RESUMO
PROBLEM: There is paucity of human data about the effects of depot medroxyprogesterone (DMPA) and norethisterone enanthate (Net-En) use on systemic immune function, which may have implications for reproductive tract infection susceptibility and transmissibility. We sought to evaluate the impact of injectable contraceptive use on T-cell responsiveness using T cells exposed in vivo and tested ex vivo. METHODS: Peripheral blood mononuclear cells were obtained from healthy, HIV-negative women after 30, 90 and 180 days of DMPA, norethisterone enanthate (Net-En) or copper intrauterine device (Cu-IUD) contraceptive use. Cells were stimulated ex vivo with phorbol myristate acetate and ionomycin, stained and analysed using flow cytometry. Mixed-effects linear models were used to evaluate change in proportions of T cells producing IFN-γ, TNF-α, IL-4 and IL-13. RESULTS: Compared with baseline, decreased proportions of IFN-γ-producing CD4+ and CD8+ T cells (p = .003, p = .006, respectively) and TNF-α-producing CD4+ and CD8+ T cells (p = .039, p = .034, respectively) were observed after 180 days of DMPA use. Decreased IL-4-producing CD4+ and CD8+ T cells (p = .045 and p = .024, respectively) were noted after 180 days of Net-En use. Decreased IL-4-producing CD4+ T cells were observed after 30 days (p = .035) and not after 180 days of DMPA use (p = .49). There were no changes in proportion of T cells producing IL-13 in DMPA users, nor any changes in IFN-γ, TNF-α and IL-13 in Net-En and Cu-IUD users. CONCLUSION: In vivo exposure of CD4+ and CD8+ T cells to typical pharmacologic concentrations of DMPA does not cause broad suppression to stimuli; however, depletion of specific cytokine-producing T cells may occur after prolonged DMPA use.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Acetato de Medroxiprogesterona/imunologia , Noretindrona/análogos & derivados , Progestinas/imunologia , Anticoncepcionais Femininos , Feminino , Humanos , Injeções , Interferon gama/metabolismo , Dispositivos Intrauterinos de Cobre , Ativação Linfocitária , Noretindrona/imunologia , Adulto JovemRESUMO
Previous reports speculated that vascular events could be related to the development of antibodies against synthetic steroids contained in oral contraceptives or other hormonal treatments. This study describes original immunoassays designed to detect antisynthetic steroid antibodies. In a first step, the assays were characterized and validated using animal-raised antisteroid antibodies. In a second step, a population of 88 oral contraceptive users, 47 of them having developed a vascular thrombosis during synthetic steroid use and 41 serving as healthy control users, were tested. Detection of antibodies against ethinylestradiol, levonorgestrel, norethisterone, cyproterone acetate, and gestodene showed that the values obtained in normal oral contraceptive users as well as thrombosis patients are very low, and show no statistically significant difference between the two groups tested. Taken together, these data indicate that the "immunological hypothesis" related to antisteroid antibodies is unlikely to explain the pathogenesis of vascular events in oral contraceptive users.
Assuntos
Anticorpos/análise , Anticoncepcionais Orais/efeitos adversos , Anticoncepcionais Orais/imunologia , Tromboflebite/etiologia , Adolescente , Adulto , Antagonistas de Androgênios/efeitos adversos , Antagonistas de Androgênios/imunologia , Anticorpos/imunologia , Anticoncepcionais Orais Sintéticos/efeitos adversos , Anticoncepcionais Orais Sintéticos/imunologia , Acetato de Ciproterona/efeitos adversos , Acetato de Ciproterona/imunologia , Etinilestradiol/efeitos adversos , Etinilestradiol/imunologia , Feminino , Humanos , Técnicas Imunoenzimáticas , Levanogestrel/efeitos adversos , Levanogestrel/imunologia , Pessoa de Meia-Idade , Noretindrona/imunologia , Norpregnenos/efeitos adversos , Norpregnenos/imunologia , Congêneres da Progesterona/efeitos adversos , Congêneres da Progesterona/imunologiaRESUMO
Anomalous cross-reactions with the dihydro- and tetrahydro-reduced metabolites of norethindrone were observed utilizing antisera raised against norethindrone-3-bovine serum albumin. Whereas displacement of 3H-norethindrone from the antiserum by the metabolites was generally minimal, where one of the metabolites was used as radiotracer, displacement by the metabolites was equal to or greater than that achieved by norethindrone. This unexpected finding was examined for its usefulness in developing a radioimmunoassay system for norethindrone metabolites in plasma. The sensitivity of the resulting standard curve was such as to permit quantitation of pg amounts of the reduced metabolites.
