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Millipore filtration of motile human spermatozoa with the use of a pore size of 5 micrometer without applying high or low pressure results in a small but statistically significant increase of Y-spermatozoa in the filtrate. Immobilized spermatozoa do not pass through the membrane.

The in vivo effect of purified human alpha1-antitrypsin and inter-alpha-trypsin inhibitor on the fertilizing ability of capacitated rabbit spermatozoa was investigated and compared with that of the trypsin-kallikrein inhibitor from bovine organs (Trasylol). Only 250 mug of alpha1-antitrypsin/5 X 10(4) sperm in 0.05 ml inhibited fertilization by more than 50%. Lower concentrations of alpha1-antitrypsin (50 mug/5 X 10(4) sperm in 0.05 mo) and inter-alpha-trypsin inhibitor in amounts of 250 mug/5 X 10(4) sperm and less had no significant effect on the fertilization rate. Comparative experiments with Trasylol in conentrations of 50 mug/5 X 10(4) sperm resulted in more than 50% fertilization inhibition, whereas 100 mug/5 X 10(4) sperm decreased the fertilization rate by approximately 90%. The differences in the antifertility effect of these acrosin inhibitors toward capacitated spermatozoa may be due to differences in molecular weight. The effective concentrations required for fertilization inhibition appear to be relatively high under the experimental conditions used.

Sperm surface antigens were prepared by detergent extraction (Hyamine-Triton, Rohm and Haas Co., Philadelphia, PA) of spermatozoa from rhesus monkeys. Heterologous antisera against the extracts were produced in female New Zealand White (NZW) rabbits by hyperimmunization. The antiserum was absorbed initially with sperm-free monkey seminal plasma and then with lyophilized tissue homogenates of liver, kidney, spleen, and pancreas from male rhesus monkeys. The unabsorbed antiserum produced at least three precipitin lines against detergent extract in double immunodiffusion tests and possessed sperm immobilization and sperm agglutination antibody activity. The absorbed antiserum showed one precipitin line against the detergent extract and retained sperm agglutination antibody activity only. The sperm agglutination antibody in the absorbed antiserum was completely removed by either epididymal or washed ejaculated rhesus monkey spermatozoa. The same results were obtained with ammonium-sulfate-precipitated immunoglobulin preparations. Immunoelectrophoresis revealed only one precipitation line with the absorbed immunoglobulin preparation. It is concluded that Hyamine-Triton extract of rhesus monkey spermatozoa contains a component which can be characterized as sperm-specific by immunoabsorption techniques and is apparently associated with sperm agglutination but not with immobilization.

Auto-immunization of the male and iso-immunization of the female with spermatozoal or testicular material can lead to a significant impairment of fertility. Interference of sperm antibodies with reproductive processes may occur by impairment of sperm migration through cervix, uterus and tubes and by blocking adherence of spermatozoa to the surface of the zona pellucida of the oocyte. A very important point to consider in this context is the immunological situation in the female genital tract which is a target organ for sex hormones. Observations in human and subhuman primates as well as in other mammalian species have shown that the levels of immunoglobulins and specific antibodies in secretions of the different compartments of the female reproductive tract are low compared to the serum in general and show typical cyclic changes under the influence of oestrogens and progesterone. Antibody levels appear to be especially low during the preovulatory period in cervical mucus and--according to observations in primates--in tubal fluid. Complement is present in borderline concentrations or not detectable. Data on levels of immunoglobulins or specific antibodies in endometrial fluid are scarce. However, their presence has been demonstrated. There are indications that a secretory immune system may be operational mainly in the cervical compartment of the female genital tract. Serum antibody levels do not reflect properly the immunological situation in secretions of the genital tract in most instances. This may be different in the male. Demonstrations of sperm antibodies in serum or even in genital secretions are not necessarily indicative of permanent sterility; the chances of conception may, however, be reduced. Antibody specificity, immunoglobulin class and quantitative relationships between spermatozoa and antibodies and hormonal influences have to be taken into consideration. Immunity to spermatozoa does not seem to be an all-or-nothing phenomenon and should be considered a relative rather than absolute cause of infertility.

