T SUBSETS AND ANTITUMOR ACTIVITY OF LYMPHOCYTES INFILTRATING HUMAN PRIMARY BRAIN GLIOMAS

Glioma-infiltrating lymphocytes (GIL) were isolated from 9 surgical biopsy specimens of primary brain gliomas using mechanical and enzymatic digestion and discontinuous density gradent centrifugation. During cultured in the presence of interleukin-2 (IL-2) for a period of four weeks, GIL were expanded 48. 4-fold on the average, even up to 118-fold. GIL activated by IL- 2 had specific cytolytic activity against autologous glioma cells. Analysis of T subsets of GIL freshly isolated showed that CD3+ cells were 71.0±11.9%, CD4+ cells 34.2±6.1% and CD3+cells 37.0±7.6%. Ability of activated GIL to produce γ-Interferon (γ-IFN) was significantly higher than that of freshly isolated GIL and autologous peripheral blood lymphocytes (PBL). The results suggest that GIL have many advantages for an adoptive immunotherapy of patients with brain gllomas and be a new type of antitumor immune effector.

Determination of CD_(30) Expression on Peripheral Blood T Lymphocyte Subsets in Patients with Hemorrhagic Fever with Renal Syndrome by FCM

To determine the CD 30 expression on peripheral blood T lymphocyte subsets in patients with hemorrhagic fever with renal syndrome (HFRS) and its clinical implications, double immunofluorescence technique and flow cytometry were used. There was no significant difference among the severe group, mild-moderate group and normal control group in the CD + 4CD - 30 T lymphocyte subset. While the CD + 4CD + 30 T cells of HFRS patients were increased and the difference between severe group and mild-moderate group or normal control group were very significant (P<0.01) and the difference between the mild-moderate group and normal control group was also significant (P<0.05). The CD + 8CD - 30 T cells were increased while the CD + 8CD + 30 T cells decreased obviously in HFRS patients, and the differences among three groups in both subsets were very significant (P<0.01). The results showed that the humoral immunity and cellular immunity are overactive in HFRS patients during acute phase. The loss of balance between T lymphocyte subsets may play an important role in the pathophysiology of HFRS and is closely correlated with the severity of the HFRS.

The Effectof BCG-PSN on T-cell Subsets and Cytokines in Vernal Conjunctiviti

The effects of BCG PSN on T cell subsets and cytokines in vernal conjunctivitis were observed. The level of total IgE was quantitatively determined before and after treatment with BCG PSN by allergen diagnostic instrument in vitro . The content of T cell subsets of peripheral blood and cytokine were determined by using indirect immune fluorescence method, and IL 4 and INF γ were quantified by ELISA. The results showed that the level of total IgE was substantially reduced ( P <0.01) after treatment in the BCG PSN group. Meanwhile, CD + 8 was decreased, CD + 4 and CD + 4/CD + 8 ratio elevated with significant differences ( P <0.05) as compared with pre treatment results. The changes in total IgE, CD + 8 ,CD + 4 and CD + 4/CD + 8 ratio after treatment also presented significant differences ( P <0.05) between BCG PSN group and routine treatment group. The level of IL 4 in serum declined ( P <0.05) after treatment in the BCG PSN group, and INF γ went up ( P <0 05). IL 4 and INF γ in serum showed significant differences ( P <0.05) between two groups after treatment. It is concluded that BCG PSN has a bi directional immunoregulating effect. It can bring CD + 4 and CD + 8 into homeostasis, thereby preventing the occurrence of anaphylaxis. At the same time, BCG PSN can restrain Th 2, decrease the synthesis of IL 4, switch the balance of Th l/Th 2 to Th 1 side, boost up the predominance of Th 1 relatively, which is propitious to perennial stabilization and recovery of vernal conjunctivitis.

