Gene targeting strategy for B-hTREM2 mice(C). The exons 1-5 and 3’UTR of mouse Trem2 gene that encode the full-length protein were replaced by human TREM2 exons 1-5 and 3’UTR in B-hTREM2 mice(C).

Strain specific analysis of TREM2 gene expression in wild-type mice and B-hTREM2 mice(C) by RT-PCR.Mouse Trem2 mRNA was detectable in wild-type mice (+/+). Human TREM2 mRNA was detectable only in homozygous B-hTREM2 mice(C) (H/H) but not in wild-type mice.

Antitumor activity of anti-human TREM2 antibody in B-hTREM2 mice(C).(A) Anti-human TREM2 antibody inhibited EMT-6 tumor growth in B-hTREM2 mice(C). Murine breast carcinoma EMT-6 cells were subcutaneously implanted into homozygous B-hTREM2 mice(C) (female, 10 week-old, n=6). Mice were grouped when tumor volume reached approximately 50-80 mm³, and treated with anti-hTREM2 antibody at doses and schedules in panel A. (B) Body weight changes during treatment. As shown in panel A, anti-human TREM2 antibody (in house) were efficacious in controlling tumor growth in B-hTREM2 mice(C), demonstrating they provide a powerful preclinical model forin vivoevaluation of anti-human TREM2 antibody. Values are expressed as mean ± SEM.

Antitumor activity of anti-human TREM2 antibody in B-hTREM2 mice(C). (A) Anti-human TREM2 antibody inhibited EMT-6 tumor growth in B-hTREM2 mice(C). Murine breast carcinoma EMT-6 cells were subcutaneously implanted into homozygous B-hTREM2 mice(C) (female, 7 week-old, n=6). Mice were grouped when tumor volume reached approximately 50-80 mm³, and treated with anti-hTREM2 antibody at doses and schedules in panel A. (B) Body weight changes during treatment. As shown in panel A, anti-human TREM2 antibody (in house) were efficacious in controlling tumor growth in B-hTREM2 mice(C), demonstrating they provide a powerful preclinical model for in vivo evaluation of anti-human TREM2 antibody. Values are expressed as mean ± SEM.

Flow cytometry analysis of tumor infiltrating lymphocytes (TILs). Tumor cells were harvested at the endpoint of experiment (n=3~5). Animals in G1 were treated with PBS and animals in G3 were treated with the anti-hTREM2 antibody. Flow cytometry analysis of the blood were performed to assess cell number and proportion changes compared to the group with no anti-hTREM2 antibody treated. The ratio of hTREM2+ CTL cells and hTREM2+ NK cells was significantly increased in the group treated with anti-hTREM2 antibodies compared to the group treated with PBS. Values are expressed as mean ± SEM. *p

Flow cytometry analysis of the proportion of lymphocytes in blood from tumor-bearing B-hTREM2 mice(C). Blood was collected at the endpoint of the experiment described in the previous page. From the results, the ratio of M1 macrophages was significantly increased in the anti-hTREM2 antibody treatment group compared to the PBS group, while the ratio of mCD86+ macrophages was significantly reduced. Values are expressed as mean ± SEM. *p