Objective To explore the effect of cytotoxic T lymphocyte-associated antigen 4 (CTLA4-Ig) fusion protein on the function of orthotopic liver allograft. Methods Orthotopic liver allograft models of rhesus monkeys were established in this study. The survival time, liver function and morphologic changes of graft were observed, respectively. The levels of IL-2 and IL-10 were detected by ELISA. Apoptosis was monitored by TUNEL.Results The average survival of control group was 6.57 d, while the average survival of CTLA4-Ig group was 14.92 d, which was statistically prolonged (Plt;0.05). Serum ALT level was highly increased, and Alb level decreased obviously in control group. While the levels of ALT and Alb kept in normal in CTLA4-Ig group. After day 3-7, the expressions of IL-2 were highly expressed in control group, while the expressions of IL-10 in CTLA4-Ig group were higher than those of control group. The severity of rejection reaction after day 3 was weaker in CTLA4-Ig group than that of control group by histological assessment. The apoptosis index after day 3 in the liver cells was highly increased in control group as compared with the CTLA4-Ig group. Conclusions CTLA4-Ig fusion protein therapy can induce immunotolerance and prolong the survival of recipients. The increasing of cytokines IL-2 or the decreasing of cytokines IL-10 may be one of the laboratory indexes in monitoring immunotolerance of transplantation.
【Abstract】Objective This study was conducted to build experimental model of orthotopic liver transplantation in rat (ROLT) with the character of acute rejection; and to study the effect of cytotoxic T lymphocyte antigen 4 immunoglobulin G (CTLA4Ig) on prevention of rejection and the induction of immune tolerance of ROLT. Methods Build model of Wistar→SD ROLT(performed by the two cuff method) with character of acute rejection.Recipients were injected with CTLA4Ig 75 μg per ROLT into abdominal cavity after 2 days of operation. Contrast was made with no treatment group, the clinical characters, the liver function, the transplantated liver pathologic character and the concentrations of TNFα in serum were observed and measured on postoperative day 7. In treatment group, all above observation were done on postoperative month 4. Above all, determination of the effect of CTLA4Ig on preventing acute rejection and inducing tolerance in ROLT was observed.Results ①Recipients (no treatment group) died one by one within 6th～14th days; pathologic character of rejection in transplantation liver could be found; ② In treatment group, on postoperative day 7 and month 4, no clinical rejection character and no pathologic character of rejection in transplantation liver were found and serum concentration of cytokins related to TNFα found lower than that of contrast group(P＜0.05), and serum concentration of ALT、AST、TBIL、DBIL found lower too(P＜0.05); But serum concentration of TP and Alb was found higher than that of contrast group(P＜0.05). Conclusion ① CTLA4Ig treatment alone inhibits the rejection responce in ROLT. ② CTLA4Ig treatment can Prevent rejection and induce immune tolerance in ROLT model with characters of acute rejection; the serum concentration of cytokins related to TNFα can probably be used as evaluation standard of rejection in ROLT rejection.
ObjectiveTo investigate the change of cellmediated immunity in gut mucosa after major hepatectomy and to study its relationship with the bacteria translocation.MethodsFortyeight Spraguedawley adult male rats were randomly allocated into two groups, the sham operation group and the operation group. Besides without the hepatectomy, the sham operation group has the same course with the operation group. Seventy percent hepatectomy rats are divided as postoperative 6 h group (n=6)，12 h group (n=6)，24 h group (n=6) and 72 h group (n=6). Sixhour, 12hour, 24hour and 72hour after operation specimens were taken from jejunoileum respectively. Immunohistochemical staining was performed on frozen sections and image pattern analysis was used. We also investigate the change of liver function. ResultsTwentyfour hours and 72 hours after 70% hepatectomy, there was a significant reduction in the number of CD3+，CD4+and CD8+ T lymphocytes in the mucosal lamina propria of the operation group compared with the sham operation group （Plt;0.05）. There was significant difference between these two groups in liver function change （Plt;0.05）.ConclusionThere is an altered pattern of intestinal mucosal T lymphocytes after major hepatectomy, then the local cellmediated immunity was depressed, which may be the cause of translocation of enteric bacteria.
Objective To explore the effect of CD3AK cells on T lymphocyte subsets and its functions of patients with primary liver carcinoma (PLC). MethodsFiftyeight patients with PLC were divided into two groups, CD3AK group (n=37), control group (n=21). Eleven blood donors were taken as normal control. All patients from the treatment group were given 2×109 CD3AK cells once a day for 5 days on the 1st day after operation, having received IL2 at a dose of 100 units per 24 hour by intravenous injection starting 30 minutes before administration of CD3AK cells. T lymphocyte subsets, IL2R, NK activity, LAK activity and proliferative activity were determined. ResultsThe CD4/CD8 ratio, expression of IL2R, proliferative activity, NK activity and LAK activity of T lymphocytes from the treatment group were significantly higher than those of control respectively.Conclusion The adoptive transfer of CD3AK cells could improve the disorder of T lymphocyte subsets and its functions and provide the patients with PLC with fresh abundant effectors against tumor.