Observation of T Lymphocyte Subsets in the Liver of Patients with Advanced Schistosomiasis and Advanced Schistosomiasis Accompanied with Hepatitis B

T lymphocyte subsets in the liver were detected by Avidin-Biotin Complex (ABC) assay in 22 patients with advanced schistosomiasis (AS) and 5 cases of AS accompanied with hepatitis B. T lymphocytes in the liver of AS patients were distributed in the peripheral layer of egg granuloma or the area near eggs in non-granuloma. No infiltrative T lymphocytes were observed in area with extensive fibrosis. There was infiltration of many T cells in the portal tract, piecemeal and focal necro- sis area as well as in hepatic sinus in AS patients accompanied with hepatitis B. CD8+ T cells (sup pressor/cytotoxic T cells, Ts/Tc) in the liver were predominant in the two groups. In AS patients, marked hepatic fibrosis, a small number of T cell infiltration and slight hepatocellular degeneration and necrosis were observed. However, obvious hepatocellular degeneration and necrosis were seen in AS patients accompanied with hepatitis B, and 3 cases of them developed active liver cirrhosis. The results indicated immune response was weak in the liver in AS patients and T. cells might be predominant in the subset of CDS+ T lymphocytes. Cellular immune response was relatively strong in AS patients accompanied with hepatitis B and the infiltrative CD8+ T lymphocytes might be mainly Tc cells.

Therapeutic Effect of Integrative Traditional Chinese and Western Medicine on 51 SARS Patients and Its Influence on Their TLymphocyte Subsets

Objective: To observe the clinical effect of integrative Chinese a nd western medicine (ICWM) in treatment of patients with severe acute respirator y syndrome (SARS) and its influence on their T lymphocyte subsets.Methods: Fifty one patients with SARS of severe type were obser ved with synchronous non randomized controlled method. They were divided into the ICWM group (29 patients) and the western medicine (WM) group (22 patients). Western medical treatment was applied to both groups, but to the ICWM group, Ch inese medicine was given additionally. The therapeutic course was 2-3 weeks for both groups. Clinical effect and changes of T lymphocyte subsets (CD4 +) aft er treatment were observed.Results: In the ICWM group, 26 patients (89.66%) were cured and 3 (10.34%) died, while in the WM group, 12 (54.55%) cur ed and 10 (45.45%) died, thus comparison of the cure rate between the two groups showing significant difference ( P <0.01). The score of clinical symptoms in the ICWM group was decreased from 7.14±5.20 scores before treatment to 1.82±3. 75 scores after treatment, while in the WM group, it lowered from 7.36±3.84 sco res before treatment to 5.17±4.17 scores after treatment, significant diffe rence shown in the comparison of the values between the two groups after treatme nt (P<0.01). Immunological function test showed that CD4 + T lymphocyte in the ICWM group rose from 361±278 cells/mm 3 before treatment to 630±454 c ells/mm 3 after treatment, showing significant difference( P <0.01 );bu t in the WM group, it merely rose from 288±186 cells/mm 3 to 376±285 cells/mm 3 in the corresponding period (P>0.05). Conclusion: ICWM could improve the clinical symptoms of SARS pa tients markedly, enhance their T lymphocyte immune function, and reduce their mortality.

Study on T lymphocyte subsets and NK cells in patients with Graves' disease combined with type 2 diabetes

Objective To investigate changes in T lymphocyte subsets and NK cells in patients with simple Graves’ disease(GD)and Graves’ disease combined with type 2 diabetes mellitus(GD/T2DM).Methods Fifteen cases of GD/T2DM were selected from our hospital from November 2001 to November 2004.Before and after therapy thyroid function,thyroglobulin antibody(TGA),thyroid microsomal antibody(TMA)and blood glucose level were measured,and T lymphocyte subsets(CD3,CD4,CD8,CD4/CD8)and NK cells(CD56)were measured by immunofluorescence double labeling monoclonal antibody and flow cytometry,respectively.At the same time,comparison was made with simple GD(15 cases),T2DM(15 cases)and healthy control(20 cases).Results Before therapy,CD4/CD8,CD4 and NK cells in GD/T2DM were less than normal,and there was no significant difference in comparison with simple GD(P<0.05).In T2DM group,only CD4/CD8 and CD4 were less than those of healthy controls(P<0.05).When thyroid function recovered after 1 to 3 months of methimazole treatment in both GD/T2DM and simple GD groups,various indexes recovered,which were more obvious in simple GD.Conclusion Immune hypofunction of GD may be the key to the immune abnormality of GD/T2DM,which is more significant than that of simple GD or T2DM.The recovery of thyroid function and immune abnormality is not consistent,and the recovery of GD is more significant than that of GD/T2DM.

