[Mechanisms influencing implantation of the embryo--the last-years' news]. / Mechanismy ovliviiující implantaci embrya--co je nového v posledních letech.

OBJECTIVE: To summarize recent knowledge concerning mechanisms which influence the implantation of embryo. DESIGN: Literature-based overview. SETTING: Institute for the Care of Mother and Child, Prague. SUBJECT OF THE STUDY: Factors influencing implantation of embryo in the uterus elicit increased interest due to study of unexplained failures of embryotransfer following the successful in vitro fertilization. Our article points to recent information about physiology and pathology of mechanisms controlling implantation, namely the factors of immunity (antibodies, cells, cytokines and other mediators) whose exact regulation on the feto-maternal interface is a crucial precondition of successful implantation. Also the genetics of early embryo, as well as the possibilities of modern endoscopic techniques offer new insight onto mechanisms of implantation. Recommendations for diagnostics and treatment of implantation failure are given in the end of the article.

[Immunologically conditioned fertility disorders in men--experience of the immunobiological department of the Institute for Maternal and Child Care]. / Imunologicky podmínené poruchy plodnosti u muzu - zkusenosti imunobiologického pracoviste Ustavu pro péci o matku a díte.

OBJECTIVE: To provide an overview of the results obtained from the more than ten-year systematic monitoring of the contribution of immunopathological mechanisms to the ethiopathogenesis of fertility disorders in men. DESIGN: A summarising retrospective study. SETTING: Mother and Child Care Institute, Prague. METHODS: The data source is a selected group of a total of 3,800 couples, who were examined in the Immunobiological Department (ID) of the Mother and Child Care Institute (MCCI) in Prague- -Podolí for fertility disorders in the past. From among the laboratory quantities, the following ones were systematically monitored: a) sperm parameters; b) cell-mediated immunity against spermatozoa; c) the presence of anti-spermatozoa auto-antibodies attached to spermatozoa; d) in a part of the clinical group, the serum concentrations of FSH, testosterone, SHBG and the free androgen index (FAI) were monitored. RESULTS: After selection was made according to defined criteria, the group comprised of a total of 1,680 men, of whom 49.4% were normozoospermic and 50.6% suffered from some form of seminal pathology. Increased cell-mediated immunity against spermatozoa was identified in 10.2% of fertile men, in 18.5% of normozoospermic men and in 66.3% of azoospermic men. In asthenozoospermic and teratozoospermic men, the increased cell-mediated immunity against spermatozoa was identified in 48.3% and 53.1% of them, respectively. The auto-antibodies attached to spermatozoa were identified in 3 out of every 42 fertile men (7%), while in asthenozoospermic men, it was a total of 21% (IgA antibodies) and 22% (IgG antibodies). As concerns the concentration of free androgens (FAI) in the serum, there was no difference among the individual subgroups of men. In oligoasthenozoospermic men, FSH was significantly higher on the average in comparison with normospermic men. CONCLUSION: The significantly higher incidence of increased cell-mediated immunity against spermatozoa in men with a pathological spermiogram in comparison with the control group (fertile men and normozoospermic men) indicates that cell-mediated immunity participates in the pathogenesis of seminal pathologies.

[Role of sperm antibodies and cellular autoimmunity to sperm in the pathogenesis of male infertility]. / Role protilátkové a bunecné autoimunity proti spermiím v patogenezi muzské neplodnosti.