To evaluate effect of recombinant human growth hormone (rhGH) on immunologic function in patients with gastrointestinal malignant tumor (GIMT). Before and 3 weeks after surgical treatment and administration of rhGH, the amount of T lymphocyte subset (T-LS) and soluble interleukin 2 receptor (sIL-2R) level were measured in 12 patients with GIMT, which were compared with 20 cases of normal control and 18 cases of GIMT treated by surgery alone. Result: ①In all GIMT patients, the serum CD＋3, CD＋4 level and the ratio of CD＋4/CD＋8 were lower than normal control and the sIL2R level was much higher; ②After operation, the serum CD＋3, CD＋4 level and the ratio of CD＋4/CD＋8 of all patients increased, the serum sIL2R level decreased; ③In patients recieved rhGH, the serum CD＋3, CD＋4 level and the ratio of CD＋4/CD＋8 were much more increased and the serum sIL-2R level much more decreased than those of surgery alone group. Conclusion: rhGH can enhance the immunologic function of patients with GIMT.
To search for the relationship between immune state and tumor metastases, CD3,CD4,CD8 and CD44 contents in 13 speciments of fine needle aspiration (FNA) from thyroid cancer patients were detected by the flowcytometry (FCM) and comparison between thyroid cancer and benign tumor was made. The result showed :in thyroid cancer group, CD＋3，CD＋4 cells and the ratio of CD＋4/CD＋8 reduced significantly (Ｐ＜0.01),and CD＋8 cell increased significantly (Ｐ＜0.01), in metastases group,this change was much significantly. CD44 expressed significantly higher in cancer group than that of the benign thyroid neoplasms, and the change was related to the tumor metastases. The results indicate that CD44 can be as a marker of tumor and indicator of metastases. The disturbance of immune system results in active expression of CD44 by tumor cells, so, lead to metastases. It is helpful to the diagnosis of thyroid cancer, assessment of metastases and management in surgery.
Objective To investigate the effects of down-regulating of Rfng gene ( 1, 3-Nacetylglucosaminyltransferases) in lung CD4 + T cells of asthmatic rat model by small interfering RNA ( siRNA) and explore the role of Rfng in pathogenesis of asthma. Methods An asthmatic rat model was established by OVA sensitization and challenge. Total T cells were isolated from lung tissue of asthmatic rats, and CD4 + T lymphocytes were purified using magnetic beads. CD4 + T lymphocytes were transfected by siRNA targeting Rfng gene. The mRNA and protein expressions of Rfng were detected by quantitative PCR and Western blot. Quantitative PCR was performed to determine the mRNA levels of Th1 /Th2 cytokines and related genes including IL-12, IFN-γ, IL-4, IL-5, T-bet, and GATA3. ELISA was performed to determine the concentrations of IL-12, IFN-γ, IL-4, and IL-5 in supernatant. Results The mRNA and protein expression of Rfng in RNAi group decreased significantly. IL-12, IFN-γ, T-bet increased and while IL-4, IL-5, and GATA3 decreased significantly. The concentrations of IL-12 and IFN-γ in the supernatant increased significantly, while IL-4 and IL-5 decreased significantly. Conclusions Down regulation of Rfng affects T cell differentiation. It is presumed that Fringe contribute to the pathogenesis of asthma.
Objective To explore the impact of recombinant human growth hormone （rhGH） on T lymphocyte subsets in patients with rheumatic heart disease during the perioperative period of heart valve replacement. Methods A total of 65 patients with rheumatic valvular heart disease who received heart valve replacement in Department of Cardiothoracic Surgery of Xiangyang Central Hospital from June 1， 2011 to March 31， 2012 were enrolled in this double-blind randomized controlled clinical study. All the patients were divided into 2 groups by random number produced by SAS software：the trial group and the control group. There were 35 patients in the trial group including 19 males and 16 females with their average age of 50.57 years， and 30 patients in the control group including 16 males and 14 females with their average age of 49.87 years. Apart from routine cardiac glycosides， diuretics， glucose-insulin-potassium solution， and postoperative anti-infective therapy， patients in the trial group also received subcutaneously injection of rhGH 5 U （1 ml）daily from 1 day before surgery to 3 days after surgery， and patients in the control group received subcutaneously injection of normal saline 1 ml as placebo. Peripheral venous blood samples were taken in the morning 2 days before surgery and 1 st， 3 rd， 7 th day after surgery respectively. Percentages of CD3+， CD4+， CD8+ were examined timely by flow cytometry and CD4+ /CD8+ ratio was calculated. Results In the control group， percentages of CD3+， CD4+ and CD4+ /CD8+ ratio on the 1st， 3rd， 7th postoperative day were significantly lower than preoperative levels， and percentages of CD8+ on the 1st and 3rd postoperative day were significantly lower than preoperative level （P＜0.05）. In the trial group， percentages of CD3+， CD4+， and CD8+ on the 1st and 3rd postoperative day were significantly lower than preoperative levels（P＜0.05）， while percentages of CD3+， CD4+， and CD8+ on the 7th postoperative day were not statistically different from preoperative levels （P＞0.05）; CD4+ /CD8+ ratio on the 1st postoperative day was significantly lower than preoperative level （P＜0.05）， while CD4+ /CD8+ ratios on the 3rd and 7th postoperative day were not statistically different from preoperative level （P＞0.05）. There was no statistical difference in preoperative T lymphocyte subsets between the trial group and the control group （P＞0.05）. The percentages of CD4+ and CD4+/CD8+ ratio in the trial group were significantly higher than those of the control group on the 1st postoperative day （P＜0.05）， while the percentages of CD3+ and CD4+ and CD4+ /CD8+ratio in the trial group were significantly higher than those of the control group on the 3rd and 7th postoperative day（P＜0.05）. Conclusion Use of rhGH can significantly increase T lymphocyte subsets expression， enhance body cellular immunity， and improve postoperative recovery of patients with rheumatic valvular heart disease during the perioperative period of heart valve replacement.