Immunohistochem ical Study of T Lym phocyte Subsets onFrozen Substituted Plastic Em bedding Section of the BoneMarrow from Patients with Myelodysplastic Syndrom e andIts ClinicalIm plication

An immunohistochemical study of T lymphocyte subsets on frozen substituted plastic embedding bone marrow sections obtained from 10 patients with myelodysplastic syndrome (MDS) was presented. The results of qualitative and quantitative immunohistochemical analysis are as follows: (1) Labile antigens of T lymphocytes were well preserved, thus allowing analysis of distribution of T lymphocyte subsets in situ ; (2) the average number of T 3, T 4 and T 8 lymphocyte of the diffuse infiltrate was about 2 %, 0.4 %, 0.5 %, respectively, of all nucleated cells in bone marrow, and T 4/T 8 of T cells were below 1.0 in patients with MDS; (3) there were cases of RAS showing T lymphocyte aggregation in bone marrow, but no patient exhibited progressive refractory anemia with excess of blasts(RAEB) and RAEB in transformation (RAEBT). These findings indicated that the immunological abnormalities are of importance in the evaluation of pathogenesis and prognosis of MDS.

T LYMPHOCYTE SUBSETS OF PERIPHERAL BLOOD AND THEIR RELATIONSHIP WITH BETA－2－MICROGLOBULIN IN MULTIPLE MYELOMA

patients with multiple myeloma (MM) were investigated for lymphocyte subsets using APAAP (Alkaline Phosphastase Anti-Alkaline Phosphatase) technique and a panel of monoclonal antibodies.Radioimmunoassay was used to examine serum beta-2-microglobulin (Sβ2-MG).The results showed that OKT3, OKT4 positive cells and OKT4/OKT8 ratio reduced significantly. OKT8 positive cells increased sharply in MM. We conclude that,in MM,all patients have severe abnormalities in the number and proportion of T cells.The level of S β2-MG increased significantly, especially in the initial and relapsing patients.But,we did not find the relationship between Sβ2-MG and T subsets.

THE EFFECT OF LOCAL RADIOTHERAPY ON T LYMPHOCYTE SUBSETS OF CANCER PATIENT

In the study,we utilized the OKT monoclonal antihuman T Iymphocyte antibodies by indirect immunoenzyme staining method to determinc T lymphocyte subsets in the peripheral blood of 72 cancer patients treated by local radiotherapy.The haematological assays were performed immediately Pre-and Postradiotherapy.In the postoperative radiotherapy group(PR group) of 40 breast cancer patients treated by prophylactic postoperative radiotherapy,after radiotherapy blood platelet,white blood cell and lymphocyte count decreased significantly.CD2,CD4 and CD8 cell counts all very significantly decreased.Proportion of CD2 and CD2 had no apparent change but proportion of CD8 increased significantly.The group appeared apparent hypoimmunity.While the complete response group(CR group) of 32 cancer paticnts treateil with local radiothcrapy,after radiotherapy blood Platelet,white blood cell and lymphocyte count had no significant decrease.CD2,CD4 and CD8 cell count had no significant decrease.Proportion of CD2,CD4 and CD8 had no apparent change.The CR group had no apparent hypoimmunity.Comparing decrease rate of immunologic parameters in these two groups,decrease rate of white blood cell,lympheeyte and T lymphocyte subsets count in CR group significantly were slightened rather than pR group.This study proved that effect of radiation-damaging tumor cell enhance antigenicity of the tumor and stimulate host cell immunity including a type of T-lymphogenesis.Induced hypoimmunity by postoperative radiotherapy should be avoided in this point of view.

