Methods of reducing large granular lymphocyte and natural killer cell levels
A NK cell, cell surface technology, applied in chemical instruments and methods, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, blood cell cancer vaccine, etc., can solve the problem of reducing LGL or NK cell levels, depletion Problems like LGL or NK cells
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Embodiment 1
[0211] Example 1: Analysis of CD94 expression on immune cells from healthy donors and from T-large granular lymphocytic leukemia (T-LGLL) and NK-cell chronic lymphoproliferative disorder (CLPD-NK) patients.
[0212] This example describes the results of experiments used to determine the level of CD94 receptor expression on immune cells obtained from healthy donors and from T-LGLL and NK-LGLL patients.
[0213] Materials and Methods
[0214] Healthy Donor and Patient Samples
[0215]Fresh healthy donor buffy coats were obtained from the Stanford Blood Center. Peripheral blood mononuclear cells (PBMCs) were isolated via ficoll-paque (GE Healthcare, Chicago, IL) and cryopreserved in Bambanker cell freezing medium (Bulldog-Bio, Portsmouth, NH). Briefly, buffy coats were diluted 1:1 in phosphate-buffered saline (PBS), and the diluted buffy coats were layered and centrifuged in ficoll at 760 g. The PBMC layer was separated and washed in PBS prior to downstream analysis. Periph...
Embodiment 2
[0242] Example 2: Analysis of NKG2A expression in immune cells from healthy donors and from patients with NK cell chronic lymphoproliferative disorder (CLPD-NK) and the effect of anti-NKG2A antibodies on ADCC activity.
[0243] This example describes the results of experiments used to determine the level of NKG2A receptor expression on immune cells obtained from healthy donors and from CLPD-NK patients. This example also shows the results of experiments measuring the effect of anti-NKG2A antibodies on antibody-dependent cellular cytotoxicity (ADCC).
[0244] Materials and Methods
[0245] Antibody-dependent cytotoxicity assay
[0246] Will be about 1x10 5 -2x10 5 Fresh or frozen PBMC were plated in tissue culture treated 96-well U-bottom plates in RPMI with 10% low IgG FBS. Cells were incubated overnight in 10-fold dilutions of a human IgG1 isotype control antibody, NKG2A Z199 fucosylated antibody, or NKG2A Z199 afucosylated antibody at concentrations ranging from 10 1 ...
Embodiment 3
[0256] Example 3: Analysis of NKG2A and CD94 expression on liver-derived cells.
[0257] This example describes the results of experiments used to determine the expression levels of CD94 and NKG2A receptors on liver-derived immune cells obtained from healthy donors.
[0258] Materials and Methods
[0259] Single and live liver-derived cells (CD45-) and lymphocyte populations (CD45 / CD4 / CD8 / CD19 / CD56+) were analyzed by flow cytometry as described in Example 1.
[0260] result
[0261] Single live liver-derived cells (CD45-) and lymphocyte populations (CD45 / CD4 / CD8 / CD19 / CD56+) were examined to screen for CD94 and NKG2A expression. As shown in Figure 8A, CD94 was highly expressed on NK cells of normal liver samples, and each cell had about 200,000 CD94 receptors. CD94 expression is also present on a subset of T cells (CD45+CD3+CD4+ / CD8+). CD94 expression was not detected on epithelial cells (CD45-) and B cells (CD45+CD3-CD19+). As shown in Figure 8B, NKG2A was only detect...
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