Epstein-Barr virus (EBV) antigen composites and dendritic cell (DC)-based vaccine, and use thereof
View Patent ↗Epstein-Barr virus (EBV) antigen composites and a dendritic cell (DC)-based vaccine, and a use thereof in preparation of a drug for controlling an EBV-associated infectious disease are provided. Patient-derived DCs are stimulated in vitro, loaded with lysates of various types of EBV-infected cells with strong immunogenicity for EBV-associated infectious diseases, and induced into mature dendritic cells (mDCs) by various cytokines and specific agonists, so as to obtain a complete DC-based vaccine with corresponding antigens. The DC-based vaccine can be injected back into the patient to activate the immune system to produce cytotoxic T cells, thereby killing EBV-infected cells, exerting an immunological effect, and improving a life quality of the patient. In addition, the DC-based vaccine can be prepared in about one week with a low cost, is safe, and shows no obvious side effects.
1. A method for treating an Epstein-Barr virus (EBV)-associated infectious disease, comprising: administering to a subject a dendritic cell (DC)-based vaccine loaded with EBV antigen composites, wherein the EBV-associated infectious disease comprises infectious mononucleosis (IM), chronic active EBV (CAEBV) infection, and EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH);
the EBV antigen composites comprise lysates of human immortalized B lymphoblastoid cell lines (B-LCLs) B95-8-LCL, GD1-LCL, M81-LCL, HKNPC1-LCL to HKNPC9-LCL, SNU-719-LCL, YCCEL1-LCL, and lysates of EBV positive infected cells C666-1, HNE1, and EB-3;
wherein a method for preparing the DC-based vaccine comprises:
(1) induction of immature DC (imDC): transferring a CD14 + cell suspension to a well plate with 2×10 6 cells/mL in each well, and then adding human recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) to a final concentration of 2,000 IU/mL and human recombinant interleukin (IL)-4 to a final concentration of 1,000 IU/mL to the well plate for induction to prepare imDC;
(2) induction of mature DC (mDC): co-cultivating the EBV antigen composites with imDC in step (1), adding tumor necrosis factor (TNF)-α to a final concentration of 2,000 IU/mL, LPS to a final concentration of 2 μg/mL and Poly(I:C) to a final concentration of 1 μg/mL to stimulate a maturation of imDC, wherein an amount of cells for producing each of the lysates is 3×10 7 ; and
(3) detection of induced mDCs: measuring an increase in cell surface markers CD11c, CD40, CD80, CD83, CD86, HLA-DR, and HLA-ABC in the mDC compared to the imDC by flow cytometry (FCM).