手术联合树突状细胞治疗肾透明细胞癌的效果

目的 观察手术联合树突状细胞治疗肾透明细胞癌的临床效果,为肾癌治疗提供依据.方法 入组患者分为A、B两组,A组(n=33)为术后接受细胞治疗组,B组(n=37)为单纯手术治疗组.所有入组患者分别于手术治疗前8周及治疗后8周采外周血行流式细胞术检测T淋巴细胞亚群(CD3+、CD4+、CD8+、CD4 +/CD8+比值),了解治疗前后机体免疫水平.结果 Ⅰ、Ⅱ期患者A、B组治疗前后T淋巴细胞亚群差异无统计学意义(P＞0.05);Ⅲ期患者A组治疗前后T淋巴细胞亚群差异有统计学意义(P＜0.05),B组治疗前后T淋巴细胞亚群差异无统计学意义(P＞0.05).A组治疗前后T淋巴细胞亚群差异有统计学意义(P＜0.05),B组差异无统计学意义.结论 手术联合树突状细胞治疗可明显提高Ⅲ期肾透明细胞癌患者术后免疫功能,对肾癌治疗具有积极意义.     

氯胺酮对兔全脑缺血-再灌注海马细胞凋亡及TNF-β和IL-10的影响

目的 研究氯胺酮对兔全脑缺血-再灌注损伤中海马细胞凋亡及肿瘤坏死因子β(TNF-β)、白细胞介素-10(IL-10)表达的影响.方法 新西兰大白兔45只,随机分为三组,每组15只.A组仅分离股动脉、股静脉和双侧颈总动脉.B组和C组采用股动脉放血,夹毕颈总动脉30min制作全脑缺血-再灌注模型.C组经股静脉推注氯胺酮4.5 mg/kg.取海马组织行HE染色、TUNEL、免疫组化染色,计数CA1区存活细胞、凋亡细胞、TNF-β和IL-10阳性细胞个数.结果 再灌注后12～72 h,C组存活细胞明显多于B组(P＜0.05),凋亡细胞明显少于B组(P＜0.01).再灌注后6～72 h,B、C组TNF-β明显高于A组(P＜0.05).再灌注后6h和72 h,C组TNF-β明显少于B组(P＜0.05).再灌注后6h,C组IL-10阳性细胞数明显多于A、B型,再灌注后72 h明显少于A、B组(P＜0.05).结论 氯胺酮可通过调节细胞免疫应答,减轻炎症反应,减少细胞坏死和凋亡,具有良好的脑保护作用.     

吞噬刺激素(Tuftsin)对肝癌细胞凋亡的影响

目的研究Tuftsin与肝癌细胞凋亡之间的关系，并对其可能作用机制进行探讨。方法昆明种小鼠30只随机分成3组：对照组(A组)、全脾切除后给予生理盐水组(B组)、全脾切除后给予Tuftsin组(C组)，比较这三组小鼠移植性肝癌生长情况。采用免疫组织化学及脱氧核糖核酸末端转移酶介导的缺口末端标记(TUNEL)技术，对三组肝癌组织中总的T淋巴细胞(即CD3^ T细胞)和凋亡细胞进行原位观察和比较。结果B组移植性肝癌重量大于A、C组的重量；B组肝癌细胞凋亡指数低于A、C组；肝癌细胞凋亡指数与CD3 T细胞数有显著正相关关系。结论全脾切除能减少肝癌细胞凋亡，促进移植性肝癌生长。Tuftsin能通过CD3^ T细胞数的增加，促进肝癌细胞凋亡。     

