RESUMEN
In order to gain more information about local humoral immune responses to HPV infection, we quantified IgG, IgM, secretory-IgA (S-IgA), and total-IgA by ELISA, and lysozyme and lactoferrin by TR-IFMA, in cervical and cervicovaginal secretions of 40 healthy women and 28 high-risk HPV infected patients (11 were HPV16+). IgG, total-IgA, and S-IgA concentrations in cervicovaginal secretions (p < 0.0001) and high IgG and total-IgA concentrations (p < 0.001 and p < 0.01, respectively) in endocervical secretions were significantly higher in HPV+ patients than in the healthy group. Since the S-IgA/total-IgA ratio was significantly lower in cervicovaginal (7.5%) and endocervical secretions (36.5%) in HPV+ women compared to the control group (p < 0.003 and p < 0.001, respectively), HPV could be responsible for an increase in local production of non-secretory IgA (monomeric and dimeric forms). IgG and total-IgA concentrations in cervicovaginal and endocervical secretions fell in the same general percentage range in both HPV16+ and HPV+ groups (80% and 15%, respectively). However, the S-IgA/total-IgA ratio was much lower in HPV16+ than in HPV+ women, in both cervicovaginal secretions (3.4%) (p < 0.003) and in endocervical secretions (23.3%) (p < 0.001). Innate immunity proteins and local S-IgA response could not stop the spread of HPV infection in spite of high lysozyme and lactoferrin concentrations. HPV16+ disturbed the local humoral immune system, which could partly explain its low clearance.
Asunto(s)
Anticuerpos Antivirales/análisis , Cuello del Útero/inmunología , Infecciones por Papillomavirus/inmunología , Vagina/inmunología , Adolescente , Adulto , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Isotipos de Inmunoglobulinas/análisis , Lactoferrina/sangre , Persona de Mediana Edad , Muramidasa/sangre , Albúmina Sérica/análisisRESUMEN
Buccal and digestive tract opportunistic infections occur frequently in patients infected by HIV. In this study, we measured lysozyme (Lz), lactoferrin (Lf), total IgA (T-IgA), and secretory IgA (S-IgA) levels to investigate nonspecific secretory immunity in HIV-infected patients with oral candidiasis. Serum, saliva, and stool samples were analyzed by time-resolved immunofluorometric assay for Lz and Lf levels and by enzyme-linked immunosorbent assay for T-IgA and S-IgA levels. Mean salivary Lf and T-IgA levels (66.50 mg/L and 0.10 g/L, respectively) and mean fecal Lf, T-IgA, and S-IgA outputs (0.87, 54.0, and 43.6 mg/d, respectively) were significantly higher in HIV-infected patients with oropharyngeal candidiasis than in HIV-infected patients without oropharyngeal candidiasis and healthy subjects. There was a modification in the molecular form rate, with a high increase in S-IgA and monomeric IgA transudation from the plasmatic compartment into salivary and digestive fluids and an increase in salivary Lf local synthesis by polymorphonuclear neutrophils. HIV infection appears to be associated with dysregulation of some of the nonspecific immune factors at the mucosal surface. Despite high saliva concentrations and high intestinal output, innate immunity was not able to stop yeast expansion in HIV-infected patients.
Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/inmunología , Candidiasis Bucal/complicaciones , Candidiasis Bucal/inmunología , Infecciones Oportunistas Relacionadas con el SIDA/sangre , Infecciones Oportunistas Relacionadas con el SIDA/metabolismo , Adulto , Candidiasis Bucal/sangre , Candidiasis Bucal/metabolismo , Heces/química , Femenino , Humanos , Inmunidad Mucosa , Inmunoglobulina A/sangre , Inmunoglobulina A/metabolismo , Inmunoglobulina A Secretora/sangre , Inmunoglobulina A Secretora/metabolismo , Mucosa Intestinal/inmunología , Lactoferrina/sangre , Lactoferrina/metabolismo , Masculino , Persona de Mediana Edad , Mucosa Bucal/inmunología , Muramidasa/sangre , Muramidasa/metabolismo , Saliva/inmunología , Saliva/metabolismo , Albúmina Sérica/metabolismo , alfa 1-Antitripsina/metabolismoRESUMEN
Several studies were carried out to characterize the humoral immune response on mucosal genital surfaces. However, the results obtained so far were particularly conflicting due to the absence of validation methods. The aim of this study was to develop and validate a quantitative ELISA method, which is sensitive and reproducible, to measure immunoglobulin and secretory immunoglobulin concentrations in various biological fluids. This quantitative, sensitive (detection limit = 1 microg/L) and reproducible (coefficient of variation < 15%) method could be of interest to study the effects of viral infections on mucosal non-specific immune response in genital tract. To explore the humoral response, serum, saliva, vaginal secretions, and cervicovaginal secretions from 18 women, 20-45 years old, were evaluated for total-IgA, secretory IgA, IgM, and IgG. Albumin level was also evaluated by immuno-nephelometry. The secretion rates of immunoglobulins were measured by calculating their relative coefficients of excretion by reference to albumin. Despite large individual variations, median immunoglobulin levels were higher in the endocervical secretions than in the cervicovaginal secretions. When we compared the rates of immunoglobulins in genital fluids, IgG prevalence was higher (80%) in cervicovaginal and endocervical secretions than IgA prevalence (12%). In contrast, digestive mucosal secretions, such as saliva, contained mostly IgA (80%). In cervicovaginal and endocervical secretions, IgG and IgM originated mainly from serum, whereas a local synthesis provided total-IgA and secretory IgA. These results allowed us to raise a possible hypothesis for the origin of immunoglobulins in the genital tract. They illustrated the peculiar feature of the female reproductive tract and the difficulty for this tissue to contribute in the mucosal associated lymphoid tissue. The low secretory-IgA and total-IgA levels could explain the particular sensitivity of the vagina and the cervix to infections.
Asunto(s)
Genitales Femeninos/inmunología , Inmunoglobulinas/inmunología , Radioinmunoensayo/normas , Saliva/inmunología , Adolescente , Adulto , Moco del Cuello Uterino/inmunología , Moco del Cuello Uterino/metabolismo , Cuello del Útero/inmunología , Cuello del Útero/metabolismo , Ensayo de Inmunoadsorción Enzimática , Exudados y Transudados/inmunología , Femenino , Genitales Femeninos/metabolismo , Humanos , Inmunidad Mucosa/inmunología , Técnicas para Inmunoenzimas , Isotipos de Inmunoglobulinas/biosíntesis , Isotipos de Inmunoglobulinas/sangre , Inmunoglobulinas/análisis , Inmunoglobulinas/sangre , Reproducibilidad de los Resultados , Saliva/química , Albúmina Sérica/metabolismo , Vagina/inmunología , Vagina/metabolismoRESUMEN
Molecular typing systems have been needed to study Candida colonization in HIV-infected patients, particularly for investigating virulence and fluconazole resistance. Three methods--electrophoretic karyotyping (EK), detection of restriction fragment length polymorphisms (RFLP) and randomly amplified polymorphic DNA analysis (RAPD)--have been most frequently used. In this study, comparative sequence analysis of the internal transcribed spacer (ITS) region of rDNA was evaluated for delineation of Candida isolates from 14 HIV-infected patients. EK, ITS sequence analysis, RFLP and RAPD resulted in 11, 10, 9 and 8 DNA genotypes, respectively, from 39 Candida albicans isolates. The 10 genotypes observed using ITS sequence analysis were defined by six variation sites in the sequence. Molecular typing of sequential oral isolates showed the persistence of the same genotype of C. albicans in nine patients, and genotype variation in one patient. EK and RAPD showed that another patient was co-infected by two distinct genotypes and ITS analysis identified one of the two genotypes as Candida dubliniensis. Comparative ITS sequence analysis is a quick and reproducible method that provides clear and objective results, and it also identifies C. dubliniensis. The discriminatory power of this new typing approach could be improved by concomitant analysis of other DNA polymorphic sequences.
