ELISA法检测不育男子精浆中抗精子IgG和IgA

将精子经Tritonx－100处理，冷冻高速离心后，经抗人全血清-SephadexG－75亲和柱层析分离，提取分子量为59KD人精子膜蛋白作为抗原，经ELISA间接法对20例生育男子和50例不育男子精浆中抗精子IgG和IgA进行了测定。结果显示：生育男子精浆中抗精子IgG为阴性；抗精子IgA阳性率为5％。不育男子精浆中抗精子IgG和IgA阳性率分别为10％、30％；抗精子IgG和IgA均为阳性者3例，均为阴性者33例；抗精子IgG阳性而IgA阴性者2例；抗精子IgA阳性而IgG阴性者12例。不育组与生育组间抗精子IgG阳性率无显著性差异（P＞0．05）；而IgA阳性率间则有显著性差异（P＜0．05）。不育组抗精子IgG和IgA阳性率间有极显著性差异（P＜0．01）。本文利用ELISA法对精浆中抗精子抗体的分类及可能来源进行了讨论。     

抗SPIM自身抗体的ELISA测定与临床应用

用ELISA 法定量检测生育、不育男性血清、精浆抗SPIM 自身抗体(SPIM-Ab)。结果表明:不育组和妻子流产组患者血清、精浆SPIM-Ab 含量均显著高于生育组(P<0.01),两组血清、精浆的SPIM-Ab 阳性率与生育组之间有显著性差异(P<0.01);精浆SPIM-Ab 含量高于5.0U/ml 时,精于密度、精子活率及精浆SPIM 活性均明显降低;SPIM-Ab 与血清、精浆抗精子抗体(AsAb)无明显关系。提示:SPIM-Ab 水平增加可能抑制SPIM 活性、与SPIM一起参与局部免疫反应,并减少精子密度和活率。     

Cytotoxic sperm antibodies inhibit sperm penetration of zona-free hamster eggs

Zona-free hamster egg sperm penetration assay was used to study the effects of cytotoxic sperm antibodies on egg penetration by the sperm of fertile and infertile men. Twenty-nine fertile and 9 infertile men did not have significant cytotoxic sperm antibodies in their serum and seminal plasma; 7 infertile men were positive for these antibodies in serum and seminal plasma. Two others were positive in sera, and 14 were positive in seminal plasma. Sperm from 18 of 23 (78%) infertile men with sperm antibodies had poor egg penetration (less than or equal to 20%) compared with only 6 of 38 (16%) nonautoimmune men (P less than 0.0001). Sperm from nonautoimmune fertile men were coated with seminal plasma and serum of autoimmune men and serum of isoimmune women, resulting in a significant decrease in hamster egg penetration. Sixteen of 21 (76%) seminal plasma samples with cytotoxic sperm antibodies reduced the control sperm penetration of hamster eggs by greater than or equal to 50%. Coating of sperm from fertile men with serum and seminal plasma samples from non-sperm-immune fertile and infertile subjects did not alter their penetration of hamster eggs. Coating of sperm from autoimmune men with cytotoxic antibody-positive autologous seminal plasma samples resulted in a significant decrease of egg penetration. The inhibitory effect of antibody-positive seminal plasma samples on egg penetration by control sperm was abrogated when the samples were preabsorbed with sperm. It is concluded that cytotoxic sperm antibodies, especially those in seminal plasma, inhibit hamster egg penetration by autologous and control sperm. This may explain in part the incidence of infertility associated with sperm antibodies.     

