Method for detecting secretory cells of peripheral blood hepatitis B surface antibody

Abstract

The invention relates to a method for detecting secretory cells of a peripheral blood hepatitis B surface antibody. According to the method disclosed by the invention, human Peripheral Blood Mononuclear Cells (PBMC) are stimulated and activated in vitro, and a pretreated Polyvinylidene Fluoride (PVDF) plate is used for detecting anti-HBs secretory cells in the PBMCs, to detect that a million of PBMCs contain one anti-HBs secretory cell. The method overcomes the detection difficulties in peripheral blood anti-HBs secretory cells, such as small quantity of the peripheral blood anti-HBs secretory cells, low detection rate, low experiment repetitiveness, complex operation, long time consumption and high difficulty, solves the problems that much peripheral blood needs to be adopted and in-vitro B cell activation methods such as CPG, CD40L and pokeweed mitogen are low in activation efficiency, and overcomes the defects of reduction of the detection rate of the anti-HBs secretory cells and the like. According to the method, R848, HBsAg and IL-2 are used for stimulating and activating peripheral blood B cells, and the HBsAg pre-coated PVDF plate is used for capturing the peripheral blood anti-HBs secretory cells, so that the blood amount for detection is reduced, and the detection sensitivity is improved.

Description

technical field [0001] The invention relates to a method for detecting special antibody-secreting cells in the technical field of biomedical engineering, in particular to a method for detecting antibody-secreting cells on the surface of hepatitis B in peripheral blood. Background technique [0002] Hepatitis B surface antibody (anti-HBs) is a sign of the body's immune protection against hepatitis B virus (HBV), and plays a very important role in the process of HBV clearance and prevention of reinfection. Anti-HBs in peripheral blood is secreted by plasma cells. After the body is stimulated by hepatitis B surface antigen (HBsAg), B cells mature and differentiate into plasma cells that secrete anti-HBs and memory B cells that do not secrete antibodies. The life span of plasma cells is short, while Memory B cells can survive for a long time without antigen stimulation, but their number is very small in peripheral blood. After the body is stimulated by HBsAg again, memory B cel...

AI Technical Summary

Problems solved by technology

[0005] (1) The number of anti-HBs secreting cells in peripheral blood is small, the detection rate is low, and the repeatability of the experiment is poor;

[0006] (2) The detection of antibody-secreting cells by the limited dilution method is cumbersome, time-consuming, and difficult, and requires a large amount of peripheral blood, making it difficult to popularize and apply;

[0007] (3) In addition to plasma cells directly secreting antibodies in peripheral blood, memory B cells need to be stimulated and activated to differentiate into antibody secreting cells. Therefore, without sufficient activation, peripheral blood anti-HBs secreting cells are extremely difficult to detect, while Existing techniques for activating and detecting peripheral blood anti-HBs secreting cells have many deficiencies, such as CPG, CD40L, pokeweed mitogen and other B cell activation methods in vitro, all have the problem of low activation efficiency and cannot fully activate B cells. Transformation into antibody-secreting cells reduces the detection rate of anti-HBs-secreting cells

Images