In silico epitope prediction, expression and functional analysis of Per a 10 allergen from the American cockroach

Tong,Xunliang; Guo,Miao; Jin,Min; Chen,Hao; Li,Yanming; Wei,Ji-Fu

doi:10.3892/ijmm.2016.2790

In silico epitope prediction, expression and functional analysis of Per a 10 allergen from the American cockroach

Authors:
- Xunliang Tong
- Miao Guo
- Min Jin
- Hao Chen
- Yanming Li
- Ji-Fu Wei
View Affiliations

Affiliations: Department of Geriatrics, Beijing Hospital, National Center of Gerontology, Beijing 100730, P.R. China, Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China, Department of Respiratory and Critical Care Medicine, Beijing Hospital, National Clinical Research Center for Respiratory Disease, Beijing 100730, P.R. China
Published online on: October 26, 2016 https://doi.org/10.3892/ijmm.2016.2790
Pages: 1806-1814
Copyright: © Tong et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cockroach (CR) allergies caused by the American cockroach hyave been recognized to be repsonsible for IgE-mediated type I hypersensitivity worldwide. Per a 10 is one of the recognized main allergens of the American CR. In a previous study, we examined another American CR allergen, Per a 9 in patients with CR allergies and examined epitope sequences in this allergen. In the present study, we aimed to examine epitope sequences in the Per a 10 allergen. for this purpose, the Per a 10 gene was cloned and expressed in Escherichia coli (E. coli) systems. Our results revealed that 9 out of 16 (56.3%) sera from patients with American CR allergies reacted to Per a10, as assessed by ELISA, confirming that Per a 10 is a major allergen of the American CR. Our results also revealed that the expression of CD63 and CCR3 on passively sensitized basophils (obtained sera of patients with American CR allergies) was increased by approximately 2.3-fold, indicating that recombinant Per a 10 is functionally active. In addition, 3 immunoinformatics tools, namely the DNAStar Protean system, the Bioinformatics Predicted Antigenic Peptides (BPAP) system and the BepiPred 1.0 server were used to predict the peptides and the results revealed 8 peptides (2-12, 55-67, 98-120, 125-133, 149-160, 170-182, 201-208 and 223-227) as potential B cell epitopes of the Per a 10 allergen. Moreover, Per a 10 was predicted to have 3 T cell epitope sequences, namely 83-92, 139-147 and 162-170. The findings of our study on the CR allergen may prove to be useful in the development of peptide-based vaccine for the prevention and/or treatment of CR allergies.

