hCD5(BALB/c)

利用同源重组，将小鼠Cd5基因进行人源化修饰。

Fig.1 Detection of CD5 expression in thymus by RT-PCR. Mouse Cd5 mRNA (162 bp) was detectable only in wild-type BALB/c mice. Human CD5 mRNA (249 bp) was detectable only in homozygous hCD5 (BALB/c) knockin mice but not in wild-type mice. Thymus RNA was extracted from 7-week-old female wild-type BALB/c (WT)(n=3) and homozygous hCD5 (BALB/c) knockin mice (HO) (n=3), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. 

Abbr. M, marker; HO, homozygous; WT, wild type.

Fig.2 Detection of CD5 expression on T cells and B cells in spleen of WT (BALB/c) mice and HO hCD5(BALB/c) mice by FACS. hCD5 was exclusively detectable in hCD5(BALB/c) humanized mice but not in wild-type (BALB/c) mice. hCD5 was exclusively detectable on T cells and B cells in hCD5(BALB/c) humanized mice, whereas mCD5 was exclusively detectable on T cells and B cells in wild-type BALB/c mice. Leukocytes were collected from spleen of wild-type BALB/c mice (female, 6 weeks old) and homozygous hCD5(BALB/c) humanized mice (female, 6 weeks old), and analyzed by flow cytometry with anti-mCD5 and anti-hCD5. 

Abbr. HO, homozygous; WT, wild type.

Fig.3 Detection of CD5 expression on T cells and B cells in blood of WT (BALB/c) mice and HO hCD5(BALB/c) mice by FACS. hCD5 was exclusively detectable in hCD5(BALB/c) humanized mice but not in wild-type (BALB/c) mice. hCD5 was exclusively detectable on T cells and B cells in hCD5(BALB/c) humanized mice, whereas mCD5 was exclusively detectable on T cells and B cells in wild-type BALB/c mice. Leukocytes were collected from blood of wild-type BALB/c mice (female, 6 weeks old) and homozygous hCD5(BALB/c) humanized mice (female, 6 weeks old), and analyzed by flow cytometry with anti-mCD5 and anti-hCD5. 

Abbr. HO, homozygous; WT, wild type.

Fig.4 Frequency of leukocyte and myeloid subpopulations in the blood by flow cytometry. hCD5(BALB/c) knockin mice showed a normal immune phenotype in blood. Blood was collected from WT BALB/c mice and HO hCD5(BALB/c) mice (6-week-old, n=3). Flow cytometry analysis of frequencies of (A) lymphocyte and (B) myeloid subpopulations in the blood. Bar indicated expressed as mean ± SEM.

Abbr.  HO, homozygous; WT, wild type.

Fig.5 Frequency of leukocyte and myeloid subpopulations in the spleen by flow cytometry. hCD5(BALB/c) knockin mice showed a normal immune phenotype in spleen. Spleen was collected from WT BALB/c mice and HO hCD5(BALB/c) mice (6-week-old, n=3). Flow cytometry analysis of frequencies of (A) lymphocyte and (B) myeloid subpopulations in the spleen. Bar indicated expressed as mean ± SEM.

Abbr.  HO, homozygous; WT, wild type.