RU2008131594A - AUTOLOGICAL VACCINE FOR TREATMENT OF ONCOLOGICAL DISEASES AND METHOD FOR ITS OBTAINING - Google Patents

Abstract

1. An autologous cancer vaccine containing lymphocytes activated by interleukin 2, dendritic cells obtained by incubating immature DCs with a tumor lysate, characterized in that it contains T-lymphocytes specifically activated by mature DCs. ! 2. A method of obtaining an autologous vaccine for the treatment of cancer, including the isolation of mononuclear cells (MNCs) from the patient’s peripheral blood, culturing MNCs in DMEM, separating cells into monocytes that attach to the substrate and lymphocytes that do not attach to the substrate, placing MNCs in the culture medium, separation of non-adherent lymphocytes and addition of IL2 to them to obtain lymphokinactivated killers (LAC cells, addition of growth factor to the remaining adherent monocytes, stimulation of maturation DC by an autologous in vitro tumor lysate and the subsequent addition of maturing factors for 1 day, characterized in that to obtain adherent monocytes, MNCs are cultured in medium supplemented with 10% FCS for one hour, after which, to obtain immature dendritic cells (nDK) the monocytes attached are cultured in a medium supplemented with a neupogen of 50 ng / ml for 48 h, after which, to obtain mature dendritic cells, the NDCs are cultured in a medium containing 2000 IU / ml of rheoperon and 50 ng / ml of betaleukin, while o cultivate LAC cells from the non-adherent fraction of MNCs in a medium containing 100 IU / ml of roncoleukin for 72 hours, and then the obtained ZDK and LAC cells are washed by centrifugation in physiological saline for 10 min at 1500 rpm, after which a joint

Claims (2)

1. An autologous cancer vaccine containing lymphocytes activated by interleukin 2, dendritic cells obtained by incubating immature DCs with a tumor lysate, characterized in that it contains T-lymphocytes specifically activated by mature DCs. 2. A method of obtaining an autologous vaccine for the treatment of cancer, including the isolation of mononuclear cells (MNCs) from the patient’s peripheral blood, culturing MNCs in DMEM, separating cells into monocytes that attach to the substrate and lymphocytes that do not attach to the substrate, placing MNCs in the culture medium, separation of non-adherent lymphocytes and addition of IL2 to them to obtain lymphokinactivated killers (LAC cells, addition of growth factor to the remaining adherent monocytes, stimulation of maturation DC by an autologous in vitro tumor lysate and the subsequent addition of maturing factors for 1 day, characterized in that to obtain adherent monocytes, MNCs are cultured in medium supplemented with 10% FCS for one hour, after which, to obtain immature dendritic cells (nDK) the monocytes attached are cultured in a medium supplemented with a neupogen of 50 ng / ml for 48 h, after which, to obtain mature dendritic cells, the NDCs are cultured in a medium containing 2000 IU / ml of rheoperon and 50 ng / ml of betaleukin, while o cultivate LAC cells from the non-adherent fraction of MNCs in a medium containing 100 IU / ml of roncoleukin for 72 hours, and then the obtained ZDK and LAC cells are washed by centrifugation in physiological saline for 10 min at 1500 rpm, after which a joint cultivation is carried out zDK with LAC cells in medium supplemented with 100 IU / ml roncoleukin for 24 hours, then the attached fraction and the non-attached cell fraction are removed and washed separately by centrifugation in physiological saline for 10 min at 1500 rpm, obtaining a vaccine.