Wolff et al (2003) [664] and Wolff et al. (2003) [858] report symptoms from two groups of 28 and 20 patients (34 in all) with cutaneous symptoms (urticaria and/or angioedema and/or exacerbation of atopic dermatitis) in 20/34, Gastrointestinal symptoms (diarrhoea, vomiting) in 5/34, respiratory symptoms (rhinitis, bronchospasm and/or asthma) in 6/34 and anaphylaxis in 5/34 patients.

Asthma and anaphylactic shock (12 patients) Morisset et al. (2003) [613]

Cutaneous, gastrointestinal, respiratory symptoms and anaphylaxis (23 patients including some previously described). The mean age of the first allergic reaction was 11.7 months. Although the main clinical manifestation was urticaria/angioedema (n = 14, 60%), anaphylaxis was the presenting symptom in seven (30%) patients; all of them were younger than 1 year. Sixteen (70%) were found to be allergic to other foods, and other atopic diseases were identified in 18 (78%) patients. Three patients 'outgrew' their allergy within 1-2 years. (Dalal et al. 2003) [861].

Cutaneous reactions (angiodema, and urticaria) were observed in 16/20 patients, rhinitis, rhinoconjunctivitis and oral or largyngeal in 5/20 patients, gastrointestinal symptoms in 8/20 patients and respiratory symptoms (2/20) (Beyer et al. 2002 [859]).

Reactions included: anaphylaxis (6 patients), urticaria (4 patients), urticaria and angiodema (4 patients), gastrointestinal (1 patient) and urticaria and exacerbation of atopic eczema/dermatitis syndrome (1 patient) (Dalal et al. 2002) [931].

Fremont et al. (2002) [941] report data on 6 patients of whom 3 showed symptoms of sesame allergy with one case of anaphylactic shock and 2 patients with atopic dermatitis and asthma who gave a positive result on challenge with sesame. Three others showed atopic dermatitis and 2 of these also had asthma but were negative on challenge but positive with SPT and RAST.

Reactions included: angiodema and urticaria in 4/10 patients, glottis oedema in 3/10 patients, 1/10 with gastrointestinal symptoms, 1/10 asthma and 1/10 shock (Pastorello et al. 2001 [159]).

Levy and Danon (2001) [399] report ten patients with cutaneous symptoms such as rash, pruritus and urticaria, angiedema, of whom two showed respiratory distress (one with cyanosis) and one had gasterointestinal symptoms .

Pajno et al. (2000) [932] report gastrointestinal symptoms, itching, largyngeal oedema, anaphylaxis in a single patient.

Asero et al. (1999) [11] report systematic urticaria, wheezing, largyngeal oedema, vomiting and abdominal pain followed by loss of consiousness in one patient about 5 minutes after eating sesame seeds.

Stern & Wuthrich (1998) [1071] report anaphylaxis in a single patient.

Kolopp-Sarda et al (1997) [387] report a group of 7 sesame allergic patients (1 with baker's asthma, 1 showing atopic dermatitis and asthma, 2 showing angioedema, 2 showing anaphylactic shock and 1 laryngeal angioedema) and 5 with only sensitization to sesame.

Kanny et al. (1996) [770] reported 9 patients showing urticaria, gasterointestinal symptoms, allergic rhinitis, asthma and anaphylaxis following ingestion of sesame or sesame oil.

Kagi and Wuthrich (1993) [933] report anaphylaxis in a single patient.

Malish et al. (1981) [934] report anaphylaxis or angioedema and urticaria in 4 patients.

Dalal et al. (2003) [861] tested 32 patients.
Dalal et al. (2002) [931] tested 78 patients.
Fremont et al. (2002) [941] tested 6 patients
Pastorello et al. (2001) [159] tested 10 patients.
Levy and Danon (2001) [399] tested 10 patients.
Asero et al. (1999) [11] tested a single patient.
Kolopp-Sarda et al (1997) [387] tested 12 patients.
Sporik and Hill (1996) [938] tested 2789 patients.
Kanny et al. (1996) [770] tested 9 patients.

