[Changes in the expression of T lymphocyte subsets in ageing mice induced by D-galactose].

AIM: To explore the changes and significance of spleen T cell subsets in ageing mice induced by D-galactose. METHODS: Ageing mice model was successfully established by 100 g/L D-galactose. The content of IFN-γ and IL-4 in serum was measured by ELISA. T cell subsets were detected by Immunofluorescence technique and flow cytometry. RESULTS: The content of IFN-γ and IL-4 in the serum was decreased of model group (P<0.01). The naive T cell related molecule, CD45RA was decreased(P<0.05). T cell activation-related molecule, CD25 was decreased(P<0.05). The Foxp3 in CD4(+);CD25(+); T cell was increased(P<0.01). CONCLUSION: In spleen of the ageing mice, the percentage of naive and active T cell are decreased, but The percentage of CD4(+); Tr subset is increased.

[Changes and effects of significant membrane molecules on thymic T cell of aging mice induced by D-galactose].

AIM: To establish subacute aging mice model by D-galactose and to explore the changes and effects of significant membrane molecules on thymic T cell. METHODS: Female Kunming mice of 8 weeks old were injected with D-galactose of 12.5 mL/(kg.d) by subcutaneous in scruff for 42 days. The animals' living conditions and biological behaviors were observed everyday.SOD activities and MDA content of serum were measured to determine whether the aging model was successfully established.On the basis of successfully establishing aging model, detect the significant membrane molecules of thymic T cell by Immunofluorescence technique and Flow Cytometer. RESULTS: During the 42 days, gradually, the model mice showed bending body, loose skin, slow action and so on.The activities of SOD in the serum were significantly decreased(P<0.01), and the content of MDA in the serum was significantly increased(P<0.01). The thymic naive T cell significant molecule, CD45RA was decreased(P<0.05). T cell activation-related molecules, CD28 and CD25 were both decreased(P<0.05), and PD-1 was significantly increased(P<0.01). The memory T cell significant molecule, CD196 was increased, but was not significantly compared to the control mice. CONCLUSION: The D-galactose subacute aging mice model was successfully established.The naive and active T cell were decreased and the memory T cell was increased in the thymic of the aging.

[The changes of spleen B cells of D-galactose-induced aging mice].

AIM: This paper is to analyze the changes of important membrane type molecules of the spleen B cells and their significance on the basis of biology identification by establishing subacute aging mice model with D-galactose. METHODS: Health Kunming mice are injected with 100 ng/L D-galactose, 0.25 mL/20 g, once per day, for 42 days consecutively, into their back necks. The animals' living conditions and biological behaviors are observed everyday and the dynamic changes of their weight are measured regularly. The biology identification of aging model mice is conducted by means of measuring the SOD viability and malondialdehyde (MDA) concentration in serum; the analysis of the activation- and memory-related important membrane type molecules of spleen cells is carried out with immune fluorescence and flow cytometry; the analysis of the concentration of IL-4 in serum is conducted by ELISA. RESULTS: Building a successful subactue aging mice model.The results of immune fluorescence and flow cytometry showed that CD20(+); was 56.8%, CD40(+); was 43.6%, CD20(+);CD45RA(+);B was 14.04%, CD40(+);CD45RA(+);B was 35.4%, CD20(+);CD86(+);B was 2.25%, CD40(+);CD86(+);B was 4.38%, CD20(+);CD196(+);B was 10.68%, and CD40(+);CD196(+);B was 10.98%. The results of ELISA showed that the average level of IL-4 in serum was 7.93 ng/L. The statistical analysis showed that the expressions of CD20, CD40, CD45RA, and CD86 of the spleen cells of the model control group and the average level of IL-4 in serum were lower than the normal control group(P<0.05)while CD196 was higher than the normal control group(P<0.01). CONCLUSION: In the body's aging process, the expressions of the activation- and memory-related important membrane type molecules of spleen cells change, activated cells reduce, memory cells increase, and the expression of IL-4 in serum drop, resulting in the immune system function disorder.

[Construction of human CXCR3B gene transfected cell line, identification and study of its biological function].

AIM: To construct recombinant human CXCR3B gene expression vector and obtain L929-CXCR3B gene transfected cell line for stably expressing human CXCR3B. METHODS: Human CXCR3B gene of full length was amplified by PCR from the plasmid pMD19-T/huCXCR3A. Then, it was inserted into eukaryotic expression vector pIRES2-EGFP to construct recombinant vector pIRES2-EGFP/huCXCR3B. The recombinant vector was transfected into L929 cells with Lipofectamine(TM); 2000, and cell clones stably expressing human CXCR3B molecule were screened by G418.We used FCM and RT-PCR to verify expression of CXCR3B from protein level and gene level. The ability of proliferation of L929-huCXCR3B under the circumstance of CXCR3B ligand called Mig was analyzed via MTT methods. RESULTS: We have constructed recombinant human CXCR3B gene expression vector and obtained L929-huCXCR3B gene transfected cell line which can stably express human CXCR3B molecule. The positive expression rate of CXCR3B on L929-huCXCR3B cells was 93&. The result of MTT assay showed that the proliferation of L929-huCXCR3B cells were inhibited when the cells were cocultured with Mig after 24 h, 48 h and 72 h, and the inhibition ratio were 41.44&, 44.01& and 24.80& respectively. CONCLUSION: Construction of L929-huCXCR3B cell line has laid a good foundation on research of the CXCR3B signal pathway and preparation of the CXCR3B monoclonal antibody.