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EP-4317423-B1 - METHOD FOR PRODUCING IMMUNE CELL COMPOSITION

EP4317423B1EP 4317423 B1EP4317423 B1EP 4317423B1EP-4317423-B1

Inventors

  • LEE, HEE JIN
  • LEE, KYU JIN
  • PARK, HYE SEON
  • GONG, Gyung Yub
  • LIM, CHAE LYUL
  • KIM, YOUNG AE
  • CHOI, JI IL

Dates

Publication Date
20260506
Application Date
20220330

Claims (15)

  1. A method for producing an immune cell composition, comprising: (a) culturing a tumor sample in a first container, wherein the tumor sample is cultured in a first composition in the first container, wherein the first composition comprises medium, serum, and 500 to 1500 IU/ml of IL-2, wherein the first container is a gas-permeable container or a container comprising gas supply system, wherein the culturing is performed for 13 to 15 days, wherein, during the culturing, a first additive composition is added to the first container once or twice or more, and the first additive composition comprises medium, serum, and IL-2; (b) obtaining a first immune cell population from the cultured product obtained from (a); (c) seeding a second composition and a second immune cell population in a second container, wherein the second immune cell population is derived from the first immune cell population, wherein the second composition comprises medium, serum, 1000 to 3000 IU/ml of IL-2, anti-CD3 antibody, and a feeder cell, wherein the second container is a gas-permeable container or a container comprising gas supply system; (d) expanding the second immune cell population to produce a third immune cell population, wherein during the expansion, a second additive composition is added to the second container once or twice or more, and the second additive composition comprises medium, serum, and IL-2, wherein the expansion is performed for 13 to 15 days; and (e) obtaining the third immune cell population from the expanded product obtained from (d) and producing the immune cell composition,
  2. The method of claim 1, in (a), 12 to 24 of the tumor samples are cultured, wherein each of the tumor sample has cross-sectional area of 1mm 2 to 4mm 2 , and wherein the tumor samples are obtained from a tumor fragment obtained from a subject.
  3. The method of claim 1, wherein a concentration of serum of the first composition is 5 vol% or less, and wherein a concentration of serum of the second composition is 5 vol% or less.
  4. The method of claim 1, wherein a volume of the first composition is 30% or less of a maximum allowable capacity of the first container, and wherein a volume of the second composition is 30% or less of a maximum allowable capacity of the second container.
  5. The method of claim 1, wherein the first additive composition is a composition having the same compositional substances and ratio as the first composition, and wherein during the culturing, the first additive composition is added to the first container two or more times.
  6. The method of claim 5, during the culturing, the first additive composition is added to the first container on the 7th, 10th, and 12th day from the start date of the culturing, wherein amount of the first additive composition added at each addition point is the same.
  7. The method of claim 1, wherein the anti-CD3 antibody is at least one selected from OKT3, Otelixizumab, teplizumab, and visilizumab, and wherein a concentration of the anti-CD3 antibody of the second composition is 10ng/ml to 100ng/ml.
  8. The method of claim 1, wherein the feeder cell is an inactivated allogeneic peripheral blood mononuclear cell.
  9. The method of claim 1, wherein the feeder cell is included in the second composition in a ratio of approximately 1:200 (number of cells of the second immune cell population:number of feeder cells) based on the number of cells in the second immune cell population.
  10. The method of claim 1, in the step of seeding, the number of cells of the second immune cell population seeded in the second container is 1E5 to 10E5.
  11. The method of claim 1, wherein the second additive composition has the same compositional substances and ratio as the second composition excluding the anti-CD3 antibody and the feeder cell, and wherein during the expansion, the second additive composition is added to the second container two or more times.
  12. The method of claim 11, during the expansion, the second additive composition is added to the second container on the 4th, 6th, 8th, and 10th days from the start date of expansion, and amount of the second additive composition added is gradually increased at each addition point.
  13. The method of claim 1, wherein the immune cell composition comprises the third immune cell population or a fourth immune cell population obtained from the third immune cell population.
  14. The method of claim 1, wherein the tumor sample is derived from a solid tumor of a subject having breast cancer, colon cancer, lung cancer, stomach cancer, kidney cancer, ovarian cancer, or skin cancer including melanoma, or is derived from a malignant pleural effusion of a subject having lung cancer.
  15. The method of claim 1, wherein the method further comprises: cryopreserving the first immune cell population; and thawing the cryopreserved first immune cell population, wherein cryopreserving the first immune cell population and the thawing the cryopreserved first immune cell population are performed after (b) obtaining a first immune cell population from the cultured product obtained from (a).