View Figures

View References

1	Bernton HS and Brown H: Insect allergy preliminary studies of the cockroach. J Allergy. 35:506–513. 1964. View Article : Google Scholar : PubMed/NCBI
2	Arruda LK, Vailes LD, Ferriani VPL, Santos AB, Pomés A and Chapman MD: Cockroach allergens and asthma. J Allergy Clin Immunol. 107:419–428. 2001. View Article : Google Scholar : PubMed/NCBI
3	Sun BQ, Lai XX, Gjesing B, Spangfort MD and Zhong NS: Prevalence of sensitivity to cockroach allergens and IgE cross-reactivity between cockroach and house dust mite allergens in Chinese patients with allergic rhinitis and asthma. Chin Med J (Engl). 123:3540–3544. 2010.
4	Thangam Sudha V, Arora N, Sridhara S, Gaur SN and Singh BP: Biopotency and identification of allergenic proteins in Periplaneta americana extract for clinical applications. Biologicals. 35:131–137. 2007. View Article : Google Scholar
5	He S, Zhang Z, Zhang H, Wei J, Yang L, Yang H, Sun W, Zeng X and Yang P: Analysis of properties and proinflammatory functions of cockroach allergens Per a 1.01s. Scand J Immunol. 74:288–295. 2011. View Article : Google Scholar : PubMed/NCBI
6	Wu HQ, Liu ZG, Ran PX, Zhou ZW and Gao B: Expression, purification, and immunological characterization of Cr PI. Protein Pept Lett. 14:881–885. 2007. View Article : Google Scholar : PubMed/NCBI
7	Mindykowski B, Jaenicke E, Tenzer S, Cirak S, Schweikardt T, Schild H and Decker H: Cockroach allergens Per a 3 are oligomers. Dev Comp Immunol. 34:722–733. 2010. View Article : Google Scholar : PubMed/NCBI
8	Tan YW, Chan SL, Ong TC, Yit Y, Tiong YS, Chew FT, Sivaraman J and Mok YK: Structures of two major allergens, Bla g 4 and Per a 4, from cockroaches and their IgE binding epitopes. J Biol Chem. 284:3148–3157. 2009. View Article : Google Scholar
9	Wei JF, Yang H, Li D, Gao P and He S: Preparation and identification of Per a 5 as a novel American cockroach allergen. Mediators Inflamm. 2014:5914682014. View Article : Google Scholar : PubMed/NCBI
10	Chen H, Yang HW, Wei JF and Tao AL: In silico prediction of the T-cell and IgE-binding epitopes of Per a 6 and Bla g 6 allergens in cockroaches. Mol Med Rep. 10:2130–2136. 2014.PubMed/NCBI
11	Yang H, Kong X, Wei J, Liu C, Song W, Zhang W, Wei W and He S: Cockroach allergen Per a 7 down-regulates expression of Toll-like receptor 9 and IL-12 release from P815 cells through PI3K and MAPK signaling pathways. Cell Physiol Biochem. 29:561–570. 2012. View Article : Google Scholar : PubMed/NCBI
12	Sookrung N, Chaicumpa W, Tungtrongchitr A, Vichyanond P, Bunnag C, Ramasoota P, Tongtawe P, Sakolvaree Y and Tapchaisri P: Periplaneta americana arginine kinase as a major cockroach allergen among Thai patients with major cockroach allergies. Environ Health Perspect. 114:875–880. 2006. View Article : Google Scholar : PubMed/NCBI
13	Yang H, Chen H, Jin M, Xie H, He S and Wei JF: Molecular cloning, expression, IgE binding activities and in silico epitope prediction of Per a 9 allergens of the American cockroach. Int J Mol Med. 38:1795–1805. 2016.
14	Sudha VT, Arora N, Gaur SN, Pasha S and Singh BP: Identification of a serine protease as a major allergen (Per a 10) of Periplaneta americana. Allergy. 63:768–776. 2008. View Article : Google Scholar : PubMed/NCBI
15	Fang Y, Long C, Bai X, Liu W, Rong M, Lai R and An S: Two new types of allergens from the cockroach, Periplaneta americana. Allergy. 70:1674–1678. 2015. View Article : Google Scholar : PubMed/NCBI
16	Goel C, Kalra N, Dwarakanath BS, Gaur SN and Arora N: Per a 10 protease activity modulates CD40 expression on dendritic cell surface by nuclear factor-kappaB pathway. Clin Exp Immunol. 180:341–351. 2015. View Article : Google Scholar :
17	Goel C, Govindaraj D, Singh BP, Farooque A, Kalra N and Arora N: Serine protease Per a 10 from Periplaneta americana bias dendritic cells towards type 2 by upregulating CD86 and low IL-12 secretions. Clin Exp Allergy. 42:412–422. 2012. View Article : Google Scholar : PubMed/NCBI
18	He W, Jimenez F, Martinez H, Harper NL, Manoharan MS, Carrillo A, Ingale P, Liu YG, Ahuja SS, Clark RA, et al: Cockroach sensitization mitigates allergic rhinoconjunctivitis symptom severity in patients allergic to house dust mites and pollen. J Allergy Clin Immunol. 136:658–666. 2015. View Article : Google Scholar : PubMed/NCBI
19	Bassirpour G and Zoratti E: Cockroach allergy and allergen-specific immunotherapy in asthma: Potential and pitfalls. Curr Opin Allergy Clin Immunol. 14:535–541. 2014. View Article : Google Scholar : PubMed/NCBI
20	Nielsen M, Lund O, Buus S and Lundegaard C: MHC class II epitope predictive algorithms. Immunology. 130:319–328. 2010. View Article : Google Scholar : PubMed/NCBI
21	Sookrung N, Khetsuphan T, Chaisri U, Indrawattana N, Reamtong O, Chaicumpa W and Tungtrongchitr A: Specific B-cell epitope of Per a 1: A major allergen of american cockroach (Periplaneta americana) and anatomical localization. Allergy Asthma Immunol Res. 6:325–332. 2014. View Article : Google Scholar : PubMed/NCBI
22	Lee MF, Chang CW, Song PP, Hwang GY, Lin SJ and Chen YH: IgE-binding epitope mapping and tissue localization of the major American cockroach allergen Per a 2. Allergy Asthma Immunol Res. 7:376–383. 2015. View Article : Google Scholar : PubMed/NCBI
