Munehiro Kato

Acrobeads test: a new diagnostic test for assessment of the fertilizing capacity of human spermatozoa.

OBJECTIVE To determine the effectiveness of the Acrobeads test for predicting the outcome of IVF. DESIGN Human spermatozoa express the CD46 molecule (membrane cofactor protein) on their heads after the acrosome reaction. CD46-positive spermatozoa formed a sperm-bead complex with immunobeads coated with anti-CD46 monoclonal antibody. In the Acrobeads test, fertilizing capacity was determined by assessing sperm-bead agglutination. SETTING Department of Obstetrics and Gynecology, Osaka University Hospital. PARTICIPANTS Thirty-seven donors of proven fertility and 88 male partners of infertile couples. MAIN OUTCOME MEASURES We carried out the Acrobeads test and a sperm penetration assay (SPA) using zona-free hamster oocytes within 3 months before IVF and we then analyzed the results in relation to IVF outcome. RESULTS The sensitivity of the Acrobeads test and SPA was 100% and 88%, respectively, whereas the specificity was 43% and 52%, respectively. The negative predictive value of the Acro-beads test was 100%, whereas that of the SPA was 73%. These results indicate that there was no significant difference between these two tests in terms of predicting IVF outcome. CONCLUSION We suggested that the Acrobeads test be used to evaluate the fertilizing capacity of human spermatozoa because we should avoid using the SPA to prevent cruelty to animals.

HLA expression on human ejaculated sperm.

ABSTRACT: The expression of human leukocyte antigen (HLA) on human ejaculated sperm has been investigated by using cellular binding radioimmunoassay (CB‐RIA) with sperm samples purified by Percoll discontinuous gradient and monoclonal antibodies specific to the frame work of HLA molecules. The results in packed sperm obtained from ten healthy males indicated that HLA class II and its subclasses, DP, DQ, and DR, were expressed at a low level on human ejaculated sperm, while HLA class I was not detected. Furthermore, we could not detect the individual variation of HLA expression on ejaculated sperm in the cases of twenty healthy males.

Preparation of oligozoospermic and/or asthenozoospermic semen for intrauterine insemination using the SpermPrep * semen filtration column †

Objective To improve the quality of oligozoospermic and/or asthenozoospermic semen by the SpermPrep (Fertility Technologies Inc., Natick, MA) semen filtration column. Design The SpermPrep column was applied for semen manipulation in oligozoospermia and/or asthenozoospermia (sperm count 6 /mL, sperm motility Setting Department of Obstetrics and Gynecology, Osaka University Hospital. Patients Twenty-one couples with long-standing infertility because of poor quality semen. Main Outcome Measure Recovery of motile sperm, sperm motility, and outcome of IUI were evaluated among three semen preparations. Results Motility was improved by the SpermPrep method in 32 of 33 cases of oligozoospermia and/or asthenozoospermia. Percentage yield of motile sperm by the SpermPrep method was significantly greater than those by the two-layer Percoll density gradient and swim-up methods (42.7±4.6 versus 22.1±3.1 and 13.8±3.5), but there is no significant difference in the sperm motility among three semen preparations. After one treatment cycle for each preparation, 2 of 21 women conceived after IUI with motile sperm separated in the SpermPrep method. Conclusions The SpermPrep method is an improved semen manipulation method for oligozoospermia and/or asthenozoospermia.

Expression of HLA class I and β2-microglobulin on human choriocarcinoma cell lines: Induction of HLA class I by interferon-γ

Abstract The trophoblast cells which contact with maternal tissue express HLA class I either weakly or not at all. This phenomenon is beneficial for the protection of the fetus from immunological rejection by the mother. However, the mechanisms of reduced expression of HLA class I are unexplained. Therefore, we utilized choriocarcinoma cells to analyse the suppressive mechanisms to HLA class I expression in trophoblasts. In this paper we reported the heterogeneity of HLA class I expression in five choriocarcinoma cell lines. The levels of surface HLA class I molecules correlate approximately with mRNA of HLA class I heavy chain, and all choriocarcinoma cells produce moderate amounts of surface and secreted β 2 microglobulin. It was also found that interferon γ upregulates HLA class I molecules and mRNA in two choriocarcinoma cell lines, NUCI and HCCM5.

Regulation of Classical HLA Class I Genes in Human Choriocarcinoma Cells by Nuclear Proteins Binding to MHC Class I Regulatory Elements

PROBLEM: The regulation of classical HLA class I genes in choriocarcinoma have been reported.

Clinical evaluation of the enzyme-linked immunosorbent assay (ELISA) kit for antisperm antibodies

A commercially available enzyme-linked immunosorbent assay (ELISA) kit for the detection of antisperm antibodies in serum was compared with standard sperm immobilization test (SIT) with the use of sera from 83 infertile women and 29 control individuals. For the ELISA, 24% of the infertile patients and 10% of the controls showed positive results, whereas 15% of the patients and none of the control were positive in the SIT. Parallel tests carried out on the same sera indicated that these methods detect a different, though often overlapping, spectrum of antibody activity. The presence and number of motile sperm in cervical mucus during postcoital tests were found to be related to the results of the SIT. On the other hand, the ELISA did not appear to be related to the quality of postcoital tests. These data indicate that care must be employed to interpret the results of this ELISA kit for the detection of antisperm antibodies.

Evaluation of acrosomal status using MH61-beads test and its clinical application**Supported in part by a Grant-in-Aid for Scientific Research (no. 01570929, 0254383) from the Ministry of Education, Science, and Culture of Japan, Tokyo, Japan.

OBJECTIVE To evaluate the acrosomal status of viable sperm by MH61-beads test and its clinical application. DESIGN Acrosomal status was evaluated using immunobeads coated with MH61 monoclonal antibody, which is specific for acrosome-reacted sperm. When viable sperm was coincubated with MH61-beads for an appropriate length of time, the formation of a sperm-bead complex was observed with a phase-contrast microscope, and the number of sperm binding to MH61-beads increased with the progression of the acrosome reaction. Using this agglutination, we developed the MH61-beads test to assess sperm function. SETTING Department of Obstetrics and Gynecology, Osaka University Hospital. PARTICIPANTS Forty-three volunteers and 20 males in our in vitro fertilization (IVF) and embryo transfer program. MAIN OUTCOME MEASUREMENT The results of MH61-beads test were compared with the percentage of acrosome-reacted sperm detected by Pisium satinum agglutinin staining on 30 volunteers, with the results of sperm penetration assay (SPA) in 43 volunteers, and with the outcome of IVF in 20 patients. RESULTS The results of MH61-beads test showed good reproducibility and correlated with the results of SPA using zona-free hamster eggs and IVF. CONCLUSIONS The MH61-beads test, the results of which reflect the acrosomal status of sperm, may provide useful information concerning the fertilizing ability of sperm.