B-hIGHA1/hCD89 mice_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen, drug development, antibody discovery

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B-hIGHA1/hCD89 mice

Strain Name	C57BL/6N-Sμ^{tm1(IGHA1)Bcgen}Gt(ROSA)26Sor^{tm1(CD11b-FCAR)Bcgen}/Bcgen	Common Name	B-hIGHA1/hCD89 mice
Background	C57BL/6N	Catalog number	112799
Aliases	IGHA1 (IgA1); CD89 (FcalphaRI; CTB-61M7.2, FCAR, Fc fragment of IgA receptor)

Protein expression analysis in T, B cells

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CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was mildly detectable in T and B cells in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Protein expression analysis in serum

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Strain specific IgA expression analysis in homozygous B-hIGHA1/hCD89 mice by ELISA. Serum were collected from 8-weeks wild-type mice C57BL/6 mice (+/+) and 8-weeks homozygous B-hIGHA1/hCD89 mice (H/H, H/+ or H/H, H/H), and analyzed by ELISA with species-specific IgA ELISA kit. hIgA was exclusively detectable in homozygous mice, but soluble IgA levels were higher in B-hIGHA1/hCD89 mice (H/H, H/+) compared to that in B-hIGHA1/hCD89 mice (H/H, H/H).

Protein expression analysis in NK cells and DCs

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CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was detectable in NK cells and DCs in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Protein expression analysis in monocytes and macrophages

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CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was detectable in monocytes and macrophages in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Protein expression analysis in neutrophils

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CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was detectable in neutrophils in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Protein expression analysis in spleen

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CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was mainly detectable in NK cells, DCs, monocytes, macrophages and neutrophils in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Analysis of leukocytes cell subpopulation in spleen

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Analysis of spleen leukocyte subpopulations by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H). 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. Percent of B cells was decreased and T cells increased in B-hIGHA1/hCD89 mice accompanied with a smaller spleen compared to that in C57BL/6 mice. Values are expressed as mean ± SD.

Protein expression analysis in T, B cells

from clipboard

CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was detectable in T and B cells in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Protein expression analysis in NK cells and DCs

from clipboard

CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was detectable in NK cells and DCs in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Protein expression analysis in monocytes and macrophages

from clipboard

CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was detectable in monocytes and macrophages in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Protein expression analysis in neutrophils

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CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was detectable in neutrophils in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Protein expression analysis in Bone marrow

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CD89 protein expression analysis in heterozygous and homozygous B-hIGHA1/hCD89 mice by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H), and then analyzed by flow cytometry with anti-hCD89 antibody. hCD89 was detectable in T, B, NK cells, DCs, monocytes, macrophages and neutrophils in B-hIGHA1/hCD89 mice (H/H, H/+) and B-hIGHA1/hCD89 mice (H/H, H/H).

Analysis of leukocytes cell subpopulation in bone marrow

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Analysis of bone marrow leukocyte subpopulations by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6 mice (+/+), heterozygous B-hIGHA1/hCD89 mice (H/H, H/+) and homozygous B-hIGHA1/hCD89 mice (H/H, H/H). Flow cytometry analysis of the bone marrow cells 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. Percent of B cells was decreased in B-hIGHA1/hCD89 mice compared to that in C57BL/6 mice. Values are expressed as mean ± SD.

Analysis of blood and urine

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Blood and urine analysis: Four 30-weeks-old male B-hIGHA1/hCD89 mice(H/H,H/H) and C57BL/6N were selected to assess the blood and urine indicators. The results showed that the levels of creatinine (CREA) and urine protein in the urine, as well as blood urea nitrogen (UREA) and creatinine (CREA), did not show significant differences compared to wild mice.

Analysis of kidney tissue sections

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Analysis of kidney tissue sections: Four 30-week-old male B-hIGHA1/hCD89 mice(H/H,H/H) and C57BL/6N were selected for kidney tissue section immunohistochemical staining analysis. The results showed a slight increase in CD11b staining in the B-hIGHA1/hCD89 mice(H/H,H/H), while IgA and C3 staining showed no significant differences between B-hIGHA1/hCD89 mice(H/H,H/H) and C57BL/6N.

FAQ

1. Q: Why do the results show that the serum IgA levels are lower in CD89 homozygous mice compared to CD89 heterozygous mice?
A: It is speculated that the increase in hCD89 expression may lead to a higher occurrence of IgA-sCD89 complexes, resulting in reduced serum hIgA levels.
2. Q: Have the expression profiles of IgA and CD89 been assessed?
A: We are about to conduct this detection using newborn mice (born on August 18).
3. Q: Is Gd-IgA present in the mice?
A: We will also check whether Gd-IgA is present in the serum of the newborn mice.
4. Q: Have the T cell and B cell populations in these mice been affected?
A: We will also perform T and B cell phenotype analysis using the newborn mice.
5. Q: This mouse does not have the IgAN phenotype. What is your next development plan?
A: The lack of phenotype in this mice may be due to low hCD89 expression levels and inconsistencies with the expression profile reported in the literature, as well as relatively low serum IgA levels. Therefore, we have developed a new mouse modle, anticipating that both hCD89 and hIgA expression levels can be improved. We expect to have heterozygous mice born in January 2025, and we plan to perform phenotype validation once the mice reach 12 weeks of age. We anticipate new progress in May 2025, and we appreciate your patience.
6. Q: Is induction required during the mouse breeding process?
A: No, it is not necessary. The article shows that α1KI-CD89 mice raised in an SPF environment can develop the disease by 12 weeks of age..