OBJECTIVE: To test the hypothesis of relationship between sperm pathology and elevated humoral and/or cell-mediated antisperm autoimmunity in male partners from infertile couples. DESIGN: Analytic study. SETTING: Department of Immunobiology, Institute for the Care of Mother and Child, Prague. METHODS: Sperm samples were evaluated according to WHO rules. Sperm-bound antisperm autoantibodies (ASA) were determined by SpermMar Test (FertiPro N. V., Sint-Martens-Latem, Belgium). For evaluation of cell-mediated antisperm autoimmunity (CMAA) the authors used their own modification of migration-inhibition test (Dimitrov et al., J. Immunol. Methods 154: 147, 1992). RESULTS: The pool of men was divided into groups according to the result of sperm examination: normozoospermia (740 men), asthenozoospermia (244), teratozoospermia (191), oligoasthenozoospermia levis (61), oligoasthenozoospermia gravis (29), oligoteratozoospermia (82), and azoospermia (54). Subgroup of fertile men (32) consisted of normozoospermic men--fathers of child younger than 3 years. Percentage of sperm-bound ASA-positive samples was significantly higher in asthenozoospermia in comparison with normozoospermia in both IgA (20.8% versus 10.6%) and IgG classes (13.8% vs 6.8%). Positivity of CMAA was significantly more frequent in group of asthenozoospermic (52%) than in normozoospermic (28.5%) and fertile (12.5%) men. CONCLUSION: Antisperm autoimmunity, namely its cell-mediated form, appears to play a significant role in impairment of spermiogenesis. Sperm-bound autoantibodies were found more frequently in asthenozoospermia, but also in some men with normozoospermia they may impair fertility.

Thrombin and trypsin receptors: the same mechanism of signalling on cellular surfaces.

Proteinase-activated receptors (PARs), ubiquitous surface molecules participating on many biological processes have been recently discovered. Specific receptors for thrombin (PAR-1 and PAR-3) and trypsin (PAR-2) are described in this review. They belong to a family of G protein-coupled receptors activated by amino acid sequence of N-terminal part of bound ligand revealed by site-specific proteolysis. PARs participate in tissue growth and differentiation, regeneration and reparation, inflammatory response regulation, malignant transformation, but even in vascular tonus and blood pressure regulation. (Fig. 5, Ref. 35.)

[Pathogenesis and therapy of multiple sclerosis]. / Patogeneza a terapie mnohotné (roztrousené) sklerózy mozkomísní.

Multiple sclerosis (MS) is one of the most frequent serious neurologic diseases. Etiologically, MS involves genetic, viral and other factors. The key pathogenic mechanisms reside in the autoimmune reaction of activated CD4+ T lymphocytes crossing the haematoencephalic barrier and attacking different epitopes of the basic protein and proteolipid of myelin sheaths. The damaging reaction involving activated macrophages, destructive inflammatory cytokines and toxic radicals leads to the development of disseminated plaques. The passage of autoimmune T lymphocytes to the brain tissue is facilitated by overexpression of adhesion molecules on endothelial cells, neural cells and immunocytes. The diagnosis of MS is based on characteristic changes in the blood and liquor, and on the results of modern methods, especially (gadolinium enhanced) MRI. The "classical" treatment is based on glucorticosteroids, azathioprine, cyclophosphamide and other chemicals which are not fully satisfactory and are accompanied by serious side-effects. Therefore, attention will be paid to three prospective biological methods of treatment: 1) peroral application of bovine myelin, its fractions, and synthetic copolymer-1, aimed at the restoration of immune tolerance; 2) injections of natural and recombinant interferon-beta, interfering with the pathogenic IFN-gamma and other cytokines; 3) systemic enzyme therapy (residing in peroral application of combinations of animal and herbal hydrolytic enzymes), which modulates adhesion molecules and suppresses the activation of autoimmune T lymphocytes. The chief results of clinical studies with respect to the effectiveness and safety of these therapeutic methods, will be summarized. (Ref. 34.)

[Practical results of monitoring pregnancy in women with systemic lupus erythematosus]. / Praktické výsledky sledování tehotenství u zen se systémovým lupus erythematodes.

BACKGROUND: The authors dealt with the urgent problem under what conditions it is possible to achieve in a woman with systemic lupus erythematosus (SLE) or another collagenosis, or secondary antiphospholipid syndrome (APS) a favourable outcome of pregnancy and the delivery of a healthy infant. METHODS AND RESULTS: The investigation comprised 23 women incl. 20 with SLE, two with the mixed form of a diffuse connective tissue disease (MCTD) and one with Sjögren's syndrome of the primary type. From the total number of 20 pregnancies six were consulted in advance with a doctor (group I-s-called planned pregnancies) and all terminated by a successful delivery. Of 11 pregnancies which were not consulted with a doctor in advance (group II-so-called unplanned pregnancies) 9 were terminated in term, however, only 5 with a successful delivery (55.5%), two women are still pregnant. Exacerbation of the basic disease during pregnancy was recorded only once and did not lead to discontinuation of the pregnancy. CONCLUSIONS: The authors provide evidence that desired pregnancy of informed women suffering from SLE or another collagenosis when assisted by a specialized medical team can lead to a successful delivery of an infant.