CD4+CD25+Treg细胞对肿瘤特异性CTL杀伤效果的影响

目的 探讨CD4+CD25+Treg细胞对肿瘤特异性细胞毒T细胞(CTL)杀伤效果的影响及机制.方法 将C57BL/6小鼠80只随机分为4组,每组20只.A组:树突状细胞(DC)与T细胞共同培养前删除CD4+CD25+Treg;B组:DC与T细胞共同培养后删除CD4+CD25+Treg;C组:DC与T细胞共同培养时不删除CD4+CD25+Treg;对照组:无DC诱导的T细胞.应用脾脏来源DC细胞诱导T细胞制备CTL,在CTL形成的不同时期采用MACS法删除CD4+CD25+Treg.应用噻唑蓝(MTY)比色法检测不同组别CTL对B16黑色素瘤细胞的杀伤效果.同时应用酶联免疫吸附试验(ELISA)法检测细胞培养液中白细胞介素(IL)-2、干扰素(IFN)-γ含量变化.结果 3组实验组CTL杀伤率明显高于对照组(P<0.05).删除CD4+CD25+Treg的A组、B组CTL杀伤率明显高于未删除的C组(P<0.05).但CTL形成的不同时期删除CD4+CD25+Treg对CTL杀伤率的影响无统计学意义(P>0.05).IL-2、IFN-γ含量变化与杀伤率呈现相同的变化趋势.结论 删除CD4+CD25+Treg细胞可明显提高CTL的杀伤效果,是消除肿瘤免疫耐受机制的新途径.     

胰腺癌患者外周血CD4-CD8-T细胞的检测及其临床意义

目的 研究胰腺癌患者外周血CD4-CD8-T细胞含量,并探讨其含量与肿瘤的临床病理类型及临床分期的相关性.方法 应用流式细胞仪分析胰腺癌患者和健康对照组外周血中CD4-CD8-T细胞含量.结果 20例胰腺癌患者CD4-CD8-T细胞占CD3+T细胞的比例为(4.13±1.81)%;20例健康对照组CD4-CD8-T细胞占CD3+T细胞的比例为(6.39±1.83)%,两组之间差异有统计学意义(P＜0.01).高、中分化的胰腺癌患者CD4-CD8-T细胞占CD3+T细胞的比例为(4.41±1.66)%;低分化胰腺癌患者CD4-CD8-T细胞占CD3+T细胞比例为(4.18±2.32)%,两组之间差异无统计学意义(P＞0.05).Ⅲ、Ⅳ期胰腺癌患者CD4-CD8-T细胞占CD3+T细胞的比例为(2.96±1.50)%;Ⅰ、Ⅱ期胰腺癌患者CD4-CD8-T细胞占CD3+T细胞的比例为(5.09±1.50)%,两组之间差异有统计学意义(P＜0.01).结论 CD4-CD8-T细胞在胰腺癌中低表达,可能促进胰腺癌进一步发生发展.     

甲状腺癌患者手术前后CD8+CD28-调节性T细胞的变化与临床意义

【摘要】〓目的〓探讨甲状腺癌患者手术前后CD8+CD28-调节性T细胞的变化与临床意义。方法〓收集接受手术治疗的双侧甲状腺癌患者30例作为观察组(A),分别收集良性甲状腺瘤患者(B)及健康体检者(C)各30例作为对照组;采用流式细胞术检测三组外周血CD8+CD28-调节T细胞百分含量及其胞内细胞因子转化生长因子β1(TGF-β1)及白介素10(IL-10)的平均荧光强度;比较术后4周甲状腺癌患者上述指标的变化。结果〓与C组比较,A及B组CD8+CD28- T细胞及TGF-β1均升高,但B与C组无统计学差异,而A组两者均显著高于C组(P<0.05);与C组比较,A及B组IL-10均显著升高,且三组间两两比较均有统计学差异(P<0.05);手术后4周复查显示A组患者CD8+CD28-T细胞、TGF-β1及IL-10三者与手术前比较均有显著性差异(P<0.05),其中以CD8+CD28-T细胞的回落最为明显,TGF-β1次之。结论〓CD8+CD28-T细胞、TGF-β1及IL-10表达增加是甲状腺癌患者的一个重要免疫学特征;TGF-β1是相对特异的一个细胞因子,在评价术后疗效较IL-10敏感。     

