography when greater than 70% of the spermatozoa were coated with antibodies (after 30-90 min of incubation). At least 50 tracks of progressively motile spermatozoa were analysed in order to obtain the mean values of the amplitude of lateral head displacement (ALH) and the velocity of progression (VSL). Serum antibodies and sperm eluted antibodies had quite consistent but opposite effects on sperm movement; serum antibodies increased ALH and decreased VSL whereas eluted antibodies decreased ALH and increased VSL. Seminal antibodies did not affect these two parameters significantly. Furthermore, seminal antibodies and sperm eluted antibodies obtained from the same ejaculates had distinct effects on ALH and/or VSL. This diversity was apparently not linked to antibody isotype or localization on the sperm membrane; it might be due to differences in the composition of the extracellular media. These results suggest a dynamic effect of anti-sperm antibodies on sperm movement, a possibility that merits further investigation.
Human Reproduction 04/1991; 6(3):405-10. · 4.47 Impact Factor
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ABSTRACT: The auto-antibody response in infertile men was investigated by means of immunoenzymatic methods, dot-immunobinding assay (DIBA), and ELISA, using, as antigens, human sperm basic nuclear proteins. Comparison was made, for the same patients, with antibody response to membrane antigens, detected by tray agglutination test (TAT), spermotoxic test (STT), and immunobead binding test (IBT). A very good agreement was observed between the two kinds of antibody responses. Thus, an ELISA or a dot-immunobinding test with sperm nuclear proteins may be considered as a simple and sensitive method for detection of auto-antibodies in infertile men. The reactivity in ELISA of various synthetic peptides corresponding to sequences of human protamines HP1 and HP2 was also studied: all the sera containing anti-nuclear antibodies do not react with synthetic peptides. This observation suggests that antibodies to sperm nuclear proteins recognize conformational epitopes that are not present on small synthetic peptides.
American journal of reproductive immunology (New York, N.Y.: 1989) 06/1989; 20(1):17-20. · 3.05 Impact Factor
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ABSTRACT: In-vitro fertilization and embryo transfer were carried out in twenty infertile couples with significant levels of anti-sperm antibodies on the ejaculated spermatozoa. Over 21 cycles (15 couples), 95 mature oocytes were collected and inseminated with spermatozoa obtained by swim-up migration after rapid dilution and washing of the ejaculates. An overall fertilization rate of 38.9% was obtained with these post-migration (PM) preparations. When greater than 70% of the inseminated spermatozoa were covered with both IgG and IgA antibodies, only 14% of the 43 oocytes were fertilized. A higher fertilization rate was obtained with the PM preparations containing less than 70% of spermatozoa coated with one or both classes of antibodies. Under these conditions 60% of the 52 oocytes were fertilized. Fertility rate correlated better with IgG than with IgA antibody levels. In order to decrease the proportion of antibody-coated spermatozoa in the inseminated populations, washed spermatozoa were immuno-adsorbed on Mage's plates before swim-up migration. Over 11 cycles (10 couples) 52 mature oocytes were inseminated with these post-migration immuno-depleted sperm preparations (PMP) containing less than 65% of antibody-coated spermatozoa: 31% of the oocytes were fertilized. This rate compares favourably to IVF results obtained with the PM preparations with greater than 70% of spermatozoa coated with both classes of antibodies. In five couples who had different IVF attempts with the two sperm preparations, the immuno-depletion resulted in a slight increase in the fertilization rates: 10% for the PM preparations versus 26% for the PMP preparations. Sperm binding to the zona pellucida was decreased in the majority of unsuccessful attempts.(ABSTRACT TRUNCATED AT 250 WORDS)
Human Reproduction 02/1989; 4(1):49-53. · 4.47 Impact Factor
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ABSTRACT: Ejaculates from 51 infertile men with significant levels of anti-sperm antibodies were submitted to processing in vitro in order to increase the number of antibody-free spermatozoa. Rapid removal and washing of spermatozoa from antibody-containing seminal fluid resulted in a variable decrease in IgG and/or IgA binding; only IgG binding was significantly reduced. Those spermatozoa were allowed to swim-up into an overlaying medium to select those with the best progression. Unexpectedly, in the post-migration samples (PM), the proportion of progressively motile spermatozoa coated with IgG and IgA antibodies increased significantly. An efficient selection of antibody-free spermatozoa was achieved prior to swim up migration by immuno-binding to polystyrene Petri plates coated with anti-human immunoglobulin antibodies. Non-adherent populations were depleted of 27-100% of IgG and IgA antibody-coated spermatozoa. After migration, the percentages of antibody-coated spermatozoa of those populations (PMP) remained low. The comparison between PM and PMP populations shows that immunobinding increases the number of motile antibody-free spermatozoa. The fertility potential of both sperm populations is currently under investigation.
Human Reproduction 02/1989; 4(1):44-8. · 4.47 Impact Factor
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ABSTRACT: Thirteen infertile women with high titres of spermagglutinating antibodies in their serum and/or in cervical mucus underwent in-vitro fertilization (IVF) and embryo transfer (ET). Fertilization occurred in 68% of the oocytes in a serum-free medium. Eight pregnancies were obtained in 22 IVF cycles (36.4%). Anti-sperm antibodies were found in the follicular fluids of 5 out of the 11 women with circulating antibodies. Fertilization results were independent of both the localization and the level of anti-sperm antibodies. From these data we can conclude that IVF-ET is a suitable treatment for long lasting female infertility linked to anti-sperm immunity.
Human Reproduction 11/1987; 2(7):599-602. · 4.47 Impact Factor
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ABSTRACT: Semen samples from 120 infertile men with suspected autoimmunity to sperm were investigated by a direct immunobead test (IBT). Fifty-three (44%) of them had 10% or more motile sperm coated with anti-IgG and/or anti-IgA immunobeads. Both classes of immunoglobulins were found to be present in 88.7% of the antibody positive ejaculates. These sperm-bound Igs were associated with sperm autoagglutination in 80% of the ejaculates and with decreased sperm penetration into cervical mucus in 97.6% of the cases. The close correlation found between the IBT results and the occurrence of antisperm antibodies in serum and in seminal plasma suggests that sperm-bound Ig's are sperm-specific antibodies. It is concluded that the direct IBT is not only a reliable screening test for sperm antibodies but is also a relevant test to determine whether these antibodies exert an influence on male fertility.
International Journal of Andrology 11/1986; 9(5):321-30. · 3.59 Impact Factor
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ABSTRACT: Ten infertile men with significant titres of antisperm antibodies were entered into a randomized double-blind study to test the efficacy of corticosteroid therapy compared to a placebo. Prednisolone was given orally at a dose of 1 mg/kg/day for 9 days repeated over 3 cycles (their wives' menstrual cycles). This treatment had no significant effect, when compared to placebo, on fertility, serum antibody levels or semen characteristics. A slight decrease in the titre of seminal antibodies was, however, observed in patients receiving prednisolone. In individuals, there were important fluctuations in antibody levels which were independent of drug administration.
International Journal of Andrology 05/1985; 8(2):111-7. · 3.59 Impact Factor
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La Revue du praticien 01/1984; 33(57):3133-8.