23	Wu CH, Lee MF and Tseng CY: IgE-binding epitopes of the American cockroach Per a 3 allergen. Allergy. 58:986–992. 2003. View Article : Google Scholar : PubMed/NCBI
24	An S, Chen L, Wei JF, Yang X, Ma D, Xu X, Xu X, He S, Lu J and Lai R: Purification and characterization of two new allergens from the venom of Vespa magnifica. PLoS One. 7:e319202012. View Article : Google Scholar : PubMed/NCBI
25	An S, Ma D, Wei JF, Yang X, Yang HW, Yang H, Xu X, He S and Lai R: A novel allergen Tab y 1 with inhibitory activity of platelet aggregation from salivary glands of horseflies. Allergy. 66:1420–1427. 2011. View Article : Google Scholar : PubMed/NCBI
26	Sanz ML, Gamboa PM, Antépara I, Uasuf C, Vila L, Garcia-Avilés C, Chazot M and De Weck AL: Flow cytometric basophil activation test by detection of CD63 expression in patients with immediate-type reactions to betalactam antibiotics. Clin Exp Allergy. 32:277–286. 2002. View Article : Google Scholar : PubMed/NCBI
27	Sainte-Laudy J, Vallon C and Guérin JC: Analysis of membrane expression of the CD63 human basophil activation marker. Applications to allergologic diagnosis. Allerg Immunol (Paris). 26:211–214. 1994.In French.
28	Yang L, Luo Y and Wei J: Integrative genomic analyses on Ikaros and its expression related to solid cancer prognosis. Oncol Rep. 24:571–577. 2010. View Article : Google Scholar : PubMed/NCBI
29	Yang L, Luo Y, Wei J and He S: Integrative genomic analyses on IL28RA, the common receptor of interferon-λ1, -λ2 and -λ3. Int J Mol Med. 25:807–812. 2010.PubMed/NCBI
30	Yang L, Wei J and He S: Integrative genomic analyses on interferon-λs and their roles in cancer prediction. Int J Mol Med. 25:299–304. 2010.PubMed/NCBI
31	Wang M, Wei X, Shi L, Chen B, Zhao G and Yang H: Integrative genomic analyses of the histamine H1 receptor and its role in cancer prediction. Int J Mol Med. 33:1019–1026. 2014.PubMed/NCBI
32	Ding Z, Yang HW, Xia TS, Wang B and Ding Q: Integrative genomic analyses of the RNA-binding protein, RNPC1, and its potential role in cancer prediction. Int J Mol Med. 36:473–484. 2015.PubMed/NCBI
33	Li X, Yang HW, Chen H, Wu J, Liu Y and Wei JF: In silico prediction of T and B cell epitopes of Der f 25 in Dermatophagoides farinae. Int J Genomics. 2014:4839052014. View Article : Google Scholar : PubMed/NCBI
34	McGuffin LJ, Bryson K and Jones DT: The PSIPRED protein structure prediction server. Bioinformatics. 16:404–405. 2000. View Article : Google Scholar : PubMed/NCBI
35	Petersen B, Petersen TN, Andersen P, Nielsen M and Lundegaard C: A generic method for assignment of reliability scores applied to solvent accessibility predictions. BMC Struct Biol. 9:512009. View Article : Google Scholar : PubMed/NCBI
36	Laskowski RA, MacArthur MW and Thornton JM: Validation of protein models derived from experiment. Curr Opin Struct Biol. 8:631–639. 1998. View Article : Google Scholar : PubMed/NCBI
37	Maganti L, Manoharan P and Ghoshal N: Probing the structure of Leishmania donovani chagasi DHFR-TS: Comparative protein modeling and protein-ligand interaction studies. J Mol Model. 16:1539–1547. 2010. View Article : Google Scholar : PubMed/NCBI
38	Burland TG: DNASTAR's Lasergene sequence analysis software. Methods Mol Biol. 132:71–91. 2000.
39	Larsen JE, Lund O and Nielsen M: Improved method for predicting linear B-cell epitopes. Immunome Res. 2(2)2006. View Article : Google Scholar : PubMed/NCBI
40	Yang X and Yu X: An introduction to epitope prediction methods and software. Rev Med Virol. 19:77–96. 2009. View Article : Google Scholar
41	Zheng LN, Lin H, Pawar R, Li ZX and Li MH: Mapping IgE binding epitopes of major shrimp (Penaeus monodon) allergen with immunoinformatics tools. Food Chem Toxicol. 49:2954–2960. 2011. View Article : Google Scholar : PubMed/NCBI
42	Karosiene E, Rasmussen M, Blicher T, Lund O, Buus S and Nielsen M: NetMHCIIpan-3.0, a common pan-specific MHC class II prediction method including all three human MHC class II isotypes, HLA-DR, HLA-DP and HLA-DQ. Immunogenetics. 65:711–724. 2013. View Article : Google Scholar : PubMed/NCBI
43	Nielsen M and Lund O: NN-align. An artificial neural network-based alignment algorithm for MHC class II peptide binding prediction. BMC Bioinformatics. 10:2962009. View Article : Google Scholar : PubMed/NCBI
44	Schmidt M and Hoffman DR: Expression systems for production of recombinant allergens. Int Arch Allergy Immunol. 128:264–270. 2002. View Article : Google Scholar : PubMed/NCBI
45	Li GF, Wang Y, Zhang ZS, Wang XJ, Ji MJ, Zhu X, Liu F, Cai XP, Wu HW and Wu GL: Identification of immunodominant Th1-type T cell epitopes from Schistosoma japonicum 28 kDa glutathione-S-transferase, a vaccine candidate. Acta Biochim Biophys Sin (Shanghai). 37:751–758. 2005. View Article : Google Scholar
46	Nair S, Kukreja N, Singh BP and Arora N: Identification of B cell epitopes of alcohol dehydrogenase allergen of Curvularia lunata. PLoS One. 6:e200202011. View Article : Google Scholar : PubMed/NCBI
47	Pomés A: Relevant B cell epitopes in allergic disease. Int Arch Allergy Immunol. 152:1–11. 2010. View Article : Google Scholar :