[Systemic enzyme therapy]. / Systémová enzymoterapie.

The paper summarizes the hitherto knowledge concerning the absorption of perorally administered enzymes in the digestive tract. After the transport through the intestinal wall the enzymes are protected against immunity recognition and they reach the affected regions via blood and lymphatic routes. A survey of enzymes taken into consideration and the essence of their action are presented. In the final part, the fields are summed up in which systemic enzymotherapy proves useful.

[Systemic enzyme therapy: problems of resorption of enzyme macromolecules]. / Systémová enzymoterapie: k problematice vstrebávání enzymových makromolekul.

Systemic enzyme therapy represents a special therapeutic approach consisting in the oral application of high doses of hydrolytic animal and plant enzyme combinations. The originally empirical method was by detailed experimental analyses and successful clinical studies transformed into a widely appreciated therapeutic method of various pathologic processes. In spite of this fact systemic enzyme therapy has been repeatedly questioned by referring to an almost hundred year old dogma claiming the unabsorbality of enzymes in the macromolecular form. The authors present arguments denying the unexceptional validity of this dogma. The histological, radiological, biochemical (chromatographical, enzymological), immunological and biological methods have convincingly proven that a part of swallowed enzymes may pass the intestinal barrier in an undamaged macromolecular form and realize their activities in the body. The most important elements able to absorb macromolecules seem to be so called "M-cells" (FAE) which cover lymphoid foci of the organized gut lymphoid tissue. Other mechanisms of enzyme resorption are under discussion. The absorbed enzymes are rapidly complexed with naturally occurring blood antiproteases. In these complexes the potential immunogenicity of enzymes is restricted and they are concentrated into pathologically affected areas of the body. Complexes in addition display important immunoregulatory activities.

[Systemic enzyme therapy in diseases of the vascular system]. / Systémová enzymoterapie chorob cévního systému.

The treatment of autoimmune and immune complex diseases of the vascular bed consists--similarly as of immunopathologic processes of other systems--in the use of risky immunosuppressive agents and antiinflammatory as well as symptomatic therapy. In the article the author informs about the possibility to use in these indications (and in addition also in other angiologic diseases) the systemic enzyme therapy, residing in the oral application of high-dosed combinations of several animal and plant proteolytic enzymes. About four tens years of positive medical empirical experience have been supported by a concentrated sophisticated research and approximately 150 clinical studies according to GCP. These revealed in most autoimmune and immune complex diseases a surprisingly high effectiveness and complete harmlessness of the enzyme therapy. After the short introduction mentioning the important indications for enzyme therapy in the field of clinical immunology, the major attention is paid to the results of enzymotherapy in angiology. Strong evidence indicates that enzymotherapy ameliorates the disturbed composition and properties of blood and vessel walls, acts preventively as well as therapeutically in thromboses, thromboflebitides and consequences of venous insufficiency; it seems be prospective in afflictions of arterial bed, including vasculitides and glomerulonephritides, also. The key feature of enzymotherapy is the immunomodulatory activity. There exists a strong evidence for the favourable modulation of pathogenic autoantibodies, inhibition of the neogenesis of immune complexes and cleavage of their deposits, normalization of the T cell system, network of cytokines, adhesion molecules and inflammatory cascades. Besides the direct peptidolytic and proteolytic effects of hydrolases, the indirect effects realized in the course of interaction between the resorbed enzymes and their natural "partners"--antiproteases (mainly alfa-2-macroglobulin)--have become a topic of intensive research. The author feels, that systemic enzyme therapy should become a regular component of the treatment of immunopathologic processes in general and of angiologic diseases specially. (Ref. 42.).

Correlation of asthenozoospermia with increased antisperm cell-mediated immunity in men from infertile couples.

