ABSTRACT
La determinación de la carga viral de HIV y la detección del ADN proviral en semen se utilizan en protocolos de fertilización para parejas discordantes (hombre HIV positivo - mujer AMPLICOR HIV-1 MONITOR tm Test versión 1.5 (Roche) en plasma seminal para estimar la carga viral. Se procesaron 31 muestras de plasma seminal, diluidas y sin diluir para evaluar la amplificación del control interno y descartar posibles inhibiciones. Luego se contaminaron 22 muestras con plasma de pacientes HIV positivos para verificar la cuantificación del ARN viral. Finalmente, se procesaron 12 muestras en paralelo por Nuclisens HIV-1 QT (Biomerieux) para descartar potenciales falsos negativos. Concluimos que el sistema evaluado es un método adecuado para cuantificar ARN viral en plasma seminal. No se observó inhibición, ni falsos negativos y los valores de carga viral y el límite de detección no se vieron modificados por la matriz diferente.
Analyses of the HIV load and presence of proviral DNA in sperm samples are used in assisted fertilization protocols for discordant couples (infected man-healthy woman). We evaluated the use of the COBAS Ampliprep/COBAS AMPLICOR HIV-1 MONITOR tm Test versión 1.5 (Roche) for viral load quantification in seminal plasma samples. We first tested 31 sperm samples for amplification of the internal control to discard potential inhibition. Seminal plasmas were analyzed directly and diluted. We then spiked 21 sperm samples with human plasma from HIV-positive patients to confirm that HIV RNA could be amplified. We also compared the results of 12 samples with NASBA (Nuclisens HIV-1 QT, Biomerieux), and confirmed lack of false negative results. We conclude that the new assay is an adequate methodology to analyze HIV load in sperm samples. We did not observed inhibition, neither false negative results and quantification demonstrated equivalent HIV loads.
Subject(s)
Humans , Male , Antiretroviral Therapy, Highly Active , DNA , Fertilization/immunology , HIV , Guidelines as Topic/methods , Reverse Transcriptase Inhibitors , Reverse Transcriptase Polymerase Chain Reaction , Semen/immunology , Nucleic Acid Amplification Techniques/methods , Viral LoadABSTRACT
As células natural killer endometriais, também chamadas células natural killer uterinas, têm recebido especial atenção no campo da imunologia reprodutiva. Teorias que consideram alterações na resposta imune como uma causa de infertilidade conjugal e de falhas nos tratamentos de reprodução assistida têm ponderado um possível envolvimento negativo das células natural killer endometriais. As células natural killer são linfócitos que podem ser identificados no sangue periférico e no endométrio, apresentando diferenças fenotípicas e funcionais importantes. As células periféricas não se alteram com a fase do ciclo menstrual e implantação, sendo que as células natural killer endometriais apresentam variações durante o ciclo menstrual e período peri-implantacional, com menores concentrações durante a fase proliferativa e aumentando na segunda fase do ciclo. A célula natural killer endometriais participam nas várias fases da implantação, invasão trofoblástica, placentação e desenvolvimento fetal e no desenvolvimento da gestação humana até aproximadamente 20 semanas.
Endometrial natural killer cells have been given special attention in reproductive immunology. The relation between the endometrial natural killer cells and alterations in the immune response as a cause of couples infertility and failure in assisted reproduction treatment have been studied in several theories. Natural killer cells are lymphocytes that may be identified in peripheral blood and endometrium, with phenotypical and functional differences between them. Peripheral natural killer cells do not change with the menstrual cycle or implantation, as opposed to endometrial natural killer cells which present lower concentration in the proliferative phase and higher concentration in the luteal phase. Endometrial natural killer cells play an important role in the implantation, trophoblastic invasion, placentation, fetal development and development of the human pregnancies up to 20 weeks of gestation.
Subject(s)
Female , Pregnancy , Abortion, Habitual/etiology , Killer Cells, Natural/cytology , Killer Cells, Natural/immunology , Embryo Implantation , Embryo Transfer , Endometrium/immunology , Endometrium/pathology , Fertilization/immunology , Pregnancy Maintenance/immunologyABSTRACT
Existen una serie de eventos inmunitarios que rodean la fertilización, un 3 por ciento de las parejas en las cuales se descarta toda explicación orgánica, hormonal o autoinmune para la infertilidad, parecen ser infértiles por mecanismos inmunológicos. Se conoce que hay anticuerpos antiespermatozoides en el suero del hombre y de la mujer, en las secreciones vaginales de la mujer y en el esperma de hombres infértiles. Se presenta un caso de infertilidad de causa no explicada, se trata de un hombre con semen normal y una mujer con ovulación normal y trompas permeables. Se evaluó el moco cervical posterior a lo cual se realizó prueba poscoital; se determinó la presencia de anticuerpos anti espermatozoides de la clase IgA e IgG adheridos al espermatozoide, en el moco cervical y plasma seminal mediante la prueba de Inmunobeads directo e indirecto. Detectamos una prueba poscoital anormal, 6 por ciento de anticuerpos antiespermatozoides de la clase IgG, asociados al espermatozoide y un 8 por ciento de anticuerpos antiespermatozoides de la clase IgG en el moco cervical. Los anticuerpos antiespermatozoides en el plasma seminal fueron negativos.
