Albanian j. agric. sci. 2014 (Special edition)

Agricultural University of Tirana

(Open Access)

RESEARCH ARTICLE

Food allergy in children (6 to 11 years old) in Tirana ANA KALEMAJ1*, MIRELA LIKA (ÇEKANI)1, ENVER ROSHI2 1

Department of Biology, Faculty of Natural Scinces, Tirana University, «Bulevardi Zogu i Parë”, No.7, Tirana, Albania.

2

Department of Public Health, Faculty of Public Health, University of Medicine, Tirana. "Kongresi i Manastirit", no.133,

Tirana, Albania. * Corresponding author e-mail: [email protected]

Abstract One hundred and twenty Albanian children from 6 to 11 years of age, descending from two different school, were considered as representative sample of Tirana’s children population. After we filled the questionnaires we estimated the level of the eosinophyle and immunoglobulin E (IgE) in the blood. In this article, we have summarized the state of knowledge about the healthy immune response to antigens in the diet and the basis of immune deviation that results in IgE sensitization and allergic reactivity to foods. Food allergies are increasing in prevalence at a higher rate than can be explained by genetic factors, suggesting a role for as yet unidentified environmental factors.The intestinal epithelium forms the interface between the external environment and the mucosal immune system, and emerging data suggest that the interaction between intestinal epithelial cells and mucosal dendritic cells is of a particular importance in determining the outcome of immune responses to dietary antigens. For more than 50 years, many children with food protein allergies and other forms of dietary protein intolerance have been treated successfully with protein hydrolysates with highly reduced allergenicity and, more recently, also with products based on amino acid mixtures. We have used the color methods and blood striche to diagnose the eosiniphilia presence. To determine the IgE there have been used the EIA kits. There is no correlation between the size of a skin prick test or the level of specific IgE and the clinical sensitivity in individual patients. Key words: immune response, food allergy, immunglobulin, antigen.

1. Introduction Food allergy is a set of events which are associated with an immune response, which is directed against an allergic food [1, 4]. The origin of food allergy varies, although it is believed that mostly it can be of an animal or vegetal origin [2, 6]. Epidemiological studies have shown that there is a prevalence of allergies towards vegetal proteins in adults and towards animal protein in children. The allergy in general is a modification of the body, which is provoked by the appearance of a substance which is able to act as an antigen and in a second contact with it, the individual becomes more sensitive in the case of anaphylaxis, or the person reacts in an expected way, in the case of immunity [2, 3, 7, 8]. An allergic reaction to food, to be developed it only affects a small population of cells which are genetically responsible for it [1, 5, 9, 13]. The food allergy is an immunopathological response to a food, or to a component of it [14, 15]. The reaction is mediated by specific antibodies called IgE in the general case of allergy, an allergy known as the first type [10, 11]. There are three other types of allergy: cytotoxic and 313

cytolysis of the immune complexes and cellular mediation, respectively of the second, third and fourth type [1, 5, 12]. Food allergens are numerous and belong to the class of proteins of globulins, albumin, as well as gliadins [12, 13]. Antigenicity of proteins lies in a tiny part of them, which varies from 8 to 16 amino acids. This area is called the epitope. There are epitopes T, which react with T lymphocytes and B epitopes, which are related to IgE and B lymphocytes [3, 7, 9]. IgE-mediated reactions comprise the majority of the allergic reactions to foods and are among the best known and studied ones. If the oral tolerance is not developed, this is a product of specific IgE-s for certain food. IgE are strictly associated with receptors which are located on the surface of mastocytes, cells fixed in the mucus or skin as well as in the circulating eosinophiles in the blood [2, 8]. These cells contain granules in their cytoplasm which have histamine and other mediators inside. When proteins allergens enter the body they get in contact with specific IgE which are related to mast cells or related to the eosinophiles [4, 7]. The conjugation of these allergens with IgE stimulates these cells to release their granule contents and leave them in the

Kalemaj et al

surrounding environment and then to synthesize other mediators as leukotrienes, cytokines, prostaglandins, etc. A quick answer comes a few minutes after the contact with the allergen and it is dedicated to histamine. As the basis of the reaction are the vasodilations, the widening of the blood vessels, muscle contraction and mucus secretion. A late response occurs after four to six hours after the contact with the allergen and it continues for several days [14, 16]. This response is caused by chemotactic mediators issued simultaneously with the immediate response, which help inflammatory cells such as neutrophils, eosinophiles to penetrate the tissue in order to produce an inflammatory response after several days. 2. Methods and Materials To conduct this research we have obtained biological samples, blood (serum), which are collected by the respective procedures and stored under proper conditions. This work was carried out in the period September 2013 - January 2014 and was conducted at the Clinical, Microbiological, Biochemical, Immunological, Laboratory of Tirana. During the period September 2013 - January 2014, 150 pupils aged 6 to 11 years have filled in the forms prepared and distributed by us. Individuals who resulted to be allergic to food were subjected to further analysis in order to determine the total IgE, in the ELISA serum and simultaneously it was defined level of eosinophiles in the blood [8]. To analyze the number of eosinophiles, firstly it must be performed the blood taking procedure.

100

% 100

93.8

80 60 40 6.2

20 0 Total

Alergyc

Negative

Figure 1: Distribution of the cases allergic to food and the negative cases

In cases of allergic to food, we had no significant differences regarding the gender. So we have had 30 cases of boys (54% of the total) and 26 cases of girls (46% of total) (Fig. 2). This tells us that there is no relation between the gender and the allergic cases, as it is also concluded from the literature.

