F Morell1, R Codina, M J Rodrigo. 1. Servei de Pneumologia, (Unit d'Immunologia), Hospital General Vall d'Hebron, Barcelona, Spain.
Abstract
BACKGROUND: Results obtained from previous experiences with soybean hull antigens suggest that the addition of glycerine to the extract increases positivity of the skin test. OBJECTIVE: The purpose of this study was to investigate the effect of this glycerine addition and the influence of different storage temperatures on the potency of soybean hull extracts. METHODS: Twenty-two asthmatic patients admitted to emergency rooms during one of the soybean-related Barcelona asthma epidemics were evaluated 2 years after the last epidemic for: (1) sensitivity to prick test with glycerinated and non-glycerinated extracts, both fresh and stored at 4 degrees C, - 20 degrees C and - 70 degrees C for 30 days, and (2) specific IgE to a fresh hull extract. All extracts were also studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: No differences in positivities were detected for prick tests performed the same day of preparation between non-glycerinated (5/22; 22.7%) and glycerinated (4/22; 18.2%) extracts. After 30 days storage, positivities of prick test with non-glycerinated extracts stored at 4 degrees C did not significantly differ from the results obtained on the first day. However, prick tests performed with glycerinated extracts stored for 30 days showed increased positivity for the extracts stored at 4 degrees C (7/22; 31%) (P < 0.05) as well as for those stored at - 20 degrees C and - 70 degrees C (9/22; 40.9%) (P < 0.001). This latter percentage is similar to that of specific immunoglobulin E detected by radioimmunoassay (10/22; 45. 4%). Also, for the glycerinated extracts only, the quantitative response showed an increase in mean diameter of the induration at 30 days at the three temperatures, even though owing to the great standard deviation and limited number of patients, it was only significant for the extract stored at - 20 degrees C (P < 0.01). SDS-PAGE gel densitometry at 30 days demonstrated a loss of protein in the bands with a molecular weight higher than 66.2 kDa for non-glycerinated extracts stored at 4 degrees C. CONCLUSIONS: The results of this study confirm previous observations with soybean hull allergen extracts and indicate that (1) storage of soybean hull extracts with glycerine addition for 30 days at the concentration studied (1:100 w/v) determines a significant increase in the positivity of skin tests, and (2) glycerine addition stabilizes soybean hull extracts at any of the temperatures studied.
BACKGROUND: Results obtained from previous experiences with soybean hull antigens suggest that the addition of glycerine to the extract increases positivity of the skin test. OBJECTIVE: The purpose of this study was to investigate the effect of this glycerine addition and the influence of different storage temperatures on the potency of soybean hull extracts. METHODS: Twenty-two asthmatic patients admitted to emergency rooms during one of the soybean-related Barcelona asthma epidemics were evaluated 2 years after the last epidemic for: (1) sensitivity to prick test with glycerinated and non-glycerinated extracts, both fresh and stored at 4 degrees C, - 20 degrees C and - 70 degrees C for 30 days, and (2) specific IgE to a fresh hull extract. All extracts were also studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: No differences in positivities were detected for prick tests performed the same day of preparation between non-glycerinated (5/22; 22.7%) and glycerinated (4/22; 18.2%) extracts. After 30 days storage, positivities of prick test with non-glycerinated extracts stored at 4 degrees C did not significantly differ from the results obtained on the first day. However, prick tests performed with glycerinated extracts stored for 30 days showed increased positivity for the extracts stored at 4 degrees C (7/22; 31%) (P < 0.05) as well as for those stored at - 20 degrees C and - 70 degrees C (9/22; 40.9%) (P < 0.001). This latter percentage is similar to that of specific immunoglobulin E detected by radioimmunoassay (10/22; 45. 4%). Also, for the glycerinated extracts only, the quantitative response showed an increase in mean diameter of the induration at 30 days at the three temperatures, even though owing to the great standard deviation and limited number of patients, it was only significant for the extract stored at - 20 degrees C (P < 0.01). SDS-PAGE gel densitometry at 30 days demonstrated a loss of protein in the bands with a molecular weight higher than 66.2 kDa for non-glycerinated extracts stored at 4 degrees C. CONCLUSIONS: The results of this study confirm previous observations with soybean hull allergen extracts and indicate that (1) storage of soybean hull extracts with glycerine addition for 30 days at the concentration studied (1:100 w/v) determines a significant increase in the positivity of skin tests, and (2) glycerine addition stabilizes soybean hull extracts at any of the temperatures studied.