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.
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 explore the effects of corticosteroid on peripheral blood T lymphocyte subsets in patients with coronavirus disease 2019 (COVID-19).MethodsThis was a retrospective study and 376 patients were included in the study. The patients were classified into three type: moderate type (118 patients), severe type (215 patients), critical type (43 patients). Six critical patients died. T lymphocyte subsets were analyzed and compared among these patients. In severe patients, T lymphocyte subsets were compared between no corticosteroid therapy patients (178 patients) and patients who were treated with corticosteroid for 3 to 5 days (37 patients).Results(1) In contrast with those in moderate patients, in severe patients total lymphocytes [(1359.2±597.9)×106 vs. (1703.7±702.4)×106/L, LSD-t=4.786, P<0.001], total T lymphocytes [(949.2±454.0)×106 vs. (1235.5±555.7)×106/L, LSD-t=5.175, P<0.001] and CD8+ T cells [(336.8±189.8)×106 vs. (461.7±242.8)×106/L, LSD-t=5.332, P<0.001] decreased significantly, and CD4+/CD8+ ratio (1.81±0.92 vs. 1.64±0.74, LSD-t=1.574, P=0.116) was increased. In contrast with those in severe patients, in critical patients CD4+/CD8+ ratio (2.23±1.24 vs. 1.81±0.92, LSD-t=2.627, P=0.009) increased and CD8+ T cells [(232.5±159.8)×106/L vs. (336.8±189.8)×106/L, LSD-t=2.867, P=0.004] decreased significantly, total lymphocytes [(1161.1±583.7)×106/L vs. (1359.2±597.9)×106/L, LSD-t=1.772, P=0.077], total T lymphocytes [(790.5±419.3)×106/L vs. (949.2±454.0)×106/L, LSD-t=1.846, P=0.066] also decreased but without significant difference. There was no significant difference between dead and survived critical patients. (2) In severe type, in contrast with no corticosteroid therapy patients, 37 patients were therapy with corticosteroid for 3 to 5 days, and their total T lymphocytes [(770.6±480.3)×106 vs. (986.3±440.7)×106/L, t=2.666, P=0.008] and CD4+/CD8+ ratio (1.30±0.73 vs. 1.91±0.92, t=3.771, P<0.001) were decreased significantly.ConclusionsIn COVID-19 patients, lymphocytes, T lymphocytes and CD8+ T cells are decreased, but CD4+/CD8+ ratio is increased, and these changes are positively related to the severity of the disease. After corticosteroid therapy, the increase of CD4+/CD8+ ratio is relieved, but T lymphocytes are decreased further.
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 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.
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 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 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.
ObjectiveTo understand the interplay between tumor-associated macrophages (TAMs) and T lymphocytes, as well as the effect on the pathogenesis of hepatocellular carcinoma (HCC) again, so that providing new ideas and methods for the immunotherapy of HCC.MethodSearched the literatures about the interplay between TAMs and T lymphocytes in HCC to analyze and summarize the relationship between TAMs and T lymphocytes in HCC.ResultsWhile TAMs and T lymphocytes themselves regulate the process of tumorigenesis and development, they also had a mutual regulatory mechanism to further promote the development of HCC.ConclusionsThere is an interaction between TAMs and T lymphocytes, and this interaction forms a vicious circle to a large extent and promotes the development of HCC. Recognizing and making rational use of this interaction can provide new ideas and methods for the future immunotherapy of HCC.