CRISPR/Cas9-Mediated Knock-in in K562 and Raji Cell Lines for Tracking Immune Cell Killing Assay

Glioblastomas (GBM) are the most aggressive tumors affecting the central nervous system in adults, causing death within, on average, 15 months after diagnosis. Immunocompetent in-vivo models that closely mirror human GBM are urgently needed for deciphering glioma biology and for the development of effective treatment options. The murine GBM cell lines currently available for engraftment in immunocompetent mice are not only exiguous but also inadequate in representing prominent characteristics of human GBM such as infiltrative behavior, necrotic areas, and pronounced tumor heterogeneity. Therefore, we generated a set of glioblastoma cell lines by repeated in vivo passaging of cells isolated from a neural stem cell-specific Pten/p53 double-knockout genetic mouse brain tumor model. Transcriptome and genome analyses of the cell lines revealed molecular heterogeneity comparable to that observed in human glioblastoma. Upon orthotopic transplantation into syngeneic hosts, they formed high-grade gliomas that faithfully recapitulated the histopathological features, invasiveness and immune cell infiltration characteristic of human glioblastoma. These features make our cell lines unique and useful tools to study multiple aspects of glioblastoma pathomechanism and to test novel treatments in an intact immune microenvironment.

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A Novel Peptide Derived from Ginger Induces Apoptosis through the Modulation of p53, BAX, and BCL2 Expression in Leukemic Cell Lines

Planta Medica ◽

10.1055/a-1408-5629 ◽

2021 ◽

Author(s):

Chawalit Chatupheeraphat ◽

Sittiruk Roytrakul ◽

Narumon Phaonakrop ◽

Kamolchanok Deesrisak ◽

Sucheewin Krobthong ◽

...

Keyword(s):

Cell Viability ◽

Cell Lines ◽

Mononuclear Cells ◽

Raji Cell ◽

Leukemic Cell ◽

B Cell Lymphoma ◽

Leukemic Cells ◽

Peripheral Blood Mononuclear ◽

Normal Peripheral Blood ◽

Peptide Fraction

AbstractDespite the efficacy of chemotherapy, the adverse effects of chemotherapeutic drugs are considered a limitation of leukemia treatment. Therefore, a chemotherapy drug with minimal side effects is currently needed. One interesting molecule for this purpose is a bioactive peptide isolated from plants since it has less toxicity to normal cells. In this study, we extracted protein from the Zingiber officinale rhizome and performed purification to acquire the peptide fraction with the highest cytotoxicity using ultrafiltration, reverse-phase chromatography, and off-gel fractionation to get the peptide fraction that contained the highest cytotoxicity. Finally, a novel antileukemic peptide, P2 (sequence: RALGWSCL), was identified from the highest cytotoxicity fraction. The P2 peptide reduced the cell viability of NB4, MOLT4, and Raji cell lines without an effect on the normal peripheral blood mononuclear cells. The combination of P2 and daunorubicin significantly decreased leukemic cell viability when compared to treatment with either P2 or daunorubicin alone. In addition, leukemic cells treated with P2 demonstrated increased apoptosis and upregulation of caspase 3, 8, and 9 gene expression. Moreover, we also examined the effects of P2 on p53, which is the key regulator of apoptosis. Our results showed that treatment of leukemic cells with P2 led to the upregulation of p53 and Bcl-2-associated X protein, and the downregulation of B-cell lymphoma 2, indicating that p53 is involved in apoptosis induction by P2. The results of this study are anticipated to be useful for the development of P2 as an alternative drug for the treatment of leukemia.