46

Girls

54

Boys 40

45

50

55

Figure 2: Distribution of the cases allergic to food according to gender (%) 21

3. Results and discussion First of all in this case study we have delivered questionnaires to 900 pupils aged 6 to 11, in Tirana schools. In the questionnaire, along with the general characteristic data, there were also data on the allergies to different food noticed by the pupils or their families. After collecting the questionnaire the data were processed. It resulted in 6.2% positive (56 cases out of 900 cases in total) (Figure 1). This percentage coincides with previous studies, where the result of this food allergy cases has been about 5%.

314

10

7

9 4

5

25 20 15 10 5 0

Figure 3: Distribution of the allergy cases related to various food.

Based on chart 3, we see that in most cases children of this age are allergic to eggs. In numerous books we read about the cases of allergies which are

Food allergy in children (6 to 11 years old) in Tirana

caused mainly by the protein of egg yolk. Further foods that cause allergies are sea products such as fish or shellfish. After them come peanuts and tree nuts (various kinds of dried fruits) and in the end, there are fewer cases which are allergic from soya. The latter happens because soy-based foods are still not very usable by the Albanian population, especially by children of this age.

. 38

34

40 21

%

20

7

0 165-175 176-186 187-197 197-207 UI/ml UI/ml IgE UI/ml UI/ml

4. References

Figure 4: Distribution of the Total IgE level in positive cases.

Individuals who were allergic to various foods, serum was taken which was subjected to ELISA test to check the levels of total IgE. At intervals of IgE 165-175 UI / ml, 7% resulted positive out of the total cases, at intervals of IgE from 176 to 186 UI / ml, 21% resulted positive, at intervals of IgE from 187 to 197 UI / ml, 34% resulted positive, and in the intervals of IgE from 197 to 207 UI / ml, 38% of the cases resulted positive to food allergy. In positive cases of allergies to food, we studied as well the level of eusinophiles. Eosinophiles level has shown that in all the allergy cases there is an increase of them, indicating that high eosinophilia is one of the main parameters of allergies caused by food

(Fig 5).

1. Bruijnzeel KC, Ortolani C, Aas K, BindslevJensen C, Bjorksten B, Moneret-Vautrin D Bengtsson U, BindslevJensen C, Bjorksten B, Host. Adverse reactions to food. European academy of allergology and clinical immunology subcommittee. Allergy 1995;50:623–35. 2. Commission of the European Communities. Commission directive 96/4 EC of 16 February 1996 amending directive 91/321/EEC on infant formulae and follow-on formulae. Official Journal of the European Commission 1996;39:12–16. 3. De Jong MH, Scharp-van der Linden VTM, Aalberse RC, Oosting J, Tijssen JGP, de Groot CJ. Randomised controlled trial of brief neonatal exposure to cows’ milk on the development of atopy. Arch Dis Child 1998;79:126–30. 4. Halken S, Host A. How hypoallergenic are hypoallergenic cow’s milk based formulas? Allergy 1997;52:1175–83. 5. Halken S, Host A. Prevention of allergic disease. Exposure to food allergens and dietetic intervention. Pediatr Allergy Immunol 1996;7(suppl 9):102–7.

62% 38% 0% 20% 40% 1 level of eosinophils

Almost in all the cases of children who are allergic to various foods, levels of IgE increase, which otherwise is called allergic immunoglobulin, we notice also an increase of eosinophiles as polymorph nuclear leukocytes, which emit the contents of granules that help in the emergence of allergic signs. Certain syndromes occur in children or individuals who are allergic to different food, fruits or vegetables either. One of the first signs is edema in lips and difficulty in swallowing. Other symptoms, for almost all the positive cases are those of abdominal pain, vomiting and diarrhea. Also in the respiratory organs we have asthma or hoarseness of voice. Hence, allergic reactions to food include almost all the major organs as the digestive, respiratory, etc.

6. Host A. Cow’s milk protein allergy and intolerance in infancy. Some clinical, epidemiological and immunological aspects. Pediatr Allergy Immunol 1994;5(suppl 5):1–36.

60% 80% 2 level of eosinophils

7. Host A, Husby S, Osterballe O. A prospective study of cow’s milk allergy in exclusively breast-fed infants. Acta Paediatr Scand 1988;77:663–70.

Figure 5: Distribution of eosinophiles level in positive cases of food allergies 1 Level of eosinophils (0.6-0.7 x 10³/mm

8. Lika (Çekani) M., Bërxholi K. Reaksionet e mbindjeshmërisë. Imunologjia. 2007. 154-163.

cells3); 2 Level of eosinophils (0.8-0.9 x 103/mm3 cells) 315

Kalemaj et al

9. Lucas A, Brooke OG, Cole TJ, Morley R, Bamford JTM. Food and drug reactions, wheezing, and eczema in preterm infants. Arch Dis Child 1990;65:411–15.

13. Saarinen K, Juntunen-Backman K, Järvenpää AL. Early feeding of cow’s milk formula—a risk for cow’s milk allergy [abstract]. J Pediatr Gastroenterol Nutr 1997;24:461.

10. Lee YH. Food-processing approaches to altering allergenic potential of milk-based formula. J Pediatr 1992;121:S47

14. Strobel S. Neonatal oral tolerance. AnnNYAcad Sci 1996; 778:88–102. 15. Szepfalusi Z, Nentwich I, Gerstmayr M, et al. Prenatal allergen contact with milk proteins. Clin Exp Allergy 1997;27:28–35.

11. Moneret VD. Modifications of allergenicity linked to food technologies. Allerg Immunol (Paris) 1998;30:9–13. 12. Rigo J, Salle BL, Picaud JC, Putet G, Senterre J. Nutritional evaluation of protein hydrolysate formulas. Eur J Clin Nutr 1995;49(suppl 1):S26– 38.

316

16. Witteman AM, van Leeuwen J, van derzee J, Aalberse RC. Food allergens in house dust. Int Arch Allergy Immunol 1995;107:566–8.