EFFECTS OF MICROWAVE TREATMENT ON THE LYMPHOCYTE SUBSETS IN CASES OF CHRONIC LYMPHEDEMA OF EXTREMITIES

Objective To determine the immune status of T lymphocyte subsets of patients with chronic lymphedema of extremities and the effects of microwave treatment. Methods Patients with lymphedema of extremities (n = 20) and normal volunteers (n = 10 ) were studied by fiuorescein isothiocyanate (FITC) or phycoerythrin (PE)-conjugated monoclonal antibo dies (MoAbs) and dual color flow cytometry to examine the changes of lymphocyte phenotype in them before and after treatment. Results The percentage of CD+4 T lympcutes and CD+4 /CD+8 T cell ratios in lymphedema group were less than those in control group before treatment. They increased significantly and restored to nearly normal level after microwave treatment. The percentage of CD +8 and HLA-DR + T lymphocytes in lymphedema group also demonstrated significant decline after treat- ment. Conclusion Disorder of T lymphocyte subsets existed in patients with chronic lymphedema of extremities, and microwave treatment can improve the states and enhance the cellular immunity of the patients.

Association of T lymphocyte subset counts with the clinical features of colorectal cancer

Background:Colorectal cancer(CRC)is a common gastrointestinal malignancy.The T lymphocyte subsets are important in the develop-ment,invasion and metastasis of tumors,including CRC.Nevertheless,limited research has explored the relationship between T cell subpopu-lations and the clinical characteristics of CRC.This study compared the T lymphocyte subsets in patients with CRC and healthy individuals,and assessed the relationship between these values and clinical characteristics.Methods:Peripheral blood was collected from 100 patients with CRC and 54 healthy individuals.The numbers of CD3^(+)T,CD4^(+)T,and CD8^(+)T lymphocytes,NK cells,and the CD4^(+)T/CD8^(+)T ratio in peripheral blood were measured using flow cytometry,and were compared between CRC patients and healthy individuals.Spearman's correlation analysis was performed to investigate the relationship between the T lymphocyte subsets in patients diagnosed with CRC and the levels of carcinoembryonic antigen(CEA)and thymidine kinase 1(TK1).Receiver operating characteristic(ROC)curves were utilized to evaluate the potential utility of the T lymphocyte counts in predicting lymph node metastasis,vas-cular infiltration,and high Ki-67 expression.Results:The CRC patients had lower counts of CD3^(+)T,CD4^(+)T,and CD8^(+)T lymphocytes compared to the healthy population(P<0.05).However,no significant differences were observed in the CD4^(+)/CD8^(+)ratio or NK cells(P>0.05).Notably,the CD3^(+)T,CD4^(+)T,and CD8^(+)T lym-phocyte counts were higher in patients with stageⅠ-Ⅱdisease,no lymph node metastasis,no vascular invasion,and low Ki-67 expression than in those with stageⅢ,lymph node metastasis,vascular invasion,and high Ki-67 expression(P<0.05).There was a negative association be-tween the CD3^(+)T,CD4^(+)T,and CD8^(+)T lymphocyte counts and CEA and TK1 levels in patients with CRC.The ROC curves demonstrated that CD3^(+)T,CD4^(+)T,and CD8^(+)T lymphocyte counts had significant predictive value for lymph node metastasis,vascular infiltration,and high Ki-67 expression.Conclusions:The peripheral blood CD3^(+)T,CD4^(+)T,and CD8^(+)T lymphocyte counts are related to the clinical traits of patients with CRC and can predict the prognosis of the disease.

