Cell line overexpressing human CD303 antigen
The invention relates to a cell line expressing the gamma chain of the FcεRI receptor and human CD303 antigen, characterized in that said cell line is transfected in a stable manner by an expression vector comprising a nucleic acid molecule coding for human CD303 antigen and having strong expression of human CD303 on the surface thereof, e.g. at least 10000 molecules of human CD303 per cell, as well as a vector or vector kit that can be used to co-express the gamma chain of the FcεRI receptor and human CD303 antigen, and different uses of the cell line of the invention.
1. A cell line expressing the gamma chain of Fcε receptor I (FcεRI) and human CD303 antigen, wherein said cell line is stably transfected with an expression vector comprising a nucleic acid molecule encoding human CD303 antigen and has high expression of human CD303 on its surface, wherein said cell line has at least 10,000 human CD303 molecules per cell and said cell line is:
a) a cell line of human plasmacytoid dendritic cells (pDCs) stably transfected with an expression vector comprising a nucleic acid molecule encoding human CD303 antigen; or
b) a human lymphocyte cell line, stably transfected with an expression vector comprising a first nucleic acid molecule encoding human CD303 antigen and a second nucleic acid molecule encoding the gamma chain of human FcεRI, or with two expression vectors each comprising a nucleic acid molecule encoding human CD303 antigen and the gamma chain of human FcεRI, respectively.
2. The cell line according to claim 1 , wherein said cell line is a cell line stably transfected with a vector comprising a nucleic acid molecule encoding human CD303 antigen, selected from the CAL-1 line or a human pDC line obtained from blastic plasmacytoid dendritic cell neoplasm (BPDCN) cells from a human patient by a method comprising the following steps:
obtaining primary malignant BPDCN cells from peripheral blood of a patient diagnosed with BPDCN according to WHO classification; and
cloning surviving cells after long-term culture to obtain a long-lasting cell line in culture.
3. The cell line according to claim 1 , wherein said cell line is a Jurkat cell line, stably transfected with an expression vector comprising a first nucleic acid molecule encoding human CD303 antigen and a second nucleic acid molecule encoding the gamma chain of human FcεRI, or with two expression vectors each comprising a nucleic acid molecule encoding human CD303 antigen and the gamma chain of human FcεRI, respectively.
4. The cell line according to claim 1 , wherein said cell line has on its surface between 20,000 and 60,000 human CD303 molecules per cell.
5. A method for testing ADCC activity of an antibody against human CD303 antigen, comprising the following steps:
a) contacting the cell line according to claim 1 with an antibody against human CD303 antigen and effector cells;
b) incubating the cells obtained in step a) for a suitable time to allow lysis of the cells; and
c) measuring degree of lysis of the incubated cells.
6. A kit for characterizing anti-CD303 antibodies comprising the cell line according to claim 1 .
7. The cell line according to claim 4 , wherein said cell line has on its surface between 25,000 and 50,000 human CD303 molecules per cell.
8. The cell line according to claim 2 , wherein said cell line has on its surface between 20,000 and 60,000 human CD303 molecules per cell.
9. The cell line according to claim 8 , wherein said cell line has on its surface between 25,000 and 50,000 human CD303 molecules per cell.
10. The cell line according to claim 3 , wherein said cell line has on its surface between 20,000 and 60,000 human CD303 molecules per cell.
11. The cell line according to claim 10 , wherein said cell line has on its surface between 25,000 and 50,000 human CD303 molecules per cell.
12. The method of using the cell line according to claim 5 , wherein said activity of an antibody against human CD303 antigen is selected from ADCC, CDC, cytokine secretion, apoptosis or phagocytosis.
13. The method of using the cell line according to claim 5 , wherein said human effector cells are from a subject suffering from BPDCN or from an autoimmune disease.
14. The kit according to claim 6 , further comprising at least one reagent selected from effector cells, polyvalent IgG, and reference anti-CD303 antibodies.