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With approximately 90 million cases annually, infection with Chlamydia trachomatis is the most prevalent sexually transmitted bacterial disease in the world. Considering that these infections are often asymptomatic and cause major complications like acute pelvic inflammatory disease, ectopic pregnancy, infertility or infant pneumonia, the estimated costs for diagnosis and treatment in the USA amounts to 2.2 million US dollars for each 500 cases. Therefore, there is a high need for correct, quick and cost-effective diagnosis and treatment of this urogenital tract infection. New innovative therapies provide good results with regard to efficacy and patients' compliance. The success rates of treatments are at least 95%. However, the occurrence of antibiotic resistance should not be ignored and new treatment schemes must be developed. The state-of-the-art of diagnosis and treatment of chlamydial infections as well as the pathophysiology is discussed in this review. In conclusion, infections with C. trachomatis is an important public health problem, especially in third world and developing countries, and more socio-economic studies linking secondary prevention of chlamydial infections, infertility and adverse pregnancy outcome are needed to understand more of its aetiology. In addition, diagnosis and treatment should be improved. Data in men revealed that past infections but not present infections are more related to male infertility. There is still controversial results. In future studies, function of the seminal vesicles and evaluation of the antioxidant capacity should be taken into account when role of C. trachomatis infection on male fertility is assessed.

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PROBLEM: Asymmetric IgG antibodies (AAb) possess a mannose-rich oligosaccharide residue bound to one of the Fab regions, making them unable to activate immunoeffector mechanisms. The proportion of asymmetric antibodies is increased after prolonged immunization with particulate antigens like cellular spleen cells. During pregnancy, AAb were found in serum and bound to placenta with specific activity to paternal antigens. No previous reports about the status of AAb in recurrent spontaneous abortion (RSA) patients have been published to date. Therefore, the aim of the present study was to analyze the percentage of asymmetric IgG molecules in serum samples of (a) healthy pregnant and non-pregnant women, (b) pregnant women with a history of RSA, and (c) non-pregnant RSA patients receiving paternal lymphocyte immunotherapy (LIT) or intravenous gammaglobulin therapy (IVIgs). METHOD OF STUDY: A previously-described differential ELISA technique was used to determine the percentage of IgG that was of the asymmetric type. RESULTS: During normal pregnancy, there was an increase in the percentage of high ConA affinity IgG serum molecules with a major increase at the second trimester. Pregnant RSA patients at the second trimester had lower values. When evaluating non-pregnant RSA patients who received LIT, it was observed that the immunized patients expressed a higher percentage of asymmetric IgG antibodies. The pregnant patients who received IVIgs had a percentage of AAbs comparable to normal pregnant patients. Additionally, the presence of IgG asymmetric molecules was confirmed in commercial gammaglobulin preparations. CONCLUSION: Results suggest a protective role of AAb during pregnancy.