A series of inmune events exists that surround the fertilization, 3 percent of copules in which all the organic, hormonal or autoimmune explanation is discarded for the infertility, seem to be infertile because of immunological mechanisms. It is known the existence of anti spermatozoa antibodies in the serum of the man and the woman, in vaginal secretions of the woman and in the sperm of infertile men. A case of infertility of nonexplained cause appears, it is about a man with normal semen and a woman with normal ovulation and permeable tubes. The cervical snot was subsequently evaluated to which a postcoital test was done; the presence of anti spermatozoa antibodies of the IgA and IgG class was determined associated to the spermatozoon, in the cervical snot and seminal plasma by means of the test of direct and indirect Inmunobeads. It was detected an abnormal postcoital test, 6 percent of anti spermatozoa antibodies of the IgG class, associated to the spermatozoon and 8 percent of anti spermatozoa antibodies of the IgGG class in the cervical snot. The anti spermatozoa antibodies in the seminal plasma were negative.
Subject(s)
Humans , Male , Female , Fertilization/immunology , Immune System Diseases , Reproduction/immunology , Case ReportsABSTRACT
Antifertility effects of naturally occuring antisperm antibody [ASA] in infertile couples and studies on experimental immunization of various animals with sperm antigens represent ASA as an immunocontraceptive target. The effects of different factors on sperm immunogenecity and ASA production have been studied and different results have been reported. In this study, whole sperm immunization was evaluated. In this experimental study, whole mice sperm with different adjuvants i.e. complete Freund's adjuvant [CFA], incomplete Freund's adjuvant [ICFA], cholera toxin subunit- beta [CTS-beta] were administrated to mice by different routes; Intramuscular [IM], Subcutaneous [SC], Intranasal [IN], Intra peritoneal [IP], Intrarectal [IR], Intravaginal [IVA] and oral. Control groups were inoculated with phosphate buffer saline [PBS] plus corresponding adjuvant. Immunization was carried out on day 0,7,14,28 and ASA titers were detected by indirect immunofluorescence [IFA] technique. The results were compared between control and experimental groups by Mann Whitney and Fisher exact tests. The number of positive mice for ASA in IM, IN and SC experimental and control groups were significantly different [P=0.01, P=0.01, P=0.04 respectively]. However, there were no significant differences between the IR, IVA, and oral experimental and control groups. No differences were observed between ASA in vaginal washing of all groups. Due to high mortality, the IP group was excluded from the study. It can be concluded that whole sperm antigen can induce immune response in female mice by IM. SC and IN routes, but not through IAV, IR and oral administration routes
Subject(s)
Animals, Laboratory , Animals , Mice , Antibody Formation , Fertility/immunology , Fertilization/immunology , ImmunizationSubject(s)
Humans , Male , Female , Pregnancy , Infertility, Female/immunology , Infertility, Male/immunology , Spermatozoa/immunology , Acrosome/immunology , Fertilization/immunology , Infertility, Female/diagnosis , Infertility, Female/etiology , Infertility, Male/diagnosis , Infertility, Male/etiologyABSTRACT
Effects on fertility of female mice after hyper-immunization with human lactate dehydrogenase-C4 have been studied at (50 + 25 x 4) and (50 + 50 x 4) micrograms doses of protein for two consecutive cycles of pregnancy. Results of heterologous immunization are described in relation to the role of LDH-C4 in conception and contraception. (i) Low dose of Immunization produced higher antibody (Ab) titer as tested before mating followed by higher infertility (70%) than by a higher dose of immunogen (46%), taking pregnancy as the end point. (ii) However, the results were reversed after second mating. For example, low dose of immunogen produced insignificant infertility, whereas high dose of immunogen gave 80% infertility. Similar order was shown by humoral Ab titer before mating i.e., high infertility was associated with high Ab titer and viceversa. (iii) It shows that the LDH-C4 induced infertility is reversible as is evidenced by low dose of immunization. (iv) In contrary to infertile dams, immunized fertile dams delivered a significantly higher litter size as compared to non-immunized control dams. It is concluded that immunity due to LDH-C4 is beneficial for embryo survival suggesting an immuno-suppressive nature of LDH-C4. Nonetheless, infertility ensues when immune-suppression is overcome by its own immune-activation.