RESUMEN
OBJECTIVES: To assess anti-ovarian antibodies (AOA) in serum samples at various times of in vitro fertilization (IVF) attempts to determine whether ovarian stimulation could result in the production of such autoantibodies in women. PATIENTS AND METHODS: Prospective study on 134 patients and 138 IVF cycles using a classical long protocol. For each attempt, four serum samples were obtained, respectively, at the onset of downrelation (S1), end of downregulation (S13), after 7 days of follicular stimulation (S21) and the day of follicular puncture (SP). Five hundred and fifty two samples were tested with an enzyme-linked immunosorbent assay for three isotypes (IgG, IgA, IgM) of AOA. RESULTS: In the whole group, mean concentrations of AOA for each isotype were compared group by group: S1-S13, S1-S21, S1-SP, S13-S21, S13-SP, S21-SP. Not any significant difference was observed whatever the isotype considered. DISCUSSION AND CONCLUSION: This study shows the absence of influence of endogenous or exogenous ovarian stimulation by gonadotropins on anti-ovarian autoimmunity.
Asunto(s)
Autoanticuerpos/sangre , Fertilización In Vitro , Ovario/inmunología , Adulto , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Estudios ProspectivosRESUMEN
The purpose of this study was to identify autoantigens contained in human ovary extracts. Serum samples from 36 infertile women with anti-ovary antibodies as detected with an ELISA technique were tested in Western blot against human ovary extracts. A reactive protein with a molecular mass matching that of the FSH was detected in 34 cases. These serum samples also reacted strongly in Western blot and ELISA with purified FSH and, in immunofluorescence, with pituitary cells. Using the Pepscan approach, with overlapping peptides matching the amino acid sequence of the human FSH beta-chain, several immunoreactive regions were evidenced. The 78-93 amino acid sequence of the human FSH beta-chain appeared as one of the major epitopes. Synthetic peptides of this region were prepared and demonstrated to react with human serum samples from women with anti-ovary antibodies. These data demonstrate that FSH can be an autoantigen, recognized by autoantibodies associated with infertility.
Asunto(s)
Autoantígenos/química , Enfermedades Autoinmunes/inmunología , Hormona Folículo Estimulante/inmunología , Infertilidad Femenina/inmunología , Fragmentos de Péptidos/inmunología , Autoanticuerpos/sangre , Western Blotting , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Epítopos/química , Femenino , Técnica del Anticuerpo Fluorescente , Hormona Folículo Estimulante/química , Humanos , Ovario/inmunología , Fragmentos de Péptidos/químicaRESUMEN
BACKGROUND: Peanut-containing food products may induce severe clinical reactions in sensitized subjects, and high levels of antipeanut IgE have been reported in the literature. Immunotherapy, proposed for the prevention of severe accidents, is often ill-tolerated and only partly efficient. This could be due to the spontaneous development of polyisotypic antipeanut antibodies. OBJECTIVE: To appreciate the presence and reactivity of other isotypes other than IgE of peanut-specific antibodies in serum samples from peanut-sensitized subjects. METHODS: Serum samples were obtained from 20 non-sensitized subjects and 23 sensitized patients divided in three groups according to their response to peanut oral challenge (no response or response to high or low doses, respectively). Peanut-specific IgG, IgG subclasses, IgA and IgM were assayed using an ELISA, and their reactivity against peanut proteins tested using Western Blot. RESULTS: A large dispersion of antipeanut antibody levels was observed in the three groups of patients, high levels of IgG, IgG1, IgG4 and IgA usually correlating with highly positive radioallergosorbent test (RAST). Such high levels were observed at onset in four patients who underwent peanut immunotherapy who had side effects and poor efficiency. Western blotting demonstrated that the polyisotypic response observed was directed to several peanut antigens, including the major allergens, Ara h1 and Ara h2. CONCLUSION: Peanut-sensitized patients who spontaneously develop specific IgE, display polyisotypic-specific antibody responses, whatever their response to oral challenge. This might explain the poor efficiency of peanut rush immunotherapy attempts.