Description

[Technical Field] The present application relates to a method for producing an immune cell composition. More particularly, the present application relates to a method for producing an immune cell composition comprising tumor-infiltrating lymphocytes (TILs). [Background Art] Cancer immunotherapy is one of the methods of treating cancer and this method activates the immune system of the human body to treat cancer. As a type of immunotherapeutic used in cancer immunotherapy, there is immune cell therapy, which is a cell therapy method in which immune cells in the body are collected and reinforced or modified using a genetic engineering technique, and then put back into the body. One of such cell therapy methods is a method using tumor-infiltrating lymphocytes (TILs), and here, the tumor infiltrating lymphocytes (TILs) refer to lymphocytes gathered around cancer cells. The therapy using TILs includes a process of collecting naturally-occurring T cells infiltrating a patient's tumor and then activating and expanding the T cells. A large number of activated T cells that have been activated and expanded in a large number through the above process are injected back into the patient, resulting in destruction of the tumor. In other words, the process of culturing and expanding TILs is essential for cancer therapy using tumor infiltrating lymphocytes. However, in this process, the isolation TILs from tumor tissue, culture and expansion are complicated, and there is a difficulty in expanding TILs with a higher yield. Therefore, a more efficient process for TIL culture and expansion is required, and the present application relates to a method for producing an immune cell composition with a high yield. [Disclosure] [Technical Problem] The present application is directed to providing a method for efficiently producing an immune cell composition with a high yield, compared to a conventional production method with a low yield. In one embodiment, the present application provides a method for efficiently culturing and expanding immune cells. In one specific embodiment, the present application provides a method for efficiently culturing and expanding tumor-infiltrating lymphocytes (TILs). [Technical Solution] One aspect of the present application provides a method for producing an immune cell composition. Another aspect of the present application provides an immune cell composition including TILs. Still another aspect of the present application provides a use of an immune cell composition including TILs. [Advantageous Effects] A method for producing an immune cell composition according to the present application has higher efficiency than that of a convention method for producing an immune cell composition. Specifically, compared to the conventional method for producing an immune cell composition, a low concentration of IL-2 is used in the method of the present application, and immune cells may be obtained in high yield. [Description of Drawings] FIGS. 1 to 4 illustrate Examples of a process of producing an immune cell composition.FIG. 1 illustrates a process of producing a semi-product (an intermediate product) (a first immune cell population produced and acquired by isolating an immune cell population from a sample and culturing).FIG. 2 illustrates a process of producing a final product (an immune cell composition produced by acquiring a third immune cell population expanded from a second immune cell population) from a cryopreserved semi-product.FIG. 3 illustrates a process of producing a finished product from a tumor sample, wherein the process is an example that includes a cryopreservation step.FIG. 4 illustrates a process of producing a finished product from a tumor sample, wherein the process is an example that does not include a cryopreservation step.FIG. 5 shows the experimental results obtained by measuring a tumor-infiltrating lymphocyte (TIL) expansion yield.FIGS. 6 and 7 show the experimental results for optimizing an IL-2 concentration in the production of TILs.FIG. 6 shows the experimental results for optimizing an IL-2 concentration in TIL culture.FIG. 7 shows the experimental results for optimizing an IL-2 concentration in TIL expansion.FIG. 8 shows the experimental results for the cell proliferation rate of TILs according to the volume of a composition used in culture.FIG. 9 shows the experimental results obtained by measuring the number of culture cells per tumor tissue in TIL culture.FIG. 10 shows the results of confirming the percentage of T cells in cultured and expanded TILs.FIG. 11 shows a representative example of a process for isolation/culture of TILs from a tumor sample and mass culture (expansion) of TILs.FIG. 12 shows the results of confirming the percentages of CD8+ T cells and CD4+ T cells in TILs (sample No. BC16208) expanded from a cryopreserved and thawed semi-product.FIG. 13 shows the results of confirming the type of T cells in the CD4+ T cells and CD8+ T cells identified in FIG. 12.FIG. 14