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。

A Comparative Study on the Effect of BCG-PSN and Thymopeptides on T-lymphocyte Subsets of Normal and Immunosuppressed Mice

To compare the effects of polysaccharide nucleic acid fraction of bacillus calmette guerin (BCG PSN) and thymopeptides on T lymphocytes of normal and immunosuppressed mice, CD4 + and CD8 + T lymphocyte subsets of single nucleic cell in thymus, spleen and peripheral blood were detected successively by flow cytometry after application of BCG PSN and thymopeptides. Meanwhile, CD4 +/CD8 + ratio was also calculated. The results showed that both BCG PSN and thymopeptides could decrease the proportion of CD4 + CD8 + T lymphocyte subsets in the thymus, at the same time increase CD4 + T lymphocyte, CD8 +T lymphocyte proportion in the three tissues. The fluctuation in amplitude was greater in thymopeptides group than that in BCG PSN group. It is concluded that acting location of thymopeptides is in thymus, its stimulating action is stronger than that of BCG PSN, while BCG PSN not only accelerates the differentiation in thymus, but also has some direct stimulation to peripheral CD4 +T lymphocytes, and can maintain CD4 +/CD8 + ratio within normal range. So, BCG PSN is safer.

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.

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.

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.

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.

Breeding and preliminarily phenotyping of a congenic mouse model with alopecia areata

In the current study, the alopecia areata gene was introduced into the C57BL/6 （B6） mouse through repeated backcrossing/intercrossing, and the allelic homozygosity of congenic AAtJmice （named B6.KM-AA） was verified using microsatellites. The gross appearance, growth characteristics, pathological changes in skin, and major organs of B6.KM-AA mice were observed. Counts and proportions of CD4＋ and CD8＋ T lymphocytes in peripheral blood were determined by flow cytometry. Results show that congenic B6.KM-AA mice were obtained after 10 generations of backcrossing/intercrossing. B6.KM-AA mice grew slower than B6 control mice and AA skin lesions were developed by four weeks of age. The number of hair follicles was reduced, but hair structures were normal. Loss of hair during disease progression was associated with CD4＋ and CD8＋ T lymphocytes infiltration peri- and intrahair follicles. No pathological changes were found in other organs except for the skin. In the peripheral blood of B6.KM-AA mice, the percentage of CD4＋ T cells was lower and percentage of CD8＋ T cells higher than in control mice. These findings indicate that B6.KM-AA mice are characterized by a dysfunctional immune system, retarded development and T-cell infiltration mediated hair loss, making them a promising new animal model for human alopecia areata.

Expression of chemokine receptors CCR3, CCR5 and CXCR3 on CD4^＋ T cells in CBA/J×DBA/2 mouse model, selectively induced by IL-4 and IL-10, regulates the embryo resorption rate

Background Chemokines and their receptors have been a research focus in transplantation immunology. Chemokines and their receptors play a role in lymphocyte recruitment and differentiation process. This study aimed to observe whether IL-4 and IL-10 may regulate the expression of chemokine receptors CCR3, CCR5 and CXCR3 on CD4^＋ T cells in CBA/J×DBA/2 mouse model and to explore the role of CCR3, CCR5, CXCR3 in immune tolerance in pregnancy. Methods The mouse model of spontaneous abortion （CBA/J×DBA/2） and the normal pregnant mouse model （CBA/J×BALB/c） were used. CBA/J×DBA/2 mice were injected with IL-4 （CBA/J×DBA/2-IL-4）, IL-4 and IL-10 （CBA/J×DBA/2-IL-4＋IL-10）, or normal saline （CBA/J×DBA/2-NS） as a control. The expression of CCR3, CCR5 and CXCR3 on CD4^＋ T cells from mouse peripheral blood was measured by the double-labelled FCM method, and the embryo resorption rate was also examined. Results The embryo resorption rate in the CBA/J×DBA/2 group without any treatment was significantly higher than that in the CBA/J×BALB/c group （17.9% vs 3.7%, P 〈0.01）. The embryo resorption rate in the CBA/J×DBA/2 group immunized with IL-4 or IL-4 together with IL-10 was significantly decreased, compared with that in the control and NS groups respectively. CCR3 expression on CD4^＋ T cells in the CBA/J×DBA/2 group without any treatment was significantly lower than that in the CBA/J×BALB/c group （0.3738±0.3575 vs 1.2190±0.2772, P 〈0.01）; both CCR5 （3.0900±1.5603 vs 1.2390±0.6361, P〈0.01） and CXCR3 （2.4715±0.9074 vs 0.9200±0.5585, P 〈0.01） expressions on CD4^＋ T cells of the CBA/J×DBA/2 group without any treatment were significantly higher than those of the CBA/J×BALB/c group. Significant up-regulation of CCR3 and down-regulation of CXCR3 were found in the CBA/J×DBA/2 group treated with IL-4 （CCR3： 2.0360±0.6944, CXCR3： 1.3510±0.5263, P〈0.01） or IL-4 and IL-10 （CCR3： 1.8160±1.0947, CXCR3：1.0940±0.7168, P〈0.01）. Because of the CCR5, IL-4 and IL-10 （1.9400±0.8504 vs 3.0900±1.5603, P 〈0.05）, but IL-4 alone （2.5310±1.3595 vs 3.0900±1.5603, P 〉0.05） treatment significantly decreased the expression of CCR5 in CBA/J×DBA/2. Conclusions The abnormal expression of CCR3, CCR5 and CXCR3 on CD4^＋ T cells may play an important role in the pathogenesis of spontaneous abortion. The pregnancy immune tolerance may be induced through selective induction of CCR3, CCR5 and CXCR3 expressions by IL-4 together with IL-10.