The lack of discriminant seminal and endocrine variables in the partners of fertile and infertile women

Seminal and endocrinological investigations were done on 50 fertile men whose wives became pregnant spontaneously (Group I), on 100 men from our fertility clinic whose wives conceived after treatment of the couple (Group II) and in 100 infertile men whose wives failed to become pregnant even after 5-year treatment of the couple (Group III). Patients with azoospermia were excluded. While there were no significant differences in the mean sperm density and total sperm count of the men from all three groups, mean sperm motility and sperm morphology were significantly better (p less than 0.001) in both Group I and Group II men than in the infertile Group III men. The mean serum LH and FSH concentrations were significantly lower in Group I than in the two other groups. However, there were no differences between Groups II and III. The mean serum concentrations of prolactin, testosterone, and estradiol were similar in all three groups. Based on semen and hormone data, stepwise multiple discriminant analysis could allocate 80% of the Group I men correctly, whereas only 50% of Groups II and III could be correctly classified. Although overall differences in sperm motility and morphology as well as in serum gonadotrophin concentrations were noted, no specific seminal or hormonal variable could be identified as being of value in predicting the fertility of an individual man.     

Seminal immune response in infertile men with leukocytospermia: effect on antioxidant activity.

OBJECTIVE: To investigate the incidence of leukocytospermia and relation to T helper cytokines, tumor necrosis factor alpha (TNFalpha) and interleukin-4 (IL-4), antisperm antibodies and antioxidant activity. Design: Semen samples from 176 infertile men and 24 fertile controls were investigated. METHODOLOGY: The protocol included tubal patency test, hysterosalpingography and laparoscopy and dye test and ovulation through mid-luteal phase progesterone for the wives. The husbands had semen analysis, cytomorphology evaluation and semen culture. Seminal TNFalpha and IL-4, antisperm antibodies, total antioxidant activity, superoxide dismutase and zinc were determined. RESULTS: Leukocytospermia occurred in 44.3% of the infertile men compared to 12.5% of the fertile men (P<0.01). Thirty-six (20.5%) men had pathogenic bacterial organisms which constituted 46.2% of those with leukocytospermia. Sperm parameters were worse with leukocytospermia in terms of sperm count (P<0.01), total motility progressive motility (P<0.01), morphology, asthenozoospermia, sperm membrane integrity and antisperm antibodies. TNFalpha and IL-4 had an inverse relationship; the expression of TNFalpha was higher with leukocytospermia and bacteriospermia (P<0.001), while IL-4 was higher in fertile controls (P<0.005). Incidence of antisperm antibodies was higher with leukocytospermia. Total antioxidant activity, superoxide dismutase and zinc were lower with leukocytospermia. CONCLUSION: Leukocytospermia impairs sperm function through reduced antioxidant activity and enhanced T helper 1 modulation.     

Identification of human sperm antigens reacting with antisperm antibodies from sera and genital tract secretions.

OBJECTIVE: To identify sperm antigens reacting with antisperm antibodies relevant in human infertility. DESIGN: The reactions of separated sperm antigens with antibodies present in sera and genital tract secretions from infertile and fertile females and males were examined by immunoblotting techniques. SETTING: The patients were followed in an outpatient setting of a hospital clinic. PATIENTS: One hundred consecutive infertile males and females, referred for determinations of antisperm antibodies, comprised the study group. Fifty hospital and faculty employees with proven fertility served as a control group. RESULTS: A high proportion of sera from fertile and infertile humans contained antibodies reacting with at least one sperm antigen. However, two discrete bands of antigenic proteins with molecular weights of 44 and 72 kd reacted significantly more frequently with serum antibodies from infertile females than from fertile females. No apparent correlation could be demonstrated between any particular antigen and serum antibodies from infertile males. Nevertheless, antigenic proteins of 62 kd were identified as the major sperm antigens reacting with antibodies present in seminal plasmas from infertile males. CONCLUSIONS: The major sperm antigens reacting with systemic antibodies differ from the antigens recognized by local antisperm antibodies. Sperm antigens exhibiting relative molecular weights of 62 kd are major antigens reactive with local antisperm antibodies from infertile humans.     

