case_images: PMC11410408_01
case_images: (a) Redness and swelling of eyelids induced after ingesting the fried fish sausages. (b) Results of skin prick tests with the eyeshadow, the fish sausages, carmine, and carminic acid. Skin prick tests with (A) the pink eyeshadow 1% in saline (mean diameter of wheal, 4.4 mm), (B) the fish sausages 10% in saline (7.4 mm), (C) the carmine 0.1% in phosphate buffered saline (PBS), (4.9 mm), and the carminic acid 0.1% in PBS, (3.6 mm) were positive. The mean diameter, which was induced by 10 mg/ml histamine hydrochloride as positive control, was 6.6 mm, whereas the mean diameter, which was induced by saline as negative control, was 1.1 mm. The elicited response was considered positive if the largest allergen-induced wheal diameter was >=3 mm.
case_images: CRIDM2024-1057957.001.jpg
case_images: PMC11410408_01_CRIDM2024-1057957.001.jpg
case_images: fig1
case_images: Tokyo, Japan), which contained carmine, on several occasions for 3 years. Two months ago, she developed itching, wheals, and swelling on her whole body, especially her eyelids, and dyspnea immediately after ingesting fried pink fish sausages (Maruha Nichiro, Tokyo, Japan), which contained cochineal (Figures 1(a)). The skin prick test (SPT) indicated a positive result for the eyeshadows 1% in saline (mean diameter of wheal, 4.4 mm) (Figure 1(b)).
case_images: (a) SDS-PAGE of the fish sausage, the eyeshadow, the carmine, and the carminic acid. M, molecular mass standard; lane 1, eyeshadow extract; lane 2, fish sausage extract 1 mg/mL in phosphate buffered saline (PBS); lane 3, the carmine extract, 20 mug/mL in PBS; and lane 4, the carminic acid, 20 mug/mL in PBS. (b) IgE immunoblotting using the fish sausage, the eyeshadow, the carmine, and the carminic acid. M, molecular mass standard; lanes 1 and 5, the eyeshadow extract; lanes 2 and 6, the fish sausage extract; lanes 3 and 7, the carmine extract; and lanes 4 and 8, the carminic acid extract. We used the patient serum for lanes 1-4 and the control pooled serum from subjects without food allergy for lanes 5-8. The patient's IgE antibody was bound to the eyeshadow protein bands at the molecular weight of approximately 86, 114, and 130 kDa, which were the same as that of the fish sausage protein bands. In addition, IgE immunoblotting using the carmine showed the broad band at 86-130 kDa, whereas IgE immunoblotting using the carminic acid showed no band. These results indicated that the eyeshadow and the fish sausage could contain the same causative allergen as the carmine. On the other hand, IgE immunoblotting with the control pool serum shows no bands. (c) Evaluation of the allergenicity of the eyeshadow and the carmine after the chelation with EDTA. M, molecular mass standard; lanes 1-3, the eyeshadow extract; lanes 4, 6, and 7, the carmine extract, 20 mug/mL; lanes 5, 8, and 9, the carmine extract, 40 mug/mL. The eyeshadow and carmine extracts were applied to lanes 2, 6, and 8, respectively, after preincubation in the presence of the chelating agent, 5 mM ethylenediaminetetraacetic acid (EDTA). In addition, the eyeshadow and the carmine extracts were applied to lanes 3, 7, and 9, respectively, after preincubation in the presence of the chelating agent, 25 mM EDTA. The three protein bands at the molecular weight ranging 86-130 in the eyeshadow and the carmine extracts were diminished through preincubation in the presence of EDTA in a concentration-dependent manner. These results suggest that the causative allergen in the eyeshadow and the carmine lost its allergenicity through chelation with EDTA.
case_images: CRIDM2024-1057957.002.jpg
case_images: PMC11410408_01_CRIDM2024-1057957.002.jpg
case_images: fig2
case_images: In IgE-immunoblotting using the eyeshadow and fish sausage, the patient serum IgE was bound to three protein bands at approximately 86, 114, and 130 kDa (Figures 2(a) and 2(b)). In addition, IgE immunoblotting using the carmine showed a broad band at 86-130 kDa, which were consistent with those using the eyeshadow and fish sausage, whereas there is no band using CA. The results indicated that the causative allergens of the carmine contained in eyeshadow could be dechelated by EDTA, reducing its allergenicity (Figure 2(c)).