白细胞介素2、白细胞介素10及人类白细胞抗原G与肝移植的急性排斥

背景:移植肝急性排斥反应是导致移植物功能损伤的主要危险因素,目前诊断主要靠肝组织穿刺活检,带来感染、出血等风险.如何无创、简单、准确地进行诊断成为当前该领域的热点.目的:观察分析白细胞介素2、白细胞介素10、人类白细胞抗原G的表达与人类肝移植急性排斥之间的关系.方法:用酶联免疫方法检测59例肝脏移植受者外周血白细胞介素2、白细胞介素10、人类白细胞抗原G的表达情况.将受者按照有无急性排斥反应分为急性排斥组和非排斥组;并将非排斥组按肝功能情况分为肝功能正常组与异常组.结果进行统计学分析,并绘制ROC曲线,比较曲线下面积及其临界值的敏感性和特异性.结果与结论:急性排斥组外周血中白细胞介素10、人类白细胞抗原G表达均低于与非排斥组(P=0.032,0.002),而排斥组中自细胞介素2高于非排斥组(P=0.002).肝功正常组与异常组间自细胞介素10、人类白细胞抗原G表达差异无显著性意义(P=0.525,0.084),而肝功能异常组白细胞介素2表达高于肝功能正常组(P=0.02).ROC曲线下面积为人类白细胞抗原G>白细胞介素2>白细胞介素10.结果提示,白细胞介素2、白细胞介素10、人类白细胞抗原G的表达都与排斥相关,三者中人类白细胞抗原G为诊断急性排斥的最佳指标.     

异基因外周血干细胞移植患者人类白细胞抗原B27的表达

目的:分析白血病患者行异基因外周血干细胞移植后人类白细胞抗原B27的表达特点,及其与慢性移植物抗宿主病的关系。方法:选择2003-09/2006-06在广西医科大学第一附属医院行异基因外周血干细胞移植术的45例白血病患者,按《血液病诊断和疗效标准》进行疾病的诊断和分型。全部患者均知情同意并签署知情同意书。男27例,女18例,22～51岁。其中急性髓细胞白血病13例、急性淋巴细胞白血病17例、慢性粒细胞白血病15例。供者均为白细胞抗原配型6个位点全相合的同胞兄妹。急性髓细胞白血病和慢性粒细胞白血病患者采用BU/CY方案:马利兰4mg/(kg·d)×4d,环磷酰胺60mg/(kg·d)×2d;急性淋巴细胞白血病患者预处理采用TBI/CY方案:全身照射5.0Gy/d×2d,环磷酰胺60mg/(kg·d)×2d。供者采用粒细胞集落刺激因子皮下注射动员干细胞。采用美国Baxter CS-3000plus血细胞分离机采集外周血干细胞,每位供者采集0, 1, 2d3次。回输单个核细胞、粒-单核系集落形成单位、CD34 细胞。采用流式细胞术动态检测45例白血病患者异基因外周血干细胞移植前后人类白细胞抗原B27表达的变化。结果:45例患者全部进入结果分析。①45例患者均获得造血重建,经荧光标记PCR短串联重复序列DNA位点分析,全部患者移植后血细胞的DNA与供者血细胞DNA完全一致。发生急性移植物抗宿主病4例,慢性移植物抗宿主病14例,出血性膀胱炎3例,肝静脉闭塞病4例。②移植后发生慢性移植物抗宿主病患者人类白细胞抗原B27表达水平明显低于无移植物抗宿主病患者[移植后180d:(30.13±10.24)%,(39.61±10.89)%;移植后峰值:(33.53±10.36)%,(45.76±12.09)%,P<0.05]。③移植前未缓解白血病患者人类白细胞抗原B27表达水平则明显高于移植前达到完全缓解患者[(27.39±10.31)%,(16.62±8.58)%,P=0.005],移植后疾病复发患者复发时人类白细胞抗原B27表达水平明显高于其移植后完全缓解时[(27.76±10.09)%,(15.69±6.98)%,P=0.015]。移植前未缓解和移植后复发患者人类白细胞抗原B27表达水平与肿瘤细胞占骨髓非红系细胞的百分比呈正相关关系(r=0.756,P=0.000)。结论:移植前未缓解和移植后复发白血病患者人类白细胞抗原B27表达与肿瘤细胞占骨髓非红系细胞的百分比呈正相关,人类白细胞抗原B27表达水平随着肿瘤细胞百分比的增高而升高。     