大鼠腹主动脉瘤形成过程中T细胞及CD4+CD28-T细胞的动态变化及意义

目的 探讨T淋巴细胞及其亚群在腹主动脉瘤发病中的作用.方法 50只Wistar大鼠随机分成A、B、C、D、E5组,每组10只,A组为正常对照组,B、C、D、E组分别为术后3、7、14、28 d组,运用猪胰弹力酶灌注法建立腹主动脉瘤模型.免疫组化方法检测上述各组大鼠腹主动脉局部T、B淋巴细胞及巨噬细胞的表达水平,流式细胞术检测各组大鼠外周血中CD4+ CD28-T淋巴细胞亚群的分布变化.结果 大鼠腹主动脉局部T、B淋巴细胞及巨噬细胞表达的变化:术后7dT细胞显著升高(40±15,P＜0.05),B细胞及巨噬细胞有升高,但差异无统计学意义(P＞0.05);术后14 d,3种细胞表达升高最为显著(P＜0.05);术后28 d,3种细胞的表达较术后14 d明显下降.术后各组大鼠外周血CD4+、CD28-T细胞均有升高(P＜0.05),以术后7d比例最高(8.1％±2.1％,P＜0.05).结论 T淋巴细胞是腹主动脉瘤形成过程中最重要的炎症细胞之一,T淋巴细胞的浸润及其亚群分布变化所致的免疫及炎症反应在腹主动脉瘤形成早期可能起着重要的触发及调节作用.     

电刺激对脑缺血/再灌注大鼠脊髓背角细胞凋亡的影响

目的 观察穴位电刺激对脑缺血/再灌注大鼠脊髓背角细胞凋亡的影响. 方法 采用Longa线栓法建立大鼠左侧大脑中动脉缺血/再灌注模型,将80只大鼠按随机数字表法分为对照组(C组)和电刺激组(E组)(每组40只).E组予针刺患侧曲池、足三里穴位,C组不予干预.两组于造模后l d(T1)、3 d(T2)、7 d(T3)、14 d(T4)4个时间点随机选取10只大鼠,应用TUNEL法检测脊髓背角细胞凋亡,免疫组织化学检测caspase-3蛋白表达. 结果 两组组内比较,脊髓背角细胞凋亡率及caspase-3蛋白表达阳性细胞率,T2、T3和T4时都比T1时高(P＜0.05或P＜0.01);组间比较,E组T2、T3、T4时细胞凋亡率及caspase-3蛋白表达阳性细胞率均比C组降低,差异有统计学意义(P＜0.05),T1时两组细胞凋亡率及caspase-3蛋白表达阳性细胞率差异无统计学意义(P＞0.05). 结论 穴位电刺激可以降低脊髓背角细胞凋亡.     

外周血γδT细胞在胰腺癌裸鼠过继免疫治疗中的作用

目的探讨外周血γδT细胞在胰腺癌裸鼠种植瘤体内过继免疫治疗中的作用。方法用胰腺癌细胞株Cap1注入BALB/c裸鼠皮下建立胰腺癌裸鼠种植瘤模型。裸鼠随机分为3组,每组10只。A组为γδT细胞治疗,B组为αβT细胞治疗,C组给以无血清RPMI1640作为对照。从胰腺癌患者外周血中提取淋巴细胞,体外特异性扩增γδT细胞后注入裸鼠腹腔内,观察种植瘤裸鼠生存时间、肿瘤体积和肿瘤病灶坏死程度。结果A组成瘤率9/10,B、C组均为8/10(P>0.05)。当肿瘤生长到16、37和52d时,A组与B、C组肿瘤体积差别具有统计学意义(P值分别为0.019和0.022)。120d时C组8只种植瘤裸鼠全部死亡,B组有5只死亡,A组有3只死亡,3组之间死亡率差异具有统计学意义(P=0.023);KruskalWallis检验生存时间存在统计学差异(P=0.036)。γδT细胞治疗组中有4只肿瘤病灶出现坏死脱落,B和C组中各有1只肿瘤出现坏死;肿瘤生长到1个月后,B组和C组中裸鼠发生头部、眼眶、尾部、会阴部和对侧腹股沟区转移,A组仅1例发生对侧腹股沟部转移;A组中有1例裸鼠种植瘤脱落消失,而B组和C组均无肿瘤脱落、坏死消失发生。结论外周血γδT细胞对胰腺癌裸鼠种植瘤具有显著的抑瘤作用。     