To investigate whether cell-mediated immunity (CMI) against sperm and/or antisperm circulating antibodies are associated with poor semen quality, a leukocyte migration inhibition factor (LMIF) assay and an enzyme-linked immunosorbent assay (ELISA) were performed in groups of men from infertile couples, men from fertile couples and sperm donors. Twenty-five of 102 men (25%) revealed positive CMI against sperm and 10 (10%) had positive antisperm antibody titers in their sera. Fifteen of 28 asthenozoospermic men (53%) from infertile couples revealed positive antisperm CMI. The incidence of antisperm CMI was significantly increased (P < 0.05) in the infertile men with asthenozoospermia compared with the men from the other two groups (men from fertile couples and sperm donors). No significant differences between migration indices were seen when such a comparison was done for oligoasthenoterato- and teratozoospermics. The results indicate that increased antisperm CMI is associated with asthenozoospermia in a significant number of men from infertile couples. The importance of these findings is discussed.

Outlooks of systemic enzyme therapy in rheumatoid arthritis and other immunopathological diseases.

Systemic enzyme therapy (SET) represents a specific therapeutic approach consisting in peroral application of blends of animal and plant hydrolytic enzymes. A significant part of the swallowed enzymes (about 25%) is resorbed in the intestine in functionally active form. After being complexed with natural antiproteases, enzymes set concentrated in wounds, inflammation sites and immunopathological foci. SET has many important indications in traumatologic, thrombotic, infectious, inflammatory, immunopathologic and even tumorous processes. Rheumatoid arthritis, Bechterew's disease, activated arthrosis and extraarticular rheumatism represent important and sensitive targets of SET. In situ and in vivo studies continue to elucidate selective interferences of absorbed proteolytic enzymes with the crucial pathogenic mechanisms of rheumatoid processes.

Regulatory role of T lymphocytes and NK cells in tumor allograft development.

In the present study, the respective roles of T cells and their subpopulations as well as of NK (natural killer) cells in antitumor immune responses were followed using the SaI (H-2a) allograft model. The development of this tumor in B10 (H-2b) mice was evaluated after pretreatment of the recipients with xenogeneic antithymocyte serum (ATS). Anti-Thy 1.2, anti-Lyt 2.2 and anti-L3T4 monoclonal antibodies were used in order to determine T lymphocyte phenotypes and to assess the frequency of TC/S and TH subpopulations at various periods of tumor development. Rabbit polyclonal anti-asialo GM1 antiserum was used for the identification of NK cells. In a previous work it was suggested that the first week following transplantation, the cells predominantly involved in the growth regulation of SaI belong to the TS subclass. Our results based on the use of anti-Lyt 2.2 monoclonal antibodies have further supported this finding. The application of anti-Thy 1.2 on the 3rd and 5th day has hampered a secondary tumor growth while anti-Lyt 2.2 was effective when given on day 5. The depletion of Lyt. 2.2+ cells on day 3 resulted in the inhibition of both primary and secondary tumor development. On the other hand, when anti-Thy 1.2 was applied on day 7 after transplantation, the primary and secondary tumor growth was strikingly enhanced. It appears that Thy 1.2+ lymphocytes display at this period effector functions and contribute, in conjunction with macrophages, to subsequent tumor regression. The depletion of L3T4 cells on days 3 and 5 after tumor inoculation has resulted in primary tumor growth enhancement. This suggests that cells of the L3T4+ phenotype display at this time helper functions contributing to CTL proliferation and maturation. A further indication, supporting the possible suppressor effect of L3T4+ cells, counts from the finding that anti-L3T4 treatment results in an inhibition of secondary tumor growth. The anti-asialo GM1 treatment has not enhanced, at least significantly, primary tumor development but has partially or totally inhibited the growth of secondary tumors. It appears that cells of the GM1+ (NK cells) phenotype do not participate in any substantial way in the early phases of SaI tumor development in ATS treated allogeneic recipients.

[Dynamics of the distribution of lymphocyte subpopulations in the human menstrual corpus luteum]. / Dynamika zastoupení subpopulací leukocytu v lidském menstruacním zlutém telísku (lmCL).

