Effect of testis nondescent or orchidopexy on antisperm antibodies and testis histology in rats.

OBJECTIVE: To examine effects of nondescent of normal testis and of various orchidopexy techniques on antisperm antibody (ASA) production and histologic testicular lesions. DESIGN: Experimental cohort study. SETTING: Laboratories of surgical research and biology of reproduction, academic medical centers. PATIENT(S): Lewis rats, immature and adult. INTERVENTION(S): Eighteen-day-old rats (6 groups): intra-abdominal stay of testis after closure of inguinal canal, classic dartos pouch orchidopexy, orchidopexy by testis fixation through tunica albuginea, orchidopexy by transparenchymal testicular fixation, sham operation, and bilateral vasectomy. Adult rats (1 group): transparenchymal testicular fixation. MAIN OUTCOME MEASURE(S): The ASA--antiacrosome and antitail--were measured by indirect immunofluorescence in sera collected preoperatively, on 50th and 120th day in immature rats, and 90 days after surgery in adult rats. Testicular histology was also examined at the end of sera collection. RESULT(S): Neither intra-abdominal testicular localization nor orchidopexies induced significant ASA. Testicular nondescent and fixation (transparenchymal or transtunical) caused hypospermatogenesis; dartos pouch was harmless. Bilateral vasectomy produced significantly increased ASA, but no significant testicular lesions. Contralateral testes were unaffected. CONCLUSION(S): Intra-abdominal testicular stay and orchidopexy do not elicit autoimmune response to sperm; histologic testicular lesions might not be associated with ASA. In operated cryptorchids, ASA are probably due to other reason than testicular heat or orchidopexy trauma.

Evidence that P36, a human sperm acrosomal antigen involved in the fertilization process is triosephosphate isomerase.

P36 is one of the immunodominant sperm antigens identified by antibodies eluted from the spermatozoa of infertile men. In a previous study, we isolated and characterized this auto-antigen as a glycoprotein with several isoforms. Specific rabbit antibodies were produced to investigate sperm topography and the role of P36 in the fertilization process and we showed that P36 is present on the equatorial segment of acrosome-reacted spermatozoa and is involved in sperm-binding and the penetration of zona-free hamster oocytes. In the present study, we demonstrated, by means of immunofluorescence and electron microscopy, that P36 is present all over the acrosomal membranes of non-reacted spermatozoa. We also investigated the role of P36 in the acrosome reaction and sperm binding to the zona pellucida (ZP). The exposure of capacitated spermatozoa to rabbit anti-P36 antibodies had no effect on primary fixation to the ZP, but inhibited secondary binding to the ZP and the Ca2+ ionophore-induced acrosome reaction. These results suggest that P36, an acrosomal antigen, is involved in several steps of the fertilization process. On two-dimensional Western blots, human anti-sperm antibodies (ASA) and rabbit anti-P36 antibodies recognized five to six isoforms of P36, all 36/37 kDa in size, with a pI between 5.1 and 5.7. Two major spots were identified as human triosephosphate isomerase (TPI) by MALDI-TOF mass spectrometry. Anti-TPI antibodies were shown to react with the isoforms recognized by human and rabbit anti-P36 antibodies. We also demonstrated the presence of TPI in human sperm heads. Further studies are underway to establish whether there is a sperm-specific isoform of TPI and its role in sperm function.

Evidence of genotypic resistance diversity of archived and circulating viral strains in blood and semen of pre-treated HIV-infected men.

OBJECTIVE: To study the genetic diversity of drug-resistant HIV strains present in blood and in semen, especially those archived in peripheral blood mononuclear cells (PBMC) and non-sperm cells (NSC). METHODS: Paired blood and semen samples were collected from twenty heavily pre-treated HIV-infected men. HIV RNA in blood plasma (BP) and seminal plasma (SP), as well as proviral DNA in PBMC and NSC were quantified and used for resistance genotyping. Phylogenetic analysis of protease gene clones was used to explore the diversity of the viral quasi-species. RESULTS: Median BP HIV RNA, PBMC proviral DNA, SP HIV RNA and non-sperm cell proviral DNA loads were respectively: 4.77, 3.65, 3.16 and 1.77 log10 copies per ml or per 10 cells. Resistant HIV strains were found in the BP and PBMC of all the patients, in the SP of 14 patients, and in the NSC of five patients. Overall, the blood and genital compartments exhibited different genotypic resistance patterns in six patients (30%), with additional resistance mutations in the semen of four patients. Phylogenetic analysis of clones of HIV protease gene showed that viral strains in SP originated not only from passive diffusion from BP, but also from local production in semen. The storage of archived proviruses differed according to the anatomic reservoir. CONCLUSION: HIV resistant strains are frequent (70%) in the semen of heavily pre-treated men, and the diversity of genotypic resistance pattern confirms HIV compartmentalization. Thus, the risk of sexual transmission of resistant strains can only be partly predicted by standard tests applied to BP.