维生素C对TNF-α及血清剥夺诱导的人髓核细胞凋亡的作用

目的探讨维生素C(Vitamin C,Vit C)在TNF-α及血清剥夺诱导人椎间盘髓核细胞凋亡中的作用及机制。方法取脊柱矫形手术患者的髓核组织,消化法获得正常髓核细胞,取第3代细胞按不同处理因素分为Vit C作用组(A组)、TNF-α诱导组(B组)和血清剥夺组(C组),每组再分为以下亚组:A组分为A1组(基础培养液)、A2组(100μg/m L Vit C)、A3组(200μg/m L Vit C);B组分为B0组(对照组)、B1组(100 ng/m L TNF-α)、B2组(100μg/m L Vit C+100 ng/m L TNF-α)、B3组(200μg/m L Vit C+100 ng/m L TNF-α);C组分为C0组(对照组)、C1组(FBS浓度由8%降低为2%)、C2组(2%FBS+100μg/m L Vit C)、C3组(2%FBS+200μg/m L Vit C)。各组作用24 h后,采用流式细胞术检测各组膜联蛋白V、碘化丙啶双染后髓核细胞凋亡率;实时荧光定量PCR检测p53(基因编码相对分子质量为53×103的蛋白质)、脂肪酸合成酶(fatty acid synthase,FAS)、半胱氨酸天冬氨酸蛋白酶3(Caspase3)、Ⅰ型胶原、Ⅱ型胶原、Sox9和糖胺多糖(Aggrecan)m RNA表达。结果 A组:与A1组比较,A2、A3组的Vit C均能降低髓核细胞凋亡及p53、FAS、Caspase 3 m RNA表达(P0.05),但A2、A3组间比较差异无统计学意义(P0.05);A2、A3组的Vit C可促进髓核细胞Ⅰ型胶原、Ⅱ型胶原、Aggrecan、Sox9 m RNA表达,且A3组促进作用显著大于A2组(P0.05)。B组:与B0组比较,B1组TNF-α促进了髓核细胞凋亡,增加了髓核细胞p53、FAS、Caspase 3 m RNA表达(P0.05);与B1组比较,B3组的Vit C能降低髓核细胞凋亡及p53、FAS、Caspase 3 m RNA表达,而B2组的Vit C则增加其凋亡及p53、FAS、Caspase 3 m RNA表达,差异均有统计学意义(P0.05)。C组:与C0组比较,C1组2%FBS能诱导髓核细胞凋亡,但降低了髓核细胞p53、FAS、Caspase 3 m RNA表达(P0.05);与C1组比较,C3组的Vit C能降低髓核细胞凋亡及p53、FAS、Caspase 3 m RNA表达,而C2组的Vit C则增加其凋亡及p53、FAS m RNA表达,差异有统计学意义(P0.05)。结论 Vit C能延缓或降低髓核细胞凋亡、促进细胞外基质合成与分泌,200μg/m L Vit C可延缓或降低100 ng/m L TNF-α和2%FBS诱导的凋亡,而100μg/m L Vit C则促进其诱导的凋亡。     

