Dendritic cell obtained by antigen pulsing
A technology of dendritic cells and dendritic cells, applied in animal cells, virus antigen components, vertebrate cells, etc., can solve problems such as heavy economic burdens, and achieve the effect of improving immune response and immune response
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Embodiment 1
[0172] The experimental examples of the present invention will be specifically described below, but the present invention is not limited to the following experimental examples.
experiment example 1
[0173] (Experimental example 1) Preparation of dendritic cells
(1) Use a heparin (Novo-heparin injection 1000: manufactured by Aventis) syringe (Terumo SyringeSS-50ESZ: manufactured by Terumo) from each subject (five healthy volunteers who have repeatedly used HB vaccine experience as the test subjects) Blood collection 40-60mL. Fully mix the collected blood with the same amount of phosphate buffered saline (PBS) in a clean bench. Then, mononuclear cells were isolated by density gradient centrifugation using Ficoll-Conray solution (specific gravity 1.077). At room temperature, centrifuge at 1800rpm-3000rpm for 30 minutes, and take out the middle layer after centrifugation as mononuclear cells.
[0174] (2) At the same time, autoserum was obtained from each subject according to the following procedure. First, 10 mL of blood was collected from each subject with a heparin-free syringe (Terumo Syringe SS-10ESZ: manufactured by Terumo Corporation). Then, the collected blood wa...
experiment example 2
[0181] (Experimental Example 2) Dendritic cells were pre-excited with HB vaccine.
(1) The dendritic cells obtained in Experimental Example 1 were divided into two. Add HB vaccine (r-HB vaccine "Mitsubishi": manufactured by Mitsubishi Pharma Co., Ltd.) at a final concentration of 2.5-5 μg / mL to 1 aliquot of dendritic cells (ingredients: HBs antigen 20 μg / mL, ethylmercury thiosalicylic acid Sodium 0.01w / v% / mL, Aluminum Potassium Sulfate 5.2g / mL), cultivated for 8-24 hours, thus obtained HB vaccine pre-excited dendritic cells containing HBs antigen. (Hereafter, this cell is referred to as "HB vaccine-primed dendritic cell"). In a dedicated culture room at 37°C, 5% CO 2 Concentration, cultured under the condition of 75% air. Another set of dendritic cells was cultured without HB vaccine as a negative control.
[0182] (2) After culturing, HB vaccine-pre-excited dendritic cells were recovered from the culture dish. PBS was added to the recovered cells, centrifuged at 1500 rpm...
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