Influence of different doses of X-ray irradiation combined with allogeneic lymphocyte infusion to establish a mouse model of aplastic anemia
10.19405/j.cnki.issn1000-1492.2025.08.004
- VernacularTitle:不同剂量 X 射线联合异体淋巴细胞输注对建立再生障碍性贫血小鼠模型的影响
- Author:
Xiaoxiao Zhou
1
;
Caixian Xu
2
;
Guiqin Wang
3
;
Qiang Hong
4
;
Qianshan Tao
1
;
Cong Li
1
;
Huiping Wang
5
;
Zhimin Zhai
1
Author Information
1. Dept of Hematology, The Second Afiliated Hospital of Anhui Medical University, Hefei 230601
2. Dept of Hematology, The Second Afiliated Hospital of Anhui Medical University, Hefei 230601;Cellzoy Biotech Co. , Ltd , Hefei 230601
3. Cellzoy Biotech Co. , Ltd , Hefei 230601
4. School of Basic Medicine , Anhui Medical University,Hefei 230032
5. Dept of Hematology, The Second Afiliated Hospital of Anhui Medical University, Hefei 230601Dept of Hematology, The Second Afiliated Hospital of Anhui Medical University, Hefei 230601
- Publication Type:Journal Article
- Keywords:
aplastic anemia;
allogeneic lymphocyte;
X⁃ray;
peripheral blood;
survival rate;
bone marrow;
immunity
- From:
Acta Universitatis Medicinalis Anhui
2025;60(8):1387-1394
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of different doses of X ⁃rays irradiation combined with allogeneic lymphocyte infusion on the establishment of aplastic anemia in mice.
Methods:Forty BALB/c mice were randomly divided into four groups : the 3 Gy group (n = 9) , the 4 Gy group (n = 9) , the 5 Gy group (n = 10) , and the con⁃trol group (n = 12) . In the 3 Gy , 4 Gy , and 5 Gy groups , the experimental mice were exposed to corresponding do⁃ses of X ⁃ray and then intravenously infused with 0. 2 mL mixed suspension of the thymus and spleen cells from DBA/2 mice , at a concentration of 1 × 107 cells/mL , within 4 hours after irradiation. The control group did not un⁃dergo X ⁃ray irradiation and infused with an equivalent volume of physiological saline instead. Blood samples were collected from the orbital venous plexus of BALB/c mice and analyzed using an animal automated hematology analy⁃zer to measure peripheral blood parameters , including red blood cells ( RBC) , white blood cells ( WBC) , and platelets (PLT) . The general condition of mice was monitored daily , and survival rates were recorded for each group. At the experimental endpoint , the tibias were harvested for hematoxylin and eosin (HE) staining , while the femurs were used to prepare bone marrow smears for morphological examination. For the 5 Gy group , T ⁃cell subsets(ELISA) at the endpoint.
Results :In the 3 Gy group , pancytopenia was observed , but platelet recovery occured rapidly , returning to normal levels by day 17 post⁃modeling. No deaths occurred during the observation period. At myeloid⁃to⁃erythroid (M/E) ratio , and no significant morphological abnormalities were noted in cells at any devel⁃with hematopoietic cells. In the 4 Gy group , pancytopenia persisted throughout the observation period. The survival rate was 90% . Endpoint analysis showed hypocellular marrow by morphological examination. HE staining indicated minimal fatty infiltration in the bone marrow tissue. In the 5 Gy group , pancytopenia was observed , though erythro⁃cyte counts returned to normal levels by day 24. The survival rate during the observation period was 50% . Endoint analysis revealed vacuolization of marrow particles and reduced hematopoietic cells with predominantly non⁃hematopoietic cells in bone marrow morphology. HE staining demonstrated severe fatty infiltration in the bone mar⁃row tissue , with scarcity of immature cells and hematopoietic precursor cells. Flow cytometry analysis showed a de⁃creased proportion of CD4 + T cells (% ) and an increased proportion of CD8 + T cells (% ) . ELISA confirmed elevated secretion of negative hematopoietic regulators : interferon⁃gamma ( IFN⁃γ) and tumor necrosis factor⁃alpha (TNF⁃α ) .
Conclusion:Combined administration of varying radiation doses with allogeneic lymphocyte infusion consistently induced peripheral blood cytopenia in mice , characterized by reductions in RBC , WBC , and PLT counts. Integrated analysis of bone marrow morphology , histopathological assessment via HE staining , and immuno logical parameters confirmed that a mouse model of aplastic anemia can be successfully established using 5 Gy X ⁃ ray irradiation coupled with infusion of 2 × 106 allogeneic lymphocytes.
- Full text:2026040323014742196不同剂量X射线联合异体淋巴细胞输注对建立再生障碍性贫血小鼠模型的影响_周晓晓.pdf
