Strain specific B2M expression analysis in wild-type B-NDG mice and homozygous B-NDG hB2M/HLA-A11.1 mice plus by flow cytometry. Splenocytes, blood cells and bone marrow were collected from wild-type B-NDG mice and homozygous B-NDG hB2M/HLA-A11.1 mice plus. Protein expression was analyzed with anti-mouse B2M antibody and anti-human B2M antibody by flow cytometry. Mouse B2M was detectable in wild-type mice but not in homozygous B-NDG hB2M/HLA-A11.1 mice plus. Human B2M was exclusively detectable in homozygous B-NDG hB2M/HLA-A11.1 mice plus but not in wild-type mice.

Strain specific HLA-A expression analysis in wild-type B-NDG mice and homozygous B-NDG hB2M/HLA-A11.1 mice plus by flow cytometry. Splenocytes, blood cells and bone marrow were collected from wild-type B-NDG mice and homozygous B-NDG hB2M/HLA-A11.1 mice plus. Protein expression was analyzed with anti-mouse H-2Kb/H-2Db antibody and anti-human HLA-A,B,C antibody by flow cytometry. Mouse H2-K1/H2-D1 was detectable in wild-type mice but not in homozygous B-NDG hB2M/HLA-A11.1 mice plus. Human HLA-A was exclusively detectable in homozygous B-NDG hB2M/HLA-A11.1 mice plus but not in wild-type mice.

Frequency of leukocyte subpopulations in bone marrow, spleen and blood by flow cytometry. Bone marrow, splenocytes and blood cells were isolated from B-NDG mice and homozygous B-NDG hB2M/HLA-A11.1 mice plus (male,7-week-old, n=3). A. Flow cytometry analysis of the immune cells was performed to assess the frequency of leukocyte subpopulations. Frequencies of T cells, B cells, NK cells, DCs, neutrophils, monocytes and macrophages in B-NDG hB2M/HLA-A11.1 mice plus were similar to those in B-NDG mice, demonstrating that humanization of HLA-A11.1 does not change the frequency or distribution of these cell types in bone marrow, spleen and blood when compared with those in B-NDG mice. Values are expressed as mean ± SEM.