Dalal et al. (2003) [861] found 30/32 positive results of which 23 were clinically relevant.
Dalal et al. (2002) [931] found 16 positive SPT results (in three cases the SPT was negative with the commercial extract, but was positive with the extract from brown sesame seeds).
Fremont et al. (2002) [941] found 6/6 patients SPT positive of which 3 were clinically relevant.
Pastorello et al. (2001) [159] reported positive SPTs for 10/10 patients.
Levy and Danon (2001) [399] reported 10/10 positive SPTs.
Asero et al. (1999) [11] reported a 20X15 mm wheal with both fresh sesame and with an extract. Poppy seed caused a 10X10 mm wheal.
Kolopp-Sarda et al (1997) [387] reported 9/12 positive SPTs.
Sporik and Hill (1996) [938] found 531/2789 children sensitized to sesame.
Kanny et al. (1996) [770] report 4 positive skin prick tests, one 1.5 mm wheal and 4 negative with commercial extract. All 6 patients tested with crude sesame were positive including the patients with 1.5 mm wheal and 3 who were negative to commercial extract.

Commercial sesame seed extracts were used with the CAP systems. One method for making an extract for immunoblotting is given below.

Sesame seeds were ground in a coffee grinder, followed by mortar and pestle, until a smooth paste was achieved. The paste was defatted by washing with at least 20 volumes (w/v) of cold acetone and dried overnight at 4°C. Dried acetone powder was stored at –80°C. Protein was extracted from the defatted, dried, powdered seed pulp by agitating with phosphate buffered saline and a protease inhibitor cocktail without EDTA (Roche) overnight at 4°C. After centrifugation at 2500g for 15 minutes at 4°C, supernatant was collected, filtered, and centrifuged at 12,000g for 3 minutes (Beyer et al. 2002 [859]).

Detection of IgE specific to sesame seed protein extract, was performed with Pharmacia CAP System using a fluorocounter, according to the fluoroimmunological method (FEIA) described by Leimgruber et al. (1991) [935] with a commercial batch of sesame seed extract (Pharmacia) (Wolff et al. 2003 [858]).

Antibodies to sesame were measured using the Phaebas CAP System (Pharmacia). Concentrations of ≥0.35 kUA/l were considered positive (Dalal et al. 2003 [861]).

Dalal et al. (2003) [861] tested sera from 14 patients.
Wolff et al. (2003) [858] tested sera from 28 patients diagnosed as allergic to sesame.
Beyer et al. (2002) [859] tested sera from 20 patients.
Fremont et al. (2002) [941] used sera from 6 sensitized patients, 3 with clinical symptoms.
Pastorello et al. (2001) [159] tested sera from 10 patients.
Asero et al. (1999) [11] tested sera from a single patient.
Kolopp-Sarda et al (1997) [387] tested sera from 10 patients.
Kanny et al. (1996) [770] tested sera from 8 patients.
Alday et al. (1996) [775] tested sera from a patient with occupational asthma rather than food allergy.

Dalal et al. 2003 [861] found 12/14 patients had specific anti-sesame IgE (0.38-24.7 kU/L). All 14 were diagnosed as cases with IgE-mediated sesame allergy.

Wolff et al. (2003) [858] found that sera from 3/28 patients were negative (< 0.35 kU/L) and 25/28 sera were positive (0.42->100 kU/L).

Beyer et al. (2002) [859] found that the 10 sera had specific anti-sesame IgE (3.83 to >100 kU/L).

Fremont et al. (2002) [941] found 15.4, 10.7 and 11.5 kU/L in patients reacting to sesame and 23.6, 7.74 and 1.03 kU/L in patients without symptoms clearly due to sesame.

Pastorello et al. (2001) [159] found that the 10 sera had specific anti-sesame IgE (0.8 to >100 kU/L).

Pajno et al. (2000) [932] reported that the total serum IgE was 22.3 kU/l but no specific anti-sesame IgE was detected.

Asero et al. (1999) [11] report 29 kU/L sesame specific IgE by RAST.

Kanny et al. (1996) [770] report that only 2/8 patients showed specific IgE to sesame with 13.2 kU/l and >100 kU/l. The total serum IgE was 14 - 1485 kU/l for the 5 sera tested.

Malish et al. (1981) [934]- Four patients with histories of systemic anaphylaxis or angioedema and urticaria after ingestion of sesame seed or sesame oil-containing products were studied for anti-sesame seed extract IgE by the radioallergosorbent test (RAST). Three of four were found to be positive. RAST inhibition after density-gradient ultracentrifugation of sesame seed extract indicates that the most active allergens appear to have molecular weights between 8,000 to 62,000 daltons

Wolff et al. (2003) [858] used 1D SDS–PAGE with a 6% stacking gel and 18% separation gel. Samples were heated to 100 °C for 10 min with SDS and beta-mercaptoethanol.

