Objective To explore the changes and interrelationship of serum interleukin-12 (IL-12) and T lymphocyte subset in patients with primary hepatic carcinoma (PHC). Methods Serum IL-12 level was determined by ELISA in 36 patients with PHC. The peripheral blood T lymphocyte subset was assessed with flow cytometry. The distribution and changes of T lymphocyte subset in the tumor tissue were detected by immunohistochemistry analysis. Results The numbers of the CD＋4 T cell were reduced and of the CD＋8 T cell increased either in peripheral blood or tumor tissue, and showed the trend of the ratio (T4/T8) declined progressively with the aggravation of the state with PHC. IL-12 and T4/T8 had significant interrelationship.Conclusion IL-12 is an important antitumor factor of the patients with PHC. T lymphocyte subset plays a great role in the process of antitumor.
ObjectiveTo investigate the method for generating anchor chemric T lymphocytes that can target tumor associated glycoprotein-72 (TAG72) antigen and analyze their repressive effects on proliferation of TAG72 positive hepatocarcinoma cells. MethodsFirstly, peripheral blood mononuclear cells (PBMCs) from healthy volunteers were isolated. And then, CD8+ T cells were isolated from PBMCs via magnetic activated cell sorting (MACS). These lymphocytes were transfected with recombinant vector, anti-TAG72-scFv-CD28-pcDNA3, through Lipofectamine2000 to gernerate anchor chimeric TAG72-specific CD8+ T cells. SMMC7721 (TAG72 positive) hepatocarcinoma cells were co-cultured with chimeric T lymphocytes and their cell cycles were analyzed by flow cytometry (FCM). ResultsAnchor chmeric T lymphcytes targetting TAG72 recognized TAG72 positive SMM7721 cells and repressive effects on their proliferation were observed by flow cytometry. ConclusionAnchor chmeric T lymphcytes targetting TAG72 on tumor surface can specifically recognize TAG72 positive hepatocarcinoma cells and may exert repressive effect on their proliferation.
Objective To observe the effects of Thymosin α1 （Tα1） on acute rejection after liver transplantation and immune function of T cells. Methods Twenty recipients of liver transplantation due to primary hepatic carcinoma were divided into two groups: Tα1 group (n=10) and control group (n=10). Tα1 group received subcutaneous injection of Tα1 1.6 mg on the first day after liver transplantation and then twice a week for at least one month. Both Tα1 group and control group took same immunodepressants. Core biopsies were carried to compare the incidence rate of acute rejection between Tα1 group and control group. Peripheral T cellular immune function in these two groups was detected on 1 d before, 1 week, 2 weeks and 1 month after transplantation. Results There was not significant difference of incidence rate of acute rejection between Tα1 group and control group (Pgt;0.05). In the Tα1 group, CD4+, CD8+ lymphocyte cell counts and the CD4+/CD8+ ratio were significantly higher than those in the control group in 2 weeks and 1 month after transplantation (P＜0.05). Conclusion Use of Tα1 in recipients who also takes rountine immunosuppressants dose not increase the risk of occurring acute rejection after liver transplantation. Tα1 can significantly increase CD4+, CD8+ counts and CD4+/CD8+ ratio, which shows that Tα1 may improve recipients’ cellular immune function.
ObjectiveTo investigate the inhibitory effect of the inhibition of antigen-presentation attenuators (iAPA)-based dendritic cells (DC) and cytotoxic T lymphocytes (CTL)-iAPA-DC/CTL on SMMC-7721 xenograft in nude mice. MethodsUsing the human hepatic carcinoma cell line SMMC-7721 on nude mice to establish a transplanted tumor model of human hepatocellular carcinoma (HCC).Twelve nude mice were divided into two groups randomly: normal saline control group (control group) and iAPA-DC/CTL group (n=6, each).After four times treatment with iAPA-DC/CTL (once a week), all mice were sacrificed.Tumor growth was calculated by measuring the long/short diameters and the tumor growth curve was delineated.The tumors were weighed and the tumor inhibition rate was calculated.In addition, the histopathological examination was conducted. ResultsThe SMMC-7721 xenograft model was successfully established in 85.71% (12/14) of all mice.The tumor volume was (3 661.48±322.59) mm3 and (2 725.36±252.65) mm3 in control group and iAPA-DC/CTL group, respectively.The tumor growth was significantly inhibited in iAPA-DC/CTL group (t=5.62, P < 0.05).The tumor weight was (1.97±0.21) g and (1.38±0.14) g in control group and iAPA-DC/CTL group, respectively.The tumor weight in iAPA-DC/CTL group was significantly reduced (t=5.73, P < 0.05), and the tumor inhibition rate was 29.95%.After immunohistochemical staining T lymphocyte counts was 0 cell/HPF and (54.24±4.31) cells/HPF in control group and iAPA-DC/CTL group, respectively.The number of T lymphocytes in iAPA-DC/CTL group was significantly increased (t=25.02, P < 0.05). ConclusioniAPA-DC/CTL could effectively inhibit the growth of subcutaneously implanted HCC.
【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.
