1.The Production and Characterization of Monoclonal Antibody to Human Chorionic Gonadotropin(hCG)
Chinese Journal of Immunology 1985;0(06):-
Splenocytes from BALB/c mice immunized with human chorionic gonadotropin(hCG) were fused with myeloma cells NS-1, and 7 hybridomas producing monoclonal antibodies to hCG were established. By using RIA to measure the binding characteristics of these monoclonal antibodies to ~(125)I-labelled hCG, their subunits, and 4 glycoprotein pitiutary hormones, 7 antibodies had been devided into 3 groups. Of these groups one(4 antibodies included) recognized intact hCG and ?-subunit, a second(one antibody only) recognized intact hCG and ?-subunit, and the third(2 antibodies included) bound to the hCG molecule only, The precipitation after double diffusion in agar with hCG antigen demonstrated that 4 antibodies of group 1 recognized 3 epitopes, and 2 antibodies of group 3 recognized different epitopes. The cooperation between pairs of antibodies specific to different epitopes in binding hCG antigen showed that most of these antibodies cooperated each other. The antibody No 10 was showed to be the best, cooperating with every other antibody,where as antibody No 22 was cooperated only with a few antibodies.The coorperation was related to the differency of epitopes recognized, but seemed not to bear relationship with the titre of antibodies.
2.Prevention of experimental autoimmune disease by T-cell vaccination
Qiang TIAN ; Lan TAIN ; Jufen PAN
Chinese Journal of Immunology 1985;0(02):-
In this paper, we described that using wistar rat rather than Lewis rat, an animal model ofexperimental autoimmune encephalomyelitis (EAE) was established. 12 out of 17 rats showedtypical clinical syndroms and pathological changes, and also developed significant delayed-typeskin hypersen-sitivity reaction to the pathogenic antigen-homogenate of guinea pig spinal cord.The cytotoxicity assay demonstrated that the T-cells of EAE rats can specifically lyse the braintissue of normal rats with specific release rate of 15.1l?t 2.79% which is positive correlativewith clinical severity of EAE. The T cell vaccine was prepared by activating EAE T-cells withConA and then treating with glutaraldehyde. Normal rats were inoculated with this T cell vac-cine and challenged with pathogenic antigen one week later. The result showed that all seven ratsgot complete resistance to EAE. T cells from vaccinated animals can specifically lyse the EAE Tcells with specific release rate of 9.38?1.68%.These results indicated: (a)wistar rat can be used for EAE animal model; (b)The incidence ofEAE is associa ted with the auto-reactive T cellsof EAE animals; (c)Inocula tion of this T cellvaccines can prevent the development of EAE, and this protective effect is also mediated by Tcells.
3.The inhibition effect of human chorionic gonadotropin(hCG) on mRNA expression of cytokines initiated inflammatory reaction
Hong BAI ; Jufen PAN ; Xiaoqing JIA ; Al ET
Chinese Journal of Immunology 1985;0(03):-
Objective:To investigate the effect of hCG on the gene expression of cytokines(TNF??IL 1 and IL 6) that initiate inflammatory reaction protected against bacteria.Methods:Human PBMC were cultured with LPS(22 ?g/ml) and different doses of hCG(100,50,25,12 5,6 25,3 125 U/ml) at 37℃,5%CO 2 for 4 h,mRNA of cells harvested were isolated,then run RT PCR with primer of TNF??IL 1 and IL 6 separately.The PCR product were quantitated by using MIAS 2000 analyzer,calculated the OD value.Results:Compared with the control group,the result showed that hCG in a dose range of 50~6.25 U/ml have significantly suppressive effect on TNF? mRNA expression ( P
4.Bone marrow mesenchymal stem cells from systemic lupus erythematosus mice have reduced osteogenic and adipogenic abilities
Guangping RUAN ; Jinxiang WANG ; Jianyong YANG ; Jufen LIU ; Xuemin CAI ; Rongqing PANG ; Yanbo LV ; Xinghua PAN
Chinese Journal of Tissue Engineering Research 2014;(1):1-6
BACKGROUND:There are less studies addressing whether bone marrow mesenchymal stem cells from systemic lupus erythematosus patients are different from healthy people.
OBJECTIVE:To compare the multi-differentiation capacity of bone marrow mesenchymal stem cells isolated from systemic lupus erythematosus model mice and normal control mice.
METHODS:Bone marrow mesenchymal stem cells of systemic lupus erythematosus model mice and C57BL mice were isolated and cultured fol owed by osteogenic and adipogenic differentiations, respectively. Differentiation abilities of two kinds of mouse bone marrow mesenchymal stem cells were observed.