Beyer et al. (2002) [859] used 1D and 2D separation. Samples for 2D separation were focussed on immobilized pH gradient strips with tributylphosphine and then modified by iodoacetamide before SDS-PAGE.

Fremont et al. (2002) [941] used both 1D and 2D separation. 1D SDS–PAGE used either 4-20% gradient or 12% separation gels. Samples were heated to 100 °C for 5 min with SDS and beta-mercaptoethanol. Samples for 2D separation were focussed reduced by beta-mercaptoethanol with 3/10 ampholyte before SDS-PAGE on a 12% gel.

Pastorello et al. (2001) [159] used 1D SDS–PAGE with a 6% stacking gel and 7.5-20% separation gel. Samples were heated to 100 °C for 5 min with SDS and beta-mercaptoethanol.

Kolopp-Sarda et al (1997) [387] used 1D SDS–PAGE with a 10-15% gradient separation gel. Samples were reduced with beta-mercaptoethanol and SDS.

Wolff et al. (2003) [858] electrophoretically transferred proteins to a nitrocellulose membrane. After blocking non-specific sites, the nitrocellulose membrane was washed, cut into strips and separately incubated overnight at 4 °C with each patient's serum (diluted 1:100). The membrane was then washed and incubated with anti-human IgE peroxidase conjugate (Sigma). IgE binding was located by incubation in SuperSignal West Pico chemiluminescent substrate working solution (Pierce, U.S.A.).

Beyer et al. (2002) [859] transfered proteins to Immobilon-P membrane (Millipore) and then stained with Coomassie blue. 20 individual sera were added, diluted from 1:4 to 1:10. Phosphatase-labeled goat anti-human IgE was added and immunoblots were developed with 5-bromo-4-chloro-indolyl-phosphatase/nitroblue tetrazolium.

Pastorello et al. (2001) [159] electrophoretically transferred proteins to a nitrocellulose membrane (Amersham Pharmacia Biotech). After blocking unoccupied protein binding sites, the nitrocellulose was then cut into strips, which were incubated overnight with each patient’s serum diluted 1:4. To reveal IgE binding to allergens, each strip was incubated for 6 h with ¹²⁵I-labeled anti-human IgE antiserum (Pharmacia) diluted 1:5, followed by washing and exposure to X-ray film at −70°C for 5 days.

Kolopp-Sarda et al (1997) [387] electrophoretically transferred proteins to a nylon membrane. The membrane was incubated for 30 minutes with the ten sera (diluted 1:100). The membrane was then washed, cut into 3 strips and incubated with goat anti-human IgG, IgA or IgE peroxidase conjugate (Dako). Binding was revealed by diaminobenzidine containing buffer.

Wolff et al. (2003) [858] used sera from 24 of the 28 subjects with sesame-specific IgE. A 14 kDa protein belonging to the 2S albumin family (Ses i 2) was recognised by 22 of the 24 sera used.

Beyer et al. (2002) [859] detected allergens of 7, 34, 45, and 78 kd by 2D-immunoblotting that were recognized by 30%, 60%, 75%, and 50% of the patients respectively. However, the 78 kDa protein only bound IgE weakly. The proteins were partially sequenced and identified as a 2S albumin, a seed maturation protein, a 7S globulin, and a homologue of an embryonic abundant protein from soybean (the 2S and 7S were named Ses i 2 and Ses i 3).

Fremont et al. (2002) [941] report that 10 of 15 1D bands were recognized by anti-sesame IgE and that the results from all 6 patients, 3 with symptoms and 3 with positive RAST and SPT but without symptoms, were similar. Bands from 12.5-57.5 kDa were recognised with the strongest bands at 12, 22 and 23.5 kda. The bands at 32-36 kDa appeared weaker in the 1D gels. However, the 2D-gels showed 22/35 proteins binding IgE with the strongest binding between pH 5.5 and 6.0 and 32-36 kDa.

Pastorello et al. (2001) [159] reported IgE binding to proteins of 9 kDa (10/10, 100%), 30 kDa (8/10, 80%), 14.4 kDa (7/10, 70%), 38.7 kDa and 43 kDa (6/10, 60%), 18 kDa and 53 kDa (5/10, 50%). Partial sequence of the 9 kDa allergen showed that it was a 2S albumin and this was named Ses i 1.

