1.Donor dendritic cells treated with B7- 1, B7- 2 antisense oligonucleotide prolonged mouse cardiac allograft survival
Xiaoyan LIANG ; Zongyou CHEN ; Shiguang QIAN ; Shunong LI
Chinese Journal of Pathophysiology 2000;16(12):1249-1254
AIM:To investigate the effect of donor bone marrow derived dentritic cell (DC) treated with B7 - 1, B7 - 2 antisense oligonucleotide on mouse heart allografe survival time and its mechanism. METHODS: There were 7 groups of C57BL/10J (B10) mouse bone marrow DCs which were treated by 400 nM antisense oligonucleotide target to B7 -1, B7 -2 mRNA (AS B7- 1/2), B7- 1 mismatch oligo control ,B7- 2 mismatch control(mASB7- 1/2), lipofeetamine only and non-treatment, respectively. Each group of DC were named as ASB7- 1 DC, ASB7- 2 DC, mASB7 - 1 DC, mAS B7 - 2DC, and Lipo DC, respectively. RESULTS: Flow cytometer results shown that AS B7- 1/2 can inhibit B7- 1 (CD80)and B7- 2 (CD86) molecule express on DC surface, while control groups have no effects. To observe their tolerogenicity in mouse cardiac allograft model, B10→C3H heterotopic heart transplantation were performed. Recepients were received 2 x 106 of DC injection 7 days before transplantation. Results showed that both AS B7 - 1 DC and AS B7 - 2 DC can prolong mouse cardiac allograft survival time to (18.6 + 0.89) days and (23.67 + 10.73) days, respectively, compared with IL - 4 DC [ (6.22 + 0.97) days ( P < 0.01 ) ]. Two mismatch control groups can slightly prolong while oligo DC has no effect. For understanding its mechanism, each group of DC was used as stimulator to stimulated C3H spleen T cell. Results suggested that AS B7 - 1DC and AS B7 - 2 DC had less allo - stimulate function, including MLR and generation CTL and IL - 2 production than IL - 4 DC but control groups have no effect. CONCLUSION: Donor bone marrow derived DC treated with AS B7 - 1 oligo and AS B7 - 2 oligo expressed lower level of CD80 and CD86, respectively. These cells can induce allogeneic T cells anergy in vitro and markedly prolong mouse heart allograft survival time in vivo.
2.Donor dendritic cells treated with B7-1,B7-2 antisense oligonucleotide prolonged mouse cardiac allograft survival
Xiaoyan LIANG ; Zongyou CHEN ; Shiguang QIAN ; Shunong LI
Chinese Journal of Pathophysiology 2000;0(12):-
AIM:To investigate the effect of donor bone marrow derived dentritic cell (DC) treated with B7-1, B7-2 antisense oligonucleotide on mouse heart allografe survival time and its mechanism. METHODS: There were 7 groups of C57BL/10J (B10) mouse bone marrow DCs which were treated by 400 nM antisense oligonucleotide target to B7-1, B7-2 mRNA (AS B7-1/2), B7-1 mismatch oligo control ,B7-2 mismatch control(mASB7-1/2), lipofectamine only and non-treatment, respectively. Each group of DC were named as ASB7-1 DC, ASB7-2 DC, mASB7-1DC, mAS B7-2DC, and Lipo DC, respectively.RESULTS: Flow cytometer results shown that AS B7-1/2 can inhibit B7-1(CD80)and B7-2 (CD86) molecule express on DC surface, while control groups have no effects. To observe their tolerogenicity in mouse cardiac allograft model, B10→C3H heterotopic heart transplantation were performed. Recepients were received 2?106 of DC injection 7 days before transplantation. Results showed that both AS B7-1DC and AS B7-2 DC can prolong mouse cardiac allograft survival time to (18.6?0.89) days and (23.67?10.73) days, respectively, compared with IL-4 DC [(6.22 ?0.97) days(P
3.Macrophage IL-33 expression in macrophages of human chronic periapical periodontitis and apical cyst
DAI Qian ; YU Yuanling ; HUANG Shiguang
Journal of Prevention and Treatment for Stomatological Diseases 2019;27(5):300-303
Objective:
To observe the expression of interleukin-33 (IL-33) in macrophages of chronic periapical periodontitis and apical cyst tissue, and to provide a basis for the study of the pathogenesis of IL-33 in periapical diseases.