人类白细胞抗原配型不合造血干细胞移植病人的护理

[目的]探讨亲缘人类白细胞抗原（HLA）配型不合造血干细胞移植（HSCT）的护理。[方法]对12例血液病病人进行亲缘HLA至少1个位点不合的HSCT。预处理分别采用改良白消安（Bu）/环磷酰胺（Cy）＋抗胸腺淋巴细胞球蛋白（ATG）,氟达拉宾（Flu）＋Cy＋ATG,阿糖胞苷（Ara-C）＋Bu＋Flu＋ATG方案。[结果]病人移植后造血均恢复顺利,除1例骨髓增生异常综合征伴骨髓纤维化病人植入失败自身恢复外,均达完全供者型植入。10例发生急性移植物抗宿主病（aGVHD）（10/12）,其中Ⅰ度4例,Ⅱ度3例,Ⅲ度1例,Ⅳ度2例,Ⅱ度～Ⅳ度的aGVHD累积发生率为51.4%,Ⅲ度～Ⅳ度aGVHD累积发生率为16.8%。巨细胞病毒（CMV）感染5例,累积发生率41.8%,其中1例为CMV间质性肺炎。[结论]亲缘HLA配型不合HSCT是治疗血液病的有效方法之一,但需警惕移植后GVHD及机会性感染的发生。精心的护理能有效降低各种并发症的发生及临床病死率。     

人类白细胞相关抗原G在移植免疫中的作用及其研究进展

人类白细胞相关抗原G(human leukocyte antigen-G,HLA-G)是一类非典型的人白细胞抗原I类分子(human lym-phocyte antigen I,HLA-I),最早发现其表达在母胎界面的绒毛膜滋养层层细胞表面,蛋白产物有HLA-G膜结合型和可溶性分子型。国内外多项研究表明HLA-G分子在免疫耐受状态的形成及维持中具有重要作用。现就其基因及蛋白产物表达特点、诱导免疫耐受的机制及临床移植应用研究做简要综述。     

肾移植患者抗人类白细胞抗体配型与术后急性排斥反应及移植肾存活的关系

背景:群体反应性抗体可介导超急性排斥反应,导致群体反应性抗体阳性致敏患者肾移植成功率和移植物存活率均低于非致敏患者.目的:根据人类白细胞抗原交叉反应组配型标准为群体反应性抗体阳性致敏肾移植患者选配合适供者,观察移植后急性排斥反?发生率及移植肾存活情况.设计:病例观察.单位:南方医科大学珠江医院.对象:选择1997-01/2003-12在南方医科大学珠江医院器官移植科施行肾移植136例群体反应性抗体阳性的致敏受者,男41例,女95例,年龄(45±9)岁.初次肾移植115例,2次移植18例,3次移植2例,4次移植1例.所有受试对象均对检测项目知情同意,实验经过医院伦理委员会批准.LAT莱姆德抗原板和LAT-Mix混合抗原板购自美国One Lambda公司.SMT72R抗人类白细胞抗原-1类单克隆抗体湿板和人类白细胞抗原-Ⅱ类DNA分型试剂购自美国One Lambda公司.方法:以酶联免疫吸附试验动态监测患者手术前后IgG型抗人类白细胞抗原抗体水平及其特异性.应用抗人类白细胞抗原-Ⅰ类单克隆抗体湿板进行供、受者人类白细胞抗原-Ⅰ类抗原分型,微量序列特异性引物法进行供、受者人类白细胞抗原-Ⅱ类基因分型,根据美国器官共享网制定的人类白细胞抗原交叉反应组配型标准和Ⅱ类抗原可接受性错配原则进行供、受者选配.评估抗原交叉反应组配型原则下患者移植术后急性排斥反应发生率及移植肾1,3,5年存活率进行评价.主要观察指标:①致敏患者手术前后抗人类白细胞抗原抗体水甲及特异性.②供受者人类白细胞抗原组织配型.③肾移植术后急性排斥反应发生率及移植肾存活率.结果:纳入136例PRA阳性致敏患者均进入结果分析,无脱落者.①致敏患者中104例存在抗人类白细胞抗原-Ⅰ类IgG抗体,76例存在抗人类白细胞抗原-Ⅱ类IgG抗体,44例同时存在抗人类白细胞抗原-Ⅰ类和Ⅱ类IgG抗体.②按照传统人类白细胞抗原配型标准供、受者人类白细胞抗原0,1,2,3,4个抗原错配者分别为7,26,47,39和17例;按照人类白细胞抗原交叉反应组配型标准供、受者人类白细胞抗原抗原0,1,2,3,4个抗原错配者分别为31,53,36,16和0例.③按人类白细胞抗原交叉反应组配型原则,人类白细胞抗原无错配者的急性排斥反应发生率明显低于2个和3个抗原错配者,差异有统计学意义(P<0.05),人类白细胞抗原无错配者移植肾1,3,5年存活率高于2个和3个抗原错配者,差异有统计学意义(P<0.05).结论:①人类白细胞抗原交叉反应组配型标准能提高肾移植供受者的相配率.②良好的人类白细胞抗原配型可降低肾移植术后急性排斥反应的发生率,提高移植肾的存活率.     