Asunto(s)
Candida albicans/clasificación , Candida/clasificación , Infecciones por VIH/complicaciones , Infecciones Oportunistas Relacionadas con el SIDA/microbiología , Candida/genética , Candida albicans/genética , Candidiasis Bucal/microbiología , ADN de Hongos/análisis , ADN Espaciador Ribosómico/análisis , Humanos , Cariotipificación , Técnicas de Tipificación Micológica , Polimorfismo de Longitud del Fragmento de Restricción , Técnica del ADN Polimorfo Amplificado Aleatorio , Análisis de Secuencia de ADNRESUMEN
The aim of this study was to explore anti-Candida albicans systemic and mucosal humoral responses against Candida virulence antigens such as somatic antigen and secreted aspartic proteases (Saps) in HIV-infected patients with oral candidiasis. Twenty-eight subjects were included in the study: 11 HIV-positive patients without oral candidiasis (group A), 6 HIV-positive patients with oral candidiasis (group B) and 11 HIV-negative healthy controls (group C). Total IgA, IgG and IgM concentrations and antibodies to C. albicans (somatic antigen, Sap1, Sap6) were measured in serum and saliva. We developed a time-resolved immunofluorometric assay with biotin and europium-labeled streptavidin for this purpose. Salivary total IgA, IgG and IgM concentrations were higher in group B. IgA, IgG and IgM anti-C. albicans antibodies (against somatic antigen, Sap1, Sap6) were higher in saliva and serum from patients from group B compared with patients from group A and controls. Our results suggest that, in oral candidiasis, HIV-infected patients have a high mucosal response, specifically directed against C. albicans virulence antigens, such as somatic antigen, Sap1 and Sap6.
Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/inmunología , Anticuerpos Antifúngicos/sangre , Anticuerpos Antifúngicos/metabolismo , Candida albicans/inmunología , Candidiasis Bucal/complicaciones , Candidiasis Bucal/inmunología , Saliva/inmunología , Adulto , Antígenos Fúngicos , Ácido Aspártico Endopeptidasas/inmunología , Candida albicans/enzimología , Candida albicans/patogenicidad , Estudios de Casos y Controles , Femenino , Infecciones por VIH/inmunología , Humanos , Inmunidad Mucosa , Inmunoglobulina A/sangre , Inmunoglobulina A Secretora/metabolismo , Inmunoglobulina G/sangre , Inmunoglobulina G/metabolismo , Inmunoglobulina M/sangre , Inmunoglobulina M/metabolismo , Masculino , Persona de Mediana EdadRESUMEN
Oropharyngeal candidiasis (OPC), mainly caused by Candida albicans, is commonly observed in HIV-infected patients. Secreted aspartic proteinases (Saps) are virulent agents involved in adherence to the mucosal surface and in tissue invasion. The immune secretory response to these agents was investigated in 15 HIV-infected patients, during oral yeast colonization and episodes of oropharyngeal candidiasis (OPC), in a 1-year longitudinal study. We developed an avidin-biotin-amplified immunofluorometric assay for the detection of specific immunoglobulins G, A, and M against somatic, Sap2 and Sap6 antigens. We report increases in anti-somatic, anti-Sap2, and anti-Sap6 salivary antibodies in patients with OPC. Over the 1-year period, not only OPC episodes but also variations in yeast colonization levels were correlated with variations in salivary anti-Sap6 antibody levels. Our results show the ability of HIV-infected patients to produce high levels of salivary antibodies; however, these antibodies were not efficient in limiting candidal infection, probably because of cellular cooperation deficiency and the enhanced virulence of the infecting strain.
Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/inmunología , Anticuerpos Antifúngicos/análisis , Ácido Aspártico Endopeptidasas/inmunología , Candida albicans , Candidiasis/inmunología , Proteínas Fúngicas , Infecciones por VIH/inmunología , Mucosa Bucal/inmunología , Saliva/microbiología , Infecciones Oportunistas Relacionadas con el SIDA/microbiología , Adulto , Antígenos Fúngicos , Avidina , Biotina , Candida albicans/inmunología , Candidiasis/microbiología , Candidiasis Bucal/inmunología , Candidiasis Bucal/microbiología , Femenino , Técnica del Anticuerpo Fluorescente , Infecciones por VIH/complicaciones , Humanos , Inmunoglobulina A Secretora/análisis , Inmunoglobulina G/análisis , Inmunoglobulina M/análisis , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Enfermedades Faríngeas/inmunología , Enfermedades Faríngeas/microbiología , Saliva/inmunologíaRESUMEN
Apolipoprotein E (apo E) is postulated to be a major lipid carrier protein in the brain involved in brain development and repair. Multiple sclerosis (MS) is a major demyelinating disease characterized by destruction of myelin and marked alteration of myelin cholesterol and lipid metabolism. We have determined serum and cerebrospinal fluid (CSF) apo E concentrations using an original time-resolved immunofluorometric assay and calculated intrathecal apo E concentration. Apo E concentrations were determined in 13 control subjects and 129 neurological patients: 34 definite MS patients, 25 with Guillain-Barré syndrome (GBS), 32 with amyotrophic lateral sclerosis (ALS) and 38 with other neurological diseases. Seven clinical parameters (sex, age, age at MS onset, duration of the disease, course, clinical status and disability score) were considered in MS patients. Significant (P < 0.01) decrease in CSF apo E was observed in MS, linked to a decrease in intrathecal apo E. The decreased CSF apo E concentration in MS patients occur independent of the apo E genotype. Apo E is considered as a neurotrophic factor in the brain. Any decrease in intrathecal apo E synthesis would thus contribute to progression of neurological diseases, such as MS.
