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SYNTHESIS OF RECOMBINANT APIS MELLIFERA ALLERGENS API M 7 AND APIM 10 IN ESCHERICHIA COLI

Aistis Navickas1, Laima Čepulytė1, Vytautas Rudokas2, Rasa Petraitytė-Burneikienė1

1 Department of Eukaryote Gene Engineering, Institute of Biotechnology, Life Sciences Center, Vilnius University, Lithuania

2 Department of Immunology, Institute of Biotechnology, Life Sciences Center, Vilnius University, Lithuania

[email protected]

Between 30% and 40% of the world’s population suffers from allergies [1]. Allergies can cause a range of symptoms, from mild to moderate, such as itching or vomiting, to severe reactions like anaphylaxis [2]. Anaphylaxis is dangerous because it can be life-threatening if the allergic person cannot receive timely treatment, such as an epinephrine injection [3]. Various allergens, including foods, pollen, and dust mites, can trigger allergic reactions. Among these, the venoms of hymenopteran insects, such as bees and wasps, are one of the most common causes of anaphylactic shock [2, 3].

Allergy diagnostic methods are rapidly improving. This is evident by the shift towards using specific allergens in in vitro tests, which are replacing the traditional use of allergen extracts in in vivo tests, as well as expanding the diagnostic capabilities of in vitro testing. This change has several advantages for the following reasons. Firstly, allergen extracts can lead to inaccurate results due to their varying compositions. Secondly, since these extracts come from natural sources, there is a risk of cross-reactivity. Finally, conducting allergy tests in vitro, rather than in vivo, eliminates the risk of triggering allergic reactions in patients. This is highly beneficial when working with potentially dangerous allergens, such as the aforementioned hymenopteran insect venoms that can cause anaphylactic shock [4].

Recombinant Api m 7 and Api m 10 allergens, naturally found in European honey bees (Apis mellifera), were synthesized during this study. European honey bees are the most common and the most studied species of bees [5]. Up to 80% of individuals allergic to honey bee venom are sensitive specifically to the 7\(^{th}\) allergen, and between 49% to 62% are sensitized to the 10\(^{th}\) of the 12 known Api m allergens [6]. In this study, the synthesis of recombinant Api m 7 and Api m 10 allergens was induced in different E. coli strains and under different temperatures to optimize the synthesis. The research was funded by the Vilnius University Science Promotion Fund, project code MSF-JM-14/2023.

[1] R. Pawankar, G. W. Canonica, S. T. Holgate, and R. F. Lockey, White book on allergy: Executive summary 2011–2012. World Health Organization, 2011.

[2] V. Cardona et al., “World Allergy Organization Anaphylaxis Guidance 2020,” World Allergy Organization Journal, vol. 13, no. 10, p. 100472, 2020. doi: 10.1016/j.waojou.2020.100472.

[3] D. Fischer, T. K. V. Leek, A. K. Ellis, and H. Kim, “Anaphylaxis,” Allergy, Asthma, and Clinical Immunology, vol. 14, Suppl. 2, p. 54, 2018. doi: 10.1186/s13223-018-0283-4.

[4] R. Valenta et al., “Allergen Extracts for In Vivo Diagnosis and Treatment of Allergy: Is There a Future?,” J. Allergy Clin. Immunol. Pract., vol. 6, no. 6, pp. 1845–1855.e2, 2018. doi: 10.1016/j.jaip.2018.08.032.

[5] J. Gould, “Meet our prime pollinators,” Nature, vol. 521, pp. S48–S49, 2015. doi: 10.1038/521S48a.

[6] M. Burzyńska and D. Piasecka-Kwiatkowska, “A Review of Honeybee Venom Allergens and Allergenicity,” Int. J. Mol. Sci., vol. 22, no. 16, p. 8371, 2021. doi: 10.3390/ijms22168371.