Antibodies to A19 and Bw35 complexes of human leukocyte antigens are present in infertile subjects with sperm antibodies

Sera and secretions from 100 couples with unexplained infertility were tested for sperm antibodies by cytotoxicity and passive hemagglutination and also for antibodies to human leukocyte antigen (HLA) by cytotoxicity assays. Lymphocytes of the study subjects were typed for 61 HLA-A and B alleles. Thirteen of 30 (43%) men with sperm autoimmunity also had HLA antibodies in their serum and/or seminal plasma samples, in contrast to 2 of 35 (6%) nonautoimmune males (P = 0.0003). Twenty-five of 35 (71%) sperm antibody-positive infertile women had HLA antibodies in their sera and/or secretions, while only 7 of 31 (23%) women without sperm antibodies were positive (P = 0.00007). Antibodies to HLA-A19 (A26, A29, AW30, AW31, AW32, AW33, and AW34) and Bw35 (B5, B15, B17, and B18) complexes were present in 19 of 22 (86%) infertile men and 44 of 48 (92%) infertile women positive for HLA antibodies (P less than 0.01). The presence of antibodies to HLA-A19 and/or Bw35 in the infertile subjects did not correlate with the presence of HLA-A19 and/or Bw35 in the husbands. The presence of antibodies to HLA-A19 and/or Bw35 in the cervical mucus of the infertile women correlated with their presence in the seminal plasma of their husbands. It is suggested that antibodies to sperm antigens cross-reactive with HLA-A19 and/or Bw35 may be relevant to infertility.     

Differences in blood and semen oxidative status in fertile and infertile men,and their relationship with sperm quality

Oxidative stress plays a fundamental role in the aetiology of male infertility by negatively affecting sperm quality and function. Assessment of blood and seminal plasma oxidative profiles might be a valuable tool to improve evaluation of sperm reproductive capacity and functional competence. This study examined the lipid-soluble antioxidant profile and levels of lipid peroxidation both in blood and seminal plasma samples of infertile and fertile males, in relation to semen parameters. Total antioxidant capacity (TAC) and vitamin E concentrations were significantly (P < 0.05) lower in seminal plasma of infertile men compared with fertile subjects; concurrently, a significant accumulation of malondialdehyde was found in infertile patients (P = 0.032 compared with controls), which was negatively correlated with sperm motility and morphology. In blood samples, infertile men presented lower concentrations of TAC, carotenoids and vitamin E than fertile subjects; TAC and carotenoids were positively correlated with sperm motility, morphology and concentration. Finally, blood TAC and vitamin E concentrations were positively correlated with the corresponding seminal values, confirming the close relationship between blood and semen antioxidants. All these results indicated the possibility of using not only seminal antioxidants but also blood antioxidants as biochemical markers to support sperm quality evaluation.Oxidative stress induced by reactive oxygen species (ROS) has been widely recognized as one of the major causes of male infertility; indeed, excessive ROS production can negatively impact sperm quality and function. The assessment of blood and seminal plasma oxidative profiles has been suggested as a valuable tool to improve the evaluation of sperm reproductive capacity and functional competence in infertile men. With this in mind, in the present study we examined the lipid soluble antioxidant profile (carotenoids and vitamins A and E) and the levels of lipid peroxidation (malondialdehyde; MDA) both in blood and seminal plasma samples of infertile and fertile males, in correlation with semen parameters namely motility, morphology and concentration. As a result, we obtained evidence that the total antioxidant capacity (TAC) and the concentrations of vitamin E of seminal plasma samples were significantly lower in infertile men than in fertile subjects; at the same time, a significant accumulation of MDA was found in infertile patients. MDA, in turn, negatively correlated with sperm motility and morphology, thus confirming that oxidative damage to lipids impairs sperm quality. In blood samples, infertile men presented lower TAC and lower concentrations of carotenoids and vitamin E than fertile subjects; interestingly, TAC and carotenoid concentrations were positively correlated with sperm motility, morphology, and concentration, confirming the close relationship between blood antioxidants and sperm quality. In conclusion, all these results suggested that the examination of blood and semen oxidative profiles might furnish useful information on sperm quality and function in infertile men.     