外周血CD8^(+)T淋巴细胞亚群检测对过敏性鼻炎临床疗效的评估价值

目的探讨外周血CD8^(+)T淋巴细胞亚群检测对过敏性鼻炎(AR)临床疗效的评估价值。方法前瞻性选取2021年3月~2022年2月温州市中西医结合医院耳鼻咽喉科门诊治疗的120例AR患者和40例健康志愿者,分别设为研究组和对照组,比较两组外周血CD8^(+)T细胞、CD8^(+)CD28^(+)T细胞、CD8^(+)CD28-T细胞比率;研究组接受鼻用激素联合口服抗组胺药物治疗方案,根据治疗效果分为有效组(n=107)和无效组(n=13),比较有效组和无效组治疗前CD8^(+)T细胞、CD8^(+)CD28^(+)T细胞、CD8^(+)CD28-T细胞比率;Pearson相关性分析法分析外周血CD8^(+)T淋巴细胞亚群水平与鼻部症状总评分(totalnose symptomscore,TNSS)的相关性;以受试者工作特征(ROC)曲线分析治疗前CD8^(+)T淋巴细胞亚群对AR临床疗效的预测价值。结果研究组治疗前外周血CD8^(+)CD28^(+)T细胞比率高于对照组,CD8^(+)CD28-T细胞比率低于对照组(P<0.05)。研究组总有效率为89.17%;有效组治疗前CD8^(+)CD28^(+)T细胞比率低于无效组,CD8^(+)CD28-T细胞比率高于无效组(P<0.05);Pearson相关性分析显示,CD8^(+)CD28^(+)T细胞比率与TNSS评分呈正相关(r=0.324,P=0.006),CD8^(+)CD28-T细胞比率与TNSS评分呈负相关(r=-0.541,P=0.000)。ROC曲线结果显示,CD8^(+)T细胞、CD8^(+)CD28^(+)T细胞、CD8^(+)CD28-T细胞比率评估AR治疗无效的曲线下面积(AUC)(95%CI)分别为0.647(0.555~0.732)、0.683(0.592~0.765)、0.911(0.845~0.955),其中CD8^(+)CD28-T细胞的评估效能优于CD8^(+)T细胞、CD8^(+)CD28^(+)T细胞比率(P<0.05)。结论AR患者外周血CD8^(+)CD28^(+)T细胞比率升高,CD8^(+)CD28-T细胞比率降低,CD8^(+)T细胞比率相对稳定,治疗前CD8^(+)CD28-T细胞比率可作为预测AR患者的临床疗效的参考指标。

CD4^+CD25^(high) Regulatory Cells in Peripheral Blood of NSCLC Patients

The proportion and changes of CD4^＋CD25^high regulatory T cells （Trs） in peripheral blood of non-small cell lung cancer （NSCLC） patients were analyzed and their clinical significance explored. The peripheral blood was collected from 61 patients with NSCLC and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets （CD3^＋, CD4^＋ and CD8^＋） and CD4^＋CD25^high Tr cells. The results showed that the proportion of CD4^＋CD25^high Tr cells in NSCLC group was significantly higher than in control group [（4.36 ±2.07） % vs （2.04±1.03） %, P〈0.01]. The proportion of CD4^＋CD25^ high Tr cells in late stage was higher than that in early stage [stages Ⅰ ＋Ⅱ （2.264±0.6） %; stage Ⅲ（3.284± 1.38） %; stage IV （6.06 4±4.08） %] （P〈0.05）. Kaplan-Meier survival analysis revealed that the prognosis of the patients who had higher proportion of CD4^＋CD25^high Tr cells in peripheral blood was worse （P=0.0026）. In conclusion, the relative increase in CD4^＋CD25^high Tr cells in peripheral blood may be related to im- munosuppression and tumor progression in patients with NSCLC. This finding suggests that CD4^＋CD25^high Tr cells in peripheral blood of NSCLC may be positive for prognosis analysis. The use of depletion of the CD4^＋CD25^high Tr cell therapy to treat NSCLC patients may be an effective strategy.

Advanced Glycation End Products Promote Differentiation of CD4^+ T Helper Cells toward Pro-inflammatory Response

This study investigated the effect of advanced glycation end products（AGEs） on differentiation of na ve CD4＋T cells and the role of the receptor of AGEs（RAGE） and peroxisome proliferator-activated receptors（PPARs） activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin（BSA） with glucose. Human na ve CD4＋T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin（sh） RNA knock-down experiment, na ve CD4＋T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-XTM293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4＋T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T（Treg） cells was determined by a [3H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from na ve CD4＋T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in na ve CD4＋T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4＋T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα; PPARγ agonist, PGJ2, inhibited the effect of AGEs on na ve CD4＋T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4＋T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity. ＋

