Studies of serum-free culture medium in the generation of lymphokine activated killer cells

Muul, Linda; Nason-Burchenal, Kathryn; Hyatt, Cornelia; Schwarz, Susan; Slavin, D.; Elaine, Paula; Rosenberg, Steven A.

doi:10.1016/0022-1759(87)90265-1

Cited by 27 publications

(2 citation statements)

References 13 publications

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“…T lymphocytes freshly isolated from cancer patients rarely exhibit anti-tumor cytotoxic activity [l-31. Nevertheless, lytic activity against autologous tumor cells has been developed in vitro among these cells after a mixed lymphocyte-tumor cell culture and expansion with IL2 [4][5][6][7][8][9][10][11][12][13][14][15][16].This suggested that some tumor cells express epitopes which activate autologous T lymphocytes and induce their clonal expansion in v i m , while similar activatiodexpansion or function of these lymphocytes was impaired in vivo for unknown reasons. The success of immunotherapeutic stra-[I 77461 * This work was supported by funds from INSERM, by grant no.…”

Section: Introductionmentioning

confidence: 99%

Selective expansion of a specific anti‐tumor CD8⁺ cytotoxic T lymphocyte clone in the bulk culture of tumor‐infiltrating lymphocytes from a melanoma patient: Cytotoxic activity and T cell receptor gene rearrangements

et al. 1990

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In vitro culture of metastatic melanoma fragments with 150 units of recombinant interleukin 2 resulted in the successive expansion of CD4+ and then CD8+ tumor-infiltrating lymphocytes (TIL) throughout a 2-month period. TIL cultured for 43 days and consisting of 95% CD8+ and 10% CD4+ T lymphocytes were cloned by limiting dilution (LD). Thirteen CD8+ and thirty-one CD4+ clones were obtained, indicating that the frequency of clonogenic CD8+ proliferative T lymphocytes was much lower than that of their CD4+ homologues. When LD was performed in the presence of autologous melanoma cells the frequency of CD8+ clones was increased by factor 4. The DNA from TIL of day 43 bulk culture and of six CD8+ clones was hybridized with T cell receptor (TcR) beta and gamma probes. Identical configuration of the nonfunctional gamma and functional beta TcR genes was found in "bulk culture" and cloned TIL. The CD8+ clones therefore derived from a clonal population of CD8+ cells which had expanded in vitro before the LD. All the CD8+ clones tested were strongly cytotoxic for autologous melanoma cells but did not kill autologous fibroblasts or concanavalin A blasts, or any of the 10 allogeneic tumor targets tested, including 5 melanomas, 2 breast cell lines, 1 neuroblastoma, K-562 and the Epstein-Barr virus-transformed cell line used as a feeder. Furthermore, specific killing was inhibited by monoclonal antibodies against CD3, CD8, TcR alpha/beta and against class I major histocompatibility complex antigens indicating that these cytotoxic T lymphocyte clones recognized autologous tumor cells through the TcR, in an HLA class I-restricted manner. These data show that it is feasible to obtain tumor-specific cytotoxic T lymphocytes from melanoma TIL with a simple culture technique and that a single clone could be expanded to more than 10(10) cells which should allow testing of immunotherapeutic potential of these cells by adoptive transfer into melanoma patients.

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Section: Introductionmentioning

confidence: 99%

Selective expansion of a specific anti‐tumor CD8⁺ cytotoxic T lymphocyte clone in the bulk culture of tumor‐infiltrating lymphocytes from a melanoma patient: Cytotoxic activity and T cell receptor gene rearrangements

et al. 1990

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“…In the 1980s, the availability of recombinant IL-2 for in vitro use brought the breakthrough of ACT trials [19–21]. Using IL-2 as a T cell growth factor in culture promoted the ability to expand human lymphocytes to larger scales and for longer periods.…”

Section: Initial Act Studiesmentioning

confidence: 99%

Focus on Adoptive T Cell Transfer Trials in Melanoma

Hershkovitz

Schachter

Treves

et al. 2010

Clinical and Developmental Immunology

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Adoptive Cell Transfer (ACT) of Tumor-Infiltrating Lymphocytes (TIL) in combination with lymphodepletion has proven to be an effective treatment for metastatic melanoma patients, with an objective response rate in 50%–70% of the patients. It is based on the ex vivo expansion and activation of tumor-specific T lymphocytes extracted from the tumor and their administration back to the patient. Various TIL-ACT trials, which differ in their TIL generation procedures and patient preconditioning, have been reported. In the latest clinical studies, genetically engineered peripheral T cells were utilized instead of TIL. Further improvement of adoptive T cell transfer depends on new investigations which seek higher TIL quality, increased durable response rates, and aim to treat more patients. Simplifying this therapy may encourage cancer centers worldwide to adopt this promising technology. This paper focuses on the latest progress regarding adoptive T cell transfer, comparing the currently available protocols and discussing their advantages, disadvantages, and implication in the future.