Relationship between expression of chemokine receptors CCR3, CCR5 and CXCR3 on CD4^＋ T cells and spontaneous abortion in mice

Background Previous studies have shown that local immune cells in the feto-maternal interface are recruited from peripheral blood, and that chemokines and their receptors play an initial and key role in this recruitment process. In this study, we aimed to determine whether spontaneous abortion is associated with the expression of chemokine receptors CCR3, CCR5, and CXCR3 on CD4^＋ T cells. Methods Peripheral blood, spleen, and thymus were collected from the spontaneous abortion mouse model CBA/JxDBA/2 （SA group, n=14）, the normal pregnant mouse model CBA/JxBALB/c （NP group, n=13）, and normal non-pregnant CBA/J mice （NNP group, n=11）. The number of chemokine receptors CCR3, CCR5, and CXCR3 expressed on CD4^＋ T cells was measured by double-label flow cytometry （FCM） method. Results In peripheral blood, the SA group had significantly lower CCR3 expression （P 〈0.01） and higher CCR5 and CXCR3 expression （P 〈0.01） on CD4^＋ T cells than did the NP group. But comparing these chemokines between the SA and NNP groups, there was no significant difference （P 〉0.05）. In spleen, the SA group expressed significantly lower CCR3 expression （P 〈0.01） and higher CCR5 and CXCR3 expression （P 〈0.05） on CD4^＋ T cells than did the NP group. When compared with the NNP group, the SA group had significantly higher CCR3 expression （P 〈0.01）, but was not statistically different with regards to the other two chemokines （P 〉0.05）. In thymus, the SA group had significantly lower CCR3 expression （P 〈0.05） and higher CXCR3 expression （P 〈0.05） on CD4^＋ T cells than the NP group, with no significant difference in CCR5 expression （P 〉0.05）. Compared with the NNP group, the SA group had higher CCR3 expression （P 〈0.01）, but there was no statistical difference in CXCR3 and CCR5 expression （P 〉0.05） between the two groups. Conclusion The abnormal expression of CCR3, CCR5 and CXCR3 on CD4^＋ T cells may play an important role in the pathogenesis of spontaneous abortion.

CD4^＋T细胞对损伤面神经元的保护作用

面神经是脑神经中最易受损的神经之一。面神经断伤将引起一定数量的面神经元死亡或萎缩；而神经元数量的减少，功能的降低必然影响神经功能的恢复。在面神经损伤后免疫反应机制的研究中，研究者发现CD4^＋T细胞可以维持受损面神经元存活，并再次激活萎缩的神经元。本文就该领域的研究进展做一综述。

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.

EB病毒诱导基因3在Th细胞免疫反应中的作用

EB病毒诱导基因3（epstein．Barrvirusinducedgene3，EBl3）是在EB病毒转化的B细胞培养上清液中发现的与细胞因子IL一12同源的可溶性细胞因子受体，在体内以同源二聚体、IL．27异源二聚体和IL一35异源二聚体3种形式存在。在免疫应答反应中，EBl3参与调节效应性T辅助细胞Thl、Th2和Thl7细胞的活化与分化，发挥抗感染和抗变应性炎症等调节作用。最近发现EBl3是调节性T细胞中Foxp3转录因子的下游靶分子，抑制调节性T细胞的分化和其免疫抑制活性，并与多种疾病的发病机制和免疫状态密切相关，是治疗感染性疾病、变应性疾病以及自身免疫性疾病的新靶点。