Methods :
The apical tissues of 20 normal control group, 15 chronic periapical periodontitis group and 15 apical cyst group were collected for HE staining and optical microscopy respectively. CD14 was used as the marker of macrophages and double immunofluorescence staining was used to observe the changes of periapical tissues under fluorescence microscopy. The expression of IL-33 in CD14-positive macrophages was observed.
Results:
The macrophage density (cell/mm 2) of IL-33 and CD14 positive expression in normal control group, chronic periapical periodontitis group and root cyst group were(23.81 ± 5.16,62.97 ± 8.54,119.83 ± 14.61) respectively, and there were significant differences among the three groups(F=87.17,P < 0.01). The density of IL-33 and CD14 positive macrophages in root cyst group was significantly higher than that in chronic periapical periodontitis group and control group(P < 0.01).
Conclusion
IL-33 and CD14 positive macrophages increased in normal apical tissue, chronic periapical periodontitis tissue and apical cyst tissue in turn.
4.Expression and relevance of long non-coding RNA MIAT in CD4+T cells in peripheral blood of gastric cancer patients
Min SHA ; Jingyu QIAN ; Chuanmeng ZHANG ; Zhiyi CHENG ; Yabin ZHU ; Shiguang JIN
Chinese Journal of Laboratory Medicine 2022;45(9):921-929
Objective:To explore the expression of long-chain noncoding RNA (lncRNA) and myocardial infarction-associated transcription (MIAT) in Leukocyte differentiation antigen (CD)4+T cells in peripheral blood of gastric cancer patients and its value of clinical application.Methods:Peripheral blood CD4+T cells were collected from 124 patients with gastric cancer, 90 benign gastric diseases patients and 80 healthy controls enrolled in Taizhou People′s Hospital from January 2019 to April 2021. The expression levels of MIAT and N6-methyladenosine(m6A) binding to MIAT promoter in CD4+T cells were detected by real-time fluorescent quantitative polymerase chain reaction (qPCR) and Chromatin immunoprecipitation (ChIP)-qPCR, respectively. Spearman test was used to analyze the correlation between MIAT and clinicopathological features, as well as between MIAT and regulatory T cell levels. The receivor operating characteristic curve (ROC) of the subjects was used to evaluate the MIAT expression level in the auxiliary diagnostic value of gastric cancer.Results:The relative expression levels of MIAT in the gastric cancer patients, the benign gastric diseases patients, and the healthy controls were 2.849 (2.131, 4.062), 1.511 (0.916, 1.855) and 0.963 (0.729, 1.432), respectively. The difference among the three groups was statistically significant ( H=158.25, P<0.001). The relative expression level of MIAT in the gastric cancer patients was significantly higher than the levels in the benign gastric diseases patients and healthy controls. The difference was statistically significant ( Z=100.63, 145.14, P<0.001). The binding activity of m6A to MIAT promoter in patients with early stage (stage Ⅰ and Ⅱ) and end stage (stage Ⅲ and Ⅳ) gastric cancer was 8.590±1.483 and 4.274±0.425, respectively. The difference was statistically significant ( t=6.255, P=0.002). Furthermore, the binding activity of m6A to MIAT promoter in the gastric cancer patients was significantly lower than that in patients with benign gastric diseases (17.267±3.106) and healthy controls (27.637±3.945) ( t=-7.331,-12.832, P<0.001). The relative expression of MIAT in CD4+T cells in peripheral blood of the gastric cancer patients had no significant difference in age(χ2=0.000, P=1.000), gender(χ2=0.000, P=1.000), CEA (χ2=0.648, P=0.421) and CA199(χ2=1.554, P=0.213), but had significant difference with tumor size expression(χ2=9.443, P<0.01), TNM stage(χ2=23.571, P=0.002) and lymph node metastasis (χ2=45.248, P<0.01). In addition, there was a significant positive correlation between the relative expression of MIAT in CD4+T cells and Treg level ( r2=0.76, P<0.001). The diagnostic efficacies of MIAT in CD4+T cells, CEA and CA199 in the gastric cancer patients were analyzed by ROC curve. When compared with patients with benign gastric diseases, the areas under the curve were 0.879, 0.635 and 0.611, respectively. When compared with healthy patients, the areas under the curve were 0.953, 0.784 and 0.598, respectively. Conclusions:The level of MIAT in CD4+T cells in peripheral blood of patients with gastric cancer is significantly higher than the levels in patients with benign gastric diseases and the healthy controls, which may be related with the decreased activity of m6A binding to the promoter of MIAT. The level of MIAT in CD4+T cells may be a relevant biomarker for the diagnosis and prognosis of gastric cancer.