RESULTS AND CONCLUSION:Passaged bone marrow mesenchymal stem cells from C57BL mice were long spindle-shaped and evenly distributed, while bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice showed slow growth and were relatively smal er than those from C57BL mice. After osteogenic induction, the amount of calcium salt and calcium nodules were significantly less in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice than from normal control mice. PCR detection showed that expressions of Runx2, alkaline phosphatase and osteocalcin were also significantly decreased in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice. After adipogenic induction, the number of lipid droplets in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice was significantly less than the control group, and PCR detection also showed significantly decreased expression of adipogenic genes, including PPARγ2 and lipoprotein lipase. These findings suggest that the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice exhibit lower osteogenic and adipogenic capacities than those from normal C57BL mice, and also have the impaired multi-differentiation ability.
5.Role of human umbilical cord mesenchymal stem cells:cell transplantation, immuoregulation and target cells
Guangping RUAN ; Xiang YAO ; Jufen LIU ; Fan SHU ; Jie HE ; Jianyong YANG ; Rongqing PANG ; Xinghua PAN
Chinese Journal of Tissue Engineering Research 2014;(41):6714-6718
BACKGROUND:Umbilical cord as childbirth waste has wide variety of sources and can be easily obtained, without any ethical and legal restrictions. Therefore, human umbilical cord mesenchymal stem cells break al the restrictions originated from other sources of mesenchymal stem cells. OBJECTIVE:To review the application of human umbilical cord mesenchymal stem cells in cartilage diseases, neuroglioma, ischemic brain injury, lung disease, liver disease and models of myocardial infarction. METHODS:The PubMed and Wanfang databases were searched by the first author using the keywords of“human umbilical cord mesenchymal stem cells, disease models, celltherapy”in English and Chinese, respectively. Seventy-three articles were searched and final y, 35 articles were included in result analysis. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells have multilineage differentiation capacity similar to bone marrow mesenchymal stem cells. Compared with bone marrow mesenchymal stem cells, human umbilical cord mesenchymal stem cells have lower immunogenicity. Human umbilical cord mesenchymal stem cells show certain curative effects on cartilage disease, neuroglioma, ischemic brain injury, lung disease, liver disease and myocardial infarction, indicating that human umbilical cord mesenchymal stem cells can be used for celltransplantation to treat various diseases.
6.Stem cells protein microarray analysis after chicken egg-white extract induced cells
Guangping RUAN ; Xiang YAO ; Jufen LIU ; Jinxiang WANG ; Zian LI ; Yuanyuan HU ; Xinghua PAN
Chinese Journal of Immunology 2015;(9):1217-1220
Objective:To analyze the chicken egg-white extracts were co-cultured with cells whether elevated stem cells protein,whether the cells transformation into stem cell.Methods:Four kinds of cells,making a common culture,a 50% chicken egg-white extract co-cultured for 3 days,cells were collected and frozen at -80 degrees,sending the company to do stem cell protein microarray.Results:C57-BMSC has three proteins occurred statistically significant change , TS-UC-MSC has one proteins occurred a statistically significant change ,293T has one protein occurred a statistically significant change ,and 293T-GFP has one protein occurred a statistically significant change.Conclusion:50% chicken egg-white extract co-cultured cells,the cells occurred the phenomenon of transformation into stem cells.
7.Different components of chicken egg-white extracts affect cell cycle and apoptosis
Guangping RUAN ; Xiang YAO ; Jufen LIU ; Jinxiang WANG ; Rongqing PANG ; Xinghua PAN
Chinese Journal of Tissue Engineering Research 2015;(46):7451-7455
BACKGROUND:If an extract can prolong the S phase and reduce the percentage of apoptosis after co-culture with cels, it is proved that the extract is able to promote cel proliferation. OBJECTIVE: To prove the effects of chicken egg-white extracts with different components on the proliferation, cel cycle and apoptosis of 293T cels. METHODS: An ultrafiltration unit was used to separate chicken egg-white extracts into different components that were > 10 ku, < 10 ku, > 3 ku and < 3 ku to co-culture with cels for 3 days. Then, cel proliferation, cel cycle and cel apoptosis were detected. RESULTS AND CONCLUSION:Chicken egg-white extract components of < 10 ku and < 3 ku could promote cel proliferation, increase the percentage of S-phase cels and reduce the percentage of apoptosis. In conclusion, chicken egg-white extract components that are < 10 ku and < 3 ku can promote cel proliferation, as wel as increase the percentage of S-phase cels and reduce apoptosis percentage.