Epididymal cysts and/or hypoplastic testes have been found in 31.5 per cent of 308 men exposed to diethylstilbestrol in utero, compared to 7.8 per cent of 307 placebo-exposed controls. Analyses of the spermatozoa have revealed severe pathological changes (Eliasson score greater than 10) in 134 diethylstilbestrol-exposed men (18 per cent) and 87 placebo-exposed men (8 per cent). Further investigation of the 26 diethylstilbestrol-exposed men with testicular hypoplasia has revealed that 65 per cent had a history of cryptorchidism. Only 1 of the 6 placebo-exposed controls with testicular hypoplasia had a history of testicular maldescent. Although none of our Diekmann's lying-in study group has had carcinoma to date one must keep in mind the reported increased risk of testicular carcinoma in testes that are or were cryptorchid. A 25-year-old man who was not part of the study group was treated recently by us for a testicular carcinoma ( mixed anaplastic seminoma plus embryonal cell carcinoma) and he had a history of diethylstilbestrol exposure in utero and cryptorchidism.

The concentrations of spermatozoa, fructose, IgG, IgA, albumin lactoferrin, transferrin, secretory piece of IgA, Beta 1C/Beta 1A-globulin (C'3-component of complement), ceruloplasmin and fibrinogen were evaluated in human split ejaculates and/or in whole human seminal plasma. The concentrations of spermatozoa, IgG, IgA, albumin and transferrin decreased from the first portion of the split ejaculate to the last, indicating that these proteins originate mostly from secretions other than the seminal vesicles. By contrast, the highest amounts of fructose and lactoferrin were present in the final portion of the split ejaculates, showing their seminal vesicle origin. No secretory piece, IgM, Beta 1C/Beta 1A-globulin, ceruloplasmin of fibrinogen could be detected in human semen. An unidentified antigen was found that has a relatively high molecular weight and shows Beta 1-mobility on immunoelectrophoresis.

Twenty-two synthetic proteinase inhibitors were tested for their inhibitory properties towards human acrosin. p-Nitrophenyl-p1-guanidino benzoate (NPGB) was the most effective (K1 value of 1-5 X 10(-8) M), producing a non-competitive type of inhibition in contrast to all other inhibitors which showed a competitive type of inhibition. The Michaelis constant for human acrosin on BAEE at pH 8-1 was calculated to be 4-25 X 10(-5) M.

The in utero effects of diethylstilbestrol on the human male genital tract are reported in our followup study of male offspring of mothers treated with diethylstilbestrol. Anatomical and functional abnormalities were significantly greater in male patients exposed to diethylstilbestrol compared to male controls whose mothers were all participants in a prospective, randomized double-blind study on the effects of diethylstilbestrol on pregnancy at our hospital during the early 1950s. Epididymal cysts, hypotrophic testes and capsular induration of the testes were among the more common genital lesions found in more than 25 per cent of 159 male patients exposed to diethylstilbestrol compared to a 6.8 per cent incidence in 161 male controls. Spermatozoal analysis revealed severe pathological changes (Eliasson score more than 10) in 32 per cent of 31 patients exposed to diethylstilbestrol and 0 per cent of 20 male controls. Abnormal findings on physical examination combined with severe sperm abnormalities (Eliasson score more than 10) were found in 23 per cent of the male patients exposed to diethylstilbestrol versus none of the male controls. Cytologic examinations revealed no malignant cells from urine samples before and after prostatic massage or ejaculation, prostatic fluids and aspirates from epididymal cysts.

The acrosomal proteinase of human spermatozoa was characterized and differs from other human proteinases. The enzyme has optimal activity at pH 8.0, is inactive below pH 5.0 or above pH 10.5, requires calcium for maximum activity, hydrolyzes fibrinogen, gelatin, and benzoylarginine ethyl ester, and is inhibited by various synthetic and natural proteinase inhibitors. The molecular weight was estimated to be 30,000. Human spermatozoa also possess a proteinase inhibitor that is similar to one of the proteinase inhibitors from human seminal plasma.