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。

Analysis of CD8^+CD28^-T-suppressor cells in living donor liver transplant recipients

BACKGROUND:Human CD8 + CD28 - T-suppressor(Ts) cells have been considered to indicate a reduced need for immunosuppression in pediatric liver-intestine transplant recipients and recipients of deceased heart-kidney transplants.However,in adult-to-adult living donor liver transplantation(A-A LDLT)little information is available and the clinical significance is still unknown. METHODS:Flow cytometry was used to detect the population of CD8+CD28 -Ts cells present in peripheral blood in A-A LDLT recipients(n=31),patients with end- stage liver disease(n=24)and healthy controls(n=19). Meanwhile,we tested the graft function and trough levels of immunosuppression in recipients.The clinical and follow- up data of 31 transplant recipients were analyzed. RESULTS:Compared with diseased controls(P=0.007) and healthy individuals(P=0.000),a notable expansion of CD8 + CD28 - Ts cells was found in recipients of A-A LDLT.This was associated with graft function,levels of immunosuppression and rejection episodes. CONCLUSIONS:To monitor the CD8 + CD28 - Ts cells levels is important to evaluate the immune state of recipients. Meanwhile,it is also important to promote expansion of CD8+CD28 -Ts cells in recipients of A-A LDLT,not only to sustain good graft function and decrease the dosage of immunosuppressants,but also to reduce the occurrence of rejection.

Changes of CD8+CD28-T regulatory cells in rat model of colitis induced by 2,4-dinitrofluorobenzene

AIM:To determine the changes of CD8+ T subsets especially CD8+CD28-T regulatory cells in rat model of experimental colitis induced by 2,4-dinitrofluorobenzene (DNFB). METHODS:The rat model of experimental colitis was induced by enema with DNFB.Ten days later,colonic intraepithelial and splenic lymphooltes were isolated from colitis animals (n=16) and controls (n=8).The proportion of CD8+ T cells,CD8+CD28+ T cells and CD8+CD28-T regulatory cells were determined by flow cytometry. RESULTS:The model of experimental colitis was successfully established by DNFB that was demonstrated by bloody diarrhea,weight loss and colonic histopathology.The proportion of CD8+ T cells in either splenic or colonic intraepithelial lymphocytes was not significantly different between colitis animals and controls (spleen:34.6±7.24 % vs 33.5±9.41%, colon:14.0±8.93 % vs 18.0±4.06 %,P>0.05).But CD8+CD28- T regulatory cells from colitis animals were significantly more than those from controls (spleen:11.3±2.26 % vs 5.64±1.01%, colon:6.50±5.37 % vs 1.07±0.65 %,P<0.05).In contrast, CD8+CD28+ T cells from colitis animals were less than those from controls (spleen:23.3±6.14 % vs 27.8±9.70 %,P=0.06; colon:7.52±4.18 % vs 16.9±4.07 %,P<0.05).The proportion of CD8+CD28-T regulatory cells in splenic and colon intraepithelial CD8+ T cells from colitis animals was higher than that from controls (spleen:33.3±5.49 % vs 18.4±7.26 %, colon:46.0±14.3 % vs6.10±3.72 %,P<0.005). CONCLUSION:Experimental colitis of rats can be induced by DNFB with simplicity and good reproducibility.The proportion of CD8+CD28-T regulatory cells in rats with experimental colitis is increased,which may be associated with the pathogenesis of colitis.

Colon cancer and the immune system:The role of tumor invading T cells

Colon cancer is still one of the leading causes of cancer death worldwide. Although the host immune system has been shown to react against tumor cells, mainly through tumor infi ltrating lymphocytes and NK cells, tumor cells may utilize different ways to escape anti-tumor immune response. Tumor infi ltration of CD8+ and CD4+ (T-bet+) effector T cells has been attributed to a beneficial outcome, and the enhancement of T cell activation through T cell receptor stimulation and co-stimulatory signals provides promising strategies for immunotherapy of colon cancer. Growing evidence supports a role for the Fas/FasL system in tumor immunology, although the mechanisms and consequences of FasL activation in colon cancer are not completely understood. In animal models, depletion of regulatory T cells (CD4+ CD25+ T cells) can enhance the anti-tumor immune response under certain conditions. Taken together, recent insights in the immune reaction against colon carcinoma have provided new approaches to immunotherapy, although much remains to be learned about the exact mechanisms.

