Natural killer cell clearance in mice

Summary

NK cells are usually injected intravenously; in vivo clearance can be achieved by intraperitoneal injection of antibodies against NK cells, such as PK136 monoclonal antibody, anti-asialo GM1 antibody, and anti-Ly49 antibody, etc. The specificity of various antibodies is different. The specificity of various antibodies is different, with the following characteristics: ① PK136 monoclonal antibody (anti-NK1.1 antibody) specifically clears NK1.1* cells.PK136 monoclonal antibody (IgG2a) specifically recognizes mouse NK1.1+ cells, and NK cells bound by the antibody are specifically cleared in the presence of mouse complement.NK1.1 is expressed in a lineage-specific manner, only in C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6, C57BL/6 and C57BL/6. NK1.1 expression is strain-specific and is only expressed in C57BL/6, C57BL/10, or H2b, H2d, and H2q homozygous mice, whereas BALB/c, AKR, CBA, SJL, and C3 H mice do not express NK1.1. Since NKT cells express NK1.1, NKT cells are also eliminated when NKT cells are cleared by the PK136 monoclonal antibody. (ii) Anti-asialo GM1 antibody, which recognizes the glycolipid salivaryl ganglioside asialo GM1 expressed on the surface of NK cells. anti-asialo GM1 antibody is able to effectively clear NK cells in mice and rats, but high doses of anti-asialo GM1 antibody can also interact with mouse monocytes, embryonic thymocytes, and some T cells. Therefore, some of the monocytes and T cells can be deleted when this antibody is applied at high concentrations. (iii) Anti-Ly49 antibody, Ly49 is a receptor expressed on mature NK cells and is a marker of mature NK cells, but NKT cells also express Ly49, therefore, application of anti-Ly49 antibody can only remove mature NK cells and Ly49 + NKT cells.

Principle

The basic principle of natural killer cell clearance is the removal of NK cells by intraperitoneal injection of antibodies against NK cells, such as PK136 monoclonal antibody, anti-asialo GM1 antibody and anti-Ly49 antibody.

Operation method

Natural killer cell clearance in mice

Principle

The basic principle of natural killer cell clearance is the removal of NK cells by intraperitoneal injection of antibodies against NK cells, such as PK136 monoclonal antibody, anti-asialo GM1 antibody and anti-Ly49 antibody.

Materials and Instruments

Reagents:
Mouse (note the germline specificity of PK136 antibody above), PK136 monoclonal antibody or anti-asialo GM1 antibody or anti-Ly49 antibody, fluorescently labeled DX5 monoclonal antibody (pan-NK cell marker) or NK1.1 antibody.
Instruments:
Syringe, incubator

Move

The basic process of natural killer cell clearance in mice can be divided into the following steps:

(1) PK136 monoclonal antibody (200-500 μg/each), or anti-asialo GM1 antibody (200 μg/each), or anti-Ly49 antibody (10 μg/each) was drawn up in a 1 mL syringe and injected intraperitoneally into mice. A control group was also set up.

(2) One to two drops of peripheral blood were collected from mice 1, 2 and 3 days after injection to prepare peripheral blood mononuclear cell suspension.

(3) The mice were stained with fluorescently labeled DX5 monoclonal antibody or NK1.1 antibody, and the effect of NK cell clearance was detected by flow cytometry. Generally, the onset of action is 1-3 days.

Caveat

(1) The potency of the antibody may vary from batch to batch, and the effective dose of the antibody should be determined by pre-testing.(2) The antibody used should be sterile to avoid infection of experimental animals.(3) The recovery time of NK cells after clearance is related to the dose of antibody used, the higher the dose the slower the recovery.

For more product details, please visit Aladdin Scientific website.

https://www.aladdinsci.com/