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En 1994 en un estudio realizado por nuestro grupo en colabaración con la Dra. Fuksman sobre 1.200 placentas correspondientes a embarazos de alto riesgo, se halló la presencia de villitis en el 5.6 por ciento de las mismas. La histopatología detectada en ese momento fue deciduitis linfocitaria y aumento de fibrina perirvellositaria asociada con hipoirrigación e infarto placentario. Hallamos que en el 55 por ciento de las placentas con villitis los recién nacidos presentaban RCIU con respecto al 10 por ciento de los controles, con un PA de 32 por ciento en las villitis y el 83 por ciento en los controles (3). En ese material se estudiaron 68 placentas con villitis y 68 placentas sin villitis como grupo control. En 1996 demostramos en ese mismo material mediante la técnica de anticuerpos monoclonales, sobre cortes de placenta estudiando las subpoblaciones linfocitarias de las villitis, que el 50 por ciento eran CD4 (linfocitos helper), 18 por ciento CD8 (linfocitos supresoreslcitotóxicos) y 10 por ciento Leu19 (Natural Killer) pero lo significativo y anormal es que hallamos que el 65 por ciento de los linfocitos expresaban antígenos de histocompatibilidad clase II DR (40). En 1998 Jacques y Col publicaron datos similares. En 1999 comunicamos que en el informe histopatológico de material de legrado de pacientes abortadoras de causa inmunológica la descripción de villitis en un 20 por ciento de los casos. Estudios realizados en colabaración con la Dra. Zenclussen con ese material nos permitió publicar recientemente la presencia de altos niveles de Interleuquina 6(IL-6) y receptor de IL-6 en suero. El objetivo de este estudio es investigar en placentas de pacientes abortadoras recurrentes la expresión de IL-6 y sus receptores gp80 y gp130 en trece muestras de material de raspado de abortos del primer trimestre mediante la técnica de inmunofluorescencia. Como control se utilizaron cortes de placentas de embarazos normales a término. Nuestros hallazgos muestran la presencia de depósitos de IL-6 y de receptores de IL-6 con un patrón granular para las tres moléculas especificamente en el sinciciotrofoblasto mientras que fue negativo para tres en el citotrofoblasto. En los cortes de placentas normales no se hallaron en ningún caso dichos depósitos. Concluímos de todos los hallazgos antes sintetizados...(AU)

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En 1994 en un estudio realizado por nuestro grupo en colabaración con la Dra. Fuksman sobre 1.200 placentas correspondientes a embarazos de alto riesgo, se halló la presencia de villitis en el 5.6 por ciento de las mismas. La histopatología detectada en ese momento fue deciduitis linfocitaria y aumento de fibrina perirvellositaria asociada con hipoirrigación e infarto placentario. Hallamos que en el 55 por ciento de las placentas con villitis los recién nacidos presentaban RCIU con respecto al 10 por ciento de los controles, con un PA de 32 por ciento en las villitis y el 83 por ciento en los controles (3). En ese material se estudiaron 68 placentas con villitis y 68 placentas sin villitis como grupo control. En 1996 demostramos en ese mismo material mediante la técnica de anticuerpos monoclonales, sobre cortes de placenta estudiando las subpoblaciones linfocitarias de las villitis, que el 50 por ciento eran CD4 (linfocitos helper), 18 por ciento CD8 (linfocitos supresoreslcitotóxicos) y 10 por ciento Leu19 (Natural Killer) pero lo significativo y anormal es que hallamos que el 65 por ciento de los linfocitos expresaban antígenos de histocompatibilidad clase II DR (40). En 1998 Jacques y Col publicaron datos similares. En 1999 comunicamos que en el informe histopatológico de material de legrado de pacientes abortadoras de causa inmunológica la descripción de villitis en un 20 por ciento de los casos. Estudios realizados en colabaración con la Dra. Zenclussen con ese material nos permitió publicar recientemente la presencia de altos niveles de Interleuquina 6(IL-6) y receptor de IL-6 en suero. El objetivo de este estudio es investigar en placentas de pacientes abortadoras recurrentes la expresión de IL-6 y sus receptores gp80 y gp130 en trece muestras de material de raspado de abortos del primer trimestre mediante la técnica de inmunofluorescencia. Como control se utilizaron cortes de placentas de embarazos normales a término. Nuestros hallazgos muestran la presencia de depósitos de IL-6 y de receptores de IL-6 con un patrón granular para las tres moléculas especificamente en el sinciciotrofoblasto mientras que fue negativo para tres en el citotrofoblasto. En los cortes de placentas normales no se hallaron en ningún caso dichos depósitos. Concluímos de todos los hallazgos antes sintetizados...