Human mesenchymal stromal cells enhance the immunomodulatory function of CD8＋CD28- regulatory T cells

One important aspect of mesenchymal stromal cells （MSCs）-mediated immunomodulation is the recruitment and induction of regulatory T （Treg） cells. However, we do not yet know whether MSCs have similar effects on the other subsets of Treg cells. Herein, we studied the effects of MSCs on CD8＋CD28- Treg cells and found that the MSCs could not only increase the proportion of CD8＋CD28- T cells, but also enhance CD8＋CD28-T cells＇ ability of hampering naive CD4＋ T-cell proliferation and activation, decreasing the production of IFN-γ by activated CD4＋ T cells and inducing the apoptosis of activated CD4＋ T cells. Mechanistically, the MSCs affected the functions of the CD8＋CD28- T cells partially through moderate upregulating the expression of IL-10 and FasL. The MSCs had no distinct effect on the shift from CD8＋CD28＋ T cells to CD8＋CD28- T cells, but did increase the proportion of CD8＋CD28- T cells by reducing their rate of apoptosis. In summary, this study shows that MSCs can enhance the regulatory function of CD8＋CD28- Treg cells, shedding new light on MSCs-mediated immune regulation.

Characterization of myelin oligodendrocyte glycoprotein (MOG)35–55- specific CD8^+ T cells in experimental autoimmune encephalomyelitis

Background:The pathogenesis of multiple sclerosis(MS)is mediated primarily by T cells,but most studies of MS and its animal model,experimental autoimmune encephalomyelitis(EAE),have focused on CD4^+ T cells.The aims of the current study were to determine the pathological interrelationship between CD4 and CD8 autoreactive T cells in MS/EAE.Methods:Female C57BL/6 mice(n=20)were induced by myelin oligodendrocyte glycoprotein(MOG)35-55 peptide.At 14 days after immunization,T cells were isolated from the spleen and purified as CD4^+ and CD8^+ T cells by using CD4 and CD8 isolation kits,and then the purity was determined by flow cytometric analysis.These cells were stimulated by MOG35–55 peptide and applied to proliferation assays.The interferon-gamma(IFN-γ)and interleukin(IL)-4 secretion of supernatant of cultured CD4^+ and CD8^+ T cells were measured by enzyme-linked immunosorbent assays(ELISA).For adoptive transfer,recipient mice were injected with MOG35–55-specific CD8^+ or CD4^+ T cells.EAE clinical course was measured by EAE score at 0–5 scale and spinal cord was examined by staining with hematoxylin and eosin and Luxol fast blue staining.Results:CD8^+ CD3^+ and CD4^+ CD3^+ cells were 86%and 94%pure of total CD3^+ cells after CD8/CD4 bead enrichment,respectively.These cells were stimulated by MOG35–55 peptide and applied to proliferation assays.Although the CD8^+ T cells had a generally lower response to MOG35–55 than CD4^+ T cells,the response of CD8^+ T cells was not always dependent on CD4.CD8^+ T cell secreted less IFN-γand IL-4 compared with CD4^+ T cells.EAE was induced in wildtype B6 na?ve mice by adoptive transfer of MOG35–55-specific T cells from B6 active-induced EAE(aEAE)mice.A similar EAE score and slight inflammation and demyelination were found in naive B6 mice after transferring of CD8^+ T cells from immunized B6 mice compared with transfer of CD4^+ T cells.Conclusion:Our data suggest that CD8^+ autoreactive T cells in EAE have a lower encephalitogenic function but are unique and independent on pathogenic of EAE rather than their CD4^+ counterparts.

CD8＋ Tc-lymphocytes immunodeviation in peripheral blood and airway from patients of chronic obstructive pulmonary disease and changes after short-term smoking cessation

Background Cigarette smoke induces an acute but persisting inflammation in peripheral blood and airway in chronic obstructive pulmonary disease （COPD）,and CD8＋ Tc-lymphocytes are considered as a key role in this process.We aimed to investigate the Tc-lymphocytes immunodeviation in system and local airway of COPD patients and changes of the immunodeviation after short-term smoking cessation.Methods Peripheral blood （PB） and bronchoalveolar lavage fluid （BALF） were collected from 42 patients （14 COPD patients,16 smokers with normal lung function and 12 nonsmokers）,while PB and induced sputum （IS） were obtained from other 19 patients （10 quitting smokers and 9 continuing smokers） at baseline and follow-up respectively of 4-week smoking cessation.Percentages of CD8＋ Tc-lymphocytes （%CD3＋） and Tc1/Tc2 ratios were measured by flow cytometry.Results Percentages of CD8＋ Tc-lymphocytes were higher in COPD patients than those in smokers and nonsmokers in both PB and BALF.Tc1/Tc2 ratio in PB and in BALF from COPD patients was greater than that from smokers and nonsmokers and negatively correlated with FEV1%pre.When comparing the ratios between PB and BALF,significantly positive correlation was found in COPD patients.Furthermore,after 4-week smoking cessation,percentages of CD8＋ Tc-lymphocytes in PB and IS in quitting smokers were decreased compared to that in baseline and continuing smokers,whereas Tc1/Tc2 ratios were not influenced.Conclusions CD8＋ Tc1-trend immunodeviation profiles occurred in both system and local airway of COPD patients.This exceptional immunodeviation could not be relieved by short-term smoking cessation.