Abstract: Objective To investigate the influence of cardiopul monary bypass(CPB) to the cellular immune function of T lymphocyte. Me th ods Among 500 patients operated from March 2006 to September 2006，30 patients with rheumatic heart disease were selected randomly as the CPB group, which would replace mitral valve; 30 patients with congenital patent ductus arte reriosus as the nonCPB group, which would ligate ductus arteriosus without CPB . The blood was sampled before operation, at the end of CPB or operation, and 24 hours after operation. After T lymphocyte was seperated, the quantum o f T lymphocyte, apoptosis of T lymphocyte, ability of T lymphocyte to kill tumou r cell were measured. Results The quantum of T lymphocyte i n CPB group at the end of CPB was decreased than that before operation （50.9% ±6.8% vs. 58.5%± 9.1%,Plt;0.05）; apoptosis of T lymphocyte at the end of CPB and 24 hou rs after operation were increased than that before operation （6.5%±2.2% vs. 0. 9%±1.1%, 5.6%±1.8% vs. 0.9%±1.1%;Plt;0.01）; ability to kill tumour cell b reakdown in CPB group at the end of CPB and 24 hours after operation was decrea sed than that before operation （30.4%±6.0% vs. 37.3%±8.6%, 29.0%±4.9% vs . 37 .3%±8.6%;Plt;0.05）. Ability to kill tumour cell breakdown in CPB group was lower than that in nonCPB group at the end of CPB （30.4%±6.0% vs. 33.6%±5. 3%, Plt;0.05）. Conclusion CPB can depress the cellular im mune function,which causes temporary immune depression to the body.
Objective To observe the changes in the peripheral blood T lymphocyte subsets and the histomorphology of the transplanted tissues in the rabbits in the early stage after transplantation of the tissue engineered boneconstituted by the biologically-derived scaffold and to confirm the feasibility of the biologicallyderived materials as a scaffold in the bone tissue engineering. Methods Forty-eight healthy New Zealand rabbits (weight, 2.0-2.5 kg) with a 1-cm defect were equally and randomly divided into 4 groups: Groups A-D. The partial demineralized freeze-dried bone (PDFDB), the tissue engineered bone constructed by the osteoblasts derived from the lactant rabbit periosteum as a seeding cell, the xenogeneic cancellous bone undergoing the antigen self-digestion, partial demineralization and freeze-driedprocess as a scaffold, and the fresh xenogeneic allografting bone were respectively transplanted into the segmental defects of the rabbit radii in Groups A-D.To examine the effects of the 4 different materials, the flow cytometry was used to observe the changes in the T lymphocyte subsets in the rabbit peripheral blood at 1, 2, and 4 weeks after the operations and to examine the osteogenesis achieved by the 4 materials, the histological observations were also performed at 2, 4, 8, and 12 weeks after the operations. Results Two weeks after the tissue engineered bone transplantation in Group B, the osteoblasts and chondroblasts were found in the apertures of the scaffold, the new bone formation could be observed, the osteoclasts could be seen in the peripheral zone, and some of the netlike frameworks were destroyed and absorbed. Four weeks after the operation, the histological observation revealed that the osteocartilagionous callus turned into a woven bone. The peripheral blood T lymphocyte subsets of CD4+ and CD8+ were significantly greater in number 1-2 weeks after the operations and in Groups A and B than before the operations and in the other groups (.Plt;0.05)；4 weeks after the operations the T lymphocyte subset of CD4+ was only slightly greater in number than before the operations, but with no statistically significant difference (Pgt;0.05). In Group C, the increase of the T lymphocyte subsets of CD4+ and CD8+ was not significant after the operation (Pgt;0.05). The T lymphocyte subsets of CD4+ and CD8+ were significantly greater in number 1, 2 and 4 weeks after the operations and in Group D than before the operation and in the other groups (Plt;0.05). Conclusion The tissue engineered bone constructed by the partial demineralized freezedried bone as a scaffold does not cause a serious immunologic rejection in the early stage after the transplantation and does not affect its good ability to repair the bone defect. The biologicallyderived bone canbe used as a scaffold in the bone tissue engineering.