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M Kurpisz

Publications and source records attributed to M Kurpisz.

At least 73 records · Page 4Linked to original sources
PubMed1993

[Molecular basis of capacitation and acrosomal reaction].

There has been described molecular events towards preparation of spermatozoa for fertilization as capacitation (process known to be characterized by increased mobility of membrane components on sperm surface) and acrosomal reaction (fusion of spermatozoon plasmalemma with outer acrosomal membrane with following destruction of acrosomal compartment and enzyme release). There have been also described hypotheses concerning activity of exposed during acrosomal reaction receptors for gamete binding and especially sperm receptors located on egg's zona pellucida. The role of acrosomal enzymes in penetration of zona pellucida by sperm as well as inhibition of enzymatic action were discussed in aspect of contraceptive potential.

Acrosome↗
PubMed1992

The cytokines effect on EBV-immortalized human B cells producing antisperm antibody.

We have analyzed the antisperm antibody production of autoimmunized male subjects using Epstein-Barr virus (EBV) immortalization of B lymphocytes. We evaluated the influence of several in vitro culture variants applied prior to EBV infection on the frequence of antibody-producing cells and affinity of secreted antibodies. The following variants were applied: a) polyclonal antigenic stimulation of lymphocytes with PWM, b) PWM (pokeweed mitogen) + IL-2 + interferon gamma and c) PWM + IL-2 + interferon gamma + sperm antigenic extract. The variants where the cytokines were added did not increase the frequency of EBV-infected antibody-producing cells as comparing to EBV infection previously amplified by the use of polyclonal activator. Furthermore the cytokine activation either in combination with mitogen or in vitro secondary antigenic sensitization (prior to EBV transformation) did not seem to have beneficial effect on affinity of antibodies produced by EBV-infected cells in comparison to straight EBV infection. On the other hand, the attempt to promote an immunoglobulin secretion (IgM) by previously obtained human-human antisperm hybridomas by adding of IL-2 was quite successful.

Antibody Affinity↗
PubMed1992

A ligand-epitope in vitro analysis of major histocompatibility determinants expressed on B and T lymphocytes.

Human histocompatibility leucocyte antigen (HLA)-specific monoclonal antibody probes were used to determine the affinity constant, and cell-surface density of HLA class I and class II determinants. The measurements were estimated for single-cell units of B-lymphoblastoid cell line (B-LCL) and cloned activated T cells in different functional states. Each HLA subset showed unimodal affinity constant values for the interaction with the corresponding HLA-specific antibodies. Such values ranged between 2.2 x 10(7) M-1 (class I) and 4.0 x 10(7) M-1 (class II) for different histocompatibility epitopes. In both B and T cells there was a rank order of epitope expression, class I being highly expressed (5 x 10(6) epitopes/cell) followed by DR, DQ and DP, (1.1-3.0 x 10(6) epitopes/cell). Suppressive clones carrying functionally defined stimulating determinants previously designated 'DY' carried similar numbers of DR, DQ and DP binding sites to DY- non-suppressive clones, but showed selective increases of class II determinants reactive with broad class II-specific antibodies. The results are discussed in the context of the functional consequences of different patterns of HLA epitope expression in immune responses.

Antibodies, Monoclonal↗
PubMed1990

Major histocompatibility complex (MHC) protein analysis by optimised two-dimensional electrophoretic methods.

Human histocompatibility molecules HLA-Class I and Class II (DR, DQ, DP) were analysed using three two-dimensional protocols: nonequilibrium pH gradient electrophoresis (NEPHGE), isoelectric focusing-acidic gradient (IEF-AG) and isoelectric focusing-basic gradient (IEF-BG). The three methods differ in their carrier ampholyte combinations and electrophoretic conditions. They provide different pH gradients and therefore different electrofocusing profiles. The NEPHGE protocol was adequate for separating proteins across a broad range of pI mobilities, i.e. 4.4 pH units between the acidic and the basic end. In contrast, the IEF-AG and the IEF-BG protocols gave a separation power across a narrow pH range, 1.9 and 1.7 pH units respectively. Thus, whereas the NEPHGE protocol provides a tool for a global major histocompatibility complex (MHC) antigen profile analysis, the IEF-AG and -BG allows one to investigate subcomponents of the individual MHC chains. For example, NEPHGE analysis of the HLA Class I heavy chain revealed a single spot. However, IEF-BG revealed the presence of six equidistantly spaced spots spanning a short pH gradient with identical molecular weight. Similar improved resolution was seen for the HLA-DR, DQ, and DP molecules. The IEF acidic gradient was adequate for separating the alpha chain; the IEF basic gradient gave better resolution of the beta chains. This data provides a baseline set of conditions for both analytical and preparative MHC protein studies prior to amino acid sequencing.

Alleles↗
PubMed1990

Relevance of immuno-contraceptive vaccines for population control. II. Gamete antigens.

Some new information on the sperm and seminal plasma antigens which can induce responses which decrease fertility has been obtained. This recent development has not, however, brought the understanding of the subject to a stage where the production of a contraceptive antisperm vaccine for the female can be discussed realistically. Many laboratories are continuing the active investigation of the potentials of zona pellucida antigens for contraceptive approach. Also, passive immunization with human monoclonal antibodies promises to be an effective and safe method for short-term contraception and early pregnancy termination.

Amino Acid Sequence↗
PubMed1989

Mouse monoclonal antibodies against human sperm: evidence for immunodominant glycosylated antigenic sites.