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Protection mechanisms of freely suspended animal cells (CRL 8018) from fluid‐mechanical injury. Viscometric and bioreactor studies using serum, pluronic F68 and polyethylene glycol

Jd¹,

McIntire

et al. 1991

Biotech & Bioengineering

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We use bioreactor and viscometric studies to examine the mechanism by which three additives, fetal bovine serum (FBS), pluronic F68, and polyethylene glycol (PEG), protect the freely suspend CRL-8018 cells from damage due to interactions with bubbles in agitated bioreactors. In bioreactor studies, the protective effect of an addictive could be due to either changes in the ability of the cell resist shear (biological mechanism) or to changes in the medium properties that effect the level or frequency of forces experienced by the cells (physical mechanism). Bioreactor studies show that protection by all three addictives occurs whether the cells are grown in the presence of the addictives (long exposure) or the addictives are added to medium after the cells were exposed to detrimental agitation intensity (short exposure). In the viscometric studies, exposure of cells to laminar shear in the absence of gas-liquid interfaces assesses only the ability of the cells to resist a constant level of shear in a medium with or without an additive. Viscometric studies show that prolonged exposure to FBS makes the cells more shera tolerant, but that short (30-120 min) exposure to FBS does not affect their shear tolerance. We thus conclude that the protective effect of FBS in bioreactors id of both physical and biological nature. The biological contribution is metabolic in nature rather than fast acting. Viscometric studies show that either long or short exposure of the cells to either F68 or PEG does not make the cells more shear tolerant. WE therefore conclude that the protective effect of F68 and PEG does not make the cells more shear tolerant. We therefore conclude that the protective effect of F68 and PEG in bioreactors is physical in nature.

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Studies of serum-free culture medium in the generation of lymphokine activated killer cells

Cited by 27 publications

References 13 publications

Selective expansion of a specific anti‐tumor CD8⁺ cytotoxic T lymphocyte clone in the bulk culture of tumor‐infiltrating lymphocytes from a melanoma patient: Cytotoxic activity and T cell receptor gene rearrangements

Selective expansion of a specific anti‐tumor CD8⁺ cytotoxic T lymphocyte clone in the bulk culture of tumor‐infiltrating lymphocytes from a melanoma patient: Cytotoxic activity and T cell receptor gene rearrangements

Focus on Adoptive T Cell Transfer Trials in Melanoma

Protection mechanisms of freely suspended animal cells (CRL 8018) from fluid‐mechanical injury. Viscometric and bioreactor studies using serum, pluronic F68 and polyethylene glycol

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Studies of serum-free culture medium in the generation of lymphokine activated killer cells

Cited by 27 publications

References 13 publications

Selective expansion of a specific anti‐tumor CD8+ cytotoxic T lymphocyte clone in the bulk culture of tumor‐infiltrating lymphocytes from a melanoma patient: Cytotoxic activity and T cell receptor gene rearrangements

Selective expansion of a specific anti‐tumor CD8+ cytotoxic T lymphocyte clone in the bulk culture of tumor‐infiltrating lymphocytes from a melanoma patient: Cytotoxic activity and T cell receptor gene rearrangements

Focus on Adoptive T Cell Transfer Trials in Melanoma

Protection mechanisms of freely suspended animal cells (CRL 8018) from fluid‐mechanical injury. Viscometric and bioreactor studies using serum, pluronic F68 and polyethylene glycol

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Product

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Selective expansion of a specific anti‐tumor CD8⁺ cytotoxic T lymphocyte clone in the bulk culture of tumor‐infiltrating lymphocytes from a melanoma patient: Cytotoxic activity and T cell receptor gene rearrangements

Selective expansion of a specific anti‐tumor CD8⁺ cytotoxic T lymphocyte clone in the bulk culture of tumor‐infiltrating lymphocytes from a melanoma patient: Cytotoxic activity and T cell receptor gene rearrangements