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Anti Asialo GM1 (Rabbit) Application Data
Evaluation of cytotoxic activity against murine lymphoma YAC-1 cells
Each antibody was reconstituted in 1 mL of distilled water and diluted 2-fold in RPMI 1640 medium. For NK cell activity, 200 µL of anti-asialo GM1 (Product No. 986-10001 or Product No.014–09801) antibodies were administered intraperitoneally. Two days after administration of anti-asialo GM1 antibody, the control group (Group No. 2) and the antibody administration group (Group No. 3 and 4) were administered 100 μg of Poly I:C intraperitoneally.
Spleens were collected 16 to 18 hours after polyI:C administration. Cells were prepared from the collected spleens and cytotoxicity (NK cell activity) was measured against CFDA-labeled YAC-1 cells using flow cytometry.
The evaluation was conducted under the following two conditions.
Effector cells (spleen cells) : Target cells (YAC-1 cells) = 100:1 or 50:1
| No. | Group | Administration (i.p.) | |
|---|---|---|---|
| 1 | Negative control | Vehicle (RPMI 1640) |
Vehicle (RPMI 1640) |
| 2 | Control | Poly I:C (100 μg/mouse) |
|
| 3 | Antibody | pAb (Product No. 986-10001) |
|
| 4 | pAb (Product No. 014-09801) |
||
Result: In the anti-asialo GM1 antibody administration groups, NK cell activity was significantly lower than in the comparison control under both conditions. There was no significant difference in NK cell activity between Group No.3 and No.4.
Evaluation of NK cell removal
The number of NK cells in the spleen collected from the mice was measured by immunostaining.
The spleen cells were stained with CD335 (NKp46) and CD3ε. CD335-positive/CD3ε-negative cells were detected as NK cells. The flow cytometer analysis diagram shows one individual from the control group and one individual from antibody administration group.
| No. | Group | Administration (i.p.) | |
|---|---|---|---|
| 1 | Negative control | Vehicle (RPMI 1640) |
Vehicle (RPMI 1640) |
| 2 | Control | Poly I:C (100 μg/mouse) |
|
| 3 | Antibody | pAb (Product No. 986-10001) |
|
| 4 | pAb (Product No. 014-09801) |
||
Result: Compared to the percentage of NK cells in the control group (Group No. 2), the percentage of NK cells in the antibody-treated group was significantly lower. There was no significant difference in the percentage of NK cells between Group No. 3 and 4.
The ability of the two types of anti-GM1 antibodies to remove NK cells is considered equivalent.
References
- Kasai M. et al.: Eur. J. Immunol., 10, 175(1980).
A glycolipid on the surface of mouse natural killer cells - Habu S. et al.: J. Immunol., 127, 34(1981).
IN VIVO EFFECTS OF ANTI-ASIALO GM1 I. Reduction of NK Activity and Enhancement of Transplanted Tumor Growth in Nude Mice - Nishikado, H. et al.: J. Immunol., 186(10), 5766(2011).
NK Cell-Depleting Anti-Asialo GM1 Antibody Exhibits a Lethal Off-Target Effect on Basophils In Vivo
