ObjectiveTo investigate the levels of regulatory T cells (Treg) and FoxP3 gene in patients with gastric cancer before and after operation. MethodsTwenty patients with definite diagnosis of gastric cancer and 15 healthy volunteers were selected. The levels of Treg and T cell subsets in peripheral blood were determined by detecting of CD4 and CD25 with immunefluorescence stain and flow cytometry, the expressions of FoxP3 mRNA in these Treg were detected by RTPCR technique. The expression of FoxP3 protein in the gastric cancer tissue was measured by immunohistochemistry assay. ResultsThe percentage of Treg cells in total CD4+ T isolated from the patients with gastric cancer was higher than that of healthy volunteers 〔(19.39±5.58)% versus (9.91±3.23)%, Plt;0.01〕, and it markedly decreased after operation 〔（13.50±5.93）% versus (19.39±5.58)%, Plt;0.05〕. The FoxP3 mRNA expression in the patients with gastric cancer was also higher than that of healthy volunteers (0.86±0.03 versus 0.64±0.02, Plt;0.01), and decreased after operation (0.73±0.04 versus 0.86±0.03, Plt;0.05). The percentage of CD4+T cell in mononucleocytes of peripheral blood of patients with gastric cancer was significantly lower than that of healthy volunteers (Plt;0.01), but the difference was not significant between before and after operation. FoxP3 protein expressed in cytoplasm of 13 patients with gastric cancer, in which bly positive in 2 cases, middle positive in 6 cases, weakly positive in 5 cases. FoxP3 protein didn’t express in cytoplasm of 7 patients with gastric cancer. ConclusionsTreg may have a significant effect on the onset and development of gastric cancer through immunosuppressive effect. Tumor tissue is an important initiating agent on Treg proliferation.
Objective To investigate the suppression effect and mechanism of Astilbin on lung allograft rejection in rats, in order to know the function of Astilbin on rats’ lung acute rejection. Methods The model of rat left lung transplantation was set up. Sixty lung transplanted rats were divided into two groups randomly, control group: rats were fed with normal saline 1ml per day, experimental group: rats were fed with Astilbin 1mg/kg per day. Survival time, transforming rate of T cells in spleen, activity of interleukin 2 (IL-2) in spleen lymph cells and apoptosis of T cells were observed. Changes in ultrastructure of pulmonary arteries were observed by electron microscope. Results The survival time in experimental group was prolonged than that in control group (25.4±2.1 d vs. 13.4±1.2 d;t=2.042, Plt;0.05）. Transforming rate of T cells of spleen in experimental group was significant lower than that in control group (23 465.8±8 783.4 cpm vs. 74 567.3±12 874.6 cpm; t=2.284，Plt;0.05）.Activity of IL-2 of spleen lymph cells in experimental group was significant lower than that in control group (425±2.65U/ml vs. 23.46±1.82U/ml; t=3.165, Plt;0.01).Effectively derive apoptosis of activated T cells in acute rejection were observed in experimental group, the ultrastructure of pulmonary arteries showed attenuated injury in experimental group. Conclusion Astilbin decreased the IL-2 concentration in plasma and induced the apoptosis in activated T cells, then suppressed the acute rejection of lung allograft and prolonged the survival period of lung transplantation rats.
Objective To investigate the clinical implication on expression of HLA class I in breast cancer tissures.Methods The expression of HLA class I in 271 patients with breast cancer that underwent radical operation was examinedby using immunohistochemically， and the correlation between the expression of HLA class I and clinicalpathological characteristics and prognosis of breast cancer was analyzed. Results The b positive expression of HLA class I in breast cancer tissures was observed in 92 patients （33.9%）， the expressions of HLA class I in 179 patients （66.1%）were downregulation. The expression of HLA class I expression in breast cancer tissures was significantly associated with the axillary lymph node metastasis， TNM stage （P＜0.05）， other lymph node metastasis， and vascular invasion （P＜0.05）. The disease free survival rate of patients with positive expression of HLA class I was higher than that expression downregulation of HLA class I （P＜0.05）. Conclusion The examination of HLA class I expression is useful for the prediction of tumor progression and recurrent risk of breast cancer via the antitumor immune system.