Asunto(s)
Apolipoproteínas E/sangre , Esclerosis Múltiple/sangre , Adulto , Anciano , Apolipoproteínas E/líquido cefalorraquídeo , Apolipoproteínas E/genética , Femenino , Fluorometría , Humanos , Técnicas Inmunológicas , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/genética , Concentración Osmolar , Polimorfismo Genético/genéticaRESUMEN
The aim of the study was to investigate the systemic and, for the first time, the intestinal humoral events in the susceptible Balb/C mouse strain after oral administration of Echinococcus multilocularis eggs. Thirty-one mice were divided into three groups; W-2, W-8 and control group. Each mouse of the W-2 and W-8 groups was orally infected with 1,500 E. multilocularis eggs, two weeks and eight weeks before sacrifice respectively. Control group mice received phosphate buffer saline. Measurement of anti-E. multilocularis and non-specific IgG, IgA and IgM, and of a transudation marker, albumin, were performed in serum and intestinal washings by a time-resolved immunofluorometric assay. These results were complemented by microscopic examination of the intestinal mucosa. This infection model is well-suited to the study of mucosal immunity during alveolar echinococcosis. It showed a major specific intestinal response in the early stage of the disease whereas the systemic response predominated later in the disease. Histopathological studies and calculation of the relative coefficient of excretion of Ig also confirmed that the presence of the parasite, even during a short period, was responsible for a local immunological and inflammatory response and for a change in mucosal permeability. Mucosal immunity could thus play a role in tolerance induction against E. multilocularis that could be a prerequisite for the subsequent development of the larvae in the liver, and for the occurrence of the parasitic disease, alveolar echinococcosis.
Asunto(s)
Equinococosis/inmunología , Intestinos/inmunología , Intestinos/parasitología , Ratones Endogámicos BALB C/inmunología , Ratones Endogámicos BALB C/parasitología , Óvulo/inmunología , Animales , Anticuerpos Antihelmínticos/sangre , Anticuerpos Antihelmínticos/metabolismo , Equinococosis/sangre , Inmunidad Mucosa , Inmunoglobulina A/sangre , Inmunoglobulina A/metabolismo , Inmunoglobulina G/sangre , Inmunoglobulina G/metabolismo , Inmunoglobulina M/sangre , Inmunoglobulina M/metabolismo , Mucosa Intestinal/inmunología , Mucosa Intestinal/patología , Ratones , Albúmina Sérica/análisisRESUMEN
Enhanced serum IgA concentrations are common in alcoholic liver cirrhosis, but functional differences between IgA subclasses and their relation with interleukin-6 (IL-6) have not been described. Distinct immunoregulatory mechanisms may exist that selectively affect one subclass. This possibility prompted us to investigate the distribution of IgA1 and IgA2 subclasses in the serum of 25 heavy alcohol drinkers (alcohol: 80 to 200 g per day) without clinical disorders, in comparison with 35 patients affected by alcoholic liver cirrhosis, 29 viral hepatitis patients and 33 social drinkers as a control group. Mean (+/- SD) IgA2 concentration (0.56 +/- 0.31 g/l) was significantly increased (p < 0.01) in heavy alcohol drinkers, with an IgA2/IgA1 ratio of 0.33 +/- 0.12, while the mean total IgA concentration was similar to the control group. Mean IgA1 and IgA2 concentrations were significantly increased (p < 0.001) in alcoholic liver cirrhosis patients (6.13 +/- 4.52 g/l and 1.83 +/- 1.93 g/l respectively, with an IgA2/IgA1 ratio of 0.32 +/- 0.19) and viral hepatitis patients (3.66 +/- 2.59 g/l and 0.69 +/- 0.67 g/l respectively, with an IgA2/IgA1 ratio of 0.21 +/- 0.14) High serum IL-6 concentrations (34 +/- 33 ng/l) were correlated with elevated IgA1 and IgA2 concentrations only in patients with alcoholic liver cirrhosis. IgA2 subclass and IgA2/IgA1 ratio could therefore be used as markers of chronic alcohol abuse directly related to the extent and duration of the alcohol abuse and the effectiveness of alcohol withdrawal.