Recent evidence suggests that immunoendocrine interactions play a definitive role during development and regression of the human menstrual corpus luteum (hmCL). We studied the distribution of immune cells within individual structures of hmCL during various stages of its development. Immunoperoxidase-stained ultra-thin frozen sections were evaluated, using light microscopy fitted with an image analysis system. The results suggest that monocytes/macrophages and MHC class II positive cells are the most prominent immune cells within the hmCL throughout its lifespan. Both cell types are concentrated within the trabeculae. In addition, MHC class II positive cells are abundant also within the granulosa-luteal layer. T helper/inductor (Th/i) and T cytotoxic/suppressor (Tc/s) cells were detected only in minor amounts within the thecal trabeculae of mature tissue. Possible links between the occurrence and functional roles of the immune cells studied are discussed.

[Cell-mediated immune reactivity to sperm in disorders of human reproduction]. / Imunitní reaktivita proti spermiím zprostredkovaná bunkami (BI) v poruchách lidské reprodukce.

Besides the pathological anti-sperm humoral immunity, pathological anti-sperm cell-mediated immunity is considered as a crucial facet of the disturbances of human reproduction (male and female infertility, recurrent abortions, endometriosis, late EPH gestosis, fetal hypotrophy). A precise and objective method is designed, based on a one-step agarose Leukocyte Migration Inhibition Factor assay. The migration areas are evaluated by a computer-assisted image analysis system. Optimal concentrations of leukocytes and sperm, as well as technical conditions are described. The Radius Migration Indexes and Area Migration Indexes are computed and expressed as a Migration Index percentage for each patient or control. Preliminary clinical results indicate a highly significant association between leukocyte migration inhibition and cases of "immunopathological" infertility and repeated fetal loss.

A quantitative objective method for the evaluation of anti-sperm cell-mediated immunity in humans.

Anti-sperm cell-mediated immunity (CMI) is considered as a crucial facet of infertility in patients of both sexes. A precise and objective method is designed, based on a one-step agarose leukocyte migration inhibition factor (LMIF) assay. The migration areas are evaluated by a computer-assisted image analysis system. Optimal concentrations of leukocytes and sperm, as well as technical conditions are described. The radial migration indexes (RMI) and area migration indexes (AMI) are computed and expressed as a migration index (MI) percentage for each patient or control. Preliminary clinical results indicate a highly significant association between migration inhibition and cases of 'immunopathological infertility'. The method described is considered a promising tool for a rapid and quantitative evaluation of a suspected anti-sperm CMI in infertile and recurrently aborting patients.

[Systemic lupus erythematosus and pregnancy. Present status]. / Systémový lupus erytematodes a tehotenstvt. Soucasný stav problému.

The authors summarize contemporary views on the relationship between pregnancy and systemic lupus erythematosus (SLE) in women of reproductive age. It is known that pregnancy influences in a marked way the basic disease and conversely the disease has an impact on the course of pregnancy. Risks ensuing from these in errelations for women--mothers with SLE and the foetus are exacerbation (flare-up) of the basic disease, loss of the foetus and damage of the foetus by so-called "lupus neonatorum" with skin symptoms and complete cardiac A-V block. The contemporary state of knowledge makes it possible under certain conditions to achieve by therapeutic interventions the development and delivery of a healthy foetus. Among these views it is most important for conception to occur in these patients with SLE during a period without signs of active SLE. To ensure a successful course of pregnancy in women with SLE is the task of a team of appropriate specialists.

Development and distribution of the white blood cells within various structures of the human menstrual corpus luteum examined using an image analysis system.

PROBLEM: Emerging evidences suggest that immunoendocrine interactions play definitive roles during development and regression of the human menstrual corpus luteum (hmCL). We have studied the distribution of immune cells within individual structures of hmCL during various stages of its development. METHOD: Immunoperoxidase-stained ultra-thin frozen sections were evaluated using light microscopy fitted with an image analysis system. RESULTS: The results suggest that monocytes/macrophages and MHC class II positive cells are the most prominent immune cells within the hmCL throughout its whole lifespan. Both cell types are concentrated within the trabeculae. In addition, MHC class II positive cells are abundant also within the granulosa-luteal layer. T helper/inductor (Th/i) and T cytotoxic/suppressor (Tc/s) cells were detected only in minor amounts within the thecal trabeculae of mature tissue. CONCLUSIONS: Possible links between the occurrence and functional roles of the immune cells studied are discussed.