Protein expression of NKP46 in spleen

Analysis of spleen leukocyte subpopulations by FACS. Blood cells were isolated from female CB-17 SCID and B-hNKP46 mice(CB-17 SCID) (n=3, 6-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hNKP46 mice(CB-17 SCID) were similar to those in the CB-17 SCID mice, demonstrating that NKP46 humanized does not change the overall development, differentiation or distribution of these cell types in blood. Values are expressed as mean ± SD.

Protein expression analysis via immunohistochemistry(IHC)

IHC of mouse ileum (upper panel) and spleen (bottom panel). 

Image 2. Mouse ileum, huNKP46 IHC, approximately 200x. (2A) CB-17 SCID mouse. Labeling for huNKP46 is limited to nonspecific staining. The superficial epithelium of the mucosa (M) and smooth muscle of the tunica muscularis externa (TM) exhibit light nonspecific blushing. The submucosa (SM) is indicated. (2B) B-hNKP46 mice(CB-17 SCID). Small numbers of scattered round cells in the lamina propria (LP) exhibit moderate to strong membranous to cytoplasmic labeling (black arrows). There is light background nonspecific labeling similar to that in (2A). The mucosa (M), submucosa (SM), and tunica muscularis externa (TM) are indicated. 

Image 4. Mouse spleen, huNKP46 IHC, approximately 200x. (4A) CB-17 SCID mouse. There is light nonspecific background labeling of erythrocytes/ serum in the red pulp. Red pulp (RP) is indicated. (4B) BhNKP46 mice(CB-17 SCID). Moderate to large numbers of scattered round cells exhibit moderate to strong membranous to cytoplasmic labeling (black arrows). Red pulp (RP) is indicated.

In vivo efficacy of anti-human NKP46-based Ab in B-hNKP46 mice(CB-17 SCID)

Antitumor activity of anti-human NKP46-based Ab in B-hNKP46 mice(CB-17 SCID). (A) Anti-human NKP46-based Ab inhibited BxPC-3 tumor growth in B-hNKP46 mice(CB-17 SCID). BxPC-3 cells (5E6) were subcutaneously implanted into B-hNKP46 mice(CB-17 SCID) (male, 7-8 week-old, n=4). Mice were grouped when tumor volume reached approximately 150-200 mm³, at which time they were treated with anti-human NKP46-based Ab with doses and schedules indicated in panel A. (B) Body weight changes during treatment. As shown in panel A, anti-human NKP46-based Ab was efficacious in controlling tumor growth in B-hNKP46 mice(CB-17 SCID). Values are expressed as mean ± SEM. (All antibodies were provided by the clients)

In vivo efficacy of anti-human NKP46-based Ab-individual tumor growth curves 

In vivo efficacy of anti-human NKP46-based Ab in B-hNKP46 mice(CB-17 SCID)

Antitumor activity of anti-human NKP46-based Ab in B-hNKP46 mice(CB-17 SCID). (A) Anti-human NKP46-based Ab inhibited BxPC-3 tumor growth in B-hNKP46 mice(CB-17 SCID). BxPC-3 cells (5E6) were subcutaneously implanted into B-hNKP46 mice(CB-17 SCID) (male, 7-8 week-old, n=6). Mice were grouped when tumor volume reached approximately 100-150 mm³, at which time they were treated with anti-human NKP46-based Ab with doses and schedules indicated in panel A. (B) Body weight changes during treatment. As shown in panel A, anti-human NKP46-based Ab was efficacious in controlling tumor growth in B-hNKP46 mice(CB-17 SCID). Values are expressed as mean ± SEM. (All antibodies were provided by the clients)

In vivo efficacy of anti-human NKP46-based Ab-individual tumor growth curves