Clinical Evaluation of Integrative Chinese and Western Medicine in Treating SARS

Objective:To evaluate the effective treatment with integrative Chinese and western medicine (ICWM) of severe acute respiratory syndrome (SARS). Methods: The 65 cases of SARS patients were treated with ICWM and 37 cases with western medicine (WM) alone as the control in our hospital from March 11 to April 30, 2003. The results were analyzed, with T subset profile tested for all the 102 patients. Results: Sixty-five cases of SARS patients treated with ICWM included 52 mild cases of whom 51 cases were cured (98.1%) and no patient died. As to the 13 severe cases in this group, 9 were cured (69. 2%) and 2 died (15. 4%). Of the 37 cases treated with WM alone, 16 out of the 18 common cases were cured (88. 9%) and 10 out of the 19 severe cases were cured (52. 6%), with the other 9 died (47. 4%). Comparison between the two groups (P=0. 061) showed the difference was close to the value for statistical significance. But considering that the number of samples was small, more cases are needed for further study. The immune function examination: in cases of the common type, after ICWM treatment, CD4 + T lymphocytes were increased from 360±247 cell/mm3 to 563±479 cell/mm3 (P<0. 05). In the group treated with WM alone, CD4+ T lymphocytes were increased from 331 ±193 cell/mm3 to 772±354 cell/mm3 (P<0. 05). As to the severe cases after ICWM treatment, CD4+ T lymphocytes increased from 352±279 cell/mm3 to 525±490 cell/mm3 (P>0. 05). In the group of severe cases treated with WM alone, CD4+ T lymphocytes reduced from 229±69 cell/mm3 to 205±108 cell/mm3 (P>0. 05). While after ICWM treatment, CD4+ T lymphocytes were significantly higher than that of the group treated with WM alone (P< 0. 05). Conclusion: Compared with the group treated with WM alone, ICWM can significantly improve the prognosis, reduce the mortality as well as improve the immune function of SARS patients.

Effect of Early Intensive Insulin Therapy on Immune Function of Aged Patients with Severe Trauma

This study examined the effect of intensive insulin therapy on immune function and inflammatory factors at the early phase after severe trauma. At day 1, 3, 5, 7 after admission, subsets of CD4＋ helper T lymphocytes （Th1/Th2） and human leukocyte antigen （HLA）-DR expression on CD14＋ monocytes were flow cytometrically measured. Levels of cytokines, including tumor necrosis factor-alpha （TNF-α）, interleukin-6 （IL-6）, interleukin-10 （IL-10） and other immunity markers, such as IgA, IgG, IgM, C3, C4 and C reaction protein （CRP） were examined in two groups. The results showed that TNF-α, IL-6 and CRP levels in the intensive insulin therapy group were significantly lower than those in the conventional therapy group, whereas IL-10 levels were substantially increased after intensive insulin therapy. C3 level at day 3, 5, 7 and C4 levels at day 5, 7 were lower in the intensive therapy group than in the conventional therapy group. Th1/Th2 ratios decreased gradually over time in both groups, and were much lower at day 3, 5, 7 in intensive therapy group. There were significant differences among day 3 to day 7 after admission in HLA-DR expression in CD14＋ monocytes. It was concluded that the intensive insulin therapy could decrease pro-inflammatory cytokines and increase anti-inflammatory cytokines in the elderly suffering from severe trauma, at the same time, with complement recovery being delayed. Moreover, intensive insulin therapy promoted immune suppression and, therefore, measures need be taken to address the issue.