亚高温盆腔区域热疗对原位膀胱肿瘤大鼠免疫状态影响的研究

目的 研究亚高温盆腔区域热疗致原位膀胱肿瘤大鼠发生热休克反应后机体免疫状态的改变.方法 经尿道膀胱灌注N-甲基亚硝基脲(MNU)制备大鼠膀胱肿瘤动物模型35只,随机分入短期组,包括肿瘤对照组(A组)、单次热疗组(B组)、间隔24 h再次热疗组(C组)、间隔72 h再次热疗组(D组)和长期组,包括肿瘤对照组(E组)、间隔24 h再次热疗组(F组)、间隔72 h再次热疗组(G组).用内生场热疗系统从体外加热大鼠盆腔至41 ℃.热疗后对肿瘤、引流淋巴结、脾脏中免疫细胞的变化进行病理研究,检测血清鼠 IFN-γ含量.结果 透射电镜观察到B、C、D组肿瘤组织中有较多中性粒细胞、巨噬细胞;F、G组中性粒细胞聚集,巨噬细胞吞噬凋亡小体.短期组中B、C、D组膀胱肿瘤组织、淋巴结及脾脏中S-100蛋白阳性细胞率均高于A组(P<0.01).膀胱肿瘤组织及脾脏中CD8阳性细胞率、血清鼠 IFN-γ含量A、B、C、D组间差异均无统计学意义(P=0.657、0.440、0.523).长期组中F、G组的各项数据均高于E组(P<0.01).结论 随着亚高温盆腔区域热疗次数的增加肿瘤鼠的免疫细胞重新分布,有利于肿瘤抗原提呈及效应细胞攻击肿瘤.     

腰椎椎间盘突出症和椎间盘源性疼痛的免疫病理学研究

目的 探讨腰椎椎间盘突出症和椎间盘源性疼痛的免疫病理学改变及其异同点.方法 收集71例手术切除的椎间盘髓核标本,分为:腰椎椎间盘突出(A组)30例,腰椎间盘脱出或游离(B组)22例,两组均行椎间盘髓核摘除术;腰椎椎间盘源性疼痛(C组)10例,均经椎间盘造影证实并行前路手术切除;腰椎爆裂骨折(D组)9例,行前路手术切除伤椎远侧相对正常的椎间盘髓核.组织形态学观察各组椎间盘髓核细胞的病理变化;应用免疫组化技术检测髓核CD68阳性巨噬细胞和CD45RO阳性T细胞,并行统计学分析.结果 组织形态学观察:D组髓核细胞形态大小一致,分布均匀,未见明显细胞基质退变及炎性细胞浸润;其他各组均见髓核细胞空泡样变、形态大小不一、分布不均;A组和B组髓核组织边缘可见大量炎性细胞浸润、局灶性小血管增生,以B组更明显;C组细胞基质退变严重,髓核组织边缘可见散在炎性细胞浸润,未见明显血管增生.免疫组化检测:CD68阳性率,B组(63.6%)＞C组(40.0%)＞A组(26.7%)＞D组(0%),各组间差异有统计学意义(P＜0.05).CD45RO阳性T细胞均出现在CD68阳性巨噬细胞同一部位的连续切片上,阳性率B组(59.1%)＞A组(13.3%),C组和D组为阴性,各组间差异有统计学意义(P＜0.05).结论 腰椎椎间盘突出症髓核边缘有明显的炎症和自身免疫反应;椎间盘源性疼痛髓核边缘有散在炎性细胞和较多巨噬细胞,但小血管增生和T淋巴细胞浸润不明显,提示有炎性反应,但自身免疫反应不如椎间盘突出症明显.

