The antibody against a nuclear autoantigenic sperm protein can result in reproductive failure / 亚洲男科学杂志(英文版)

To study whether the antibody against the testis form of the nuclear autoantigenic sperm protein (tNASP) could result in reproductive failure, we successfully cloned and expressed a 339-bp cDNA fragment of mouse tNASP (mtNASP). Using mouse as a model, recombinant mtNASP (rmtNASP) and a synthetic peptide, human tNASP(393-408) (htNASP(393-408)), were investigated for their antifertility effect. Active immunization with rmtNASP or the synthesized peptide raised high antibody titers in the immunized mice. Sperm-egg binding and fusion assay were carried out in 8-10-week-old BALB/c mice. Sperm-egg binding and in vitro fertilization of mouse oocytes were inhibited by co-incubation of zona-free mouse oocytes with capacitated mouse spermatozoa in the presence of varying concentrations of the antisera against rmtNASP. There was a significant antifertility effect in animals immunized with rmtNASP or the synthesized peptide. The effect on fertility in the mice immunized with the synthesized peptide was reversible. Our data indicate that active immunization with rmtNASP antigen may induce a strong antibody response that causes an inhibition of fertility.

Inhibition of mouse acrosome reaction and sperm-zona pellucida binding by anti-human sperm membrane protein 1 antibody / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To investigate the possible functions of human sperm membrane protein (hSMP-1) in the process of fertilization.</p><p><b>METHODS</b>A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 (anti-rhSMP-1) antibodies. Sperm acrosome reaction and sperm-zona pellucida (ZP) binding assay were carried out in 10-week-old BALB/c mice.</p><p><b>RESULTS</b>Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 (P < 0.05).</p><p><b>CONCLUSION</b>The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.</p>

Progress in the study of immunocontraceptive antigens / 中华男科学杂志

Fertility management is a global issue of medical, economic, and social consequence. Although many methods have been devised to inhibit reproduction, more acceptable alternatives are still needed. Regulation by immune intervention is a promising technology as applied to human beings. The objective of this review is to indicate several immunocontraceptive antigens.

Cloning, expression and purification of human nuclear autoantigenic sperm protein (hNASP) and preparation of its polyclonal antibody / 中华男科学杂志

<p><b>OBJECTIVE</b>To acquire the purified human nuclear autoantigenic sperm protein (hNASP) and its polyclonal antibody for investigating the possible functions of hNASP involved in fertilization.</p><p><b>METHODS</b>The coding sequence of hNASP gene was amplified from human testis RNA with specific primers, and the PCR product was cloned first into pMD-18T and then into pET-28a ( + ) after restriction digestion with BamH I and Hind III. The fusion protein was expressed in E. coli BL21 (DE3) after induction with IPTG. The recombinant protein NASP was purified from the supernatant with Ni2 -NTA His-bind resin under native conditions.</p><p><b>RESULTS</b>The results of DNA sequencing and SDS-PAGE analysis showed the protein to be what we had hoped to acquire. ELISA showed that we had acquired rabbit anti-hNASP serum with high titer.</p><p><b>CONCLUSION</b>High purity recombinant hNASP protein could be obtained with the above-mentioned prokaryotic expression method, and so could the rabbit anti-hNASP serum with high titer and high specificity.</p>

Establishment and application of the approach to detecting two biovars of Ureaplasma urealyticum in human semen / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish the approach to detecting two biovars of Ureaplasma urealyticum (Uu) in human semen and to investigate the relationship between the two biovars of Uu infection and the quality of human semen.</p><p><b>METHODS</b>Based on the 16S-23S rRNA intergenic spacer region, three pairs of primers were designed, the species specific primer and two biovars primers (Parvo primer and T960 primer). The two biovars of Uu were detected in the semen from 949 men by semen culture and PCR assay. Meanwhile, semen routine analyses were performed.</p><p><b>RESULTS</b>In the 949 subjects, 199 were Uu positive both in Uu liquid culture and PCR assay (199/949, 21.1%), of which 136 (136/199, 68.3%) were Parvo biovar, 54 (54/199, 27.1%) T960 biovar, and 9 (9/199, 4.5%) both Parvo and T960 biovars. Compared with the Parvo and the negative groups, human sperm viability was significantly decreased (P < 0.05 ) in the Uu T960 infection group. The difference of sperm motility and density had no statistic significance.</p><p><b>CONCLUSION</b>A significant correlation has been found between Uu T960 biovar infection and human sperm viability</p>