Asunto(s)
Alcoholismo/inmunología , Inmunoglobulina A/sangre , Adulto , Consumo de Bebidas Alcohólicas/inmunología , Biomarcadores/sangre , Estudios de Casos y Controles , Femenino , Hepatitis Viral Humana/inmunología , Humanos , Interleucina-6/sangre , Cirrosis Hepática Alcohólica/inmunología , Masculino , Persona de Mediana EdadAsunto(s)
Apolipoproteínas E/biosíntesis , Esclerosis Múltiple/metabolismo , Adulto , Anciano , Apolipoproteínas E/líquido cefalorraquídeo , Estudios de Casos y Controles , Estudios de Evaluación como Asunto , Femenino , Fluoroinmunoensayo , Humanos , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/líquido cefalorraquídeo , Sensibilidad y EspecificidadRESUMEN
Paired sera and cervicovaginal secretions from 35 HIV-1-infected women representing different CDC stages of HIV infection were evaluated for total IgA, IgA1 and IgA2, for IgA, IgA1 and IgA2 to gp160, and for albumin. Age-matched healthy women (n = 45) served as controls. The secretion rates of total IgA, IgA1 and IgA2 were evaluated by calculating their relative coefficients of excretion by reference to albumin. In HIV-infected women, total IgA1 and IgA2 in sera and in cervicovaginal secretions increased proportionately as early as stages II + III and more markedly at stage IV. By contrast, the secretion rates of total IgA IgA1 and IgA2 were markedly reduced in AIDS women, the IgA2 secretion rate decreasing significantly as early as stages II + III. This apparent discrepancy was probably the result of increased transudation of serum-borne immunoglobulins into the vaginal cavity, since albumin levels in cervicovaginal secretions increased significantly according to the stages of disease. HIV-reactive IgA antibodies in serum, as in cervicovaginal secretions, were principally found within the IgA1 subclass. In women at stage IV, a high local production of IgA1 to gp160 occurred in spite of the impairment of cervicovaginal IgA synthesis, probably because of marked genital HIV replication at advanced stages.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/metabolismo , Cuello del Útero/metabolismo , Inmunoglobulina A/biosíntesis , Inmunoglobulina A/sangre , Vagina/metabolismo , Adolescente , Adulto , Especificidad de Anticuerpos , Femenino , Productos del Gen env/inmunología , Proteínas gp160 de Envoltorio del VIH , Humanos , Inmunoglobulina A/inmunología , Precursores de Proteínas/inmunología , Albúmina Sérica/análisisRESUMEN
BACKGROUND/AIMS: An alteration of the secretory immune response has been forwarded to explain frequent and chronic mucosal infections in patients with acquired immunodeficiency syndrome (AIDS). The aim of this study was to explore the intestinal immunoglobulin (Ig) secretions in patients with AIDS and their relationships to cryptosporidiosis. METHODS: Patients with AIDS and enteric cryptosporidiosis (n = 12), other enteric infections (n = 10), and no identifiable enteric pathogen (n = 10) and human immunodeficiency virus-seronegative controls (n = 18) were studied. The number of intestinal IgA and IgM plasma cells of the duodenal lamina propria mucosa and total and anti-Cryptosporidium IgA, IgM, and IgG were measured in serum and feces. RESULTS: Although not significantly increased, the number of IgA and IgM plasma cells was greater in patients with AIDS (n = 20) than in controls (n = 5). In feces, total IgA outputs and specific anti-Cryptosporidium IgA levels were significantly higher in patients with AIDS and cryptosporidiosis than in the two other groups of patients with AIDS (P < 0.05 and P < 0.01, respectively) and controls (P < 0.001 and P < 0.01, respectively). Total fecal IgM output and specific anti-Cryptosporidium IgM coproantibodies were increased only in the Cryptosporidium-infected patients relative to the controls (P < 0.05). CONCLUSIONS: Despite the development of pathogen-specific mucosal antibody responses, patients with AIDS and cryptosporidiosis fail to clear the parasite.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/inmunología , Anticuerpos Antiprotozoarios/análisis , Cryptosporidium/inmunología , Mucosa Intestinal/inmunología , Mucosa Intestinal/metabolismo , Síndrome de Inmunodeficiencia Adquirida/complicaciones , Adulto , Animales , Formación de Anticuerpos , Recuento de Células , Criptosporidiosis/complicaciones , Electroforesis en Gel de Poliacrilamida , Heces/química , Femenino , Humanos , Immunoblotting , Inmunoglobulinas/análisis , Masculino , Persona de Mediana Edad , Células Plasmáticas/patología , alfa 1-Antitripsina/metabolismoAsunto(s)