Oral immunization with sperm antigens: possible therapy for sperm antibodies?

Young adult male CD-1 mice were given intraperitoneal injections (IP) of saline (controls) and pooled sperm or seminal plasma of two autoimmune infertile men and two nonautoimmune fertile men (n = 40 per treatment). Other mice received only an oral challenge with the same antigens (oral controls; n = 20 per treatment). Three weeks after the booster challenge (day 36), 20 mice in each group were orally immunized with the antigens, whereas the other 20 were not (IP controls). Cytotoxic antibody titers (immunoglobulin M) to human sperm were significantly higher in mice IP immunized with sperm or seminal plasma from autoimmune infertile men or orally immunized with autoimmune men's sperm, in contrast to the controls. Oral challenge with sperm or seminal plasma of autoimmune infertile men after the IP immunization with the same resulted in significantly decreased cytotoxic sperm antibody titers (P less than 0.001 versus oral or IP controls in sperm immunization; P less than 0.001 versus IP controls in seminal plasma immunization). Fertility was unaffected by any mode of immunization. It is concluded that, in mice, sperm and seminal plasma antigens from autoimmune infertile men are more immunogenic than those from nonautoimmune fertile men, and oral challenge with the former after an IP establishment of cytotoxic sperm immunity desensitizes the immune mice. These findings may have practical implications in the diagnosis and immunotherapy of infertile men with cytotoxic sperm antibodies.     

Occurrence of serum antisperm antibodies in patients with cystic fibrosis

OBJECTIVE: To determine if acquired obstruction of the vas deferens in men with cystic fibrosis (CF) induced the development of antisperm antibodies with genital tract obstruction similar to other men. DESIGN: Serum antisperm antibodies were assayed by an indirect immunobead test and an indirect immunofluorescence assay. Both homologous (human sperm/human zona) and heterologous (human sperm/zona-free hamster ova) sperm/egg interactions were evaluated in the presence of serum antisperm antibodies from patients with CF. SETTING: Cystic Fibrosis Clinic at the University of Oklahoma Health Sciences Center, a tertiary care referral center. PATIENTS: Fifteen CF patients (10 male and 5 female), 3 non-CF antisperm antibody-positive infertile patients (2 male and 1 female), 20 fertile controls (7 males and 13 females), and 9 fertile sperm donors were used. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Serum antisperm antibody levels in patients with CF. In those patients with antisperm antibodies, determine effect of these sperm antibodies on sperm/egg interactions and complement-mediated events. RESULTS: Sera from 3 (30%) of 10 men with CF demonstrated immunoglobulin (Ig)G, IgA, and/or IgM antisperm antibodies, whereas sera from all 5 CF women and the 20 control sera were negative for antisperm antibodies. The maximal titers for IgG, IgA, and IgM antisperm antibody were 1:8, 192, 1:256, and 1:64, respectively. The immunobead binding, which was restricted to the sperm head and tail-tip or the midpiece and tail-tip, correlated with the indirect immunofluorescence pattern. Antisperm antibody-positive sera from men with CF impaired both the binding and penetration of human zonae and the penetration of hamster ova by human sperm. CONCLUSIONS: Similar to other men with congenital or acquired obstruction of their genital tract, antisperm antibodies may occur in some men with CF. Antisperm antibodies may contribute to immune sperm dysfunction in some men with CF by activated complement-mediated events and interfering with sperm/egg interactions.     