Asunto(s)
Arachis/efectos adversos , Arachis/inmunología , Hipersensibilidad a los Alimentos , Isotipos de Inmunoglobulinas/sangre , Adolescente , Adulto , Especificidad de Anticuerpos/inmunología , Western Blotting , Niño , Preescolar , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , MasculinoRESUMEN
An immunoreactive role of lactic acid bacteria established in animals has seldom been investigated in humans. In a large-scale clinical study, children from day-care centers received either yoghurt (Y), milk fermented by yoghurt symbiosis and Lactobacillus casei (DN 114 001) (YC), or gelified milk (GM) as diet supplements during two 30-day supplementation periods separated by one 30-day period without supplementation. Feces samples were collected before, during, and after the 2nd supplementation period. Proteins were extracted in a buffer containing enzymatic inhibitors. IgA levels were assessed and adjusted to the weight of feces samples. Specific IgA to lactic acid bacteria strains (Streptococcus thermophilus 8901A, 8902A; Lactobacillus bulgaricus; Lactobacillus casei) present in Y and YC were assayed in ELISA and adjusted to individual IgA levels. Mean levels of fecal IgA were within reported ranges for pediatric populations of similar age. IgA levels decreased significantly but transiently in children receiving Y, and increased significantly in children receiving GM, but did not vary in the group of children who were given YC. Specific IgA to the 4 strains tested increased significantly during the supplementation period only in the group of children receiving GM, while it was transient and not significant in children receiving YC. No variation was noted in children given Y Specific IgA to lactic acid bacteria can be assayed in feces. Supplementation with fermented milks might induce a mucosal tolerance to environmental flora.
Asunto(s)
Heces/química , Fermentación , Inmunoglobulina A/análisis , Lactobacillus/inmunología , Leche/microbiología , Yogur/microbiología , Animales , Preescolar , Francia , Humanos , Lactante , Lacticaseibacillus casei/inmunología , Lacticaseibacillus casei/metabolismo , Streptomyces/inmunologíaRESUMEN
BACKGROUND: We conducted a prospective study to determine the prevalence of activated protein C resistance and anticardiolipin antibodies in leg ulcers, whatever venous, arterial or arteriovenous. PATIENTS AND METHODS: One hundred fifteen patients hospitalized for leg ulcers, without antiphospholipid syndrome were included. The vascular abnormalities were studied by clinical examination, Doppler, duplex Doppler and, when required, arteriography. Activated protein C resistance was isolated by a "classic" test (normalized APTT ratio in PCa presence or absence) and by a "second generation test" (by preliminary dilution with deficient factor V plasma). All patients with abnormal results on the second test were screened for the factor V Leiden (by PCR amplication with use of restriction enzymes). Anticardiolipin antibodies were investigated with an ELISA method with Harris standards as reference, in which the positive threshold was established at 20 units. RESULTS: Among these 115 patients, 50 venous (43.5 p. 100), 23 arterial (20 p. 100), 42 arteriovenous (36.5 p. 100) leg ulcers were identified. Activated protein C resistance was isolated in 12 cases (10.4 p. 100) (heterozygous carriers): 7 venous ulcers, 3 arteriovenous, 2 arterial. Anticardiolipin antibodies were measured at significant level in 49 cases (42.6 p. 100): 21 venous ulcers, 18 arteriovenous, 10 arterial. DISCUSSION: In this study, there was no statistical difference between the activated protein C resistance prevalence in leg ulcers when compared with Lorraine population (p=0.27). Factor V Leiden or anticardiolipin antibodies abnormalities were isolated in 56 cases (48.7 p. 100) without statistical difference between the 3 types of ulcers. Finally, the pathophysiology of venous, arterial and arteriovenous leg ulcers remains complex, suggesting several coagulation perturbations.