Objective To investigate the expression and clinical significance of T lymphocyte subsets, natural killer (NK) cells and CD19+ B cells in the elderly with primary immune thrombocytopenia (ITP) before and after treatment. Methods The elderly ITP patients diagnosed and treated in the Songjiang Hospital Affiliated to Shanghai Jiaotong University School of Medicine (preparatory stage) between January 2014 and June 2019 were retrospectively selected as the observation group. The healthy elderly in the same period were selected as the control group. According to the treatment, the observation group was divided into effective group and ineffective group. The expression levels of T lymphocyte subsets (CD3+, CD4+, CD8+ and CD4+/CD8+), NK cells and CD19+ B cells were observed and analyzed. Results A total of 75 subjects were included, including 35 in the observation group and 40 in the control group. The total effective rate was 85.71% (30/35). Before treatment, the expression levels of T lymphocyte subsets (CD3+, CD4+ and CD4+/CD8+) in the observation group were lower than those in the control group (P<0.05). There was no significant difference in other indexes between the two groups (P>0.05). After treatment, except for CD8+, the expression levels of T lymphocyte subsets (CD3+, CD4+ and CD4+/CD8+) in the observation group were higher than those before treatment (P<0.05). The expression levels of NK cells and CD19+ B cells were lower than those before treatment (P<0.05). The expression levels of T lymphocyte subsets (CD3+, CD4+ and CD4+/CD8+) in the effective group were higher than those before treatment (P<0.05), while the expression level of CD19+ B cells was lower than that before treatment (P<0.05). There was no significant difference in other indexes before and after treatment (P>0.05). There was no significant difference in the expression levels of T lymphocyte subsets (CD3+, CD4+, CD8+ and CD4+/CD8+), NK cells and CD19+ B cells in the ineffective group before and after treatment (P>0.05). Conclusions T lymphocyte subsets are abnormal in elderly ITP patients. The immune abnormality of T lymphocyte may be one of the reasons for elderly patients with ITP. With the improvement of therapeutic effect, immune cell subsets have also been improved.
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.
Objective To study the immunological tolerance induced by blocking the second signal of T cell with extrinsic cytotoxic T lymphocyteassociated antigen 4 immuno globlin(CTLA4-Ig). Methods Fifty-four BALB/C mice, inbred strains, were employed as recipients of bone allografts, using a model of heterotopic muscle pouch. The 54 mice were divided into 3 groups and18 for each group. The first group, in which the donor was C57BL/6 with intraperitoneal injection ofL6(as a control), was named AL group. The second group,also C57BL/6 with injection CTLA4-Ig, was named AC group. The third group,homologous BALB/C with injection of PBS buffer solution, was named AB group.The serum antibody, lymphocyte proliferation of the second stimulation by splenic cell and bone supernatant of donor, the analysis of lymphocyte subsets, a regraft experiment and histology were determined 2, 4 and 6 weeks after transplantation. The second transplantation was to regraft C57BL/6(BC group) and C3H(BHgroup) mice respectively after first 12 mice being transplantated with C57BL/6 and injected with CTLA4-Ig as to detect donor-specificity of immunological tolerance. Results Compared with AB group, AL group created more intensive immune rejection: CD4 T cell subsets(Plt;0.05), the serum antibody(Plt;0.05) and lymphocyteproliferation of the second stimulation by splenic cell and bone supernatant ofdonor (Plt;0.01 and 0.05) were significantly increased. However, the results of AC group showed that CTLA4-Ig significantly inhibited the immune rejection: CD4T cell subsets(Pgt;0.05), the serum antibody (Pgt;0.05), and lymphocyte proliferation of the second stimulation(Pgt;0.05) were similar to those of AB group. Histological observation of AC group showed that lymphocyte infiltration disappeared,cartilage and new bone formed, and bone marrow cavities emerged. A regraft experiment showed that CD4 T cell subsets (Plt;0.05) and lymphocyte proliferation of the second stimulation by splenic cell and bone supernatant of donor(Plt;0.05), BC group was significantly lower than those of BH group. So theimmunological tolerance induced by CTLA4-Ig was of donor-specificity. Conclusion The immunological tolerance induced by CTLA4-Ig was prolonged for 6 weeks. This study provides a brand-new path for bone transplantation, which can be helpful to other organ transplantation.
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.
ObjectiveTo evaluate the relationship between CT lesion changes in COVID-19 patients and different subgroups of T lymphocytes, providing reference information for assessing patient conditions, predicting outcomes, and evaluating treatment efficacy. MethodsClinical and imaging data of confirmed COVID-19 patients admitted to the Chongqing Public Health Medical Center from January 24 to March 15, 2020, were collected. Based on the absorption characteristics of lesions in CT images, patients were categorized into three groups: Group A (obviously continuously absorbed), Group B (stable-slow absorption), and Group C (progressive absorption). The relationship between CT changes and T lymphocyte subgroups was analyzed according to lesion absorption. ResultsA total of 47 patients were included, with 18 in Group A, 14 in Group B, and 15 in Group C. At different stages—admission, during treatment, and at the end of treatment—the levels of T lymphocytes were observed as follows: Group A>Group B>Group C. When lesions were absorbed, the average count of CD4+ T lymphocytes was (544.43 ± 163.34) cells/μl; when lesions showed little change or increased, CD4+ T lymphocyte levels decreased to varying degrees. During treatment, both Group A and Group B showed CD4+ T lymphocyte levels returning to above normal levels, with an average increase of 134 cells/μl in Group A, which was lower than that in Group B (192 cells/μl) and Group C (149 cells/μl). Finally, T lymphocyte levels reached normal in all groups, but Group A levels were higher than those in Groups B and C (P<0.05). Upon follow-up, the average CD4+ T lymphocyte count was (544.43 ± 163.34) cells/μl in 52 cases of lesion absorption, (339.06 ± 145.98) cells/μl in 31 cases of minimal change, and (230.50 ± 95.24) cells/μl in 16 cases of lesion progression, with statistically significant differences among the three groups (P<0.05). ConclusionsThe increase in lung lesions in patients indicates poor immune function, necessitating enhanced immune regulation. Conversely, if a decrease in T lymphocyte levels is detected during the course of the disease, attention should be given to the risk of lesion progression, and timely CT re-examinations should be conducted to monitor changes in lesions.