Breast Milk Siga Anti-inflammatory Effects

Objective: To establish enzyme-linked immunosorbent assay(ELISA) for quantitatively detecting secretory A(SIgA) in human milk. The concentrations of IgG,IgM,IgA and SIgA in the milk of mothers were detected at different conditions by using immunoprecipitation and the established ELISA.Analysis the effect to influence immunoglobulin concentration in order to prevent the infection of newborns.Methods: The ELISA for quantification detection sIgA in human milk was established by using monoclonal anti-secretory component IgA antibody(SCMcAb) as coating antibody and evaluated its specificity and sensitivity. To collect colostrum and mature milk in mothers at different conditions.The concentrations of IgG,IgA,IgM and sIgA in 60 cases of colostrum and mature milk were detected using immunoturbidimetric assay and the established sIgA ELISA. To analyze the concentrations of Ig in milk in mothers at different conditions.Results: 1. The optimized experimental conditions for sIgA ELISA were as follows: The coating solution of SCMcAb was 10μg/ml. The content of enzyme conjugation was 1:5000. The dilution of human milk was 1:1000. The sensitivity of the assay was 0.31μg/ml. The average intra-assay CV and analytical recovery were 8.3% and 98.8%, respectively. The linear detection was 5-0.3μg/ml (r=0.9882, P<0.001). The specificity identification test showed that it did not react with the IgG and IgA. 2. The levels of IgG,IgA,IgM and sIgA in colostrum were significantly higher than that in mature milk (P<0.001).The levels of IgG,IgA,IgM and sIgA were not significantly difference between the old materals and the young materals, in the normal labor and caesarean section of 60 mothers. The levels of IgA and sIgA in colostrum were significantly difference between the city and the country (P<0.05), IgG and IgM were not different between groups (P>0.05). The levels of IgA,IgG and sIgA in good nutrition were significantly higher than that of in bad nutrition. 3. Although the levels of IgA and sIgA in human milk were significantly difference, their levels were linear regression (r=0.856 P<0.001,y=0.9056+0.5097x).Conclusion: A precise method of ELISA for sIgA quantitative analysis in human milk was developed and it can be used in clinic. The IgA is dominant in human milk. The influence factors of sIgA and IgA in human milk are nutritin and condition. Objective: Investigation the specific secretory immunoglobulin A antibody in different breast milk from 81 mothers and analysis the kinds of sIgA antibody against pathogens. This study provides information on the possible protective role of human milk sIgA antibody for newborns.Methods: Selection 10 pathogens as the study objects :RV,EPEC,SF ST,HP,CJ,.Cox,Echo,Aden,RSV. Cell-cultured RV SA11 was concertrated and purified. LB-cultured EPEC. SF and ST bacteria protein were distilled. The ELISA for detection specific sIgA against 10 pathogens were established by using purified RV SA11,distilled EPEC,SF,ST and commercial CoxB, Echo, Aden, RSV, HP, CJ as coating antigens. Monpclonal anti-human secretory component(SCMcAb)antibody was selected as detection antibody. The specific sIgA against 10 pathogens in 81 cases of colostrum and mature milk were detected by the established sIgA ELISA assay. Analysis bacteria protein of the 9 positive cases of EPEC sIgA by using western bolt.Results: 1. The positive rates of RV, RSV, Aden, SF, EPEC, CJ, HP, Cox and Echo in colostrum were 34.5%, 23.5%, 18.5%, 14.8%, 11.1%, 9.8%, 8.6%, 6.2%, 3.7% respectively , and that of in mature milk were 19.8%, 7.4%, 6.2%, 3.7%, 2.5%, 2.5% respectively. 2. The positive rates and the levels of sIgA against pathogens in colostrum were significantly higher tham that in mature milk (P<0.05). 3. The positive rates of RV, RSV, Aden sIgA were not different between lying-in woman in the city and in the country (P>0.05). The positive rates of SF and EPEC sIgA in the country were significantly higher than that in city (P<0.05). 4. The 9 cases positive EPEC sIgA can recognize three bands EPEC protein between 66KD and 97KD.Conclusion: There are specific sIgA against RV, RSV, Aden, SF, EPEC, CJ, HP, Cox and Echo in human milk. The specific sIgA in human milk are considered the important for the protection of newborns against pathogens.