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 summarize the clinical, radiological and pathological characteristics of acquired immune deficiency syndrome (AIDS) combined with Pneumocystis carinii pneumonia (PCP), so as to improve the clinicians' understanding of the disease.
MethodsThe clinical data of 50 AIDS patients combined with PCP admitted between February 2006 and May 2015 were retrospectively analyzed, including medical history, physical signs, laboratory examination, chest high resolution CT (HRCT), pathological characteristics, treatment and prognosis, etc.
ResultsThe clinical features of AIDS patients combined with PCP included cough, dyspnea and fever, without obvious positive signs in the lung.The patients were divided as a mild group, a moderate group and a severe group according to the levels of PaO2.There was significant difference among three groups in serum albumin level [(23±3) g/L vs. (30±5) g/L and (28±6) g/L, P < 0.01].There were no significant differences among three groups in CD4+ T lymphocyte and lactate dehydrogenase (LDH) (P > 0.05).The typical chest radiograph feature of HRCT was ground-glass shadows in both lungs, and may be associated with reticular shadows or "gravel sign" and cyst.Of 50 patients, 16 patients were diagnosed via pathology of transbronchial lung biopsy(TBLB) and only 5 patients were diagnosed via silver staining of the bronchoalveolar lavage fluid (BALF).The other patients were clinically diagnosed.100% of the patients were treated with sulfamethoxazole (SMZco), 64%with caspofungin, and 72% with glucocorticoid.All the patients relieved with no death in hospital.
ConclusionWhen a patient got cough, dyspnea and fever, especially ground glass on HRCT in both lungs, AIDS combined with PCP should be highly considered, and diagnostic treatment with SMZco and CD4+ T lymphocyte measurement should be conducted as soon as possible, so as to reduce misdiagnosis and mortality.
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.
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.
ObjectiveTo investigate the inhibitory effect of T lymphocyte transplantation of EphrinAl-Caspase-3 on the growth of breast cancer.MethodsSix-week-old BALB/c nude mice were used to inoculate breast cancer cells to construct a nude mouse model of breast cancer. They were randomly divided into 3 groups according to random number table: PBS group received intratumoral injection of 10 μL PBS, and negative control group received intratumoral injection of 1×106 T lymphocytes uninfected with adenovirus, 1×106 EphrinAl-Caspase3-T lymphocytes were injected intratumorally into the infected group, and the tumors size (0, 3, 6, 9, 12 and 15 d) were measured with vernier calipers every 3 days until end of experiment. The content of EphrinAl-Caspase-3 in the tissues of the nude mice was measured. The presence of T lymphocytes expressing green fluorescent protein and the ratio of Caspase-3-positive and Ki-67-positive cell were observed by pathological examination.ResultsOn the day 0 and day 3, there were no significant difference in tumor volume between the 3 groups (P>0.05). On the 6th day and later, the difference between the infected group and the PBS group/negative control group were statistically significant (P<0.05), but there were no significant difference in tumor volume between the PBS group and negative control group at each time point (P>0.05). The presence of scattered green fluorescent protein-labeled EphrinAl-Caspase-3-T lymphocytes was observed in the tumor tissues of the infected group, while the presence of green fluorescent protein were not detected in the PBS group and the negative control group. In the infected cells, ratio of Caspase-3-positive cell was up-regulated and ratio of Ki-67-positive cell was down-regulated. The expression of EphrinAl-Caspase-3 could be detected on the 3rd day in the infected group, and at the peak on the 6-day, then the amount of secretion gradually decreased. The expression of EphrinAl-Caspase-3 were not detected in the PBS group and the negative control group at each time point.ConclusionEphrinAl-Caspase-3 can significantly inhibit the growth of breast cancer cells and promote apoptosis.
ObjectiveTo compare the clinical recovery and immune response between laparoscopic-assisted and open D2 gastrectomy for advanced gastric cancer.
MethodsThe clinical data of 53 patients with advanced gastric cancer from January 2012 to October 2013 were studied prospectively. According to random number table, patients were randomly divided into laparoscopic-assisted group(LA group, n=27) and open operation group(OO group, n=26). Operative time, blood loss, time to passage of flatus, time to resume soft diet, after bed time, postoperative hospital stay, and number of retrieved lymph nodes were compared respectively between the two groups. The changes in CD3, CD4+, CD8+, IgG, IgA, IgM, and CRP were examined respectively by using flow cytometry and immunoturbidimetric assays on the preoperative day 1, and on the postoperative day 1 and 7.
