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H Skrabei

Publications and source records attributed to H Skrabei.

11 recordsLinked to original sources

Detection of human spermatozoal peptides after conjugation to 125I-labelled human serum albumin.

Human spermatozoal peptides, liberated during autolysis of the cells, were fractionated by gel-filtration chromatography and thin-layer chromatography. After conjugation to 125I-labelled human serum albumin, all fractions were assayed with rabbit antihuman spermatozoa antiserum. The reactions indicated the presence of a number of antigenic peptides in the spermatozoal autolysate and one of the fractions revealed a significantly higher antibody binding capacity than all other fractions assayed (P less than 0.05). Release of immunogenic peptides during autolysis of the spermatozoal membrane could be important in the aetiology of spermatozoal iso- and autoantibodies.

Humans↗

Spermatozoal hapten gained via autolysis.

Using autolysis different spermatozoa surface peptides were solubilized. The resulting low molecular weight compounds were further separated by gel-chromatography on Biogel P-4, thin-layer chromatography on Cel 400 and thin-layer-electrophoresis. After the conjugation of the compound with cytochrome C a conformation-independent determinant was detected with immuno-electrophoresis. Out of the 17 fractions tested only fraction 3, with a molecular weight of 1800 daltons revealed a precipitation reaction with human sperm-immobilizing sera and rabbit antihuman sperm-serum. All 17 fractions showed no precipitation reaction against human sperm-agglutinating sera.

Autolysis↗

Isolation of a human spermatozoal hapten "II2.2' and its reaction with naturally occurring human sperm-immobilizing sera from infertile patients.

An extract of human spermatozoa was prepared using Hyamine 2389 and Triton X-100. With gel and ion-exchange chromatography several fractions were obtained of which are reacted specifically with sperm-immobilizing antibodies of infertile females and males in an immune inhibition test. This fraction showed haptenic properties, ahd a molecular weight of 1600 and was excluded as an anticomplement factor. After conjugation with cytochrome c the hapten II2.2 formed precipitation reactions with 3 out of 4 sperm-immobilizing sera. The titer reduction in sperm-immobilizing sera after adsorption with II2.2 may represent an in vitro model for a possible treatment of infertility in cases of a humoral sensitization against spermatozoa. On the other hand, the hapten might easily be synthesized and could, after conjugation to an appropriate carrier, serve as a contraceptive vaccine.

Antigens↗

Isolation of human spermatozoa membrane antigens binding sperm-immobilizing and sperm-agglutinating antibodies.

Peptides of human spermatozoa were dissolved with Hyamine 2389 and Triton X-100 and separated by chromatography on Biogel P4 columns into seven fractions. The antigenic activities of the separated sperm-membrane fractions were tested according to their capacity to inhibit sperm agglutination and sperm immobilization (immune inhibition test) in human sera of sterile patients. Sperm-agglutinating and sperm-immobilizing activity was tested by the microtray agglutination and microtray immobilization test. A titer reduction was achieved only in sperm-immobilizing sera. Four sperm antigenic fractions revealed in the majority of the repeatedly tested sperm-immobilizing sera an inhibition of the antigen-antibody reaction. No reaction was observed after exhaustive absorption of the tested seven antigen fractions with sperm-agglutinating sera. Therefore the conclusion can be drawn that sperm-agglutinating and sperm-immobilizing antibodies react with different sperm antigens. Normal human sera without sperm antibodies served as control. As no sperm agglutination or sperm immobilization was obtained after absorption of these control sera our antigen fractions do not produce sperm agglutination or sperm immobilization.

Antigens, Surface↗

Lactate dehydrogenase--X--separation of human spermatozoa isoenzymes by extraction with detergents.

Proteins with known enzymatic properties are attractive molecules as possible agents for immunological contraception. Therefore a new technique for the selective isolation of LDH-X from human spermatozoa is described. Applying the detergents hyamine 2389 and triton X-100 LDH-X is extracted from the sperm surface membrane without the other LDH-isoenzymes. Ultrasonification of spermatozoa yields all LDH-isoenzymes together. After cell disruption the separation of LDH-X and the isoenzymes is performed by discelectrophoresis. With lactate as substrate tetrazolium staining identifies all LDH-enzymes. The technique represents an attractive method for the selective isolation of LDH-X and opens further purification steps.

Detergents↗

[Immunofluorescent-histologic localization of a placenta-antigen with antibodies from pregnancy blood (author's transl)].

Placenta-specific antibodies were isolated by the incubation of washed placental tissue with pregancy serum and then desorption with citric acid pH3. The antigen responsible for the formation of the antibody could be localized via the immunofluorescence technique together with application of the placenta antibody in the syncytiotrophoblast layer. The antibodies are not complement-fixing and belong to the IgG class.

Antibodies↗

[New progress in detection of antigens from human ejaculate in relationship with human sterility (extraction with N-cetylpyridinium chloride) (author's transl)].

In an attempt to obtain certain sub-surface spermatozoa antigens, human spermatozoa were treated with N-cetylpyridinium chloride in basic buffer, The extract was further ultrasonated and separated with gel chromatography over a Sephadex--G--100 column. The obtained fractions were lyophilized and tested against human sperm agglutinating and sperm immobilizing sera, in order to detect their antigenicity. Positive and negative controls revealed intensive precipitations and no precipitations respectively. Fraction A4 reacted intensively with sperm immobilizing sera. The final objectives of these sperm antigens solubilization studies are twofold. On the one hand we are looking for a method to lower sperm antibodies titres in sterile females, and on the other hand we are seeking an immunological contraceptive method.

Antigens↗