Assuntos
Noretindrona/imunologia , Animais , Reações Cruzadas , Feminino , Noretindrona/análise , Noretindrona/metabolismo , Coelhos , Radioimunoensaio , TrítioAssuntos
Anticoncepcionais Orais , Noretindrona/farmacologia , Adulto , Estatura , Peso Corporal , Corpo Lúteo , Estradiol/sangue , Feminino , Fase Folicular , Humanos , Menstruação , Noretindrona/sangue , Noretindrona/imunologia , Progesterona/sangue , Radioimunoensaio , Fatores de Tempo , Hemorragia UterinaRESUMO
Exposure of polystyrene tubes to solutions of glutaraldehyde was shown to provide a stable and consistent surface for immobilising an unpurified norethisterone-specific antiserum. Properties of the solid-phase antiserum were evaluated with respect to its application in developing a solid-phase radioimmunoassay for norethisterone. The assay proved to be sufficiently accurate, precise and sensitive for routine use. The technology is considered to be suited to automation.
Assuntos
Noretindrona/imunologia , Especificidade de Anticorpos , Sítios de Ligação de Anticorpos , Feminino , Glutaral/farmacologia , Humanos , Soros Imunes/farmacologia , Poliestirenos , Radioimunoensaio/métodosRESUMO
PIP: Specifities of 4 different norethisterone (Nor) antisera (coded A,B,C, and D) were evaluated and compared by cross-reaction studies to relate the antiserum specificity to the overall specificity of the radioimmunoassay (RIA), as established by plasma levels measured in women regularly taking the microdose of Nor (300 mcg/day). Using any of the 4 antisera, no significant deviation from parallelism were found among graded doses of authentic Nor and increasing volumes of plasma from women taking Nor for contraception. Cross-reaction studies preceded by chromatography to decrease plasma blanks are described, with each antiserum compared to the others for its efficacy in estimating plasma Nor values. It was concluded that 1) the significance of cross-reaction studies as well as that of a parallelism test for assessing overall specifity of the RIA is limited; 2) a single chromatography before RIA improves assay specificity but may not be sufficient to remove all interfering compounds; and 3) a comparison of direct and chromatographic procedures using several different antisera is useful for selection of the relatively most specific RIA procedure. These study results indicated that either antiserum C or D (preceded by chromatography) will yield better results than A or B.^ieng
Assuntos
Noretindrona/sangue , Adulto , Especificidade de Anticorpos , Cromatografia , Humanos , Noretindrona/imunologia , RadioimunoensaioRESUMO
PIP: A radioimmunoassay procedure is outlined for norethisterone, a synthetic progestagen. This assay uses both tritiated and iodine-125 labelled radioligands and may serve as a model for assays of synthetic steroids for which no tritiated radioligand exists. Male volunteers took a single oral dose of 10 mg of norethisterone acetate (SH 420). Plasma hormone levels were then measured at various time intervals. The degree of binding of iodine-125 labelled radioligand to antiserum even at low serum dilution was always greater than 80%. Using antinorethisterone-11 alpha-BSA serum, triated norethisterone and norethisterone-3-OCMO-iodine -125-iodohistamine radioligands give comparable results of adequate specificity, precision, accuracy and sensitivity when used to analyze crude ether extracts of the plasma samples. The chromatographic step is unnecessary for specific analysis. Iodine-125 labelled ligands may be useful for the determination of other synthetic steroids.^ieng
Assuntos
Noretindrona/sangue , Reações Cruzadas , Humanos , Radioisótopos do Iodo , Masculino , Noretindrona/imunologia , Radioimunoensaio/métodos , Fatores de Tempo , TrítioRESUMO
PIP: A radioimmunoassay for the measurement of norethindrone, utilizing an antiserum raised in sheep against norethindrone 3-(O-carboxymethyl) oximebovine serum albumin, and its application for determination of norethindrone concentrations in plasma is presented. The practical detection limit was .5 ng/ml plasma. No significant cross reaction was found for naturally occurring steroids. The cross reaction for d-norgestrel was 88% and for lynestrenol and R 2323 26%. Peripheral plasma levels of immunoreactive norethindrone were assayed at short intervals after oral administration of norethindrone in different doses to 6 human volunteers. Peak levels occurred within 1-2 hours. The plasma half-life of norethindrone was found to be about 3 hours during the first hours after ingestion and about 9 hours thereafter. A linear correlation was found between ingested dose per kg body weight and plasma peak levels. In 3 orally treated female rhesus monkeys the peak levels were lower and not as distinct as compared to that of the humans for the corresponding dose per kg body weight. The plasma half-life after the peak was about 8 1/2 hours.^ieng