人类白细胞抗原配型不合造血干细胞移植病人的护理

[目的]探讨亲缘人类白细胞抗原(HLA)配型不合造血干细胞移植(HSCT)的护理。[方法]对12例血液病病人进行亲缘HLA至少1个位点不合的HSCT。预处理分别采用改良白消安(Bu)/环磷酰胺(Cy) 抗胸腺淋巴细胞球蛋白(ATG),氟达拉宾(Flu) Cy ATG,阿糖胞苷(Ara-C) Bu Flu ATG方案。[结果]病人移植后造血均恢复顺利,除1例骨髓增生异常综合征伴骨髓纤维化病人植入失败自身恢复外,均达完全供者型植入。10例发生急性移植物抗宿主病(aGVHD)(10/12),其中Ⅰ度4例,Ⅱ度3例,Ⅲ度1例,Ⅳ度2例,Ⅱ度～Ⅳ度的aGVHD累积发生率为51.4%,Ⅲ度～Ⅳ度aGVHD累积发生率为16.8%。巨细胞病毒(CMV)感染5例,累积发生率41.8%,其中1例为CMV间质性肺炎。[结论]亲缘HLA配型不合HSCT是治疗血液病的有效方法之一,但需警惕移植后GVHD及机会性感染的发生。精心的护理能有效降低各种并发症的发生及临床病死率。     

异种大鼠神经干细胞脑内移植未导致明显的免疫排斥反应

目的 探讨异种大鼠神经干细胞脑内移植是否会导致明显的免疫排斥反应。方法 制作SD大鼠大脑中动脉夹闭致脑缺血模型，将体外培养的Wistar大鼠神经干细胞移植到脑缺血同侧或对侧的纹状体区，在移植后的不同时间杀死大鼠，研究脑内单核吞噬细胞的侵润、脾脏淋巴滤泡增殖以及脑内干细胞存活和迁移的情况。结果 移植后的干细胞在脑内大量存活并向损伤区域迁移；与未接受干细胞移植的对照组相比较，干细胞移植组大鼠并未发生明显的脑内单核吞噬细胞侵润和脾脏淋巴滤泡的大量增殖。结论 异种神经干细胞脑内移植未导致明显的免疫排斥反应。     

