Novel B cell activation model for specific antigen loading and induction of CTL

Abstract

The invention relates to a novel B cell activation model for specific antigen loading and induction of CTL, which comprises the steps of separating human peripheral blood to obtain peripheral blood mononuclear cells, separating the B cells therein by use of CD20 magnetic beads, and culturing in an incubator; removing PBMC of the B cells; after culturing the B cells, loading the HBcAg18-27 polypeptide, wherein 50mu g / ml polypeptide is added, and the loading time is 12-18 hours; after antigen loading, washing the B cells and adding into the PBMC cell without B cells, wherein the ratio of the B cells to T cells is 1:10; continuously culturing for 4 days and taking out the cultured cells; detecting the HBV HBcAg18-27 polypeptide specific CTL cells with the pentamer technology. By adopting the B cell activation model provided by the invention, the shortcomings that the content of the peripheral blood mononuclear cells is lower than 1%, the amplification efficiency is not high, the culture time is long, the culture cost is high and the like are overcome; a reliable and simple cell experiment model is established, and the defects that the dendritic cells need long-time culture and the culture amplification efficiency is low are avoided.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to a novel specific antigen load-induced CTL activated B cell model. Background technique [0002] Prior to the present invention, past methods of inducing specific CTLs using dendritic cells (DCs) had significant problems. DC cells are highly functional APCs, which can effectively mediate T cell responses and induce the production of specific CTLs. However, as a cellular vaccine, they have two major disadvantages: 1. They account for a very small portion in peripheral blood, accounting for peripheral blood mononuclear cells. 2. At present, the common method of DC culture is to induce differentiation and expansion of monocytes into DCs. The experimental process and culture system are relatively complicated, and the efficiency of differentiation and expansion in vitro is not high, and the culture time is long and the culture The cost is high, and a reliable and simple...

AI Technical Summary

Problems solved by technology

DC cells are highly functional APCs, which can effectively mediate T cell responses and induce specific CTLs. However, as a cellular vaccine, they have two major disadvantages: 1. They account for a very small part in peripheral blood, accounting for peripheral blood mononuclear cells. 2. At present, the common method of DC culture is to induce differentiation and expansion of monocytes into DC. The experimental process and culture system are relatively complicated, and the efficiency of differentiation and expansion in vitro is not high, and the culture time is long and the culture The cost is too high to establish a reliable and simple cell experimental model for the experiment

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