Abstract：

Objective To evaluate and compare the immunopathological changes of lumbar disc herniation and discogenic pain. Methods Seventy-one lumbar disc nucleuses were collected intra-operation,and they were divided into four groups. Group A: 30 cases of disc herniation, Group B: 22 cases of sequestered disc herniation, and the patients in Group A and B received discectomy; Group C: 10 cases of discogenic pain were confirmed by discography, and the painful disc was excised through anterior retroperitoneal approach; Group D: 9 cases of lumbar bust fracture who received anterior subtotal corpectomy and discectomy, and the nearly normal caudal disc was collected as control. The disc nucleuses were processed in the following methods: 1) HE stain and pathological observation; 2) Immunocytochemical test using monoclonal antibodies to CD68 and CD45RO molecule for macrophage and T lymphocytes, respectively. The positive cells were counted and analyzed with statistic method. Results 1) Pathological observation with HE stain: compare to control group, the degeneration of nucleus cell was evident in the other groups. There were obvious infiltration of inflammatory cells and focal neovascularization in group A and especially in Group B.In Group C, only diffuse inflammatory cells was found without neovascularization, but the degeneration of matrix was more severe. 2) Positive rate of CD68 cells: Group B (63.6%)＞Group C (40.0%)＞Group A (26.7%)＞Group D (0%). There were significant differences among groups (P＜0.05). 3) Positive rate of CD45RO cells:Group B (59.1%)＞Group A (13.3%), Group C and D were negative, and the positive cell were found in slice of the same site of positive CD68 cells. The differences between each group were significant (P＜0.05). Conclusion The nucleus of herniated disc has evident inflammatory and autoimmunity reaction. The nucleus of discogenic pain is infiltrated with diffuse inflammatory cells and some macrophages, without T lymphocyte and neovasularization, so the autoimmunity course is not evident.     

联合药物预处理对非协调性异种心脏移植物的影响

目的　观察联合应用环孢素A(CsA)和来氟米特 (Lef)预处理对豚鼠 大鼠非协调性异种心脏移植物的影响并探讨其作用机制。方法　移植大鼠分为 4组 :A组 (CVF ,n =10 ) ;B组(CVF CsA ,n =8) ;C组 (CVF Lef,n =5 ) ;D组 (CVF CsA Lef ,n =11) ;记录移植物存活时间 ,检测移植物病理组织学 ,用免疫组织化学检测移植物CD68、CD5 7表达情况 ,TUNEL法检测移植物细胞凋亡。结果　移植物的平均存活时间 :A组为 41h、B组为 68h、C组为 5 5h、D组为 82h。各组移植物病理表现均为急性血管性排斥反应 (AVR)改变。B、C、D组移植物炎症细胞浸润数减少 ,CD68和CD5 7表达下调 ,凋亡指数较低 ,半定量测定与A组比较差异有显著性 (P <0 .0 5 ) ;D组凋亡指数最低而CD68和CD5 7表达最弱 ,与其他各组比较差异有显著性 (P <0 .0 5 )。结论　联合应用CsA和Lef预处理可明显延长非协调性异种移植物存活时间 ,减少炎症细胞浸润 ,抑制心肌细胞凋亡 ,减轻异种AVR损害。     

Changes in the number of gut mucosal T-lymphocytes and macrophages in patients treated by external biliary drainage.

OBJECTIVE: To examine the changes in the number of T cells and macrophages in the mucosal lamina propria in the presence or absence of bile in the gastrointestinal tract. DESIGN: Clinical study. SETTING: University hospital, Japan. SUBJECTS: 6 patients with obstructive jaundice who had external biliary drainage (drainage group) and 6 patients with no signs of obstructive jaundice (control group). INTERVENTIONS: Gastrointestinal specimens were taken at the time of operation. MAIN OUTCOME MEASURES: The number of CD4+ T cells, CD8+ T cells and CD68+ macrophages in the lamina propria mucosae in each group measured immunohistochemically. RESULTS: The numbers of CD8 T cells and CD68+ macrophages in the lamina propria of the patients treated by external drainage were significantly less than in the control group (p < 0.01). However, there was no difference in the number of CD4+ T cells between the groups (p = 0.45). CONCLUSIONS: In the absence of bile, mucosal immune function fails as a result of reduced numbers of CD8+ T cells and macrophages.     

