鸡恒定链协助抗原肽结合MHCⅡ类分子并进入细胞内吞体

doi:10.11843/j.issn.0366-6964.2019.02.018

Abstract

Abstract:

This experiment was conducted to clear if chicken invariant chain (Ii) cytoplasmic/transmembrane domains-Ii_(Cyt/Tra), as immune carrier-is able to carry antigen peptide to associate with MHC classⅡ molecules and enter the endosomes in cell transport pathway. In this study, 4 gene segments, such as Ii, Ii_(Cyt/Tra), F2 (Newcastle disease virus F protein fragment) and Ii_(Cyt/Tra)/F2, were amplified and constructed by PCR. They were inserted into prokaryotic expression plasmid pET-32a and eukaryotic expression plasmid pmCherry-C1/N1 respectively, and 8 recombinant plasmids were constructed. Then they were transferred into the engineering bacteria (E. coli) and human renal cell (293 T) respectively. The binding of the target protein to the MHCⅡβ was detected by pull-down assay, and the co-localization in the eukaryotic cells with the MHCⅡβ and the localization in the endosomes were determined by laser confocal. The results showed that all the recombinant target protein molecules were expressed in prokaryotic or eukaryotes; Ii, Ii_(Cyt/Tra) and Ii_(Cyt/Tra)/F2 were able to bind to MHCⅡβ as well as to enter endosomes except F2. To sum up, the active domains, Ii_(Cyt/Tra), not only itself had the effect of associating MHCⅡβ, but also could bring antigenic peptides to bind MHCⅡβ and transport the endosome together, and finally enter the antigen presentation pathway. This provides a theoretical basis for further study of Ii as immune carrier.

CLC Number:

S852.4

CHEN Fangfang, ZHANG Xu, TAN Hongli, GUI Yaping, YU Weiyi. Chicken Ii Helps Antigen Peptide Bind MHC ClassⅡ Molecules and Enter Endosomes[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 398-405.

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Chicken Ii Helps Antigen Peptide Bind MHC ClassⅡ Molecules and Enter Endosomes

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