Thirty mouse monoclonal antibodies (MoAbs) raised against human sperm detect common antigenic determinants on human lymphocytes, erythrocytes, bacteria and endotoxin. Specific chemical, enzymatic and lectin blocking studies indicate that the sperm-associated antigens defined by these MoAbs are glycoconjugates. Further studies including reactivity of these MoAbs with organic sperm extracts indicate that the predominant carriers of these carbohydrate antigens are glycolipids and that the terminal immunodominant monosaccharide may be N-acetyl-glucosamine or N-acetylgalactosamine.

Animals↗
PubMed1989

[Interactions between the reproductive and immune systems].

There are described the mechanism of development of antisperm antibodies, the techniques of antisperm antibodies detection and the role of these antibodies in human infertility. Furthermore, some autoimmunological syndromes associated with abnormal reactivity of immunological system against components of reproductive system are discussed.

Adult↗
PubMed1988

Production of large amounts of mouse polyclonal antisera.

A simple and inexpensive method for producing larger volumes of mouse polyclonal antisera is described. Groups of BALB/c mice were immunized with different human sperm antigens classified according to molecular weight. Mice were immunized twice with the antigen, twice pristane pretreated, and then injected with a non-antibody-secreting myeloma cell line to induce ascites formation. Antibody activity of the ascites fluid approximated that of the serum. Thus, one mouse can provide about six times the amount of antibody-containing fluid usually obtained after conventional immunization. This method also may be applicable for ascites fluid production in nude mice.

Animals↗
PubMed1987

EBV transformation and microfusion as the potential source of human monoclonal antisperm antibodies.

Peripheral blood lymphocytes were obtained from vasectomized men with high serum titers of antisperm antibodies. An Epstein-Barr virus (EBV) transformation was performed either with B cells or mononuclear leukocytes. The effect of feeder cells (irradiated umbilical cord blood lymphocytes), cyclosporin A, and in vitro stimulation of lymphocytes with sperm extract on EBV transformation was evaluated. Antibody-producing cells were screened for specificity against human sperm by an enzyme-linked immunosorption assay (ELISA) one to six weeks after transformation. Using B cells or leukocyte mononuclear cells, we found that the percentage of wells containing antibody reactive against human sperm was greatest two weeks after transformation (range 3% to 7.5% positive wells). To increase and maintain antibody synthesis by these transformed cells, microfusions were performed in those wells positive for antisperm antibody using the UC 729-6 lymphoblastoid cell partner. Then resultant hybridomas were expanded, subcloned, and preliminarily characterized.

Antibodies, Monoclonal↗
PubMed1987

HLA expression on human germinal cells.

Ejaculated human seminal plasma cells obtained from 46 healthy men and 48 infertile patients were tested for expression of HLA class I and II antigens by complement-mediated cytotoxicity, cell-binding radioimmunoassay (CB-RIA), and an ELISA involving monoclonal antibodies (mAbs). In a group of healthy men tested for HLA expression on human spermatozoa, 4 of 46 were positive for class I and II HLA antigens. However, the results were negative in a second examination of the same donors, possibly on account of alterations with time in the expression of various subpopulations of cells which are antigen positive or antigen negative. In a group of infertile patients, we found positive expression of HLA class I and II antigens in 7 of 48 men. The samples contained immature and mature sperm but no contaminating leucocytes or epithelial cells.

Antibodies, Monoclonal↗
PubMed1986

Serological biochemical and functional characterisation of three different HLA-DR monoclonal antibodies derived from C57BL6 mice.

C57BL6 mice which do not express I-E gene products were immunised with EBV transformed human B cell lines to generate MoAbs. Three hybridoma supernatants which initially reacted with the immunising donor cell but not a T cell line lacking Class II antigens were further investigated. I-D SDS-PAGE patterns of molecules precipitated by the three supernatants from a cell membrane lysate were characteristic of HLA-Class II alpha and beta chains. Two-dimensional analysis established the specificity of the supernatants as HLA-DR specific. This was confirmed by the reaction patterns with Class II mutant deletant cell lines. In both ELISA and cytotoxicity one reacted with all lymphoblastoid cell lines tested, one reacted with all except two that were DR7 homozygous and the third reacted strongly only with cells that were DR3. All three antibodies were cytotoxic to both peripheral blood lymphocytes and EBV transformed B cell lines. The DR3 specific MoAb (IgG2a) was suitable as a typing reagent. The DR3 reactive MoAb specifically inhibited stimulation by a Dw3 HTC and the other two MoAbs inhibited all HTCs tested. These findings are consistent with the view that certain determinants responsible for the Dw specificities are carried on the DR molecules.

Animals↗
PubMed1986

The non-mature sperm cells: evaluation of surface markers and interaction in in vitro cultures.

The ejaculated human seminal plasma cells may be classified into subpopulations on the basis of spermatogenesis or biological features. In this paper we divided these cells into two main groups: one of them is obviously predestinated to fertilization as mature sperm cells, viable and motile ejaculated cells. Second group of unknown biological role is very heterogeneous; simplifying its variety we call them non-mature sperm cells. Morphological studies by electron microscopy and evaluation of surface markers of these cells by monoclonal antibodies did not reveal any known biological features or markers for cells of leukocyte origin. They did not express any antigenic determinants of HLA system and they did not react with autologous or allogeneic lymphocytes.

Antibodies, Monoclonal↗