Basic Information
-
Targeting strategy
-
Gene targeting strategy for B-hCD16A mice. The regulatory region and exons 1~5 of mouse Fcgr4 gene that encode the full-length protein were replaced by human FCGR3A exons 1~4 and regulatory region in B-hCD16A mice.
-
mRNA expression analysis
-
Strain specific analysis of CD16A gene expression in wild-type mice and homozygous B-hCD16A mice by RT-PCR.
Mouse Fcgr4 mRNA was exclusively detectable in wild-type mice and human FCGR3A mRNA was exclusively detectable in homozygous B-hCD16A mice (H/H).
-
Protein expression analysis in NK cells
-
Strain specific CD16A expression analysis in wild-type C57BL/6 mice and homozygous B-hCD16A mice by flow cytometry.
Splenocytes were collected from wild-type C57BL/6 mice(+/+) and homozygous B-hCD16A mice (H/H). Human CD16A was exclusively detectable in the NK cells of homozygous B-hCD16A mice.
-
Immune Cell Profile
-
Analysis of leukocytes subpopulation in B-hCD16A mice
Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hCD16A mice (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. Values are expressed as mean ± SEM.
Analysis of spleen T cell subpopulations in B-hCD16A mice
Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hCD16A mice (n=3, 6-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Values are expressed as mean ± SEM.
Analysis of leukocytes subpopulation in B-hCD16A mice
Analysis of LNs leukocyte subpopulations by FACS. LNs were isolated from female C57BL/6 and B-hCD16A mice (n=3, 6-week-old). Flow cytometry analysis of the LNs 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. Values are expressed as mean ± SEM.
Analysis of LNs T cell subpopulations in B-hCD16A mice
Analysis of LNs T cell subpopulations by FACS. LNs were isolated from female C57BL/6 and B-hCD16A mice (n=3, 6-week-old). Flow cytometry analysis of the LNs was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Values are expressed as mean ± SEM.
Analysis of leukocytes subpopulation in B-hCD16A mice
Analysis of blood leukocyte subpopulations by FACS. Blood were isolated from female C57BL/6 and B-hCD16A mice (n=3, 6-week-old). Flow cytometry analysis of the blood 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. Values are expressed as mean ± SEM.
Analysis of blood T cell subpopulations in B-hCD16A mice
Analysis of blood T cell subpopulations by FACS. Blood were isolated from female C57BL/6 and B-hCD16A mice (n=3, 6-week-old). Flow cytometry analysis of the blood was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Values are expressed as mean ± SEM.
-
Antitumor activity of anti-human CD16A-based Ab in B-hCD16A mice
-
Antitumor activity of anti-human CD16A-based Ab in B-hCD16A mice.
(A) Anti-human CD16A-based Ab inhibited MC38 tumor growth in B-hCD16A mice. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hCD16A mice (female,13 week-old, n=7). Mice were grouped when tumor volume reached approximately 70 mm3, at which time they were treated with anti-human CD16A-based Ab with doses and schedules indicated in panel B. (B) Body weight changes during treatment. As shown in panel A, anti-human CD16A-based Ab was efficacious in controlling tumor growth in B-hCD16A mice, demonstrating that the B-hCD16A mice provide a powerful preclinical model for in vivo evaluation of anti-human CD16A-based Ab. Values are expressed as mean ± SEM.
-
Poster
