Oliveira Brito Patrícia Kellen Martins, Gonçalves Thiago Eleutério, Fernandes Fabrício Freitas, Miguel Camila Botelho, Rodrigues Wellington Francisco, Lazo Chica Javier Emílio, Roque-Barreira Maria Cristina, da Silva Thiago Aparecido
Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
Curso de Pós-Graduação em Ciências da Saúde da Universidade Federal do Triângulo Mineiro, Uberaba, MG, Brazil.
PLoS One. 2017 Oct 30;12(10):e0187151. doi: 10.1371/journal.pone.0187151. eCollection 2017.
Toll-like receptors (TLR) contain N-glycans, which are important glycotargets for plant lectins, to induce immunomodulation. The lectin ArtinM obtained from Artocarpus heterophyllus interacts with TLR2 N-glycans to stimulate IL-12 production by antigen-presenting cells and to drive the immune response toward the Th1 axis, conferring resistance against intracellular pathogens. This immunomodulatory effect was demonstrated by subcutaneously injecting (s.c.) ArtinM (0.5 μg) in infected mice. In this study, we evaluated the systemic implications of ArtinM administration in naïve BALB/c mice. The mice were s.c. injected twice (7 days interval) with ArtinM (0.5, 1.0, 2.5, or 5.0 μg), LPS (positive control), or PBS (negative control) and euthanized after three days. None of the ArtinM-injected mice exhibited change in body weight, whereas the relative mass of the heart and lungs diminished in mice injected with the highest ArtinM dose (5.0 μg). Few and discrete inflammatory foci were detected in the heart, lung, and liver of mice receiving ArtinM at doses ≥2.5 μg. Moreover, the highest dose of ArtinM was associated with increased serum levels of creatine kinase MB isoenzyme (CK-MB) and globulins as well as an augmented presence of neutrophils in the heart and lung. IL-12, IFN-γ, TNF-α, and IL-10 measurements in the liver, kidney, spleen, heart, and lung homogenates revealed decreased IL-10 level in the heart and lung of mice injected with 5.0 μg ArtinM. We also found an augmented frequency of T helper and B cells in the spleen of all ArtinM-injected naïve mice, whereas the relative expressions of T-bet, GATA-3, and ROR-γt were similar to those in PBS-injected animals. Our study demonstrates that s.c. injection of high doses of ArtinM in naïve mice promotes mild inflammatory lesions and that a low immunomodulatory dose is innocuous to naïve mice.
Toll样受体(TLR)含有N-聚糖,这是植物凝集素诱导免疫调节的重要糖靶标。从面包果中获得的凝集素ArtinM与TLR2 N-聚糖相互作用,刺激抗原呈递细胞产生白细胞介素-12(IL-12),并推动免疫反应向Th1轴发展,赋予对细胞内病原体的抗性。通过给感染小鼠皮下注射(s.c.)ArtinM(0.5μg)证明了这种免疫调节作用。在本研究中,我们评估了给未感染的BALB/c小鼠注射ArtinM的全身影响。小鼠皮下注射两次(间隔7天)ArtinM(0.5、1.0、2.5或5.0μg)、脂多糖(LPS,阳性对照)或磷酸盐缓冲液(PBS,阴性对照),三天后安乐死。注射ArtinM的小鼠体重均未出现变化,而注射最高剂量ArtinM(5.0μg)的小鼠心脏和肺的相对质量减小。在接受≥2.5μg ArtinM的小鼠的心脏、肺和肝脏中检测到少量且离散的炎症病灶。此外,最高剂量的ArtinM与血清肌酸激酶MB同工酶(CK-MB)和球蛋白水平升高以及心脏和肺中嗜中性粒细胞数量增加有关。对肝脏、肾脏、脾脏、心脏和肺匀浆中IL-12、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)和IL-10的检测显示,注射5.0μg ArtinM的小鼠心脏和肺中的IL-10水平降低。我们还发现,所有注射ArtinM的未感染小鼠脾脏中辅助性T细胞和B细胞的频率增加,而T-bet、GATA-3和ROR-γt的相对表达与注射PBS的动物相似。我们的研究表明,给未感染小鼠皮下注射高剂量的ArtinM会引发轻度炎症损伤,而低免疫调节剂量对未感染小鼠无害。