层状双氢氧化物-rFel d 1-Can f 1融合变应原对小鼠模型过敏反应的预防效果分析

doi:10.11843/j.issn.0366-6964.2024.07.032

摘要/Abstract

摘要：

旨在预防猫、犬诱导的过敏反应，本研究将猫主要变应原Fel d 1与犬主要变应原Can f 1融合原核表达，纯化制备了rFel d 1-Can f 1融合变应原。将层状双氢氧化物(layered double hydroxides，LDH)与rFel d 1-Can f 1混合，制备LDH-rFel d 1-Can f 1，皮下注射法免疫BALB/c小鼠，随后利用rFel d 1-Can f 1融合变应原诱导过敏小鼠模型，通过检测小鼠应激性过敏反应的体温变化、耳朵点刺试验、肺部组织切片HE染色、血清IgE水平等指标评价LDH-rFel d 1-Can f 1对猫犬诱发过敏反应的预防效果。研究表明，LDH与rFel d 1-Can f 1质量比为7 ∶1时，LDH可以完全吸附rFel d 1-Can f 1融合蛋白。LDH-rFel d 1-Can f 1可以有效缓解应激性过敏反应引起的小鼠体温下降，显著减少过敏导致的染料渗透面积。肺部组织切片HE染色结果显示，LDH-rFel d 1-Can f 1可以缓解过敏引起的炎症细胞浸润。此外，LDH-rFel d 1-Can f 1免疫后可以诱导小鼠产生较高水平IgG，这为开发抑制宠物猫犬诱发过敏反应的疫苗奠定基础。

关键词: 融合变应原, 层状双氢氧化物, 纳米颗粒, 宠物过敏

Abstract:

To prevent allergic reactions induced by cats and dogs, we purified rFel d 1-Can f 1 fusion allergen by fusion prokaryotic expression of cat primary allergen Fel d 1 and dog primary allergen Can f 1. We mixed Layered double hydroxides (LDH) with rFel d 1-Can f 1 to prepare LDH-rFel d 1-Can f 1 and immunized BALB/c mice by subcutaneous injection. Subsequently, we used rFel d 1-can f 1 fusion allergen to induce allergic mice models. We evaluated the preventive effect of LDH-rFel d 1-Can f 1 on allergic reactions caused by cats and dogs through anaphylaxis, ear prick test, H&E staining of lung tissue sections, and serum IgE level. The results showed that LDH could completely adsorb the fusion protein of rFel d 1-Can f 1 when the mass ratio of LDH to rFel d 1-Can f 1 was 7 ∶1. The LDH-rFel d 1-Can f 1 can effectively relieve the hypothermia caused by stress allergic reaction in mice, and significantly reduce the dye penetration area caused by allergy. H&E staining of lung tissue sections showed that LDH-rFel d 1-Can f 1 could relieve inflammatory cell invasion caused by the allergy. In addition, the LDH-rFel d 1-Can f 1 can induce mice to produce higher levels of IgG, which lays the foundation for the development of vaccines that inhibit allergic reactions induced by pet cats and dogs.

Key words: fusion allergens, layered double hydroxides (LDH), nanoparticle, pet allergies

中图分类号:

彭美琪, 魏春洁, 郑安琪, 朱亦坤, 徐琳皓, 罗承慧, 韦双双, 裴业春. 层状双氢氧化物-rFel d 1-Can f 1融合变应原对小鼠模型过敏反应的预防效果分析[J]. 畜牧兽医学报, 2024, 55(7): 3143-3154.

Meiqi PENG, Chunjie WEI, Anqi ZHENG, Yikun ZHU, Linhao XU, Chenghui LUO, Shuangshuang WEI, Yechun PEI. Preventive Effect of Layered Double Hydroxide-rFel d 1-Can f 1 Fusion Allergens on Anaphylaxis in Mouse Model[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3143-3154.