早孕期蜕膜免疫细胞谱及其与妊娠结局的相关性

目的探讨早孕期蜕膜免疫细胞谱变化及稽留流产对妊娠免疫细胞谱的影响。方法选取2017年9月至2018年9月深圳中山泌尿外科医院收治的行人工流产的正常早孕患者(人工流产组,n=34),行清宫术的稽留流产患者(稽留流产组,n=17)及同期于本院IVF助孕成功的正常妇女(正常内膜组,n=27)作为研究对象。运用免疫组化及数字化分析系统定量检测正常妇女子宫内膜及流产组蜕膜组织中各免疫细胞的检测阳性率。结果早孕期妊娠前后的免疫细胞谱有如下改变:妊娠早期人工流产组蜕膜CD56⁺自然杀伤(NK)细胞、CD68⁺巨噬细胞、CD83⁺成熟树突状细胞数均显著高于正常内膜组(P<0.001),而CD8⁺T细胞、Foxp3⁺调节性T(Treg)细胞及CD1a⁺未成熟树突状细胞在两组之间则无显著差异(P>0.05)。在不良妊娠结局稽留流产组患者蜕膜组织中CD68⁺巨噬细胞、CD8⁺T细胞及Foxp3⁺调节性T(Treg)细胞数均显著高于人工流产组(P<0.05),而CD56⁺自然杀伤(NK)细胞、CD83⁺成熟树突状细胞及CD1a⁺未成熟树突状细胞与人工流产组相比无显著差异(P>0.05)。结论早孕期蜕膜中NK细胞、巨噬细胞及成熟树突状细胞的适度增加有利于母胎免疫耐受的形成,但异常高表达的蜕膜巨噬细胞、CD8⁺T细胞及Foxp3⁺Treg细胞可能与不良妊娠结局有关。     

Dendritic cell and M2 macrophages are associated with the severity of disease in IgA nephropathy patients

Objective To investigate the number and distribution of dendritic cells (DCs), macrophages and M2 macrophages in renal tissues of patients with IgA nephropathy (IgAN) and their correlation with clinicopathological parameters, and explore its role in the progression of IgAN. Methods Renal tissue samples from 42 patients aged ≥18 years with IgAN were collected by kidney biopsy in Guangdong Provincial People's Hospital from January 2018 to June 2018. The patients were divided into different groups according to Oxford classification and Lee grade classification criteria. The distribution and number of DCs (CD209), macrophages (CD68) and M2 macrophages (CD68 and CD206) were detected by immunohistochemistry. Spearman correlation test was used to analyze the correlation between the number of DCs and macrophages in renal tissues and clinical pathological parameters. Results The number of DCs in the glomeruli of the M1, T0 and C1 groups increased significantly compared with the M0, T1 and C0 groups, and the number of DCs in the renal interstitium of the T1 group increased significantly compared with the T0 group (all P＜0.05). The number of glomerular macrophages in group C1 was significantly higher than that in group C0. The number of macrophages in S1, T1 and Lee IV-V tubulointerstitial groups was significantly higher than that in S0, T0 and Lee II-III groups (P＜0.05). The number of M2 macrophages in the S1, T1 and Lee IV-V groups was significantly higher in the tubulointerstitial group than in the S0, T0 and Lee II-III groups (all P＜0.05). The blood urea nitrogen, serum creatinine and 24 h urine protein levels in the T1 and Lee IV-V groups were significantly higher than those in the T0 and Lee II-III groups, and the serum albumin levels in the Lee IV-V group were significantly lower (all P＜0.05). Spearman correlation analysis showed that the number of DCs in renal interstitium was positively correlated with the proportion of tubulointerstitial fibrosis. There were a positive correlation between the proportion of glomerular segmental sclerosis and tubulointerstitial fibrosis and renal interstitial macrophages and M2 macrophages. The number of M2 macrophages was positively correlated with serum creatinine and 24 h urine protein (both P＜0.05). Conclusions The number of DCs and M2 macrophages in kidneys are positively correlated with the clinicopathological features of IgAN patients, which indicates that they may be associated with disease progression.     

