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Cellular immunological detection kit for evaluating curative effect of vaccine and storage method thereof

A detection kit, a technology of cellular immunity, applied in the field of kits for evaluating the efficacy of vaccines

Inactive Publication Date: 2017-05-03
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The results of clinical trials have confirmed the therapeutic effect of BG on chronic hepatitis B. However, in addition to the clinical manifestation results, there is no specific immunological evaluation index or surrogate endpoint for early and mid-term evaluation of the therapeutic effect of BG. Therefore, at the immunological level However, studies on immunological surrogate endpoints for chronic hepatitis B vaccine are still lacking

Method used

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  • Cellular immunological detection kit for evaluating curative effect of vaccine and storage method thereof
  • Cellular immunological detection kit for evaluating curative effect of vaccine and storage method thereof
  • Cellular immunological detection kit for evaluating curative effect of vaccine and storage method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Embodiment 1, the clinical sample cellular immunology evaluation of hepatitis B therapeutic vaccine B gram

[0132] 1. Materials and Reagents

[0133] 10ml EDTA anticoagulant blood collection tube (BD, catalog number: 367525), freezing tube (Corning, 430659), 15ml centrifuge tube (Corning, 430791), 12-well cell culture plate (Costar, 3513), 96-well U-bottom cell culture plate (Costar, 3799), 10ml pipette (Costar, 4488), sterile 1.5ml LEP tube, flow cytometry tube, and other consumables.

[0134] (1) Sterile phosphate buffer solution (phosphate buffer solution, PBS) was purchased from Gibco Company, the product number is 20012-027. (2) Human lymphocyte separation medium (LymphoprepTM) was purchased from Axis-Shield Company, the product number is 11114547. (3) 4% paraformaldehyde (paraformaldehyde, PFA, purchased from Sinopharm Chemical Reagent Co., Ltd.): Dissolve 8g of PFA in PBS, with a final volume of 200mL, heat and stir and add a few drops of concentrated NaOH...

Embodiment 2

[0196] Example 2: Long-term stability monitoring of Eke clinical sample cellular immunology detection kit

[0197] Materials and Reagents Reference Example 1, PBMCs are derived from chronic hepatitis B patients in the B-gram treatment group, and the detection index is mainly CD8 + Expression of T cell IFN-γ.

[0198] We monitored the stability of EK Clinical Sample Cellular Immunology Detection Kit for 1 month, 3 months, 6 months and 12 months. Store the 96-well pre-coated cell stimulation plate in the kit at -20°C, and store other reagents at 4°C.

[0199] Such as Figure 7A As shown, kits stored for 1 month, 3 months, 6 months, and 12 months compared with day 0 were tested to positively stimulate wells CD8 + The expression level of IFN-γ in T cells basically did not change, and negative stimulation wells were used as controls. Such as Figure 7B , assuming that the stability of the kit is 100% at day 0, after conversion, the stability of the kits stored for 1 month, 3...

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Abstract

The invention provides a cellular immunological detection kit for evaluating the curative effect of a vaccine and a storage method thereof. The kit comprises MHC-restricted antigen peptide. The cellular immunological detection kit for evaluating the curative effect of the vaccine can utilize a therapeutic vaccine research database and specimens in a clinic trial phase and employs flow cytometry for comprehensive detection of immune cells and cytokines secreted by the immune cells, so a cellular immunological curative effect evaluation system is established. The cellular immunological detection kit for evaluating the curative effect of the vaccine has good stability, and the stability can maintain 90% or above after for storage for one year or above.

Description

technical field [0001] The invention relates to a kit for evaluating vaccine curative effect and a storage method of the kit, in particular to a kit for evaluating vaccine curative effect through cell immunology detection and a storage method for the kit. Background technique [0002] At present, the evaluation methods of preventive vaccines are relatively simple, mainly focusing on follow-up cohorts to evaluate antibody levels and crowd protection efficacy, such as influenza vaccines, hepatitis B preventive vaccines, smallpox vaccines, etc. In the comprehensive evaluation of the effect of preventive vaccines, the antibody titer produced by the vaccine is often used as a core indicator. In the past ten years, with the strengthening of detection methods, especially the development of new cellular immunity evaluation techniques, the effectiveness of preventive vaccines The evaluation has also begun to touch on T cells, an important cell of adaptive immunity. For example, in th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N33/68
CPCG01N33/505G01N33/68G01N33/56977G01N2333/02A61K39/12C12N2730/10134A61K2039/545A61K2039/55522A61K2039/57A61P31/20G01N33/4915G01N2333/70514G01N2333/70517G01N2333/70539C12N2730/10122G01N33/5304G01N33/5761G01N33/6857
Inventor 王宾李超凡周晨亮周娴王爽
Owner FUDAN UNIV
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