图/表 8

图 1

图 2

图 3

图 4

图 5

图 6

图 7

图 8

参考文献 38

1	CHAN S K , LEUNG D Y M .Dog and cat allergies: current state of diagnostic approaches and challenges[J].Allergy Asthma Immunol Res,2018,10(2):97-105. doi: 10.4168/aair.2018.10.2.97
2	YANG M S , LEE S P , KWON Y J , et al.Dog and cat allergies and allergen avoidance measures in korean adult pet owners who participated in a pet exhibition[J].Allergy Asthma Immunol Res,2018,10(2):155-164. doi: 10.4168/aair.2018.10.2.155
3	齐景翠, 赵玉林, 李伟亚, 等.郑州地区变应性鼻炎患儿变应原谱分析[J].临床耳鼻咽喉头颈外科杂志,2015,29(5):404-406.
	QI J C , ZHAO Y L , LI W Y , et al.Analysis of allergens spectrum in children with allergic rhinitis in Zhengzhou district[J].Journal of Clinical Otorhinolaryngology Head and Neck Surgery,2015,29(5):404-406.
4	邱前辉, 卢川, 韩虹, 等.广东经济发达与欠发达地区变应性鼻炎变应原调查及分析[J].临床耳鼻咽喉头颈外科杂志,2013,27(17):928-931.
	QIU Q H , LU C , HAN H , et al.Clinical survey and analysis of allergic rhinitis patients' allergens in Guangdong developed and developing regions[J].Journal of Clinical Otorhinolaryngology Head and Neck Surgery,2013,27(17):928-931.
5	SCHOOS A M M , NWARU B I , BORRES M P .Component-resolved diagnostics in pet allergy: Current perspectives and future directions[J].J Allergy Clin Immunol,2021,147(4):1164-1173. doi: 10.1016/j.jaci.2020.12.640
6	OHMAN J L JR , LOWELL F C , BLOCH K J .Allergens of mammalian origins. Ⅲ. Properties of a major feline allergen[J].J Immunol,1975,114(3):1138.
7	THOMS F , HAAS S , ERHART A , et al.Immunization of cats against Fel d 1 results in reduced allergic symptoms of owners[J].Viruses,2020,12(3):288. doi: 10.3390/v12030288
8	DE GROOT H , VAN SWIETEN P , VAN LEEUWEN J , et al.Monoclonal antibodies to the major feline allergen Fel d I. I. Serologic and biologic activity of affinity-purified Fel d I and of Fel d I-depleted extract[J].J Allergy Clin Immunol,1988,82(5 Pt 1):778-786.
9	SCHOU C , SVENDSEN U G , LØWENSTEIN H .Purification and characterization of the major dog allergen, Can f I[J].Clin Exp Allergy,1991,21(3):321-328. doi: 10.1111/j.1365-2222.1991.tb01663.x
10	NAKATSUJI M , SUGIURA K , SUDA K , et al.Structure-based prediction of the IgE epitopes of the major dog allergen Can f 1[J].FEBS J,2022,289(6):1668-1679. doi: 10.1111/febs.16252
11	MAYA-MANZANO J M , PUSCH G , VON ESCHENBACH C E , et al.Effect of air filtration on house dust mite, cat and dog allergens and particulate matter in homes[J].Clin Transl Allergy,2022,12(4):e12137. doi: 10.1002/clt2.12137
12	SPITZAUER S , SCHWEIGER C , ANRATHER J , et al.Characterisation of dog allergens by means of immunoblotting[J].Int Arch Allergy Immunol,1993,100(1):60-67. doi: 10.1159/000236389
13	KONRADSEN J R , FUJISAWA T , VAN HAGE M , et al.Allergy to furry animals: New insights, diagnostic approaches, and challenges[J].J Allergy Clin Immunol,2015,135(3):616-625. doi: 10.1016/j.jaci.2014.08.026
14	SAARELAINEN S , TAIVAINEN A , RYTKÖNEN-NISSINEN M , et al.Assessment of recombinant dog allergens Can f 1 and Can f 2 for the diagnosis of dog allergy[J].Clin Expl Allergy,2004,34(10):1576-1582. doi: 10.1111/j.1365-2222.2004.02071.x
15	KONIECZNY A , MORGENSTERN J P , BIZINKAUSKAS C B , et al.The major dog allergens, Can f 1 and Can f 2, are salivary lipocalin proteins: cloning and immunological characterization of the recombinant forms[J].Immunology,1997,92(4):577-586. doi: 10.1046/j.1365-2567.1997.00386.x
16	SMITH D M , COOP C A .Dog allergen immunotherapy: past, present, and future[J].Ann Allergy Asthma Immunol,2016,116(3):188-193. doi: 10.1016/j.anai.2015.12.006
17	WESTERA L , DRYLEWICZ J , DEN BRABER I , et al.Closing the gap between T-cell life span estimates from stable isotope-labeling studies in mice and humans[J].Blood,2013,122(13):2205-2212. doi: 10.1182/blood-2013-03-488411
18	YOUNES S A , PUNKOSDY G , CAUCHETEUX S , et al.Memory phenotype CD4 T cells undergoing rapid, nonburst-like, cytokine-driven proliferation can be distinguished from antigen-experienced memory cells[J].PLoS Biol,2011,9(10):e1001171. doi: 10.1371/journal.pbio.1001171
19	CHOO D K , MURALI-KRISHNA K , ANITA R , et al.Homeostatic turnover of virus-specific memory CD8 T cells occurs stochastically and is independent of CD4 T cell help[J].J Immunol,2010,185(6):3436-3444. doi: 10.4049/jimmunol.1001421