Asunto(s)
Resistencia a la Proteína C Activada/inmunología , Anticuerpos Anticardiolipina/sangre , Úlcera de la Pierna/inmunología , Resistencia a la Proteína C Activada/diagnóstico , Resistencia a la Proteína C Activada/genética , Anciano , Estudios Transversales , Factor V/genética , Factor V/metabolismo , Femenino , Tamización de Portadores Genéticos , Predisposición Genética a la Enfermedad/genética , Humanos , Úlcera de la Pierna/diagnóstico , Úlcera de la Pierna/genética , Masculino , Estudios Prospectivos , Factores de Riesgo , Úlcera Varicosa/diagnóstico , Úlcera Varicosa/genética , Úlcera Varicosa/inmunologíaRESUMEN
BACKGROUND: We examined the clonality of the specific T cell reaction activated with specific drug antigens in patients with a positive lymphocyte activation test. PATIENTS AND METHODS: Ten patients were investigated. Their clinical features suggested drug allergy. Six weeks after a skin reaction, skin tests and lymphocyte activation tests were run. For each lymphocyte activation test, we explored the clonality of the T cell proliferation by examining the VJ rearrangement of the TCR gamma locus using polymerase chain reaction and denaturing gradient gel electrophoresis. RESULTS: All samples gave positive lymphocyte activation tests proving in vitro polyclonal T cell proliferation in reaction to a drug. There was no difference in clonality between the pilot lymphocyte activation test and the lymphocyte activation test after stimulation with phytohemagglutinin or with the drug antigen. DISCUSSION: Our findings show that in vitro T cell proliferation induced by a specific drug antigen as disclosed by the lymphocyte activation test is a polyclonal or oligoclonal cell proliferation. This lack of production of a preferential T clone in a T cell-mediated immune drug reaction implies that many T lymphocytes can recognize the antigen and be stimulated by the same drug.
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Erupciones por Medicamentos/inmunología , Epítopos/inmunología , Activación de Linfocitos/inmunología , Subgrupos de Linfocitos T/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Células Clonales/inmunología , Femenino , Humanos , Masculino , Persona de Mediana EdadAsunto(s)
Anticuerpos Anticardiolipina/inmunología , Síndrome Antifosfolípido/diagnóstico , Complicaciones del Embarazo/diagnóstico , Embarazo de Alto Riesgo , Diagnóstico Prenatal/métodos , Anticuerpos Anticardiolipina/sangre , Síndrome Antifosfolípido/sangre , Síndrome Antifosfolípido/inmunología , Femenino , Humanos , Embarazo , Complicaciones del Embarazo/sangre , Complicaciones del Embarazo/inmunologíaRESUMEN
Autoimmunity may be involved in idiopathic premature ovarian failure (POF). However, the frequency, physiopathology and potential reversibility of autoimmune oophoritis needs clarification. Using an ELISA against whole tissue homogenate as antigen, from human ovaries at different ages, positive circulating ovarian antibodies (AOA) were found in 59% of patients with primitive idiopathic POF (27/46); 20/27 were positive for IgG isotype, 9/27 for IgM and 8/27 for IgA. Specificity of AOA was examined (i) by comparison to different control groups; mean values of the three subclasses of immunoglobins were significantly higher in POF women than in normally cycling fertile young women (n = 23) and fertile young men (n = 17), in untreated Grave's disease (n = 35) or in women with positive antinuclear factor (n = 25); and (ii) by assessing possible cross-reaction; only six out of the 27 positive sera reacted with other tissues (thyroid, pancreas, adrenal), including four clinical polyendocrinopathies. Significance of AOA was explored (i) by comparison with postmenopausal women (n = 40) and older women (n = 15), who did not have enhanced ratios of AOA, thereby excluding a potential role of high FSH values; (ii) by analysing the factors time and surgery; no relation could be found either with the duration of amenorrhoea (6 months to 21 years) or with the history of an ovarian biopsy (12/47) in the absence of any associated pelvic surgery; and (iii) by screening for other immunological factors; familial or personal autoimmune disease (8/46), HLA DR3 (10/42), HLA DR4 (11/42), associated autoantibodies (thyroperoxidase, adrenal, beta islets, parathyroid, DNA, smooth muscles) (12/42). If one positive AOA isotype was present, a second immunological factor was found in 45% of cases. Spontaneous pregnancies during oestrogen therapy occurred in four cases, including three women with positive AOA. Circulating AOA detected by an ELISA may represent a practical and suitable marker for diagnosis of POF. Its use for prognosis and rational treatment needs further evaluation.
