Superior biologic activity of the recombinant bee venom allergen hyaluronidase expressed in baculovirus-infected insect cells as compared with Escherichia coli☆☆☆★★★

Soldatova, Lyudmila; Crameri, Reto; Gmachl, Michael; Kemeny, David M.; Schmidt, Margit; Weber, M; Muêller, U.

doi:10.1016/s0091-6749(98)70179-4

Cited by 92 publications

(82 citation statements)

References 29 publications

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“…Melittin (Api m 4), a basic peptide consisting of 26 aas, is considered a minor allergen (8). Phospholipase A 2 and hyaluronidase have been cloned and expressed in bacteria and baculovirus-infected insect cells (9)(10)(11)(12), and their structures have been elucidated by x-ray crystallography (13,14). Acid phosphatase (15,16) was cloned and recombinantly expressed in insect cells very recently (17).…”

mentioning

confidence: 99%

Identification, Recombinant Expression, and Characterization of the 100 kDa High Molecular Weight Hymenoptera Venom Allergens Api m 5 and Ves v 3

Blank

Seismann

Bockisch

et al. 2010

The Journal of Immunology

113

107

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mentioning

confidence: 99%

Identification, Recombinant Expression, and Characterization of the 100 kDa High Molecular Weight Hymenoptera Venom Allergens Api m 5 and Ves v 3

Blank

Seismann

Bockisch

et al. 2010

The Journal of Immunology

113

107

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“…Numerous experiments were conducted with insect cells. For instance, human β-interferon and O-glycan core 2β-1,6-N-acetylglucosaminyltransferase (C2GnT) with high levels of activity have been successfully expressed in Sf-9 cells (Smith et al, 1983;Toki et al, 1997), and Tn cells showed high glycosylation potential for the recombinant insect glycoprotein, BvHya from the European honeybee (Soldatova et al, 1998). However, its capability to produce human-like complex type N-glycans has been a matter of controversy for many years.…”

Section: Discussionmentioning

confidence: 99%

“…The similar phenomenon was found for another recombinant bee venom allergen, hyaluronidase (BvHya) from the European honeybee. The biological activity of the BvHya expressed in E. coli was only 20%-30% of that of the BvHya expressed in the baculovirusinfected insect cells (Soldatova et al, 1998).…”

Section: Introductionmentioning

confidence: 93%

Expression of a bee venom phospholipase A2 from Apis cerana cerana in the baculovirus-insect cell

Shen

Ding

Zhang

et al. 2010

J. Zhejiang Univ. Sci. B

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Abstract:Bee venom phospholipase A 2 (BvPLA 2 ) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids. In this work, a new BvPLA 2 (AccPLA 2 ) gene from the Chinese honeybee (Apis cerana cerana) venom glands was inserted into bacmid to construct a recombinant transfer vector. Tn-5B-4 (Tn) cells were transfected with the recombinant bacmid DNA for expression. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed a double band with molecular weights of 16 and 18 kDa. Products of hexahistidine AccPLA 2 fusion protein accumulated up to 5.32% of the total cellular proteins. The AccPLA 2 fusion protein was cross reactive with the anti-AmPLA 2 (BvPLA 2 of the European honeybee, Apis mellifera) polyclonal serum. The reaction resulted in a double glycosylation band, which agrees with the band generated by the native AmPLA 2 in Western blot analysis. The PLA 2 activity of the total extracted cellular protein in the hydrolyzing egg yolk is about 3.16 μmol/(min·mg). In summary, the recombinant AccPLA 2 protein, a native BvPLA 2 -like structure with corresponding biological activities, can be glycosylated in Tn cells. These findings provided fundamental knowledge for potential genetic engineering to produce AccPLA 2 in the pharmaceutical industry.

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“…Accordingly, the vaccine should be formulated in such a way that it does not carry any risk of side effects due to IgE cross-linking on mast cells and basophils. The Api m (1/2/3) chimeric protein is comprised of fragments of major BV allergens Api m 1, Api m 2 and Api m 3 [4,5]. The three-dimensional structures of BV allergens Api m 1 and Api m 2 are stabilized by intra-chain cysteine disulfide bridges in their native forms [12,14].…”

Section: Discussionmentioning

confidence: 99%

“…Phospholipase A 2 (Api m 1) and hyaluronidase (Api m 2) represent major allergens in BV [4,5]. The cytolytic peptide melittin (Api m 3) with potent pharmacological activity is the major component in BV and is possibly responsible for many of the side effects in BV allergen immunotherapy.…”

Section: Introductionmentioning

confidence: 99%

Prevention of allergy by a recombinant multi-allergen vaccine with reduced IgE binding and preserved T cell epitopes

Karamloo¹,

Schmid‐Grendelmeier

Kussebi³

et al. 2005

Eur. J. Immunol.

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Novel approaches for the prevention of allergy are required, because of the inevitably increasing prevalence of allergic diseases during the last 30 years. Here, a recombinant chimeric protein, which comprises the whole amino acid sequences of three bee venom major allergens has been engineered and used in prevention of bee venom sensitization in mice. Phospholipase A 2 (Api m 1), hyaluronidase (Api m 2) and melittin (Api m 3) fragments with overlapping amino acids were assembled in a different order in the Api m (1/2/3) chimeric protein, which preserved entire T cell epitopes, whereas B cell epitopes of all three allergens were abrogated. Accordingly, IgE cross-linking leading to mast cell and basophil mediator release was profoundly reduced in humans. Supporting these findings, the Api m (1/2/3) induced 100 to 1000 times less type-1 skin test reactivity in allergic patients. Treatment of mice with Api m (1/2/3) led to a significant reduction of specific IgE development towards native allergen, representing a protective vaccine effect in vivo. These results demonstrate a novel prototype of a preventive allergy vaccine, which preserves the entire T cell epitope repertoire, but bypasses induction of IgE against native allergen, and side effects related to mast cell/basophil IgE FceRI cross-linking in sensitized individuals.

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Superior biologic activity of the recombinant bee venom allergen hyaluronidase expressed in baculovirus-infected insect cells as compared with Escherichia coli☆☆☆★★★

Cited by 92 publications

References 29 publications

Identification, Recombinant Expression, and Characterization of the 100 kDa High Molecular Weight Hymenoptera Venom Allergens Api m 5 and Ves v 3

Identification, Recombinant Expression, and Characterization of the 100 kDa High Molecular Weight Hymenoptera Venom Allergens Api m 5 and Ves v 3

Expression of a bee venom phospholipase A2 from Apis cerana cerana in the baculovirus-insect cell

Prevention of allergy by a recombinant multi-allergen vaccine with reduced IgE binding and preserved T cell epitopes

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