20	PARRETTA E , CASSESE G , SANTONI A , et al.Kinetics of in vivo proliferation and death of memory and naive CD8 T cells: parameter estimation based on 5-bromo-2'-deoxyuridine incorporation in spleen, lymph nodes, and bone marrow[J].J Immunol,2008,180(11):7230-7239. doi: 10.4049/jimmunol.180.11.7230
21	CHEN W Y , ZHANG B , MAHONY T , et al.Efficient and durable vaccine against intimin β of diarrheagenic E. Coli induced by clay nanoparticles[J].Small,2016,12(12):1627-1639. doi: 10.1002/smll.201503359
22	GU Z , ZUO H L , LI L , et al.Pre-coating layered double hydroxide nanoparticles with albumin to improve colloidal stability and cellular uptake[J].J Mater Chem B,2015,3(16):3331-3339. doi: 10.1039/C5TB00248F
23	CHEN W Y , ZUO H L , MAHONY T J , et al.Efficient induction of comprehensive immune responses to control pathogenic E. coli by clay nano-adjuvant with the moderate size and surface charge[J].Sci Rep,2017,7(1):13367. doi: 10.1038/s41598-017-13570-2
24	CUNHA V R R , DE SOUZA R B , DA FONSECA MARTINS A M C R P , et al.Accessing the biocompatibility of layered double hydroxide by intramuscular implantation: histological and microcirculation evaluation[J].Sci Rep,2016,6,30547. doi: 10.1038/srep30547
25	MIN J , BRAATZ R D , HAMMOND P T .Tunable staged release of therapeutics from layer-by-layer coatings with clay interlayer barrier[J].Biomaterials,2014,35(8):2507-2517. doi: 10.1016/j.biomaterials.2013.12.009
26	PEI Y C , XIAO Z P , WEI S S , et al.Studies on HBcAg-rBlo t 5-21 fusion protein vaccine that alleviates Blomia tropicalis airway inflammation[J].J Inflammat Res,2022,15,6343-6355. doi: 10.2147/JIR.S380526
27	PEI Y C , GENG S , LIU L , et al.Fel d 1-airway inflammation prevention and treatment by co-immunization vaccine via induction of CD4⁺CD25-Foxp3⁺ Treg cells[J].Hum Vaccines Immunother,2013,9(5):1019-1031. doi: 10.4161/hv.23518
28	CURIN M , WEBER M , THALHAMER T , et al.Hypoallergenic derivatives of Fel d 1 obtained by rational reassembly for allergy vaccination and tolerance induction[J].Clin Exp Allergy,2014,44(6):882-894. doi: 10.1111/cea.12294
29	ZALESKA A , EIWEGGER T , SOYER Ö , et al.Immune regulation by intralymphatic immunotherapy with modular allergen translocation MAT vaccine[J].Allergy,2014,69(9):1162-1170. doi: 10.1111/all.12461
30	VAN REE R , VAN LEEUWEN W A , BULDER I , et al.Purified natural and recombinant Fel d 1 and cat albumin in in vitro diagnostics for cat allergy[J].J Allergy Clin Immunol,1999,104(6):1223-1230. doi: 10.1016/S0091-6749(99)70017-5
31	周迎迎, 熊实秋, 刘传合.宠物变应原及相关过敏性疾病诊治的新进展[J].中华预防医学杂志,2023,57(9):1342-1347. doi: 10.3760/cma.j.cn112150-20230216-00113
	ZHOU Y Y , XIONG S Q , LIU C H .New progress in the diagnosis and treatment of pet allergens and related allergic diseases[J].Chinese Journal of Preventive Medicine,2023,57(9):1342-1347. doi: 10.3760/cma.j.cn112150-20230216-00113
32	TASANIYANANDA N , CHAISRI U , TUNGTRONGCHITR A , et al.Mouse model of cat allergic rhinitis and intranasal liposome-adjuvanted refined Fel d 1 vaccine[J].PLoS One,2016,11(3):e0150463. doi: 10.1371/journal.pone.0150463
33	DURAMAD P , HARLEY K , LIPSETT M , et al.Early environmental exposures and intracellular Th1/Th2 cytokine profiles in 24-month-old children living in an agricultural area[J].Environ Health Perspect,2006,114(12):1916-1922. doi: 10.1289/ehp.9306
34	GOVEA-ALONSO D O , GARCÍA-SOTO M J , MENDOZA-PÉREZ E S , et al.Assessing the adjuvant effect of layered double hydroxides (LDH) on BALB/c mice[J].Materials,2023,16(15):5647.
35	LI D D , XU M J , LI G T , et al.Mg/Al-LDH as a nano-adjuvant for pertussis vaccine: a evaluation compared with aluminum hydroxide adjuvant[J].Nanotechnology,2022,33(23):235102. doi: 10.1088/1361-6528/ac56f3
36	罗应, 彭美琪, 肖正泮, 等.热带无爪螨主要变应原Blo t 5和Blo t 21融合表达质粒诱导小鼠产生IgG和调节性T细胞[J].畜牧兽医学报,2022,53(4):1220-1230.
	LUO Y , PENG M Q , XIAO Z P , et al.Expression of Blomia tropicalis major allergens Blo t 5 and Blo t 21 induced the production of IgG and regulatory T cells in mice[J].Acta Veterinaria et Zootechnica Sinica,2022,53(4):1220-1230.
37	KAISER L , GRÖNLUND H , SANDALOVA T , et al.The crystal structure of the major cat allergen Fel d 1, a member of the secretoglobin family[J].J Biol Chem,2003,278(39):37730-37735. doi: 10.1074/jbc.M304740200
38	NIEMI M H , RYTKÖNEN-NISSINEN M , JÄNIS J , et al.Structural aspects of dog allergies: the crystal structure of a dog dander allergen Can f 4[J].Mol Immunol,2014,61(1):7-15. doi: 10.1016/j.molimm.2014.04.003