Enfermedad Celíaca/inmunología , Gliadina/inmunología , Inmunoglobulina A/clasificación , Inmunoglobulina A/metabolismo , Adulto , Anciano , Heces/química , Femenino , Humanos , Inmunidad Mucosa , Inmunoglobulina A/sangre , Mucosa Intestinal/inmunología , Masculino , Persona de Mediana EdadRESUMEN
Although low density lipoprotein receptors have been described on oligodendrocytes, apolipoprotein B was thought to be absent or present in only very small amounts in cerebrospinal fluid (CSF). Several immunoassays have been used for the measurement of apolipoprotein B in serum. However, the majority of methods cannot be used to measure small amounts of apolipoprotein B in CSF. In this study, we describe a highly sensitive time resolved immunofluorometric assay (TR-IFMA) using europium as label (detection limit: 0.3 microgram/l). The reliability of the TR-IFMA for the measurement of apolipoprotein B was first studied in serum. Serum and CSF apolipoprotein B concentrations were then determined in subjects free of neurological disorders and in patients with multiple sclerosis. Local intrathecal apolipoprotein B synthesis was calculated. Although the high sensitivity of the TR-IFMA allowed low amounts of apolipoprotein B in CSF to be detected (0.11 +/- 0.06; 0.12 +/- 0.06 mg/l in controls and multiple sclerosis patients, respectively), no apolipoprotein B could be detected in CSF by electroimmunodiffusion. As suggested by the blood/CSF apolipoprotein B ratio (about 6000), no apolipoprotein B synthesis was observed by both using apolipoprotein B index and formula. This indicates its probable serum origin. Moreover, there was no difference between controls and multiple sclerosis patients in CSF, serum, blood/CSF, index, and local intrathecal apoliprotein B synthesis. Finally, these results suggest that the role of apolipoprotein B in lipid transport in the central nervous system may be questionable.
Asunto(s)
Apolipoproteínas B/líquido cefalorraquídeo , Fluoroinmunoensayo/métodos , Esclerosis Múltiple/líquido cefalorraquídeo , Enfermedades del Sistema Nervioso/líquido cefalorraquídeo , Adulto , Anciano , Apolipoproteínas B/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/sangre , Nefelometría y Turbidimetría , Enfermedades del Sistema Nervioso/sangre , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Local immunological defense mechanisms in the cervicovaginal mucosa currently remain incompletely defined, especially from a quantitative point of view. Addition of an inert substance, lithium chloride (LiCl), into the washing buffer used to carry out the vaginal washing for collecting cervicovaginal secretions and measurement of its concentration with a flame absorption spectrophotometer, before and after the specimen is sampled, permits the quantification of the volume of cervicovaginal secretions collected and the approximation of the dilution factor of a soluble component introduced by the washing. Lithium, at a concentration of 10 mM, gives the best precision of measurement and has no effect on the results of the immunoassays. In a population of 27 nonpregnant women (age range, 18 to 45 years), the volume of cervicovaginal secretions collected by vaginal washing with 3 ml of LiCl-phosphate-buffered saline was 12% +/- 3.2% (mean +/- standard deviation) of the total volume and showed large interindividual variations (range, 5.6 to 18.8%); the mean dilution factor of a soluble component from the vaginal secretions was 9.9% +/- 2.8% (range, 6.3 to 18.8%). According to the date of the menstrual cycle, the mean volume of collected cervicovaginal secretions was significantly increased in the luteal phase in comparison with the follicular phase; conversely, the mean dilution factor of a soluble component was more important in the follicular than in the luteal phase. These features strengthen the need to quantify accurately the dilution factor introduced by vaginal washing when studying cervicovaginal immunity.