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PROBLEM: Recurrent spontaneous abortion (RSA) could be interpreted as the cause for the incapacity of the mother to recognize paternal antigens to produce the desired protective response. The practise of alloimmunization was introduced in an attempt to induce in the mother the production of an alloimmune response; some authors proposed an association between cytokines and RSA. The production of IL6 and its soluble receptor (sIL6R) before and after lymphocyte immunotherapy was evaluated in sera of 33 patients suffering from two or more RSA, and in sera of 47 women with normal pregnancy. METHOD OF STUDY: The immunization of RSA patients was achieved by injection of four doses of 10(5) mononuclear cells (MNC) from the husband, at weekly intervals, before pregnancy. The IL6 and sIL6R levels were measured using sandwich ELISAs and the results evaluated by Tukey-Kramer multiple comparison-tests. RESULTS: Our data show no significant differences between IL6 and sIL6R serum levels of normal pregnant women and RSA pregnant women with white-cell immunization before pregnancy. In contrast, the sera of pregnant RSA patients without allogeneic therapy show higher values. We also found significant differences between IL6 levels in non-pregnant RSA women with and without immunotherapy. CONCLUSION: These results show that the alloimmunization with paternal white cells leads the serum IL6 and sIL6R-levels to the values observed in the course of normal pregnancy, suggesting a role for IL6 and sIL6R in the modulation of the immune response's quality.

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Intra- and interprotamine cross-linking by disulphide bonds are the main factors responsible for the highly compact and stable structure of chromatin in mammalian spermatozoa. Unfixed or methanol fixed smears of human sperm and sperm suspensions from fertile donors and oligospermic patients were subjected to a reductive cleavage of disulphide bonds by using 2-mercaptoethanol (ME) or dithiothreitol (DTT). Untreated (control) and ME or DTT treated samples were stained with toluidine blue (TB) and examined in light microscopy; spectral characteristics of TB stained sperm nuclei were also analyzed. Untreated smears from fertile donors showed an orthochromatic (pale blue) staining of most sperm heads, while a variable proportion of metachromatic nuclei was found in spermatozoa from patients with oligospermia. After treatment with DTT followed by TB staining, fixed and unfixed smears showed metachromatic sperm heads. ME treatment only induced a scarce colour shift, whereas a striking metachromatic reaction and variable nuclear swelling were observed in DTT treated sperm suspensions. These results indicate that after cleavage of disulphide bonds, phosphate groups from chromatin DNA are unmasked and available for TB binding and metachromatic staining.

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The study was designed to determine whether hyperviscosity of the semen sample is related to dysfunction of the male accessory glands. It was carried out on men who consecutively attended an infertility clinic between June 1989 and June 1991, and the men were grouped according to viscosity of semen samples (normal viscosity or higher viscosity). Semen samples from 229 infertility patients were studied. From these, 155 had normal viscosity and 74 showed hyperviscosity. The effect of hyperviscosity of semen samples on seminal quality and the function of the prostate was evaluated by acid phosphatase measurement, and the seminal vesicles by measurement of corrected fructose. Sperm motility (grades II-III), sperm vitality, and corrected fructose were significantly reduced in samples with high viscosity (p < .05). A high prevalence of hyperviscosity in semen samples was associated with only hypofunction of the seminal vesicles. In fact, 36.5% of subjects with hyperviscosity showed reduced levels of corrected fructose. The same association with hyperviscosity was not observed when only hypofunction of the prostate was present, or when hypofunction of both prostate and seminal vesicles was present (P:NS). Further analysis showed that high viscosity is observed mainly when corrected seminal fructose levels were below 1.5 mg/mL x 10(6) spz/mL. It would appear that hyperviscosity affects sperm motility and is associated with hypofunction of the seminal vesicles.

