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用苏云金芽孢杆菌的Cry1Ac原毒素进行腹腔免疫可引起小鼠肠上皮中与IgG相关的Fc-γ-II和Fc-γ-III受体上调。

Intraperitoneal Immunization with Cry1Ac Protoxin from Bacillus thuringiensis Provokes Upregulation of Fc-Gamma-II/and Fc-Gamma-III Receptors Associated with IgG in the Intestinal Epithelium of Mice.

作者信息

Moreno-Fierros L, Verdín-Terán S L, García-Hernández A L

机构信息

Inmunidad en Mucosas, Unidad de Biomedicina, FES-Iztacala, Universidad Nacional Autónoma de México, Tlalnepantla, Edo. de México, México.

出版信息

Scand J Immunol. 2015 Jul;82(1):35-47. doi: 10.1111/sji.12305.

Abstract

In humans, intestinal epithelial FcRn is expressed throughout life and mediates the bidirectional transport of IgG, but in mice, it is markedly expressed in neonatal intestine. In adults, its expression is only faintly upregulated after intestinal IgG induction such as that elicited by i.p. immunization with Cry1Ac protoxin (pCry1Ac) Bacillus thuringiensis. This led us to suggest that additional Fcγ receptors (Fcγ-R) may be participating in epithelial IgG uptake. So, first we determined whether CD16/32 [an epitope shared by Fcγ-RII (CD32) and Fcγ-RIII (CD16)] was expressed in the intestinal epithelia of mice. Using confocal microscopy and flow cytometry, we detected co-localization of IgG and CD16/32 in epithelial cells, whose frequency was increased by immunization with pCry1Ac. Western blot and cross-immunoprecipitation results with anti-CD16/32 and IgG antibodies in epithelial cell extracts suggested that epithelial cells bear both Fcγ-RII and Fcγ-RIII and contained IgG associated with Fcγ-RII/RIII. Using anti-CD32 and anti-CD16 antibodies, we confirmed by Western blot, confocal microscopy and flow cytometry that both Fcγ-RII and Fcγ-RIII were expressed and suggested that upregulation occurred upon immunization in intestinal epithelia. Finally, we examined the in vitro effect of anti-CD16/32, anti-CD16 and anti-CD32 antibodies on IgG uptake and transport by intestinal epithelial cells and found that it was partially reduced. Although further studies are still required, our results suggest that Fcγ-RII and Fcγ-RIII might participate in the uptake and/or transport of IgG through the intestinal epithelia of adult mice.

摘要

在人类中,肠道上皮FcRn终生表达并介导IgG的双向运输,但在小鼠中,它在新生小鼠肠道中显著表达。在成年小鼠中,只有在肠道IgG诱导后,如通过腹腔注射苏云金芽孢杆菌Cry1Ac原毒素(pCry1Ac)引发的诱导,其表达才会轻微上调。这使我们推测可能有其他Fcγ受体(Fcγ-R)参与上皮细胞对IgG的摄取。因此,首先我们确定CD16/32[Fcγ-RII(CD32)和Fcγ-RIII(CD16)共有的一个表位]是否在小鼠肠道上皮中表达。利用共聚焦显微镜和流式细胞术,我们检测到上皮细胞中IgG与CD16/32的共定位,pCry1Ac免疫可增加其共定位频率。上皮细胞提取物的蛋白质印迹和用抗CD16/32及IgG抗体进行的交叉免疫沉淀结果表明,上皮细胞同时表达Fcγ-RII和Fcγ-RIII,且含有与Fcγ-RII/RIII相关的IgG。利用抗CD32和抗CD16抗体,我们通过蛋白质印迹、共聚焦显微镜和流式细胞术证实Fcγ-RII和Fcγ-RIII均有表达,并表明在肠道上皮免疫后会出现上调。最后,我们检测了抗CD16/32、抗CD16和抗CD32抗体对肠道上皮细胞摄取和运输IgG的体外影响,发现其摄取和运输有所部分减少。尽管仍需进一步研究,但我们的结果表明Fcγ-RII和Fcγ-RIII可能参与成年小鼠肠道上皮对IgG的摄取和/或运输。

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