【Abstract】Objective The effects of tumor infiltrating lymphocytes (TIL) on cellular immunologic function of patients with breast cancer were studied. Methods Twenty five patients with breast cancer were treated by the TIL that were isolated from tissue of tumor. T cell subgroups and natural killer cell (NK cell) activity of peripheral blood, the levels of serum soluble interleukin-2 receptor (sIL-2R) were assayed before and after treatment. Results CD3, CD4, CD4/CD8 and NK cell activity were ascended obviously, and CD8, sIL-2R were descended obviously after the treatment of TIL. Conclusion TIL can enhance the cellular immunologic function of patients with breast cancer.
Objective To summarize the clinical characteristics of pneumocystis pneumonia (PCP) secondary to interstitial lung disease (ILD) to improve the prophylaxis and management level of clinicians. Methods The clinical data of 50 patients with PCP secondary to ILD in the Department of Respiratory and Critical Care Medicine of Nanjing Drum Tower Hospital from January 2015 to December 2022 were collected. SPSS 26.0 software was used for statistical analysis. Results A total of 50 patients with PCP secondary to ILD were screened. Among the 50 patients, there were 23 males and 27 females, with a median age of 64 years old. Forty-eight cases (96%) had a history of glucocorticoid therapy with the median duration of 3 months; 31 (77.5%, 31/40) cases developed PCP in the first 6 months after glucocorticoid therapy; 34 cases had a history of glucocorticoid and immunosuppressants at the same time. None of the 50 ILD patients used drugs for PCP prophylaxis before developing PCP. The major clinical manifestations of PCP secondary to ILD were worse cough and shortness of breath or fever. Laboratory results showed 38 cases (76.0%) had peripheral blood total lymphocyte count <200/µL, 27 cases (54.0%) had CD4+ T cell count <200/µL, 34 cases (68.0%) had CD4+ T cell count <300/µL, 37 cases (74.0%) had CD3+ T cell count <750/µL, 34 cases (68.0%) had β-D-glucan test >200 pg/mL, 35 cases (70.0%) had lactic dehydrogenase > 350 U/L and 41 cases (82.0%) had type Ⅰ respiratory failure. High resolution computed tomography showed added ground-glass opacity and consolidation on the basis of the original ILD. Thirty-six cases were detected the Pneumocystis jirovecii by metagenomic next-generation sequencing with broncho-alveolar lavage fluid as the main source, and 2 cases by smear microscopy. All patients were treated with trimethoprim-sulfamethoxazole. After treatment, 29 cases were discharged with a better health condition, 10 cased died, and 11 cases left hospital voluntarily because of treatment failure or disease deterioration. Conclusions After the use of glucocorticoid and immunosuppressants, ILD patients are susceptible to life-threatening PCP. It is particularly important to make an early diagnosis. Attention should be paid to integrate the symptoms, levels of peripheral blood lymphocyte count, β-D-glucan test, lactic dehydrogenase and imaging findings to make an overall consideration. It is suggested to perform next-generation sequencing with broncho-alveolar lavage fluid at an early stage when patients can tolerate fiberoptic bronchoscopy to avoid misdiagnosis and missed diagnosis. ILD patients often develop PCP in the first 6 months after using glucocorticoid and immunosuppressants. During follow-up, peripheral blood CD4+ and CD3+ T cell count should regularly be monitored so as to timely prevent PCP.
