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.
Objective To observe the therapeutic effect of mensenchymal stem cells (MSCs) for experimental autoimmune uveitis (EAU). Methods MSCs were obtained from Wistar rats and selected by plastic adherence. Lewis rats were divided into treatment group and control group, six rats in each group. EAU models were induced by immunization with an emulsion (0.2 ml) containing 30 mu;g interphotoreceptor retinoid-binding protein derived peptide R16 and complete Freundprime;s adjuvant. The clinical manifestations of two groups were observed. Nine to 11 day after modeling, 1 ml MSCs suspension, which contained 5times;106 MSCs, were injected into the rats in treatment group via tail vein, and the rats in control group were given equal volume of phosphate buffer solution. Fifteen day after modeling, the eyes were collected to test the proportion of interferon gamma;, interleukin-17 and Foxp3 positive cells by flow cytometry. The clinical scores were analyzed by mixed linear model and statistical analysis of variance of repeated measurement data. The results of flow cytometry were analyzed using independent-sample t test. Results Six days after immunization, mild dilatation and congestion of iris vascular was observed. Nine days after immunization, mild muddy anterior chamber, myosis and absent pupillary reaction to light were observed. Twelve days after immunization, muddy anterior chamber, occlusion of pupil and dimmed or disappeared red reflex were observed, and then inflammation was slowly reduced. From 11 to 15 days after immunization, the clinical score of treatment group was lower than that in control group, the difference was statistically significant (t=2.42, 2.21, 4.16, 5.24, 4.03; P＜0.05). The results of flow cytometry showed that MSCs treatment could decrease the proportion of CD4＋T cells, Th1 cells and Th17 cells, increase the proportion of Treg cells. Conclusion MSCs treatment can ameliorate EAU, up-regulate the expression of Treg cells and down-regulate the expression of CD4＋T cells, Th1 cells and Th17 cells.
ObjectiveTo evaluate the role of CD3+CD4+T cells in patients with nosocomial infection in ICU. MethodsOne-hundred and eleven patients who admitted in ICU and in respiratory department from March to December in 2014 were recruited in the study.There were 33 patients with community-acquired pneumonia (CAP group), 31 patients without nosocomial infection (NNI group), and 47 patients with hospital-acquired pneumonia (HAP group).The counts of T cells, B cells, CD3+CD4+ T cells, CD3+CD8+ T cells, and NK cells were compared among three groups. ResultsThe comparison among the groups had no statistical significance in sex and age(P > 0.05).The three groups had statistical significance in APACHEⅡscore, CD3+CD4+T cells, T cells and B cells, but had no statistical significance in CD3+CD8+T cells, CD3+CD4+/CD3+CD8+ T cells, NK cells, white blood cells, neutrophils, procalcitonin or C reactive protein.CD3+CD4+T cells of HAP group were less than other two groups.The area under the ROC curve (AUC) was 0.660, with a threshold of 29.96%, a sensitivity of 93.8%, and a specificity of 40.4%. ConclusionCD3+CD4+ T cell is an independent predictor for nosocomial infection.
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.
Objective To study effect of carcinoembryonic antigen (CEA) positive targeted lymphocytes on gastric cancer cells in vitro and in vivo. Methods The peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy volunteers. The recombinant vector anti-CEA-scFv-CD3ζ-pcDNA3.0 was transfected into the PBMCs by lipofectamine 2000, by this means, the CEA special lymphocytes were obtained. Meanwhile, the PBMCs transfected with empty plasmid pcDNA3.0 were used as control (empty vector lymphocytes). The different lymphocytes and gastric cancer cells (CEA positive KATOⅢ gastric cancer cells and CEA negative BGC-823 gastric cancer cells) were co-cultured, then the ability to identify the gastric cancer cells and it’s effect on apoptosis of gastric cancer cells were observed at 24 h or 36 h later respectively. The CEA special lymphocytes and empty vector lymphocytes were injected by the tail vein of nude mice bearing gastric cancer cells, then it’s effect on the tumor was observed. Results ① The CEA special lymphocytes could strongly identify the KATOⅢ gastric cancer cells (identification rate was 72.3%), which could weakly identify the BGC-823 gastric cancer cells (identification rate was 7.8%). ② The apoptosis rate of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly higher than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (P=0.032), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (P=0.118). ③ The tumor volume of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly smaller than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (F=5.010, P<0.01) or the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells (F=4.982, P<0.01), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (F=1.210, P>0.05). Conclusion CEA special lymphocytes can promote cell apoptosis and inhabit tumor reproduction of CEA positive gastric cancer cells in vitro and in vivo.
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 explore the functional heterogeneity of T lymphocytes in various organs after SARS-CoV-2 infection. Methods Using the public database GEO data (GSE171668, GSE159812, GSE159556, GSE167747) and the analysis method of single-cell technology, the functional differences of T lymphocytes in various organs of patients after infection with SARS-CoV-2 were analyzed. Results Through single-cell data extraction of 16 livers, 19 hearts,2 spleens, 6 brains, 58 lungs, 21 kidneys and 5 pancreases from SARS-CoV-2 infected patients, invasion genes were relatively highly expressed in T lymphocytes of the lung and pancreas. The lung had a special ability to express the interferon signaling pathway, while the expression of other organs was relatively low; at the same time, the T lymphocytes of the lung also highly expressed fatty acid binding sites. Conclusion After SARS-CoV-2 infection, compared with other organs, the lung has a special interferon-activated signaling pathway and fatty acid binding site.