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Semen samples from 279 infertility patients attending an Immunological Centre were analysed to evaluate the relationship between the populations of leucocytes, seminal quality, antisperm antibodies, and seminal vesicle function. The most frequent finding between leucocytospermic samples was asthenozoospermia (57%), whereas in non-leucocytospermic samples normozoospermia was the most frequent finding (47%). In the samples with asthenozoospermia, granulocytes predominated, whereas in those with oligozoospermia and azoospermia a reduction in the number of macrophages and lymphocytes was observed, suggesting an obstructive process at the level of epididymis and/or vas deferens where these leucocytes are mostly produced. In the case of hypofunction of the seminal vesicles there was a predominance in granulocytes. The increased levels of each type of leucocytes affected seminal quality only when seminal vesicles were affected. Only the elevated granulocytes count was related to a decrease in sperm motility. In those samples with leucocytospermia, positive antisperm antibodies (ASA) were associated with low sperm motility, low sperm normal morphology, and low value of seminal corrected fructose, whereas, in the absence of leucocytospermia, ASA, were more related to low sperm counts. These data suggest that granulocytes were more related to seminal vesicles dysfunction and sperm motility changes, and that ASA may be observed in the presence or absence of leucocytospermia.

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A direct immunobead test (IBT) was performed on 233 men who attended an immunological centre. Thirty-four (14.6%) of these men were found to be positive (greater than 20% binding) for antisperm antibodies (ASA). IgA, IgG and IgM were the most common sperm-associated immunoglobulins. In 50% of men with ASA asthenozoospermia, teratozoospermia, leukocytospermia or hypofunction of the seminal vesicles was observed. Semen parameters were altered most frequently when IgM was present in association with IgA and/or IgG. This suggests that there is an active inflammatory process in the reproductive tract, as evidenced by leukocytospermia, and this could be responsible for the abnormal semen parameters. ASA generation could be a consequence of this process rather than being the cause of the abnormal semen quality. If ASA do affect fertility, this could take place in the female reproductive tract.

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OBJECTIVE: To determine possible relationships between number of leukocytes, function of seminal vesicles, and seminal quality. DESIGN: The study was carried out on men who consecutively attended an infertility clinic between June 1989 to June 1991. SETTING: This study was conducted in a private immunological center for infertility, a tertiary care center, The Centro Immunológico-Sección Esterilidad y Reproducción. PATIENTS: Semen samples from 280 infertility patients attending an Immunological Center for Infertility were analyzed. MAIN OUTCOME MEASURE: We evaluated the effect of leukocytospermia in the presence of normal or abnormal function of seminal vesicles on seminal quality. RESULTS: Sperm count, percent of motile sperm, and percent of sperm vitality were significantly reduced when both leukocytospermia and hypofunction of seminal vesicles were present (P less than 0.01). Leukocytospermic subjects with normal function of seminal vesicles showed similar seminal parameters to those nonleukocytspermics. The incidence of subjects with antisperm antibodies measured by direct immunobeads was significantly higher in leukocytospermic men with hypofunction of seminal vesicles. No differences in the incidence of antisperm antibodies with nonleukocytospermic samples were observed in those with both leukocytospermia and normal function of seminal vesicles. CONCLUSIONS: These data provide evidence that white blood cells were deleterious for seminal quality when seminal vesicles were also affected.

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In guinea pigs using testis homogenate with Complete Freund's Adjuvant as antigen, a close correlation between cellular immunity (CI) and orchitis injury was found. High titres of antisperm cytophilic antibodies (ACA) were found in serum simultaneously with gonadal lesions. Cytotoxic effects of ACA were investigated using a purified eluate of the antibody from macrophage surfaces and injecting it by subalbuginea route or incubating it with germinal cells or spermatozoa. The IgG2 nature of ACA was demonstrated by immunochemical techniques. Specific studies showed that the cytotoxic effect was not due to immune complexes. Ultrastructural studies of macrophage-ACA-spermatozoa complex by scanning and transmission electron microscopy showed the different stages of ACA-mediated sperm phagocytosis. "Thermal orchitis" was developed in guinea pigs as a model to induce orchitis without adjuvant. Isolation and identification of the sequential cell populations in the granuloma showed macrophages as the main cell type. T cells were seen to increase while B cells decreased and were substituted by plasma cells. Potentiality of unilateral sperm cord torsion for autosensitization against the other gonad was studied; orchiectomy of the injured gonad prevented the orchitis lesion and avoided the immune response. Infertile men were studied immunologically. Patients with genital infection or sperm duct obstruction exhibited positive antisperm autoimmunity tests. Cl alterations were detected early and persisted for many years, while humoral antisperm antibodies were not detected during more than two years. Azoospermic patients showed autosensitization in 50% of the cases. Finally, Percoll gradients were used to isolate and identify non spermatic round cells from semen samples of asthenozoospermic men.(ABSTRACT TRUNCATED AT 250 WORDS)