Infiltration of tumor-reactive transforming growth factor-beta insensitive CD8+ T cells into the tumor parenchyma is associated with apoptosis and rejection of tumor cells

BACKGROUND: TGF-beta is a potent immunosuppressant. High levels of TGF-beta produced by cancer cells have a negative inhibition effect on surrounding host immune cells and leads to evasion of the host immune surveillance and tumor progression. In the present study, we report a distinct ability of tumor reactive, TGF-beta-insensitive CD8+ T cells to infiltrate into established tumors, secrete relevant cytokines, and induce apoptosis of tumor cells. METHODS: CD8+ T cells were isolated from the spleens of C57BL/6 mice, which were primed with irradiated mouse prostate cancer cells, the TRAMP-C2 cells. After ex vivo expansion, these tumor reactive CD8+ cells were rendered TGF-beta-insensitive by infection with a retroviral (MSCV)-mediated dominant negative TGF-beta type II receptor (TbetaRIIDN). Control CD8+ cells consist of those transfected with the GFP-only empty vector and na?ve CD8+ T cells. Recipient mice were challenged with a single injection of TRAMP-C2 cells 21 days before adoptive transfer of CD8+ T cells was performed. Forty days after the adoptive transfer, all animals were sacrificed. The presence of pulmonary metastases was evaluated pathologically. Serial slides of malignant tissues were used for immunofluorescent staining for different kinds of immune cell infiltration, cytokines, and apoptosis analysis. RESULTS: Pulmonary metastases were either eliminated or significantly reduced in the group receiving adoptive transfer of tumor-reactive TGF-beta-insensitive CD8+ T cells (3 out of 12) when compared to GFP controls (9 out of 12), and na?ve CD8+ T cells (12 out of 12). Results of immunofluorescent studies demonstrated that only tumor-reactive TGF-beta-insensitive CD8+ T cells were able to infiltrate into the tumor and mediate apoptosis when compared to CD4+ T cells, NK cells, and B cells. A large amount of cytokines such as perforin, nitric oxide, IFN-gamma, IL-2, TNF-alpha were secreted in tumor tissue treated with tumor-reactive TGF-beta-insensitive CD8+ T cells. No immune cells infiltration and cytokine secretion were detected in tumor tissues treated with na?ve T cells and GFP controls. CONCLUSIONS: Our results demonstrate the mechanism of anti-tumor effect of tumor-reactive TGF-beta-insensitive CD8+ T cells that adoptive transfer of these CD8+ T cells resulted in infiltration of these immune cells into the tumor parenchyma, secretion of relevant cytokines, and induction of apoptosis in tumor cells. These results support the concept that tumor-reactive TGF-beta-insensitive CD8+ T cells may prove beneficial in the treatment of advanced cancer patients.     

Apoptosis and allograft rejection in the absence of CD8+ T cells.

BACKGROUND: The requirement for cytotoxic T lymphocytes during allograft rejection is controversial. We previously demonstrated that CD8+ T cells are not necessary for allograft rejection or for the induction of apoptosis in rat small intestinal transplantation. In this study, we examined the mechanisms of apoptosis and rejection after liver transplantation in the absence of CD8+ T cells. METHODS: Either Lewis or dark agouti rat liver grafts were transplanted into Lewis recipients to create syngeneic and allogeneic combinations. CD8+ T cells were depleted in an additional allogeneic group by treatment with OX-8 mAb on day -1 and day 1 after liver transplant. RESULTS: Apoptosis and rejection were observed in both the CD8+ T cell-depleted allogeneic and allogeneic grafts by hematoxylin and eosin staining, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining, and radiolabeled-annexin V in vivo imaging. Granzyme B and FasL were expressed in all allogeneic transplants, including those depleted of CD8+ T cells, indicating that a mononuclear cell other than a CD8+ T cell can be the source of these molecules during allograft rejection. Activation of the caspase cascade was detected in all rejecting allografts. Caspases 3, 8, and 9 were activated at similar significantly elevated levels in both allogeneic and CD8+ T cell-depleted liver grafts. CONCLUSION: These data indicate that in the absence of CD8+ T cells an alternative pathway, associated with granzyme B and FasL expression and activation of the caspase cascade, can mediate apoptosis and graft rejection.