Detection of autoimmunity to sperm: mixed antiglobulin reaction (MAR) test or sperm agglutination? A study on 537 men from infertile couples

Two different ways of testing for antisperm antibodies were compared: the mixed antiglobulin reaction (MAR) test for demonstration of antibodies of the IgG and IgA classes bound in vivo to the sperm membrane antigens and the gelatin agglutination test for detection of nonbound antisperm antibodies in serum and seminal plasma. Samples from 537 men from infertile couples were investigated. Antibodies bound to the sperm membrane were detected in 49 men (9.1%), IgG in 44 (8.2%), and IgA in 38 cases (7.1%). Sperm agglutinins were recorded in seminal plasma from 30 men (5.6%) and in serum (titer greater than or equal to 16) from 43 men (8.0%). The investigation revealed a very close correlation between the results of MAR testing and the occurrence of sperm agglutinins in serum and seminal plasma. However, if one focuses on antisperm antibodies of the IgA class, which seem to play the major role in male immune infertility, the MAR test offered the advantage that a minor group of patients with pure IgG responses could be distinguished, and rare cases with mainly or exclusively locally produced IgA antibodies could be detected.     

Antisperm antibodies in infertile and homosexual men: relationship to serologic and clinical findings

The incidence and significance of antisperm antibodies in different groups of men were evaluated by a modified enzyme-linked immunosorbent assay. In serum, 4.0% of dermatologic patients (n = 223), 9.6% of andrologic patients (n = 178), and 28.6% of homosexual men (n = 42) were positive for IgG and/or IgM antibodies. In seminal fluids, 7.3% of the andrologic patients had IgA (and IgG) antibodies to spermatozoa. Only 1 of 29 positive men had antibodies both in serum and in seminal fluid. No correlation between antisperm antibodies and IgG/IgM concentrations was found in serum, whereas in seminal plasma men with antisperm antibodies showed higher IgG/IgA concentrations than men without (IgA, 3.2 versus 1.7 mg/dl; IgG, 9.8 versus 6.3 mg/dl). It is concluded that there is a high incidence of antisperm antibodies among homosexual men, probably because of contact of spermatozoa with the immune system by passive anal intercourse. There is little correlation between antisperm antibodies in serum and seminal plasma of infertile men because of a lack of relevant antibody transfer from the serum and the formation of local antibodies in seminal plasma. Antisperm antibodies in seminal fluid are associated with elevated local IgG and IgA concentrations.     

Presence of immunoreactive beta-endorphin and calcitonin in human seminal plasma, and their relation to sperm physiology

Opioid peptides and calcitonin are found in high concentrations in the male reproductive tract. To further elucidate their role in sperm physiology, we studied semen samples from 49 infertile men and 25 men with proven fertility. beta-endorphin and calcitonin were measured in each sample by radioimmunoassay and then were correlated with seminal plasma testosterone (T) and dihydrotestosterone levels as well as sperm count, total motile sperm/milliliter, and percentage of penetrated hamster eggs. The levels of beta-endorphin (308 +/- 22 pg/ml) and calcitonin (331 +/- 32 pg/ml) in seminal plasma were 10 and 20 times higher than levels found in venous plasma (32 +/- 2 and 14.5 +/- 1.2 pg/ml, respectively) (P less than 0.001). There was no difference between the levels of beta-endorphin and calcitonin in seminal plasma of fertile and infertile men. However, seminal plasma T was significantly higher in fertile than infertile men (19.4 +/- 2 versus 11.5 +/- 1 ng/dl; P less than 0.05). No correlation could be demonstrated between either beta-endorphin or calcitonin and any of the parameters studied. In conclusion, beta-endorphin and calcitonin are produced locally in the male reproductive tract; however, their role in male reproduction remains to be elucidated.     