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In guinea pigs using testis homogenate with Complete Freunds Adjuvant as antigen, a close correlation between cellular immunity (CI) and orchitis injury was found. High titres of antisperm cytophilic antibodies (ACA) were found in serum simultaneously with gonadal lesions. Cytotoxic effects of ACA were investigated using a purified eluate of the antibody from macrophage surfaces and injecting it by subalbuginea route or incubating it with germinal cells or spermatozoa. The IgG2 nature of ACA was demonstrated by immunochemical techniques. Specific studies showed that the cytotoxic effect was not due to immune complexes. Ultrastructural studies of macrophage-ACA-spermatozoa complex by scanning and transmission electron microscopy showed the different stages of ACA-mediated sperm phagocytosis. [quot ]Thermal orchitis[quot ] was developed in guinea pigs as a model to induce orchitis without adjuvant. Isolation and identification of the sequential cell populations in the granuloma showed macrophages as the main cell type. T cells were seen to increase while B cells decreased and were substituted by plasma cells. Potentiality of unilateral sperm cord torsion for autosensitization against the other gonad was studied; orchiectomy of the injured gonad prevented the orchitis lesion and avoided the immune response. Infertile men were studied immunologically. Patients with genital infection or sperm duct obstruction exhibited positive antisperm autoimmunity tests. Cl alterations were detected early and persisted for many years, while humoral antisperm antibodies were not detected during more than two years. Azoospermic patients showed autosensitization in 50

of the cases. Finally, Percoll gradients were used to isolate and identify non spermatic round cells from semen samples of asthenozoospermic men.(ABSTRACT TRUNCATED AT 250 WORDS)

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The object of the present work was to investigate: a) The cytotoxic activity of cytophilic antibody in sera from guinea pigs sensitized with the testicular antigen; b) The presence of immune complexes. In vitro cytotoxicity induced by hyperimmune sera or by the cytophilic antibodies eluted from the corresponding macrophages, was demonstrated after incubation with either germinal cells or spermatozoa. Marked cytotoxicity was also observed in vivo when sera or antibodies were injected into the tests, as shown by sequential histologic studies. Circulating immune complexes, as tested by the 125I-C1q binding assay, were not detected in these sera. On the contrary, high levels of circulating immune complexes were found when the polyethyleneglycol precipitation test was used. It can be concluded that the cytotoxic effect induced by cytophilic antibodies of IgG2 nature, may be associated with polyethyleneglycol precipitating immune complexes.

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The object of the present work was to investigate: a) The cytotoxic activity of cytophilic antibody in sera from guinea pigs sensitized with the testicular antigen; b) The presence of immune complexes. In vitro cytotoxicity induced by hyperimmune sera or by the cytophilic antibodies eluted from the corresponding macrophages, was demonstrated after incubation with either germinal cells or spermatozoa. Marked cytotoxicity was also observed in vivo when sera or antibodies were injected into the tests, as shown by sequential histologic studies. Circulating immune complexes, as tested by the 125I-C1q binding assay, were not detected in these sera. On the contrary, high levels of circulating immune complexes were found when the polyethyleneglycol precipitation test was used. It can be concluded that the cytotoxic effect induced by cytophilic antibodies of IgG2 nature, may be associated with polyethyleneglycol precipitating immune complexes.

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