Asero et al. (1999) [11] report intense IgE binding to sesame proteins at 10, 15-20 and 30-67 kDa. Less intense bands were present at 12 and 25 kDa. Poppy seed extract bound IgE at 12 kDa. Cross-inhibition showed complete cross-reactivity of the 10-12 kDa band.

Kolopp-Sarda et al (1997) [387] report the detection of anti-sesame IgG, IgA and IgE. 1/12 patients had IgE to a 14 kD, 6/12 to a 25 kD and 2/12 to a 30 kD allergen. IgG and IgA antibodies recognized more bands. In all 8/10 samples tested for IgE recognized sesame.

Alday et al. (1996) [775] reported IgE binding to a 14 kDa allergen, with weaker binding if the allergen was reduced.

Morisset et al. (2003) [613] used stewed apples or mashed potatoes as placebo and dried biscuit powder was included to simulate crushed sesame seeds. The doses of crushed sesame seed were gradually increased from 5 to 5000 mg for solid food. Doses of sesame oil were from 1 to 30 mL.

Dalal et al. (2003) [861] - Oral challenge was used only in cases where the required combination of the strict clinical criteria together with positive SPT was not diagnostic. In two cases, an open oral challenge with incremental doses of Halva (a sesame-based confection) was performed at 20-min intervals, up to a total dose of 20 g. This was done in a hospital set-up unit, which is fully equipped for cases of anaphylaxis. The patients were observed closely during the challenge and 2 h afterwards.

Fremont et al. (2002) [941] used crushed sesame hidden in mashed potatoes. Placebo potatoes contained crushed crumb.

Pajno et al. (2000) [932] A double blind, placebo controlled food challenge (DBPCFC) was carried out in the outpatient clinic with full facilities for resuscitation. The challenge was performed with sesame seeds (5 g) masked in bread. The placebo bread was baked with hazelnut (5 g).

Kanny et al. (1996) [770] used sesame seed flour mixed in apple compote or puree. Rice flour was used for the placebo. Doses were 5 mg, 10 mg., 50 mg., 200 mg. in 30 g. of apple (or potato) on the first day. If no adverse reaction had occurred, doses of 0.5 g, 1.5 g and 5 g were given and 10 g. in 3 cases on day 2.

Morisset et al. (2003) [613] and Pajno et al. (2000) [932] were blind. Dalal et al. (2003) was open. Kanny et al. (1996) [770] were double or single blind.

The lowest reactive threshold was observed at 30 mg of sesame seed. A cumulative reactive dose inferior or equal to 65 mg of solid food characterized 8% of sesame allergy (Morisset et al., 2003 [613]).

Positive reactions were observed at 100 mg. (and 3 mls sesame oil in the same patient), 3 patients at 7 g. (one tested but negative for sesame oil) and 3 positive at 10 g.

Morisset et al. (2003) [613]-Haemodynamic modifications were observed in 8% of the oral challenges (OCs) to sesame. Respiratory symptoms were observed in 42% of sesame allergies.
Dalal et al. (2003) [861]- The main clinical manifestation was urticaria/angiedema , and anaphylaxis was the presenting symptom in seven patients; all of them were younger than 1 year.
Fremont et al. (2002) [941] report that 2/3 patients gave positive DBPCFC.
Pajno et al. (2000) [932]- About 15 min after the patient ate a small amount of sesame bread, generalized flushing and hives, abdominal pain, cough, and dyspnea appeared. She was promptly treated with subcutaneous epinephrine, intravenous corticosteroids, and intramuscular antihistamine, and the symptoms disappeared within 1 h. At the onset of symptoms, the FEV1 fell to 61% of the predicted value and returned to normal values after inhaled salbutamol and the above mentioned therapy. No increase in serum tryptase was observed (12.4 µg/ml, normal range 4.8-13.5 µg/ml).
Kanny et al. (1996) [770] report urticaria and itching with 100 mg. of sesame and urticaria with sesame oil; the responses at 7 g were urticaria, urticaria with itching and abdominal pain followed by conjunctivitis and eczema in 3 patients; the responses at 10 g were skin rash and asthma, abdominal pain followed by malaise and dyspnea, and anaphylactic shock in 3 patients. There was a negative challenge at 7 g.