Puberty does not induce serum antisperm surface antibodies in patients with previously operated cryptorchidism.

PURPOSE: We investigated serum antisperm surface antibody (ASA) prevalence at puberty, which is reported to be as high as 38% in the sera of males with cryptorchidism operated on before puberty. Operative technique impact, dartos pouch orchiopexy or testis fixation, was also examined. MATERIALS AND METHODS: We examined a total of 61 pubertal males (Tanner stage 2 or greater) divided into 3 groups. Group 1 consisted of 24 males with cryptorchidism 10 to 17.9 years old who underwent unilateral dartos pouch orchiopexy before puberty (median age 5.85). All of these cases were known to be negative for ASA preoperatively, and 20 before puberty. Group 2 consisted of 22 males with cryptorchidism 12.1 to 17.7 years old operated on previously (median age 10.35) by testicular fixation among other techniques. Group 3 consisted of 15 healthy males 12.2 to 17.3 years old. Prepubertal ASA status was unknown for groups 2 and 3. Operated testis was compared with counterpart before serum collection in group 1 and during operation in group 2. IgG IgM and IgA ASA were studied by the indirect Immunobead (BioRad, Clinisciences S.A., Montrouge, France) test. RESULTS: All sera tested were found negative in the 3 groups. Dartos pouch operation, testis fixation or even consecutive operations did not induce ASA production. Alterations in size or consistency were observed in operated testes in 10 patients in group 1, and in 8 patients in group 2. CONCLUSIONS: Our results suggest that dartos pouch orchiopexy, testicular fixation and/or intrinsic developmental alterations of the cryptorchid testis does not elicit an autoimmune response against sperm surface antigens at puberty.

Deficit in cytochrome c oxidase activity induced in rat sperm mitochondria by in vivo exposure to zidovudine.

A decreased sperm motility has been reported in men treated with nucleoside analog reverse transcriptase inhibitors (NRTI). Sperm motility is correlated with enzymatic activities of the sperm mitochondrial respiratory chain (MRC), which may be impaired by NRTI. We compared sperm and skeletal muscle MRC and citrate synthase (CS) activities, sperm adenosine triphosphate (ATP) content and sperm motility between rats exposed to zidovudine (AZT) for 10 weeks and controls. Decreased levels of CS-normalized cytochrome c oxidase (COX, the MRC complex IV) activity were observed in the spermatozoa from AZT-treated rats, with no significant decrease in ATP content or motility. In muscle absolute COX activity increased after exposure to AZT but CS-normalized COX activity remained unchanged. These results suggest that exposure to NRTI can induce MRC dysfunction earlier in spermatozoa than in skeletal muscle.

Assisted reproduction in HIV-1-serodifferent couples: the need for viral validation of processed semen.

BACKGROUND: Many HIV-infected men and women have a strong desire for a child. Assisted reproductive technologies (ART) are an option for HIV-serodifferent couples to reduce the risk of HIV transmission from an infected man to the woman. Potential HIV contamination of selected spermatozoa after semen processing is an important issue in this context. METHODS: HIV in processed semen obtained in our laboratory since 1995 were analysed. HIV RNA and DNA detection was performed in the selected spermatozoa of 125 men. HIV RNA was analysed in blood and semen plasma as well as HIV DNA in non-sperm cells. RESULTS: HIV RNA and DNA were detected in the selected spermatozoa of eight and two men (6.4% and 1.6%), respectively. HIV RNA was detected with a median load of 5 copies/10(6) spermatozoa. Six of the eight men were untreated, one was taking nucleoside analogue therapy and one was on highly active antiretroviral treatment (HAART). HIV RNA detection was more likely to be positive in selected spermatozoa of men with high seminal plasma viral load. HIV RNA was detected in 26% and 11% of selected spermatozoa fractions when the seminal plasma load was > 10,000 copies/ml and 20-10,000 copies/ml, respectively, but in none when the seminal plasma tested negative. CONCLUSION: Selected spermatozoa may be positive for HIV RNA detection even in treated patients. Viral validation of processed semen is necessary in ART programmes for serodifferent couples, particularly in men with only partially or poorly controlled HIV infection.