The Immunity Induced by Recombinant Spike Proteins of SARS Coronavirus in Balb/c Mice

The immune effect of two recombinant protein fragments of spike protein in severe acute respiratory syndrome coronavirus (SARS CoV) was investigated in Balb/c mice. Two partial spike gene fragments S1 (322-1464 bp) and S2 (2170-2814 bp) of SARS coronavirus were amplified by RT-PCR, and cloned into pET-23a prokaryotic expression vector, then transformed into competent Escherichia E.coli BL21 (DE3)(pLysS) respectively. Recombinant proteins were expressed and puri- fied by Ni2+ immobilized metal ion affinity chromatography. The purified proteins mixed with com- plete Freund adjuvant were injected into Balb/c mice three times at a two-week interval. High titer antibody was detected in the serum of immunized Balb/c mice, and mice immunized with S1 protein produced high titer IgG1, IgG2a, IgG2b and IgG3, while those immunized with S2 protein produced high titer IgG1, IgG2a, but lower titer IgG2b and IgG3. Serum IFN-γ concentration was increased significantly but the concentrations of IL-2, IL-4 and IL-10 had no significant change. And a marked increase was observed in the number of spleen CD8+ T cells. The results showed that recombinant proteins of SARS coronavirus spike protein induced hormonal and cellular immune response in Balb/c mice.

Expression of Snail in bladder urothelial carcinoma and its relationship with E-cadherin and a subset of T cell groups

Objective To investigate the expression of Snail in bladder urothelial carcinoma and evaluate its relationship with E-cadherin and a subset of T cell groups. Methods Immunohistochemical method was used to detect the expression of Snail and E-cadherin proteins in tissue

Immune‑Ageing Evaluation of Peripheral T and NK Lymphocyte Subsets in Chinese Healthy Adults

Ageing is often accompanied with a decline in immune system function,resulting in immune ageing.Numerous studies have focussed on the changes in different lymphocyte subsets in diseases and immunosenescence.The change in immune phenotype is a key indication of the diseased or healthy status.However,the changes in lymphocyte number and phenotype brought about by ageing have not been comprehensively analysed.Here,we analysed T and natural killer(NK)cell subsets,the phenotype and cell differentiation states in 43,096 healthy individuals,aged 20–88 years,without known diseases.Thirty-six immune parameters were analysed and the reference ranges of these subsets were established in different age groups divided into 5-year intervals.The data were subjected to random forest machine learning for immune-ageing modelling and confirmed using the neural network analysis.Our initial analysis and machine modelling prediction showed that na.ve T cells decreased with ageing,whereas central memory T cells(Tcm)and effector memory T cells(Tem)increased cluster of differentiation(CD)28-associated T cells.This is the largest study to investigate the correlation between age and immune cell function in a Chinese population,and provides insightful differences,suggesting that healthy adults might be considerably influenced by age and sex.The age of a person's immune system might be different from their chronological age.Our immune-ageing modelling study is one of the largest studies to provide insights into‘immune-age’rather than‘biological-age’.Through machine learning,we identified immune factors influencing the most through ageing and built a model for immune-ageing prediction.Our research not only reveals the impact of age on immune parameter differences within the Chinese population,but also provides new insights for monitoring and preventing some diseases in clinical practice.

Polarization and Apoptosis of T Cell Subsets in Idiopathic Thrombocytopenic Purpura

It is well-known that idiopathic thrombocytopenic purpura （ITP） is an acquired organ-specific autoimmune hemorrhagic disease and dysfunctional cellular immunity is considered important in the pathophysiology of ITP. However, polarization patterns and apoptosis profiles of T lymphocytes remain unclear. In this study, we investigated the polarization of T cell subsets, the expressions of apoptotic proteins Fas/FasL on the subsets and the level of anti-apoptotic gene bcl-2 and bax mRNA. It was demonstrated that the ratios of Thl/Th2 and Tcl/Tc2 in ITP children were increased obviously and that the average percentages were increased clearly for Thl and Th2, but not for Tcl and Tc2. In ITP children, the enhancing expressions were detected for FasL on Thl and Tcl and for Fas on Th2 and Tc2. With increasing level of bcl-2 mRNA and decreasing expression of bax mRNA in ITP children, the ratio of bcl-2/bax mRNA was improved obviously, which was positive correlated with the ratio of Thl/Th2. Taken together, our findings indicate that ITP is a Thl predominant disease. This polarization pattern of T cell subsets might be related to the high ratio of bcl-2/bax mRNA and the abnormal expressions of Fas and FasL on T cell subsets.