	US 12,419,912 B2
	Epstein-Barr virus (EBV) antigen composites and dendritic cell (DC)-based vaccine, and use thereof
Helen Liu, Shanghai (CN); and Ze Yin, Shanghai (CN)
Assigned to Helen Liu, Shanghai (CN); and KOUSAI Bio Co., Ltd, Shanghai (CN)
Appl. No. 18/574,764
Filed by KOUSAI Bio Co., Ltd, Shanghai (CN); and Helen Liu, Shanghai (CN)
PCT Filed Jun. 17, 2022, PCT No. PCT/CN2022/099296 § 371(c)(1), (2) Date Dec. 28, 2023, PCT Pub. No. WO2023/005486, PCT Pub. Date Feb. 2, 2023.
Claims priority of application No. 202110864886.4 (CN), filed on Jul. 29, 2021.
Prior Publication US 2024/0269171 A1, Aug. 15, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 35/15 (2025.01); A61K 40/19 (2025.01); A61K 40/46 (2025.01); A61P 31/22 (2006.01); C12N 5/0784 (2010.01)

CPC A61K 35/15 (2013.01) [A61K 40/19 (2025.01); A61K 40/46 (2025.01); A61P 31/22 (2018.01); C12N 5/0639 (2013.01)]

1 Claim

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 2×10⁶cells/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 3×10⁷; 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).