| Strain Name |
C57BL/6-H2-Eatm1(HLA-DRA1*1.1-I-Ea)Bcgen H2-Eb1tm1(HLA-DRB1*7.1-I-Eb1)Bcgen H2-Atm1Bcgen H2-Eb2tm1Bcgen/Bcgen |
Common Name |
B-hHLA-DRB1*7.1 mice |
| Background | C57BL/6 | Catalog number | 112948 |
|
Related Genes |
HLA-DRA1; SS1, DRB1, HLA-DRB, HLA-DR1B |
||
|
NCBI Gene ID |
3122, 3123 |
||
- HLA-DRA is one of the HLA class II alpha chain paralogues. HLA-DRB1 belongs to the HLA class II beta chain paralogs. This class II molecule is a heterodimer consisting of an alpha (DRA) and a beta (DRB) chain, both anchored in the membrane. It plays a central role in the immune system by presenting peptides derived from extracellular proteins. Class II molecules are expressed in antigen presenting cells (APC: B lymphocytes, dendritic cells, macrophages). The alpha chain is approximately 33-35 kDa and its gene contains 5 exons. The beta chain is approximately 26-28 kDa and its gene contains 6 exons. DRA does not have polymorphisms in the peptide binding part and acts as the sole alpha chain for DRB1, DRB3, DRB4 and DRB5.
- The exon 2 of mouse I-Ea gene that encodes the extracellular domain of I-Ea was replaced by human HLA-DRA encoding the α1 domain in B-hHLA-DRB1*7.1 mice. The exon 2 of mouse I-Eb1 gene that encodes the extracellular domain of I-Eb1 was replaced by human HLA-DRB encoding the β1 domain in B-hHLA-DRB1*7.1 mice. The mouse I-Aa, I-Ab1 and I-Eb2 were knocked out in B-hHLA-DRB1*7.1 mice.
- Human HLA-DR was exclusively detectable in homozygous B-hHLA-DRB1*7.1 mice, but not in wild-type C57BL/6 mice.
- B-hHLA-DRB1*7.1 mice provide a powerful preclinical model for in vivo evaluation of vaccines.
- Application: For example, this product is used for pharmacodynamics and safety evaluation of vaccines for cancers.
Strain specific HLA-DR expression analysis in wild-type C57BL/6 mice and homozygous B-hHLA-DRB1*7.1 mice by flow cytometry. Spleen cells were collected from wild-type C57BL/6 mice (+/+) and homozygous humanized B-hHLA-DRB1*7.1 mice, respectively, and analyzed by flow cytometry with species-specific anti-mouse I-A/I-E antibody (Biolegend, 107607), and species-specific anti-human HLA-DR antibody (Biolegend, 307610). Human HLA-DR was exclusively detectable in homozygous B-hHLA-DRB1*7.1 mice, but not in wild-type C57BL/6 mice.
Strain specific HLA-DR expression analysis in wild-type C57BL/6 mice and homozygous B-hHLA-DRB1*7.1 mice by flow cytometry. Blood cells were collected from wild-type C57BL/6 mice (+/+) and homozygous humanized B-hHLA-DRB1*7.1 mice, respectively, and analyzed by flow cytometry with species-specific anti-mouse I-A/I-E antibody (Biolegend, 107607), and species-specific anti-human HLA-DR antibody (Biolegend, 307610). Human HLA-DR was exclusively detectable in homozygous B-hHLA-DRB1*7.1 mice, but not in wild-type C57BL/6 mice.
Frequency of leukocyte subpopulations in lymph node by flow cytometry. Lymph nodes were isolated from wild-type C57BL/6 mice and homozygous B-hHLA-DRB1*7.1 mice (Female,8-week-old, n=3). A. Flow cytometry analysis of the lymph nodes cells was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Frequencies of T cells, B cells, NK cells, DCs, monocytes, macrophages, neutrophils, CD4+ T cells, CD8+ T cells and Tregs in B-hHLA-DRB1*7.1 mice were similar to those in C57BL/6 mice, demonstrating that humanization of HLA-DRB1*7.1 does not change the frequency or distribution of these cell types in lymph nodes.
B-hHLA-DRB1*7.1 mice functional validation by flow cytometry
B-hHLA-DRB1*7.1 mice functional validation by flow cytometry. The male C57BL/6 mouse (n=1) and B-hHLA-DRB1*7.1 mouse (n=1) at the age of 10 weeks were sacrificed. The splenocytes and lymph node cells were isolated and employed as antigen presenting cells (APC). Splenocytes or lymph node cells were incubated with the antigen peptides for 2hrs and washed to remove unbound peptides. These peptide-loaded splenocytes or lymph node cells were then co-cultured with engineered TCR-T cells (specifically recognizing HLA-DRB1*07:01 presented mutant peptide) overnight. On the following day, flow cytometry was performed to assess the expression of CD134 on TCR-T cells. The results clearly indicated that the control（APC without peptide loading) and the APC treated with wild-type (WT) peptides failed to elicit responses of TCR-T cells. In contrast, the mutant (Mu) peptides induced a significant up-regulation of CD134 expression on TCR-T cells.
(All results were provided by the client.)
京公网安备：