ResultsThe operative time was longer significantly in LA group than that in OO group(P < 0.05). The mean blood loss, the first flatus time, after bed time, and postoperative hospital stay in the two groups were all different statistically(P < 0.05), and all were better in LA group. However, the mean number of retrieved lymph nodes and the time to resume soft diet were not significantly different in the two groups(P > 0.05). On the day 1 and 7 after operation, the CD3, CD4+, and CD8+ significantly decreased as compared with those preoperatively in two groups(P < 0.01, P < 0.05). On the day 1 after operation, the levels of IgG, IgA, and IgM significantly decreased as compared with those preoperatively in two groups(P < 0.05). Those immunoglobulin in LA group recovered to close to the level before surgery, but in OO group sustained lower level(P < 0.05). On the day 1 and 7 after operation, CRP level significantly increased as compared with those preoperatively in two groups(P < 0.01, P < 0.05). Those changes of above index were not significantly different between the LA group and OO group on the day 1 after operation(P > 0.05). All index recovered gradually in the two groups on the day 7 after operation and were better in LA group(P < 0.05, except IgA).
ConclusionLaparoscopic radical gastrectomy for advanced gastric cancer resulted in a quicker clinical recovery and a lesser depression to the perioperative cellular and humoral immune function.
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.
ObjectiveTo evaluate the effects and mechanism of indoleamine 2, 3-dioxygenase (IDO) modified rat bone marrow mesenchymal stem cells (BMSCs) in composite tissue allograft rejection.
MethodsBMSCs isolated from Brown Norway (BN) rats (aged, 4-6 weeks) were infected by IDO[green fluorescent protein (GFP)]-lentivirus. The high expression target gene and biological activity cell line (IDO-BMSCs) were screened. IDO mRNA and protein expressions were detected by RT-PCR and Western blot. The biological activity of IDO in supernatant was detected by measuring the amount of kynurenine generation. In mixed lymphocyte reaction system, different numbers of IDO-BMSCs mixed with responding cells (peripheral blood mononuclear cell isolated from 4-6-week-old LEWIS rats, as recipient) and stimulating cells (peripheral blood mononuclear cell isolated from BN rats, as donor), with the cells ratios of 1:5:5, 1:10:10, 1:50:50, and 1:100:100 (as experimental groups 1, 2, 3, and 4, respectively). Each reaction system was blocked by 1 mmol/L 1-methyl-tryptophan (1-MT) (IDO specific inhibitor). IDO-BMSCs mixed with responding cells (1:5) as the negative control group, responding cells mixed with stimulating cells (1:1) as positive control group; and IDO-BMSCs were cultured in RPMI 1640 medium alone as blank control group. MTT assay was used to detect the T lymphocytes proliferation at 5 days. Furthermore, GFP-BMSCs (group A), IDO-BMSCs (group B), and normal saline (group C) were infused via the tail vein of allogeneic limb transplantation rats, and graft survival time and rejection were observed in each group.
ResultsThe IDO expression of BMSCs after genetic modification was higher than that before genetic modification. IDO-BMSCs could significantly improved kynurenine concentration in culture medium supernatant when compared with GFP-BMSCs (P<0.05). Before adding 1-MT, with the ratio of IDO-BMSCs to responding cells decreased, T lymphocytes proliferation rate increased in experimental groups 1, 2, and 3, showing significant differences between groups (P<0.05); there was no significant difference between experimental group 4 and the positive control group (P>0.05). After adding 1-MT, T lymphocytes proliferation rate was significantly higher than that before adding 1-MT in the other experimental groups (P<0.05) except experimental group 4 (P>0.05). In vivo, IDO-BMSCs could promote colonization in allograft, inhibit transplantation rejection, and prolong survival time of composite tissue allograft; the survival time of composite tissue allograft was (11.5±0.6) days in group A, (14.5±0.8) days in group B, and (9.0±0.3) days in group C, and it was significantly longer in group B than in groups A and C, and in group A than in group C (P<0.05).
ConclusionIDO-BMSCs can promote the survival of allogeneic composite tissue grafts in rats, and its mechanism may involve in inhibition of T lymphocytes proliferation and promotion their own colonization in allograft.
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 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.
