Artv4.01是一种众所周知的profilin蛋白,属于泛过敏原组,通常参与引发过敏性哮喘,多重过敏,和交叉敏感。由于其起源,它也被称为艾草。艾草粗提取物用于过敏原特异性免疫疗法(AIT)。重组Artv4.01(rArtv4.01)是否可以通过皮下免疫疗法(SCIT)产生体内免疫耐受性仍然难以捉摸。在这项研究中,研究rArtv4.01,Th2,Th1,Treg的体内免疫反应,检测Th17型相关细胞因子和免疫细胞表型,促进对潜在机制的探索。使用重组技术进行Artv4.01的表达和纯化。通过艾草花粉提取物的皮下致敏和鼻内刺激诱导过敏性哮喘雌性BALB/c小鼠。使用rArtv4.01和艾草花粉提取物进行无佐剂的SCIT2周。面对挑战后,免疫球蛋白E(IgE)的水平,细胞因子,和炎症细胞通过酶联免疫吸附试验(ELISA)和血清组织学检查进行评估,支气管肺泡灌洗液(BALF),和肺组织。随后在接受rArtv4.01和艾草花粉提取物的处理组之间比较这些参数。rArtv4.01蛋白表达,其具有高纯度(>90%)和致敏效力。与花粉提取物相比,rArtv4.01在减少白细胞(WBC)数量方面优于总有核细胞(跨国公司),BALF中单核细胞(MNs)和肺部炎症程度(1.77±0.99vs.2.31±0.80,P>0.05)。与模型组相比,只有rArtv4.01降低了血清IgE水平(1.19±0.25vs.1.61±0.17μg/ml,P=0.062),以及Th2型相关细胞因子的水平(白细胞介素-4(IL-4)(107.18±16.17vs.132.47±20.85pg/ml,P<0.05)和IL-2(19.52±1.19vs.24.02±2.14pg/ml,P<0.05))。研究表明,rArtv4.01在减少BALF中炎性细胞数量方面优于花粉提取物,肺炎,促炎细胞因子的水平,和血清IgE水平。这些发现证实了Artv4.01可能是过敏原特异性免疫治疗的潜在候选蛋白。
Art v4.01 is a well-known profilin protein belonging to the pan-allergens group and is commonly involved in triggering allergic asthma, polyallergy, and cross-sensitization. It is also referred to as Wormwood due to its origin. Crude wormwood extracts are applied for allergen-specific immunotherapy (AIT). Whether the recombinant Art v4.01 (rArt v4.01) can produce in vivo immunological tolerance by subcutaneous immunotherapy (SCIT) remains elusive. In this study, to investigate the in vivo immunological response of rArt v4.01, Th2, Th1, Treg, Th17 type-related
cytokines and phenotypes of immune cells were tested, facilitating the exploration of the underlying mechanisms. The expression and purification of Art v4.01 were carried out using recombinant techniques. Allergic asthma female BALB/c mice were induced by subcutaneous sensitization of wormwood pollen extract and intranasal challenges. SCIT without adjuvant was performed using the rArt v4.01 and wormwood pollen extract for 2 weeks. Following exposure to challenges, the levels of immunoglobulin E (IgE),
cytokines, and inflammatory cells were assessed through enzyme-linked immunosorbent assay (ELISA) and histological examination of sera, bronchoalveolar lavage fluid (BALF), and lung tissue. These parameters were subsequently compared between treatment groups receiving rArt v4.01 and wormwood pollen extract. The rArt v4.01 protein was expressed, which had a high purity (>90%) and an allergenic potency. Compared with the pollen extract, rArt v4.01 was superior in terms of reducing the number of white blood cells (WBCs), total nucleated cells (TNCs), and monocytes (MNs) in BALF and the degree of lung inflammation (1.77±0.99 vs. 2.31±0.80, P > 0.05). Compared with the model group, only rArt v4.01 reduced serum IgE level (1.19±0.25 vs. 1.61±0.17 μg/ml, P = 0.062), as well as the levels of Th2 type-related
cytokines (interleukin-4 (IL-4) (107.18±16.17 vs. 132.47±20.85 pg/ml, P < 0.05) and IL-2 (19.52±1.19 vs. 24.02±2.14 pg/ml, P < 0.05)). The study suggested that rArt v4.01 was superior to pollen extract in reducing the number of inflammatory cells in BALF, pneumonitis, levels of pro-inflammatory
cytokines, and serum IgE level. These findings confirmed that Art v4.01 could be a potential candidate protein for allergen-specific immunotherapy.