Asunto(s)
Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Enfermedades Autoinmunes/inmunología , Ovario/inmunología , Insuficiencia Ovárica Primaria/inmunología , Adulto , Amenorrea/inmunología , Especificidad de Anticuerpos , Terapia de Reemplazo de Estrógeno , Femenino , Antígenos HLA-DR/sangre , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Persona de Mediana Edad , EmbarazoRESUMEN
BACKGROUND: Hypersensitivity to sesame seeds is becoming increasingly frequent, probably owing to the larger use of this compound in international food. OBJECTIVES: This study investigated serum responses of 12 sesame sensitized patients (seven with food allergy, five with food sensitization), to a sesame protein extract, and attempted at identifying sesame major antigens. METHODS: Sesame protein extracts were prepared from black, white and brown sesame seeds. Electrophoretic analysis showed similar protein patterns in the three extracts, and proper preservation of the proteins integrity. The brown sesame extract was used to set-up an ELISA assay and measure serum levels of antisesame IgG, IgA, IgM and IgE in 12 samples from sesame-sensitized individuals and six controls. It also allowed to perform western blot analyses in order to investigate the molecular weight of sesame proteins recognized by IgG, IgA and IgE. RESULTS: Nineteen protein bands were observed upon polyacrylamide gel electrophoresis of the sesame protein extracts. Using this whole extract in ELISA, significant antisesame IgG, IgA and IgE-responses were observed in the serum of sensitized individuals, different from the lower signals obtained with control samples. Western blot analysis demonstrated highly polymorphic IgG and IgA responses and a more restricted IgE response pattern, suggesting that two proteins, respectively, 14 kDa and 25 kDa are mostly involved in sesame IgE-dependent hypersensitivity, the 25 kDa band presenting several characteristics of a major allergen. CONCLUSIONS: This study reports novel information on the possible involvement of a 25 kDa sesame protein in IgE-dependent hypersensitivity to sesame seeds.
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Alérgenos/análisis , Anticuerpos/sangre , Hipersensibilidad a los Alimentos/inmunología , Magnoliopsida , Proteínas de Plantas/análisis , Adulto , Femenino , Hipersensibilidad a los Alimentos/sangre , Humanos , Inmunoglobulina E/sangre , Masculino , Extractos Vegetales/química , Extractos Vegetales/inmunología , SemillasRESUMEN
The implication of autoantibodies with anticoagulant and/or so-called antiphospholipid activities, under clinical circumstances with vascular obliteration, has led to the development of various types of tests allowing their detection. The most used tests involve investigation of the presence of an anticoagulant effect and of anticardiolipin IgG. It has also been proposed that the reactivity of patient samples toward other phospholipids or proteins be tested, but it remains difficult to appreciate which tests are redundant or complementary. Here we investigated whether the dissociation or association of anticoagulant and anticardiolipin correlated with specific ELISA reactivity to five other phospholipids: phosphatidylserine, phosphatidylinositol, phosphatidic acid, phosphatidylcholine, and phosphatidylethanolamine. The study was performed with 70 samples, evenly partitioned as positive for either anticardiolipin antibodies or anticoagulant effect, or both. Our data clearly confirm that cardiolipin reactivity is an individual entity, likely to be complementary to other assays. Neither anticardiolipin nor anticoagulant levels correlated with assays investigating antibody levels toward the five other phospholipids, although higher mean levels were noted when both lupus anticoagulant and anticardiolipin antibodies are present. Individual patterns were evidenced in all groups. These data support the interest of current and further studies exploring the clinical relevance of individual reactivities to phospholipids.
Asunto(s)
Anticuerpos Anticardiolipina/metabolismo , Sitios de Unión de Anticuerpos/inmunología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Antifosfolípidos/metabolismo , Niño , Preescolar , Interpretación Estadística de Datos , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Persona de Mediana Edad , Tiempo de Tromboplastina Parcial , Ácidos Fosfatidicos/inmunología , Fosfatidilcolinas/inmunología , Fosfatidiletanolaminas/inmunología , Fosfatidilinositoles/inmunología , Fosfatidilserinas/inmunologíaRESUMEN
This case report describes a successful pregnancy and delivery in a woman presenting with ovarian autoimmunity who had previously been involved in two unsuccessful in-vitro fertilization (IVF) attempts. Ten days before a third attempt, she began a regimen of 25 mg per day of prednisolone which was continued throughout the whole IVF protocol. Ovulation was induced by human menopausal gonadotrophin. After administration of 5000 IU of human chorionic gonadotrophin, 18 oocytes were collected. At 48 h after insemination with the patient's husband's spermatozoa, four 4-cell pre-embryos were transferred. A singleton pregnancy developed and led to the birth of a girl who is doing well 10 months later.
