Immune cells capable of clearing latent infections in vivo, preparation method and application thereof
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A technology of immune cells and cells, applied in the application field of preparations
Active Publication Date: 2020-08-25
赛德特生物制药有限公司
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Since this type of cell expresses both CD3+ and CD56+ membrane protein molecules, it is also called NK cell-like T lymphocyte, which has both the strong anti-tumor activity of T lymphocytes and the non-MHC-restricted tumor killing advantages of NK cells. However, this method still has limitations
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Embodiment 1
[0057] Embodiment 1. Preparation of SIET cells
[0058] 1. Isolation of Peripheral Blood Lymphocytes
[0059] (1) One blood draw volume: 150ml, anticoagulated with heparin. 150ml of whole blood can normally get 1.5-2×10 8 cells, can plant a 75cm 2 Culture flask; the peripheral blood treated with heparin anticoagulation was divided into centrifuge tubes, centrifuged at 1800rpm for 15min, and the supernatant was discarded;
[0060] (2) Dilute the blood: AIMV culture medium is used to dilute the blood to make a lymphocyte suspension;
[0061] (3) 25ml of lymphocyte suspension was carefully placed on 12.5ml of lymphocyte separation medium, centrifuged at 1800rpm for 15min, and the lifting speed was adjusted to the lowest;
[0062] (4) Take the upper part of the plasma and keep it in a refrigerator at 4°C;
[0063] (5) Take the lymphocyte layer, place it in a 50ml centrifuge tube, add AIMV medium to a total volume of 50ml, centrifuge at 1800rpm for 5min;
[0064] (6) Discard ...
Embodiment 2
[0101] Example 2.Detection of SIET cell expansion ability
[0102] Adopt the method for embodiment 1 to cultivate, and take the SIET cells that culture time is the 0th, 3, 5, 7, 9, 12 days, detect cell proliferation situation, as figure 1 . Depend on figure 1 It can be seen that the proliferation of SIET cells cultured by the method of the present invention is better.
Embodiment 3
[0103] Example 3.SIET cell viability detection
[0104] The method of Example 1 was used to culture, and 100 μl of cells cultured for 12 days was taken, and 100 μl of 0.4% trypan blue staining solution was added. The living cells were not stained, and the dead cells were stained blue. The results are shown in Table 1.
[0107] It can be seen from Table 1 that the viability of the cells prepared by the present invention is greater than 95%, and the viability of the cultured SIET cells is better.
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Abstract
The invention provides an immune cell capable of removing recessive infection in vivo, and a preparation method and application thereof. The preparation method of the immune cells comprises the stepsthat peripheral blood mononuclear cells are separated and purified in vitro; and the peripheral blood mononuclear cells are activated by using a cell activator I; wherein the cell activator I comprises IL-2, IFN-gamma, GM-CSF, IL-4 and TNF-alpha, and is configured in normal saline; the activated cells are amplified with a cell activator II; the cell activator II comprises IL-2, IFN-gamma and IL-15, and is configured in normal saline; and the immune cells are harvested. According to the immune cell capable of removing the recessive infection in vivo, the immune can be amplified and returned tothe human body by using immune cells against bacteria, fungi, viruses, Chlamydia and Barton's small in vitro source factors, the purpose of reducing infection and eliminating hidden infection is achieved, disease occurrence is reduced, and aging is delayed.
Description
technical field [0001] The present invention relates to an immune cell capable of eliminating latent infection in the body and its preparation method and application, in particular, to an immune cell SIET (Silent Infection Elimination T cells) prepared by activating and expanding peripheral blood mononuclear cells ) and its preparation method, as well as its application in the preparation of preparations for reducing infection and clearing occult infection, belong to the field of biotechnology. Background technique [0002] Recessive infection is also called subclinical infection. It means that after the pathogen invades the human body, it only causes the body to produce a specific immune response, causing no or only slight tissue damage, so it does not show any clinical symptoms, signs, or even biochemical changes, and can only be detected by immunological examination. Find. [0003] Among them, the factors of cancer infection mainly include: hepatitis B virus (Hepatitis ...
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