大鼠小肠移植排斥反应期移植肠RANTES的表达

背景:同种异体移植排斥反应是影响移植物功能和存活的最大障碍,小肠移植排斥反应的诊断与治疗尤为困难.目的:探讨移植肠RANTES的表达在小肠移植急性排斥反应中的意义,以及他克莫司对它的影响.设计、时间及地点:随机对照动物实验,于2003-09/2005-03在解放军第四五一医院普通外科、解放军第四军医大学附属西京医院胃肠外科实验室、解放军第四军医大学基础部电子显微镜中心为大专院校的实验室完成.材料:选用健康成年雄性SD和Wistar大鼠各72只.以SD大鼠为供者,Wistar大鼠为受者,施行异位小肠移植.方法:实施大鼠异位小肠移植,按照不同品系的组合将移植大鼠分为4组(n=18):非手术对照组,Wistar大鼠作为正常对照,不实施小肠移植;同基因移植组,将Wistar大鼠的小肠移植给同品系的Wistar大鼠;异基因移植未治疗组,将SD大鼠的小肠移植给Wistar大鼠,移植后不给予免疫抑制剂治疗;异基因移植他克莫司治疗组,将SD大鼠的小肠移植给Wistar大鼠,移植后0～7 d肌肉注射他克莫司,1 mg/(kg·d).移植后3,5,7 d每组各处死6只大鼠,切墩移植肠标本进行组织病理学检查,并采用免疫荧光染色和激光扫描共聚焦显微镜技术对移植肠RANTES的表达进行连续定量测定.主要观察指标:①各组大鼠移植肠的组织病理学改变.②不同时间点移植物内RANTES阳性细胞的表达变化.③他克莫司对异基因移植大鼠移植肠RANTES表达的抑制作用.结果:72只受体大鼠均进入结果分析.异基因移植末治疗组大鼠后3,5,7 d移植肠符合轻、中、重度急性排斥反应的病理学诊断标准;他克莫司治疗组大鼠和同基因移植对照组大鼠在观察期内未发现明显排斥反应征象.异基因移植未治疗组大鼠的移植肠RANTES表达在术后均显著高于其他3组(P<0.01),其动态变化与急性排斥反应的进程呈正相关;他克莫司治疗组大鼠移植肠RANTES的表达明显低于未治疗组(P<0.01).结论:RANTES阳性细胞在小肠移植急性排斥反应中发挥了重要作用,动态检测移植肠RANTES的表达变化,可能成为小肠移植急性排斥反应有效的诊断指标之一.     

大鼠小肠移植急性排斥反应中血清白细胞介素2及干扰素γ的变化

目的:分析血清白细胞介素2、干扰素γ的水平在大鼠小肠移植急性排斥反应过程中的意义及其评估价值。方法:实验于2005-11/2006-06在解放军第四军医大学西京医院胃肠外科实验室完成。实验方法:小肠移植模型动物选用封闭群SD和Wistar大鼠42只,按随机数字表法分为3组:①对照组(n=6):SD大鼠仅作腹部开关及左肾切除术。②同基因移植组(n=18):供受体均为SD大鼠。③异基因移植组(n=18):供体为Wistar大鼠,受体为SD大鼠。同基因移植组和异基因移植组分别于术后3,5,7天处死大鼠取移植肠段行病理组织学检查,每次6只;同时取血清应用ELISA法检测血清白细胞介素2、干扰素γ水平,对照组大鼠于术后第3天取血清检测。实验评估:排斥程度评估标准分为轻、中、重3度:①轻度:肠黏膜绒毛数目减少,轻度增粗,变短,少量淋巴细胞和粒细胞浸润,少量凋亡上皮细胞。②中度:肠黏膜绒毛明显变矮增宽,黏膜上皮开始脱落,细胞浸润加重,上皮细胞凋亡数目明显增多。③重度:肠绒毛结构消失,上皮脱落,间质大量炎细胞浸润,肌层结构破坏,血管炎。结果:42只大鼠均进入结果分析,无脱失。①异基因移植组大鼠术后第3,5,7天的病理组织学检测结果分别符合轻、中、重度排斥反应,对照组和同基因移植组无明显排斥征象。②异基因移植组大鼠血清白细胞介素2、干扰素γ的表达水平在术后第3,5,7天均显著高于其他2组(P<0.01),白细胞介素2水平升高以第3天明显,第5天达到高峰,第7天即有所下降;干扰素γ水平升高较为缓慢,第7天明显升高。同基因移植组大鼠血清白细胞介素2、干扰素γ水平也高于对照组,但差异无显著性意义(P>0.05)。结论:白细胞介素2、干扰素γ在大鼠小肠移植急性排斥反应过程中发挥重要作用,血清白细胞介素2、干扰素γ水平监测可作为预测及早期诊断大鼠小肠移植急性排斥反应的重要指标。     

人类白细胞抗原抗体在非血缘脐血移植中的作用

随着非血缘脐血移植(UC BT)在临床越来越多的应用,以及相关检测技术的发展,患者体内的脐血人类白细胞抗原(HLA)抗体对UCBT疗效的影响受到广泛关注.笔者拟就近年关于HLA抗体,尤其是供者特异性HLA抗体(DSA)对UCBT影响的研究进行综述.     