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Autoanticuerpos/análisis , Fertilización In Vitro , Ovario/inmunología , Prednisolona/uso terapéutico , Resultado del Embarazo , Adulto , Femenino , Humanos , Inmunoglobulinas/análisis , Embarazo , Resultado del TratamientoRESUMEN
The partition of human serum immunoglobulins along a pH gradient of ampholynes was investigated using the recently developed method of preparative isoelectrofocusing. Each isotype was demonstrated to display a specific pI range, with limited overlapping. IgA appear to be the most acidic serum immunoglobulins while IgG are clearly basic.
Asunto(s)
Isotipos de Inmunoglobulinas/química , Adolescente , Adulto , Anciano , Femenino , Humanos , Concentración de Iones de Hidrógeno , Focalización Isoeléctrica , Punto Isoeléctrico , Masculino , Persona de Mediana EdadRESUMEN
Wheat flour is a complex organic dust likely to induce immune responses when inhaled in work environment conditions. We compared the humoral status of 159 exposed workers from 11 flour mills and one industrial bakery with that of 41 workers from a salt factory. IgG, IgA, and IgM levels of antibodies to whole flour and to gliadin were assayed using ELISA tests in serum and saliva samples. Serum levels of IgG and IgA to both antigens were significantly higher (p < 0.0001) in occupationally exposed workers. Exposed workers had significantly higher levels of salivary IgG (p = 0.005) and IgA (p < 0.0001) to whole flour and of salivary IgG (p = 0.0005) to gliadin. In both groups, similar levels of anti-gliadin salivary IgA antibodies were observed. These data suggest that occupational exposure to wheat flour triggers specific immune responses, most likely through stimulation of the mucosal immune system. The presence of significant levels of serum antibodies, however, indicates that a systemic immunologic response is also present among exposed individuals.
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Formación de Anticuerpos/efectos de los fármacos , Harina/efectos adversos , Exposición Profesional/efectos adversos , Triticum/efectos adversos , Especificidad de Anticuerpos , Ensayo de Inmunoadsorción Enzimática , Industria de Procesamiento de Alimentos , Francia , Gliadina/efectos adversos , Gliadina/inmunología , Humanos , Inmunoglobulinas/análisis , Masculino , Saliva/inmunología , Triticum/inmunologíaRESUMEN
Anti-ovary antibodies (AOA) have been detected in serum samples of women undergoing in vitro fertilization (IVF). High concentrations of these antibodies have been found in women who have had several IVF attempts and they appear to correlate with reduced chances of pregnancy. In this paper, AOA were assayed sequentially in a series of 140 IVF candidates to investigate the respective roles of hormonal stimulation and follicular puncture in inducing the autoimmune response. Serum was obtained 8 days after the beginning of ovarian human menopausal gonadotrophin (hMG) stimulation, then 15 days after follicular puncture. Significantly higher concentrations of IgG (P < 0.0001) AOA were observed in the second series of samples than in the first, suggesting that ovarian trauma and not hormonal stimulation is responsible for triggering antibody production. In the whole group, there was a negative correlation between IgM levels after puncture and oocyte numbers (P < 0.05). Among 'immune-responder' women, the concentrations of IgA AOA (P = 0.01) in the first sample, and of IgG (P = 0.01) or IgA AOA (P < 0.05) in the second, correlated with fewer oocytes after stimulation. There was no variation in the mean concentrations of AOA in women who achieved pregnancy.
Asunto(s)
Autoanticuerpos/inmunología , Fertilización In Vitro/métodos , Inmunoglobulina M/inmunología , Folículo Ovárico/cirugía , Ovario/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina G/inmunología , Menotropinas/farmacología , Oocitos/inmunología , Folículo Ovárico/efectos de los fármacos , Ovulación/inmunología , PuncionesRESUMEN
AIMS: To investigate whether autoantibodies to choroid plexus are present in human senile dementia. METHODS: Serum samples from 40 elderly people presenting with characteristic, diagnostic criteria of senile dementia of Alzheimer's type and 20 age matched healthy controls were tested by indirect immunofluorescence for the presence of autoantibodies to choroid plexuses, using frozen sections of rat or human fetal brain tissue. RESULTS: Significant labelling of choroid plexus basement membrane was observed in 17 of the 40 samples from patients with senile dementia; in the control series one sample of rat but not human plexus labelled positively (p < 0.01). CONCLUSIONS: The antibodies identified in this series of patients with Alzheimer's disease suggest that autoimmune mechanisms might be responsible for some of the changes in cerebrospinal fluid production described in this disorder.