Human LAG3-expressing Raji Cells

Human LAG3-expressing B cells

Raji-hLAG3 cells were developed from the human Raji cell line, a human B lymphocyte-derived cell line, and engineered to stably overexpress the human LAG-3 gene. Raji cells have been successfully used as target cells in human effector studies such as antibody-dependent cellular cytotoxicity (ADCC), either with peripheral blood mononuclear cells, natural killer (NK) cells, or T cell-derived Jurkat reporter cells.

Lymphocyte activation gene-3 (LAG-3) is an inhibitory immune checkpoint (IC) with significant homology to the CD4 receptor. Expression of LAG-3 has been described on activated T cells, B cells, plasmacytoid DCs, and NK cells [1]. Similar to CD4, LAG-3 interacts with MHC class II molecules, however, they do not compete. Instead, LAG3 inhibits T cell activation by transducing inhibitory signals via its intracellular domain. Inhibitory mechanisms of LAG-3 are likely to be distinct from those exerted by other ICs due to its unusual and unique cytoplasmic tail [1,2]. Notably, LAG-3 expression has been reported to be associated with tumor progression and poor prognosis, which suggests that LAG-3 contributes to immune escape mechanisms in cancer [1,2]. Therefore, LAG-3 has been proposed as a promising therapeutic target for cancer immunotherapy [2]. Additionally, LAG-3 has been demonstrated to act synergistically with PD-1 to suppress anti-tumor immunity and thus, is being tested in combination in the clinic  [2,3].

Features of Raji-hLAG3 cells:

Surface expressed markers and ICs in Raji-hLAG3 cells

• Stable overexpression of the human LAG-3 gene 
• Characterized by a number of cell-surface expressed markers including the B cell receptor (BCR), CD19, and CD20
• Constitutive expression of various immune checkpoints (ICs) such as CD27, CD70, CD80, PD-L1, and 4-1BBL

Applications for Raji-hLAG3 cells:

• Target cell line for ADCC assays using InvivoGen's Jurkat-Lucia™ NFAT-CD16 cells
• Target cell line for cell toxicity assays using NK or CAR-T cells
• For use in the development of novel human LAG-3 antagonistic mAbs

Validation of Raji-hLAG3 cells:

• Overexpression of LAG3 verified by flow cytometry
• Guaranteed mycoplasma-free

​References:

1. Maruhashi, T. et al. 2020. LAG-3: from molecular functions to clinical applications. J Immunother Cancer 8.
2. Shan, C. et al. 2020. Progress of immune checkpoint LAG-3 in immunotherapy. Oncol Lett 20, 207.
3. Yang, Z.Z. et al. 2017. Expression of LAG-3 defines exhaustion of intratumoral PD-1(+) T cells and correlates with poor outcome in follicular lymphoma. Oncotarget 8, 61425-61439.

Figures

Validation of the expression of human LAG3 by Raji-hLAG3 cells. Raji-Null (A) and Raji‑hLAG3 (B) cells were incubated with a PE-conjugated Anti-hLAG3 mAb for 30 minutes. The binding affinity was then measured using flow cytometry.

Specifications

Antibiotic resistance: Blasticidin

Growth medium: IMDM, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum (FBS; 30 min at 56 °C), Pen-Strep (100 U/ml-100 µg/ml), 100 µg/ml Normocin™

Test medium: IMDM, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated FBS, Pen-Strep (100 U/ml-100 µg/ml)

Quality control:

• Expression of human LAG3 has been verified by flow cytometry.
• The stability for 20 passages following thawing has been verified.
• Raji-hLAG3 cells are guaranteed mycoplasma-free.

Contents

• 3-7 x 10⁶ Raji-hLAG3 cells in a cryovial or shipping flask.
• 1 ml of Blasticidin (10 mg/ml). Store at 4 °C or at -20 °C.
• 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi. Store at -20 °C.

IMPORTANT: If cells are shipped frozen (i.e. in a cryovial) and are not frozen upon arrival, contact InvivoGen immediately.

Shipped on dry ice (Europe, USA & Canada)

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