Neuromuscular dysfunction in the jejunum and colon of human leukocyte antigen B27 transgenic rats

HLA-B27 transgenic rats are a model of spontaneous gastrointestinal inflammation associated with expression of human leukocyte antigen (HLA) B27 and beta(2)-microglobulin. Our goal was to investigate in vitro enteric nerve regulation and contractile activity in isolated longitudinal muscles from the jejunum and colon of HLA-B27 rats. Nontransgenic age-matched Fisher 344 rats were used as controls. Intestinal inflammation and tissue injury, quantified histologically and through tissue myeloperoxidase activity, were evident in both the jejunum and colon of HLA-B27 rats. Although resting tension and spontaneous activity of the jejunal and colonic muscles from HLA-B27 rats did not differ significantly from controls, responses to both enteric nerve stimulation or direct muscle activation were significantly inhibited. In muscles from HLA-B27 rats, electrical field stimulation (0.5 ms, 0.5-20 Hz) induced low-amplitude contractions (maximal reduction 60-65%) compared with respective controls. In the presence of atropine and guanethidine, nonadrenergic and noncholinergic contractile responses to higher frequencies of stimulation (8-20 Hz) were also of lower amplitude. These changes were accompanied by a shift in neurally mediated contractions from predominantly cholinergic in the jejunum and colon of Fisher 344 rats to predominantly nonadrenergic and noncholinergic in HLA-B27 rats. Furthermore, maximal contractions to carbachol or KCl depolarization were reduced (up to 2.7-fold) compared with respective controls. In the jejunum of HLA-B27 rats the EC(50) level for carbachol was decreased. The data indicate that gastrointestinal inflammation induced by expression of HLA-B27 is associated with hypocontractility and inhibition of enteric cholinergic control of the longitudinal muscle in both the small and large intestine.     

人类白细胞抗原新等位基因A＊2636的确认

背景:目前中华骨髓库主要采用分辨率较高的聚合酶链反应-寡核苷酸探针分型反向杂交荧光微珠法检测人类白细胞抗原HLA-A、B、DRB1,该方法在高分辨率的同时,有些检测结果难以一次性确定,可采用DNA测序技术确认.目的:识别和确认一个人类白细胞抗原新等位基因.设计、时间及地点:以DNA为观察对象的开放性实验.常规初检聚合酶链反应-寡核苷酸探针分型反向杂交荧光微珠法于2007 11在河南省红十字血液中心中华骨髓库河南分库人类白细胞抗原组织配型实验室完成,测序实验于2008-02在戴诺生物技术(北京)有限公司人类白细胞抗原实验室完成.材料:中国造血干细胞捐献者初检结果可疑为新基因的重采血样5 mL.方法:采用聚合酶链反应-寡核苷酸探针分型反向杂交荧光微珠法进行常规人类白细胞抗原分型,发现A位点反应格局异常,提示有2个无法解释的假阳性探针反应(10FP、88FP)和1个假阴性探针反应(56FN)、不能正常判读给出确切结果时,采用DNA测序技术测定人类白细胞抗原A位点外显子2,3,4(Exon2,3,4)的核苷酸序列,并与已知相近等位基因进行序列对比分析.主要观察指标:测序结果与已知人类白细胞抗原等位基因序列的比较分析.结果:聚合酶链反应-寡核苷酸探针分型反向杂交荧光微珠法基因分析结果显示,受检者人类白细胞抗原基因分型为A*02xx,26xx(10 FP、88FP 58FN),B·15(75),55xx,DRB1 08xx,11xx,A位点反应格局异常,提示有2个无法解释假阳性反应(10 FP、88FP),有1个似阴性探针反应(58FN),不能指定为任何人类白细胞抗原A位点等位基因.对本例人类白细胞抗原A基因第2,3,4外显子双向之直接测序结果分析显示:A*02010101,A*260101 292 S/C.分析测序结果峰图,292位置应为C,分析软什提示该基因改变在A*26new一侧.该序列与人类白细胞抗原A*260101进行比对,同源性99%,在第2外显子区域292位碱基发生了G→C,74位的密码了GAC>CAC最终导致了氨基酸的改变,即天冬氨酸(Asp)→组氨酸(His).结论:该序列为A位点的一个新等位基因,已在GenBank注册(注册号EU785343),并于2008-04被WHO-HLA因子命名委员会正式命名为A*2636.(HWS10005530).     