人精浆与血清中生殖激素及免疫球蛋白的相关性研究

对152例生育与不育男性,分组观察精浆和血清中五种生殖激素(ICSH、FSH、PRL、T、E_2)及三种免疫球蛋白(IgG、IgA、IgM)的变化,分析它们之间的相关性以及与精液质量的关系。结果表明:1.生殖激素在精浆与血清中含量变化不一致。精浆中ICSH与PRL呈正相关,精浆ICSH、PRL与精于活力、密度间亦呈正相关关系。2.精浆IgG与血清、精浆T呈负相关,不育者精浆IgG与精子活力亦为负相关关系。     

A reverse (antibody capture) enzyme-linked immunosorbent assay for detection of antisperm antibodies in sera and genital tract secretions

A reverse (antibody capture) enzyme-linked immunosorbent assay (ELISA) for detection of antisperm antibodies has been developed. The assay enables detection of immunoglobulin (Ig) M, IgG, IgA, or IgM, IgG, and IgA--antisperm antibodies in serum, cervical mucus, and seminal plasma samples. The reverse ELISA is more specific and sensitive than conventional ELISA in detecting human antisperm antibodies of different isotypes. Using this assay, statistically significant differences in levels of antibodies between infertile and fertile individuals were demonstrated in sera and in genital tract secretions. Studies with 143 infertile couples revealed that the presence of antibodies in sera was not necessarily reflected in individual's genital tract secretion and vice versa. These data emphasize the importance of detecting antisperm antibodies in sera as well as in genital tract secretions for correct evaluation of sperm immunity.     

溶脲脲原体感染不育者精浆α－葡萄糖苷酶活性及果糖含量的分析

本文比较了溶脲脲原体感染的不育男性与正常生育男性精浆中α－１，４－葡萄糖苷酶活性及果糖含量。结果表明，溶脲脲原体感染的不育肾性精浆中α－１，４－葡萄糖苷酶活性明显低于正常生育男性（分别为：Ｘ±ＳＤ＝２７．９９±２０．３１ｍＩＵ／ｍｌ和Ｘ±ＳＤ＝４１．８６±２１．８６ｍＩＵ／ＭＬ，ｐ＜０．０５）；溶脲脲原体感染的不育男性精浆中果糖含量（Ｘ±ＳＤ＝２３３．９６±８９．２１ｍｇ／ｄｌ）稍高于正常生育男性（Ｘ±ＳＤ＝１７９．８１±１０６．４５ｍｇ／ｄｌ），但两者之间差异无显著性（Ｐ＞０．０５）。上述结果提示，生殖道溶脲脲原体感染对附睾功能具有干扰作用，而对精囊腺的功能影响尚缺乏依据，宜进一步探索。     

精浆内抗精子抗体和免疫复合物的研究——对男性不育患者及输精管粘堵术者的观察

本文用ELISA法研究了174名男性不育患者及43名输精管粘堵术后精浆中的抗精子抗体及其种类。结果表明,精浆中抗精子抗体的检出率,在男性不育患者中为24.1%,输精管粘堵术后者为48.8%。在这些患者的精浆中可检出IgA、IgG和IgM型抗精子抗体,其中主要是IgA。同组人群血清和精浆中的抗精子抗体缺乏相关性。精浆中抗精子抗体的存在与精子活动率的下降有一定的关系。应用PEG法测定精浆中免疫复合物,在66例精子抗体阳性的患者中,阳性率为56.1%,表明局部抗精子的自身免疫反应,在一定条件下可导致免疫复合物的形成。     

A comparison of sperm agglutination and immobilization assays with a quantitative ELISA for anti-sperm antibody in serum

