IgA class entity determination study in secretion from the mucosal immune system compartments

Five separated mucosal viscous colloidal solutions were obtained from human saliva, semen, feces, colostroum and urine. These were biurate positive, 2ME resistant with antibody activity during infection. Solutions were designated as mucosal immunoglobulins [MIG]. MIG separation were performed by using 6% polyethylene glycol solution. Rabbit standard polyclonal anti SIgA was obtained through hyperimmunization of rabbit with standard SIgA. MIG solutions were of SIgA class entity as indicated by immunoelectrophoresis and radial immunoprecipition in gel. The use of PEG 6% for separation and class entity determination of MIG are being reported for the first time in this area

Purificación de IGA secretora y prepración de un antisuero específico / Purification of secretory IgA and prepation of an specific antiserum

Se purificó IgA secretora (IgAs) a partir de una mezcla de calostro humano mediante clarificación inicial y cromatografía de intercambio iónico. El suero de calostro, obtenido por acidificación y centrifugación se sometió a una cromatografía en una columna de DE-52 empleando solución tampón de fosfatos 0,01 M, pH 7,6 y posteriormente, un tampón de fosfatos 0,1 M, pH 6,4. La fracción eluida se pasó por una columna de CM-52 con un gradiente lineal de concentración con acético-acetato 0,005 M-0,5 M, pH 5,0. La pureza del producto final se demostró mediante inmunoelectroforesis y doble difusión, con el empleo de antisueros específicos. Con la IgA secretora purificada se inmunizó un grupo de conejos y se obtuvo un antisuero potente y de buena calidad

Immunoassays of secretory IgA and secretory component

1. A sandwich-type enzyme-linked immunosorbent (ELISA) is described for quantitation of secretory IgA (sIgA) in human serum, as well as an ELISA and a radioimmunoassay (RIA) for measurement of secretory component (SC) is human serum. Samples were reduced and alkylated prior to the measurement of SC. 2. Healthy individuals (N = 53) presented low levels of SC (median, 0.9 mg/l). The protein levels were significantly elevated when compared with the controls, in sera of women during the second (N = 31; median, 1.5 mg/l) and third (N = 35; median, 2.4 mg/l) and acute viral hepatitis (N = 25; median 2.4 mg/l). SC levels of women in the first trimester of pregnancy (N = 24; median, 0.5 mg/l) did not differ from the controls. 3. sIgA levels were also significantly elevated when sera of women in the third trimester of pregnancy (N = 41; median, 25.4 mg/l) and sera of patients with alcoholic cirrhosis (N = 32; median, 75.0 mg/l) or acute viral hepatitis (N = 38; median, 28.5 mg/l) were compared with controls (N = 49; median, 9.0 mg/l). women in the first (N = 25; median, 7.7 mg/l) and second (N = 29; median, 10.2 mg/l trimester of pregnancy did not present levels statistically different from the controls. 4. The results obtained for SC by RIA and ELISA were positively correlated (rs = 0.88; P < 0.001). sIgA levels determined by ELISA were also positively correlated with the results of RIA-SC(rs = 0.77;P<0.001) or ELISA-SC (rs = 0.79; P < 0.001). 5. The assays described are specific, relatively simple to perform, and can be useful for the study of the secretory system

Niveis de IgA, IgM e IgG no colostro e leite humano / Levels of IgA, IgM and IgG in the colostrum and milk human

Niveis das imunoglobulinas A, M e G foram determinados por imunodifusao radial simples, em colostro de 61 maes. Em 29 destas maes, foi feito estudo longitudinal do comportamento das imunoglobulinas, referentes aos 15e 30 dias de pos-parto. As imunoplacas para as dosagens e a SIgA de referencia para a quantificacao da imunoglobulina A foram preparadas no laboratorio. Praanalise estatistica, foram utilizados a mediana, o 1o. e 3o. quartil. Os resultados tambem foram submetidos a analise de variancia e ao teste de Tukei. As concentracoes das IgA, IgM e IgG foram significantemente mais elevadas no colostro,do que nos leites de 15 e 30 dias. As IgA e IgM nao apresentaram diferenca significante, nos leites de 15 e 30 dias; as IgG foram significantemente mais elevadas aos30 dias. A IgA foi sempre a imunoglobulina predominante no material estudado