Another advantage of targeting NKG2A is the safety of this approach, as no abnormalities have been reported in mouse strains lacking CD94 ( Vance et al. We thus concluded that the safety profile of the combination was similar to that for the two single agents.

The effector cells used were total PBMCs, purified human NK cells, nine-day culture of PBMCs with rhIL-15 or seven-day culture with rhIL-2 (PROLEUKIN) of enriched peripheral blood NK cells by CD3 depletion followed by CD56 positive selection. The variable domain of the mousified light chain was inserted in frame with a mouse Ck domain into a eukaryotic expression vector to generate the light chain.

Consistent with these data, treatment with a combination of monalizumab and cetuximab was found to be effective in the {"type":"clinical-trial","attrs":{"text":"NCT02643550","term_id":"NCT02643550"}}NCT02643550 phase II clinical trial for SCCHN. Unlike classical HLA class I molecules, HLA-E continues to be expressed on the surface of tumor cells, often even more strongly than on healthy cells, in patients with solid tumors or leukemia/lymphoma ( Benson et al. Anti-NKG2A, anti-HLA-E and anti-NKp46 Abs were used at a concentration of 5 μg/mL and anti-CD8 mAb was used at a concentration of 1 μg/mL. Tumor size was monitored with a digital caliper (Mitutoyo) every three to four days and is expressed as a volume ((length x width 2) /2).

The next day, cells were washed twice and the intracellular staining was performed for 30 min at 4°C with anti-IFN-γ-PercPCy5.NK cells were incubated at a ratio of 1:1 or 5:2 with target cell lines (SCCHN, ovarian cancer cell lines or K562 cells transfected to express HLA-E and/or PDL-1). That way you always have a machine that is tailored to the demands and opportunities that you have right now. The percentage of flu-specific CD8 + T cells was determined by staining on day 0 before coculture and on day 9 of monocyte-CD8 + T cell coculture.

We also monitored PD-1 expression, because the immune control of A20 tumors has been reported to be partially dependent on PD-1 ( Sagiv-Barfi et al.

Human PBMCs were isolated from buffy coats by gradient density centrifugation in Pancoll Lymphocyte Separating Medium. The targeting of NKG2A with a blocking antibody would therefore have the unique advantage of enhancing T and NK cell responses. We observed the generation of CD62L − CD44 + effector memory CD8 + T cells in the spleens of mice in which RMA-Rae-1β tumors were implanted and then cured by immunotherapy, but not in the spleens of untreated mice ( Figure 3D). We developed a humanized anti-NKG2A immunoglobulin G (IgG) 4-blocking mAb (monalizumab), and we describe its anti-tumor efficacy in vitro and in vivo when used as a single agent or in combination with other therapeutic antibodies, such as durvalumab, blocking PD-L1, or cetuximab, directed against the epidermal growth factor receptor (EGFR), which is expressed by tumor cells.