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Differences in Heat-induced Cell Killing as Determined in Three Mammalian Cell Lines Do Not Correspond with the Extent of Heat Radiosensitization

International Journal of Radiation Biology and Related Studies in Physics Chemistry and Medicine ◽

10.1080/09553008614551081 ◽

1986 ◽

Vol 50 (4) ◽

pp. 675-684 ◽

Cited By ~ 4

Author(s):

H.H. Kampinga ◽

J.B.M. Jorritsma ◽

P. Burgman ◽

A.W.T. Konings

Keyword(s):

Cell Lines ◽

Mammalian Cell ◽

Cell Killing ◽

Mammalian Cell Lines

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Abstract PO-064: ONC212 stimulates cytotoxic T-cell killing, increases tumor-immune cell interactions, and promotes tumor regression in combination with TLY012 in a PDAC murine model

10.1158/1538-7445.panca21-po-064 ◽

2021 ◽

Author(s):

Kelsey E. Huntington ◽

Anna Louie ◽

Young Lee ◽

Jared Mompoint ◽

Isacco Ferrarini ◽

...

Keyword(s):

T Cell ◽

Murine Model ◽

Immune Cell ◽

Tumor Regression ◽

Cell Killing ◽

Cell Interactions ◽

Cytotoxic T Cell

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Development of a novel potency assay to quantify immune cell-mediated cancer cell killing

Cytotherapy ◽

10.1016/j.jcyt.2017.02.096 ◽

2017 ◽

Vol 19 (5) ◽

pp. S41

Author(s):

A. Oumie ◽

A. Chan ◽

M. Baradez ◽

D. Marshall

Keyword(s):

Cancer Cell ◽

Immune Cell ◽

Cell Killing ◽

Potency Assay

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Search for receptors in immune cells that bind cancer cell antigens and their activation in silent cases

10.7287/peerj.preprints.27670 ◽

2019 ◽

Author(s):

Wenfa Ng

Keyword(s):

Immune System ◽

Amino Acid ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Immune Cells ◽

Immune Cell ◽

Checkpoint Inhibitors ◽

Host Cells ◽

Immune Receptor

The immune checkpoint plays an important role in keeping immune cells in check for protecting tissues and organs from attack by the body’s own immune system. Similar concepts also apply in how cancer cells managed to fool immune cells through the surface display of particular antigens that mimic those exhibited by normal body cells. Specifically, cancer cells display antigens that bind to receptors on immune cells that subsequently prevent an attack on the cancer cells. Such binding between cancer antigens and immune cell receptors can be prevented through the use of checkpoint inhibitors antibodies specific for particular receptors on immune cells; thereby, unleashing immune cells to mount an immune response against cancer cells. While demonstrating good remissions in many patients where tumours shrunk substantially after administration of checkpoint inhibitors, cases exist where an overactivated immune system cause harm to organs and tissues culminating in multiple organ failure. Analysis of such toxicity effects of checkpoint inhibitors revealed that generic nature of targeted immune receptor plays a pivotal role in determining extent of side effects. Specifically, if the target immune receptor participates in checkpoints that prevent immune cells from attacking host cells, unleashing such receptors in cancer therapy may have untoward effects on patient’s health. Hence, the goal should be the selection of immune cell receptor specific to cancer cell antigens and which does not bind antigens or ligands displayed by the body’s cells. Such receptors would provide ideal targets for the development of checkpoint inhibitor antibodies for unleashing immune cells against cancer cells. To search for non-generic receptors that bind cancer cell antigens only, a combined computational and experimental approach could be used where ensemble of surface antigens on cancer cells and available receptors on immune cells could be profiled by biochemical assays. Downstream purification of ligands and receptors would provide for both structural elucidation and amino acid sequencing useful for bioinformatic search of homologous sequences. Knowledge of the antigens’ and receptors’ structures and amino acid sequence would subsequently serve as inputs to computational algorithms that models molecular docking events between receptor and antigen. This paves the way for heterologous expression of putative ligand and receptor in cell lines cultured in co-culture format for assessing binding between ligand and receptor, and more importantly, its physiological effects. Ability of immune receptor to bind to ligands on normal cells could also be assessed. Similar co-culture studies could be conducted with cancer cells and different immune cell types to check for reproducibility of observed effect in cell lines. Finally, antibodies could be raised for candidate receptors whose inhibition would not result in systemic attack of immune cells on host cells.

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