Objective To establish a method to isolate the CD4+CD25+ regulatory T cells (Tregs) and to identify the purity and function of these cells. Methods The peripheral blood (8 mL) were collected from the great saphenous vein of 10 rhesus monkeys (4 females and 6 males, aged 4-5 years, and weighing 5-8 kg). The mononuclear cells were isolated with density gradient centrifugation. CD4+ T cells were separated by the Magnetic cell sorting (MACS) negative selection and MACS positive selection. The cell yield rate, the cell viability, and the cell purity were compared between MACS negative selection and MACS positive selection. In CD4+ MACS negative selection, the anti-biotin MicroBeads and biotin-antibody cocktai in CD4+CD25+ Tregs isolation kit non-human primate were used, and in MACS positive selection, the anti-APC MicroBeads in CD4+CD25+ Tregs isolation kit non-human primate and CD4-APC were used. The CD4+ T cells separated by positive selection were selected to obtain CD4+CD25 Tregs with CD25 MicroBeads. The purity, activity, the FoxP3 level, and the suppressive function to concanavalin A (ConA) activated autologous CD4+CD24- effective T cells (Teffs) of CD4+CD25+ Tregs were detected by flow cytometry. Results After CD4+ T cells were separated by MACS negative selection and MACS positive selection, the cell viabilities were all up to 95%, showing no significant difference (P gt; 0.05). The cell yield rate and purity of CD4+ T cells by positive selection were significantly higher than those of CD4+ T cells by negative selection (P lt; 0.05). CD4+CD25+ Tregs can be successfully isolated by MACS double positive selection. The classifying purity was 76.2% ± 8.6%; the cell activity was 93.3% ± 4.7%; and the level of FoxP3 was 74.2% ± 6.9%. The CD4+CD25+ Tregs had suppressive effect on ConA activated autologous CD4+CD25- Teffs. Conclusion MACS double positive selection can be used to isolate high-purity CD4+CD25+ Tregs from the peripheral blood of rhesus monkeys and the process does not influence the activity of CD4+CD25+ Tregs.
ObjectiveTo approach the role of CD4+CD25+ regulatory T cells in the maintenance of immunotolerance in mouse liver allograft. MethodsThe mouse orthotopic liver transplantation was performed. After the liver transplantation immunotolerance induction, antiCD25 monoclonal antibody (PC61) was injected into the recipients with a delayed timing to remove the CD4+CD25+ T cells. The percentage of CD4+CD25+ T cells and the expression of forkhead/winged helix transcription factor (Foxp3) in the recipients were examined. Furthermore, the survival time of the recipient was observed. ResultsC3H/HeJ recipients receiving DBA/2 hepatic allografts survived over 70 d as in the syngeneic liver transplantation (C3H/HeJ recipients receiving C3H/HeJ hepatic grafts). With various protocols of the delayed PC61 treatment, the CD4+CD25+ T cell was completely disappeared as observed. However, the removal of CD4+CD25+ regulatory T cells after the induction of transplantation immunotolerance did not affect the survival of hepatic allografts. ConclusionCD4+CD25+ regulatory T cells are not essential for the maintenance of spontaneous mouse liver transplantation immunotolerance.
Objective To investigate the effects of ulinastatin on Treg/Th17 and immune status in patients with severe sepsis.Methods A total of 80 patients with severe sepsis, who were hospitalized in ICU during October 2011 to July 2012, were randomly divided into a routine group and a ulinastatin group. The patients in the ulinastatin group were intravenously administered 30mg ulinastatin three times per day for 5 days in addition to routine bundle treatment. The expression of Treg, Th17 and HLA-DR were detected on the first day in ICU and 5 days after treatment. 20 healthy individuals served as controls. Results Compared with the control group, the severe sepsis group had overexpression of Treg and Th17 ( P lt;0. 01) , higher ratio of Treg/Th17( P lt;0. 01) , and decreased HLA-DR expression of CD14 monocyte ( P lt; 0. 01) . In the severe sepsis patients, ulinastatin injection reduced the abnormal expression of Treg and Th17 ( P lt; 0. 01) , decreased the ratio of Treg/Th17( P lt; 0. 01) , and improved the expression of HLA-DR ( P lt; 0. 01) more effectively compared with the routine treatment. Ulinastatin also lowered 28-day mortality of the patients with sepsis, but the difference between the ulinastatin group and the routine group was not significant. Conclusions In severe sepsis patients, there were abnormal overexpression of Treg and Th17, imbalance of Treg/Th17, and underexpression of HLA-DR which imply an immune suppression. Ulinastatin can decrease the expression of Treg and Th17, inverses the ratio of Treg/Th17, and improve the expression of HLA-DR, so as to improve the prognosis of severe sepsis patients.