Asunto(s)
Líquidos Corporales/inmunología , Cuello del Útero/metabolismo , Vagina , Adolescente , Adulto , Cuello del Útero/inmunología , Femenino , Fase Folicular/inmunología , Anticuerpos Anti-VIH/análisis , Proteína gp120 de Envoltorio del VIH/inmunología , Infecciones por VIH/inmunología , VIH-1/inmunología , Humanos , Técnicas para Inmunoenzimas , Inmunoglobulina G/análisis , Cloruro de Litio/análisis , Fase Luteínica/inmunología , Persona de Mediana Edad , Reproducibilidad de los Resultados , Irrigación TerapéuticaRESUMEN
Detection of semen anti-human immunodeficiency virus (HIV) antibodies within the cervicovaginal secretions from a non-HIV-infected woman who has had a recent sexual intercourse with an HIV-infected man is theoretically possible since the seminal fluid from all HIV-infected men contains a high titer of IgG antibodies to HIV. We report the case of an HIV-seronegative African woman whose cervico-vaginal secretions contained IgG antibodies to HIV, including antibodies to HIV-env-encoded glycoproteins. This woman had also detectable prostatic specific antigens and acid phosphatase in her cervico-vaginal secretions, establishing the persistence of semen. In order to confirm whether anti-HIV antibodies in seminal fluid could be detected in vitro when mixed with cervico-vaginal secretions, 10(-1) to 10(-6) 10-fold dilutions of seminal fluid from HIV-1-seropositive donors were realized with a pool of HIV-negative cervico-vaginal secretions as diluent. Six commercial enzyme immunoassays or rapid tests were compared for semen anti-HIV detection in the secretions. At a 10(-1) dilution of the mixture, all assays were markedly positive for all tested semens and the greatest dilutions of seminal fluid showing positivity ranged from 10(-3) to 10(-5). The IgG immunocapture assay appeared to be the most sensitive test. The rapid tests permitted the detection of semen IgG antibodies to HIV at dilutions ranging from 10(-1) to 10(-3) suggesting their potential value in emergency situations.
Asunto(s)
Coito , Anticuerpos Anti-VIH/análisis , Infecciones por VIH/prevención & control , Semen/inmunología , Vagina/inmunología , Côte d'Ivoire , Ensayo de Inmunoadsorción Enzimática , Femenino , Productos del Gen env/inmunología , Anticuerpos Anti-VIH/sangre , Infecciones por VIH/inmunología , Infecciones por VIH/transmisión , Seronegatividad para VIH , Humanos , Immunoblotting , Inmunoglobulina G/análisis , Inmunoglobulina G/sangre , Masculino , Violación , Vagina/metabolismoRESUMEN
In order to evaluate a commercial immunoglobulin G (IgG) antibody capture enzyme-linked immunoassay (ELISA) (Wellcozyme HIV1 + 2 Gacelisa; Murex Diagnostics Limited, Dartford, United Kingdom) for the detection of antibodies to human immunodeficiency virus (HIV) in vaginal secretion samples (VS) from HIV-seropositive and -seronegative women, serum samples (S) and VS were obtained from 129 African women living in the Central African Republic, a country of high HIV prevalence. Sera were tested for HIV by routine second-generation ELISA with confirmatory Western blot (immunoblot) (WB). By the Gacelisa IgG immuno-capture assay, 45 VS were positive and 84 were negative, whereas by WB, 44 VS were confirmed positive and 85 were confirmed negative. Considering WB as a reference, the IgG immunocapture assay in VS was 97.7% sensitive (43 of 44 positive samples) and 97.6% specific (83 of 85 negative samples). Of 42 HIV-seropositive women, 41 (97.6%) had S and VS that both were HIV positive (S+ VS+), and of 87 HIV-seronegative women, 83 (95.4%) had S and VS that both were HIV negative (S- VS-). Five women had discordant results for S and VS. One (S+ VS-) possibly had a false-negative VS result. Two (S- VS+) had similar indeterminate patterns for S and VS in WB. Two (S- VS+) had a typical HIV-positive pattern on WB of VS, whereas S results in WB were indeterminate in one case and negative in the other case; for both women, detection of prostatic acid-phosphatase was positive in VS, strongly suggesting recent sexual intercourse with an HIV-positive man. Because all HIV-infected men have detectable IgG antibodies to HIV in the seminal fluid, an HIV-seronegative rape victim with HIV-positive VS (S- VS+) should receive short-term antiviral therapy to prevent possible HIV transmission.