Altered CD8＋ T-cell counts as an early predictor of prognosis in critically ill immunocompromised patients with invasive pulmonary aspergillosis

Background The number of critically ill immunocompromised （CIIC） patients has increased dramatically in recent years,and they represent a high risk population for invasive pulmonary aspergillosis （IPA） infection.Host immunity should play a major role in determining the outcome and recovery of these patients.The purpose of this study was to evaluate the dynamic changes in host immune status and its potential influence on prognosis in CIIC patients with IPA.Methods We monitored the evolution of a number of key cellular and humoral parameters on days 1,3,and 10 （D1,D3 and D10） following ICU admission in sixty-two CIIC patients with microbiological evidence of IPA.We included immunoglobulins IgG,IgA and IgM,complement factors C3 and C4,and lymphocyte subgroups CD3＋,CD4＋,CD8＋,CD28＋CD4＋,and CD28＋CD8＋ T cells,CD19＋B cells,and CD3-CD16＋CD56＋ natural killer cells （NK）.Results The primary outcome was 28-day mortality.Thirty-eight （61.3％） patients died within the 28 days following ICU admission.Compared to patients who died,CD3＋,CD8＋,CD28＋CD8＋ T-cell counts on D1,D3,and D10,CD28＋CD4＋ T-cell counts on D3 and D10,and NK counts on D3 and D10 were significantly higher in survivors.Receiver operating characteristic （ROC） analysis of immune parameters predicting 28-day mortality revealed area under the curve （AUC） values of 0.82 （95％ CI 0.71-0.92）,0.94 （95％ CI 0.87-0.99）,and 0.94 （95％ CI 0.85-0.99） for CD8＋ T-cell counts for D1,D3,and D10 respectively,and 0.84 （95％ CI 0.75-0.94）,0.92 （95％ CI 0.85-0.99）,and 0.90 （95％ CI 0.79-0.99） for CD28＋CD8＋ T-cell counts for D1,D3,and D10 respectively.Kaplan-Meier survival analysis showed that CD8＋ T-cell counts ＜149.5×106 cells/L and CD28＋CD8＋ T-cell counts ＜75×106 cells/L at ICU admission were associated with lower survival probabilities in CIIC patients with IPA （both Log rank：P＜0.001）.Conclusions Low CD8＋ and CD28＋CD8＋ T-cell counts were associated with high mortality in CIIC patients with IPA.Early counts of CD8＋ and CD28＋CD8＋ T cells in CIIC patients with IPA may be valuable for predicting outcome.

Experimental Autoimmune Encephalomyelitis Inhibited by Huangqi Guizhi Wuwu Decoction via Th2 Cytokine Enhancement

Background:Huangqi Guizhi Wuwu decoction(HQGZWW)exhibits good effects when administered to treat multiple sclerosis(MS)and its animal model,experimental autoimmune encephalomyelitis(EAE).Understanding the precise mechanism of this decoction is thus important.Based on the findings of our previous study,the aim of the present study was to understand the role of antigen-specific CD8^(+)T-cells on the pathogene sis of MS/EAE when HQGZWW is administered as treatment.Methods:Myelin oligodendrocyte glycoprotein(MOG);-induced mice were administered distilled water,prednisone,and high dose or low dose HQGZWW.After purified CD4^(+)and CD8^(+)T-cells were stimulated with the MOG;peptide,proliferation and cytokine secretion assays were performed.To establish the adoptive transfer EAE model,naive mice were injected with MOG;-CD8^(+)or CD4^(+)T-cells.Results:Significant improvements in EAE score and pathology were observed in the high dose HQGZWW and prednisone groups.Compared to the low dose HQGZWW and distilled water groups,lower antigen-specific re sponses,lower levels of interferon-gamma,and higher levels of interleukin(IL)-4 and IL-10 from CD8^(+)and CD4^(+)T cells were observed in the high dose HQGZWW and prednisone groups.Finally,the EAE score was observed to be similar between the high dose HQGZWW group and prednisone group;however,this finding was not observed in the low dose HQGZWW group.Conclusion:Our findings suggest that high dose HQGZWW has similar effects on cell proliferation,cytokine secretion,and EAE score to prednisone,while low dose HQGZWW does not have such effect.The protective role of HQGZWW against EAE might thus depend on the Th2 cytokine secretion profile induced by either MOG;specific CD8^(+)or CD4^(+)T-cells.