Objective To investigate the expressions of β1, 3-N-acetyl glucosaminyl transfrases ( Fringe) ( RFNG, LFNG and MFNG) in lung tissues and lung T cells isolated from asthmatic mice, and to explore the role of Fringe in pathogenesis of asthma. Methods Asthmatic BALB/ c mouse model was established by inhalation of ovalbumin after intraperitoneal injection. Meanwhile, the control groups were established by normal saline. Lung tissues were sampled after 24 hours since the last stimulation. T cells were isolated from the lung tissues using percol and NylonFiber. The mRNA expressions of three kinds of Fringe in the lung tissues and lung T cells were examined by reverse transcription-PCR ( RT-PCR) . The protein expressions of Fringe in the lung tissues were detected by Western blot. Results The mRNA expressions of RFNG, LFNG and MFNG were detectable in the lung tissues and lung T cells. The mRNA expressions of RFNG increased in the asthmatic group compared with the control group( lung tissues: 0. 92 ±0. 35 vs 0. 51 ±0. 13, P lt; 0. 01; lung T cells: 0. 33 ±0. 06 vs 0. 18 ±0. 07, P lt; 0. 01) . LFNG mRNA had lower expression level in the asthmatic group( lung tissue: 0. 77 ±0. 32 vs 1. 61 ±0. 31, P lt; 0. 01; lung T cells: 0. 49 ±0. 19 vs 0. 71 ±0. 03, P lt;0. 01) . No difference on the mRNA expression of MFNG was found in the lung tissues( 1. 44 ±0. 29 vs 1. 70 ±0. 44, P gt; 0. 05) . MFNG mRNA expression decreased in the asthmatic group compared with the control group in the T cells( 1. 17 ±0. 04 vs 0. 68 ±0. 07, P lt;0. 05) . The results of western blot were consistent with RT-PCR results of the lung tissues. The expressions of RFNG increased in the asthmatic group( 1. 17 ±0. 04 vs 0. 68 ±0. 07, P lt;0. 05) . The expression of MFNG has no difference between two groups( 8. 10 ±0. 60 vs 9. 12 ±0. 07, P gt;0. 05) . LFNG had a lower expression in the asthmatic group( 4. 11 ±0. 38 vs 6. 41 ±0. 11, P lt; 0. 05) . Conclusion The abnormal expressions of three kinds of Fringe may be involved in the pathogenesis of asthma.
ObjectiveTo explore the effects of intraoperative lymphatic chemotherapy (LC) on immune functions of patients after esophageal carcinoma resection. MethodsPatients who underwent intraoperative LC during esophageal carcinoma resection in the Department of Thoracic Surgery of West China Hospital from March to October,2013 were prospectively included in this study, and patients who underwent esophageal carcinoma resection without intraoperative LC during the same period were also included as the control group. All the patients were divided into a pacitaxel LC group,a fluorouracil LC group,and a control group without LC. A total of 37 patients were included in this study including 25 male and 12 female patients with their age of 42-76 (61.89±7.95) years. There were 15 patients in the pacitaxel LC group,15 patients in the fluorouracil LC group,and 7 patients in the control group. Representative indexes of humoral immunity and cellular immunity in peripheral blood of all the patients were examined preoperatively and on the third and seventh postoperative day, and then compared among the 3 groups. ResultsAll the immune indexes of the 3 groups decreased after surgery to different extent. There was no statistical difference in preoperative and postoperative difference of immunoglobulin concentration between LC groups and the control group (P>0.05). CD8+ T cell count recovered more rapidly after surgery in LC groups than the control group. CD3+ T cells recovered most rapidly after surgery in the fluorouracil LC group. ConclusionLC is beneficial for the recovery of cytotoxic effects of T lymphocytes but may not promote humoral immunity for patients after esophageal carcinoma resection.