An enzyme-linked immunosorbent assay (ELISA) that quantitates antisperm antibody in serum was compared with standard sperm agglutination and immobilization assays with the use of sera from 40 normal and 292 subfertile individuals. Quantitation of the assay was accomplished by standardizing assay parameters, including the incorporation of a standard reference curve, the number of whole target sperm, the optimal dilution of serum, the selection of microtiter plate, and the time and temperatures involved in the adsorption and incubation phases. With this method, the level of antisperm antibody binding to target sperm in 40 normal fertile individuals was found to be 2.3 (+/- 1.1 standard deviation [SD]) fg immunoglobulin (Ig)/sperm. An increased mean level of 7.4 +/- 3.7 fg Ig/sperm was determined in 84 infertile patients with positive agglutination and/or immobilization tests. In 208 individuals with negative agglutination and immobilization tests the mean concentration of antisperm antibody was 2.5 +/- 1.3 fg Ig/sperm. Postvasectomy patients assayed by this method had a mean Ig binding value of 7.1 +/- 2.4 fg Ig/sperm. The infertile group with positive agglutination and/or immobilization tests had a significantly higher mean antisperm antibody level than the normal fertile group, according to the Student's t-test for independent samples (P less than 0.001). This indirect serum-based assay reproducibly quantitates antisperm antibody binding to whole target sperm, suggests the normal and abnormal levels of antisperm antibody, and correlates with standard functional assays.     

人精子中芳香化酶表达与精子功能的关系

目的:研究精子细胞色素P450芳香化酶(P450arom)的表达及其与精子功能状态和受精力的关系。方法:以人精子穿透去透明带金黄地鼠卵异种体外受精试验(SPA)检测精子受精力;以三色法染色观察精子顶体反应(AR)的发生率。采用RT-PCR,用P450arom/GAPDH光密度值的比值代表P450arom的表达水平。结果:精子P450arom表达水平与受精率有一定的相关性(生育组r=0.5622;不育组r=0.6071)。正常男性与不明原因不育症患者精子P450arom/GAPDH比值分别为0.60±0.29,0.39±0.16,有显著差异(P<0.02)。P450arom表达水平与精子AR的发生率也有一定相关性(生育组r=0.5817;不育组r=0.5535)。结论:人精子中存在着P450arom表达产物;P450arom可能与精子功能有一定关系;P450arom表达异常可能与一些不明原因不育有关。     

Zinc sulfate treatment of secondary male infertility associated with positive serum and seminal plasma anti-sperm antibody test

ObjectiveThe traditional use of corticosteroids in the treatment of infertile patients who developed anti-sperm antibodies may be associated with many systemic side effects of such administered drugs. So the use of alternatives for corticosteroids becomes mandatory in order to minimize these systemic side effects. Like the use of immune modulator zinc sulfate.DesignProspective study.SettingInstitute of the Embryo Research and Infertility Treatment, College of Medicine, AL – Nahrain University.Patients and methodsForty-eight infertile men with poor semen parameters and elevated serum and seminal plasma level of ASA were classified into three groups. The first group (n = 18 patients) received oral zinc sulfate, the second group (n = 20 patients) received oral prednisolone and the third group (n = 10 patients) received oral placebo. Drugs were administered for 45 days.Main outcome measure(1) Standard semen analysis was performed, including semen volume, sperm concentration, percent motility, sperm vitality and sperm morphology. (2) Serum and seminal plasma Anti-Sperm Antibody ELISA test.ResultsStatistical analysis of serum and seminal plasma level of ASA reveals that the level is decreased significantly (P < 0.001, P < 0.002) in zinc sulfate treatment group as compared to the control group (67.5 ± 2.64 vs. 71.6 ± 2.5, 66.94 ± 3.24 vs. 71 ± 2.54), respectively, but the decrease in prednisolone treatment group is significantly higher (P < 0.001) as compared to the zinc sulfate treatment group (57.3 ± 6.30 vs. 67.5 ± 2.64, 56.5 ± 6.83 vs. 66.94 ± 3.24), respectively. This improvement in serum and seminal plasma level of ASA, result in a significant enhancement of the seminal fluid parameters including sperm concentration, sperm motility, vitality and normal sperm morphology in both zinc sulfate and prednisolone treatment groups.ConclusionIn conclusion, infertile men with elevated level of ASA and poor basic parameters of seminal fluid characters can be treated with zinc sulfate protocol with great chance for decreasing the level of serum and seminal plasma ASA, and alleviating the serious side effects associated with corticosteroids.