hCD3EDG/hCD19/hCD20

Fig.1 Detection of hCD3/hCD19/hCD20 expression on T cells and B cells in spleen of WT C57BL/6 mice and HO hCD3EDG/hCD19/hCD20 mice by FACS. Leukocytes were collected from wild-type C57BL/6 mice (female, 11 weeks old) and homozygous hCD3EDG/hCD19/hCD20 humanized mice (female, 11 weeks old), and analyzed by flow cytometry with anti-mCD3,  anti-mCD19, anti-mCD20, anti-hCD3, anti-hCD19, and anti-hCD20. hCD3/hCD19/hCD20 was detectable in hCD3EDG/hCD19/hCD20 humanized mice.

Fig.2 Frequency of leukocyte subpopulations in blood by flow cytometry. Blood was collected from WT C57BL/6 mice and HO hCD3/hCD19/hCD20 mice (female, 11-week-old, n=3). Flow cytometry analysis of frequencies of (A) lymphocyte and (B) myeloid subpopulations in the blood. Percentages of T cells, B cells, NK cells, CD11b+myeloid cells, DCs, neutrophils, monocytes, and macrophages in the blood of HO hCD3/hCD19/hCD20 mice were similar to those in WT C57BL/6 mice. Bar indicated expressed as mean ± SEM.

Fig.3 Frequency of leukocyte subpopulations in spleen by flow cytometry. Spleen was collected from WT C57BL/6 mice and HO hCD3/hCD19/hCD20 mice (female, 11-week-old, n=3). Flow cytometry analysis of frequencies of (A) lymphocyte and (B) myeloid subpopulations in the spleen. Percentages of T cells, B cells, NK cells, CD11b+myeloid cells, DCs, neutrophils, monocytes, and macrophages in the spleen of HO hCD3/hCD19/hCD20 mice were similar to those in WT C57BL/6 mice. Bar indicated as mean ± SEM.

Fig.4 Frequency of leukocyte subpopulations in bone marrow by flow cytometry. Bone marrow were collected from WT C57BL/6 mice and HO hCD3/hCD19/hCD20 mice (female, 11-week-old, n=3). Flow cytometry analysis of frequencies of (A) lymphocyte and (B) myeloid subpopulations in the bone marrow. Percentages of T cells, B cells, NK cells, CD11b+myeloid cells, DCs, neutrophils, monocytes, and macrophages in the bone marrow of HO hCD3/hCD19/hCD20 mice were similar to those in WT C57BL/6 mice. Bar indicated as mean ± SEM.

Fig.5 In vivo B cell depletion of Bispecific Antibodies in hCD3EDG/hCD19/hCD20 Mice. B cell depletion by anti-human CD3/CD20 or CD3/CD19 bispecific antibodies in humanized mice. (A) Representative flow cytometric analysis showing B cell depletion in peripheral blood of HO-hCD3E/hCD19/hCD20 mice. (B) Quantitative summary of B cell depletion in peripheral blood of hCD3E/hCD19/hCD20 mice (female, 8 weeks old, n=3-5 per group) after a single administration. Mice received Vehicle, Glofitamab (anti-CD3/CD20), or Surovatamig (anti-CD3/CD19). The percentage of human CD45⁺cells that are B cells is plotted over time (Day-1 [baseline], Day 3, and Day 5 post-administration).