8.Cross-reactivity of tree shrew sera with various secondary antibodies:extensive application to tree shrew models of diseases
Guangping RUAN ; Xiang YAO ; Jufen LIU ; Jinxiang WANG ; Jie HE ; Jianyong YANG ; Xinghua PAN
Chinese Journal of Tissue Engineering Research 2015;(5):794-798
BACKGROUND:Tree shrew is a representative between insectivore and primates, has a high degree of evolution, is more inexpensive primates, has high use of medical biology, and has been attached by scholars. OBJECTIVE:To detect whether the commonly used secondary antibodies have immune response with tree shrew serum. METHODS:Western blot assay and enzyme linked immunosorbent assay were utilized to detect whether the tree shrew serum had cross-reacts with anti-rabbit, anti-goat, anti-human, anti-mouse, anti-rat, and anti-monkey secondary antibodies. RESULTS AND CONCLUSION:Western blot assay results indicated that tree shrew serums did not react with anti-rabbit, anti-goat, anti-human, anti-mouse, and anti-rat secondary antibodies and had cross reaction with anti-monkey secondary antibody. Enzyme linked immunosorbent assay results also indicated that tree shrew serums were cross-reactive with anti-monkey secondary antibody, but did not have cross-reactivity with the other secondary antibodies. Above data confirmed that the usual y soled secondary antibody cannot be used to immunoassay with tree shrews IgG. Only anti-monkey secondary antibody has cross-react with tree shrew serum. It is necessary to prepare anti-tree shrew IgG monoclonal and polyclonal antibodies. When no antibody is readily available at present, anti-monkey secondary antibody can be used to substitute detection, and can be widely applied in the study of tree shrew models of disease.
9.Tree shrew umbilical cord mesenchymal stem cells:isolation, cultivation and osteogenic and adipogenic differentiation
Guangping RUAN ; Lu ZHU ; Jufen LIU ; Zian LI ; Jinxiang WANG ; Rongqing PANG ; Xinghua PAN
Chinese Journal of Tissue Engineering Research 2017;21(9):1373-1377
BACKGROUND: Studies have shown that umbilical cord mesenchymal stem cells are ideal seed cells for tissueengineering research.OBJECTIVE: To isolate, culture and identify tree shrew umbilical cord mesenchymal stem cells, in order toestablish a standardized tree shrew umbilical cord mesenchymal stem cell lines.METHODS: Caesarean-isolated tree shrew umbilical cord samples were used to isolate and culture umbilical cordmesenchymal stem cells using tissue explant adherent method. Flow cytometry assay was used to detect cellsurface markers. Osteogenic and adipogenic induction media were used to induce umbilical cord mesenchymalstem cells to differentiate into osteoblasts and adipocytes.RESULTS AND CONCLUSION: The cultured umbilical cord mesenchymal stem cells expressed CD90 and CD105 with the positive rate of 99.9% and 99.8% respectively. Hematopoietic stem cell marker CD34 expression ratewas 0.0% and the endothelial cell marker CD31 expression rate was 0.7%, in line with the characteristics of umbilicalcord mesenchymal stem cell surface markers. Calcium nodules by alizarin red staining and lipid droplets by oil red Ostaining were observed in the induced cells. These experimental findings indicated that umbilical cord mesenchymalstem cells from tree shrews capable of osteogenic and adipogenic differentiation were successfully isolated and cultured.
10.Establishment of a Hybridoma Cell Line Secreting High Titer of Anti-Human Thyroglobulin Monoclonal Antibodies
Yanhua YAN ; Enjiang TIAN ; Shuyou CHONG ; Yin XUE ; Xiaoxi HU ; Jufen PAN ; Wufei ZHENG
Chinese Journal of Immunology 1985;0(05):-
A hybridoma cell line(B10)was established by fusing spleen cells of BALB/c mou- se immunized with human thyroglobulin(HTG) with SP2/0 myeloma cells. An averagefusing rate of 6.3% and antibody-secreting positive well rate of 58.3% were obtained.During the first two months, the supernatant of B10 culture had a titer of 1/128 to1/2048 measured by hemagglutination method, and the ascites was positive at 1/64000-128000 and 1/320000 respectively as measured by hemagglutination and radioimmunoass-ay.The B10 cell line is very stable and has very high activity to produce anti-HTGmonoclonal antibodies. After several times of preservation in liquid nitrogen andpassage in culture for one year,a recent determination shows that cell culture super-natant and ascites still have very high titer,being 1/4096 and 1/1048576 respectively asmeasured by hemagglutination method. The chromosome number of the B10 hybridomacell is 99.5?7.4.The success of the establishment of this cell line is briefly discussed.Attempt to establish diagnostic kit with this monoclonal antibody is being undertaken.