脾脏提取DNA与人类白细胞抗原配型实验方法

背景：人类白细胞抗原配型结果报告准确与否,直接影响供受者之间的配对选择,而配型样本的好坏则影响结果的优劣。目的：建立脾脏提取DNA进行人类白细胞抗原配型实验的实验技术平台。方法：通过从脾脏分离淋巴细胞提取DNA进行人类白细胞抗原配型实验,并同正常的和严重溶血的外周血提取DNA进行人类白细胞抗原配型的实验进行比较。结果与结论：通过脾脏和正常外周血提取的DNA经扩增、电泳后均获得了完整的DNA电泳图,脾脏比正常外周血的电泳图更清晰,而严重溶血的标本未能获得清晰完整的DNA电泳图。实验成功建立了脾脏提取DNA进行人类白细胞抗原配型实验的实验技术平台。     

Role of the low density lipoprotein receptor in regulating the content of free and esterified cholesterol in human fibroblasts.

The transfer of normal human fibroblasts from medium containing whole serum to medium devoid of lipoproteins produced a 90 percent decrease in the cellular content of cholesteryl esters and a 30 percent decrease in the free cholesterol content. When these lipoprotein-deprived cells were subsequently incubated with human low density lipoprotein (LDL), there was a 7-fold increase in the cellular content of esterified cholesterol and a 1.6-fold increase in the cellular content of free cholesterol. The concentration at which LDL produced its half-maximal effect in elevating cellular sterol content (30 mug/ml of LDL-cholesterol) was similar to the half-maximal concentration previously reported for high affinity binding of LDL to its cell surface receptor. High density lipoprotein (HDL) and whole serum from a patient with abetalipoproteinemia (neither of which contains a component that binds to the LDL receptor) did not produce a significant increase in the content of either cholesterol or cholesteryl esters in normal cells. Furthermore, in fibroblasts from patients with the homozygous form of familial hypercholesterolemia, which lack functional LDL receptors, LDL had no effect in raising the cellular content of either free or esterified cholesterol even when present in the medium at concentrations as high as 450 mug sterol/ml. It is concluded that LDL-receptor interactions constitute an important biochemical mechanism for the regulation of the cholesterol content of normal human fibroblasts. Moreover, when considered in light of current concepts of LDL metabolism in intact mammals, the present data suggest that a major function of plasma LDL may be to transport cholesterol from its site of synthesis in liver and intestine to its site of uptake in peripheral tissues.     

Role of fibronectin on the clearance and tissue uptake of antigen and immune complexes in rats.

In the present study, we have evaluated how plasma fibronectin (FN) and tissue FN can affect the clearance from the circulation and organ uptake of antigen or immune complexes (IC) that have the capacity to bind to FN. Phenylated gelatin (DNP-GL) (a FN binding antigen) and IC composed of DNP-GL and monoclonal IgGl anti-dinitrophenol (DNP) antibodies were tested. These probes were compared with DNP-bovine serum albumin (BSA) (a non-FN-binding antigen) and DNP-BSA IC formed with the same anti-DNP antibody used for the preparation of DNP-GL IC. We found evidence that DNP-GL, but not DNP-BSA, formed complexes with soluble FN in vitro and the data strongly suggest that DNP-GL-FN complexes form in vivo. The formation of complexes with plasma FN aided in the clearance of DNP-GL from the circulation, as shown by the facts that DNP-GL was removed from the circulation much faster than DNP-BSA and that complexes of DNP-GL with plasma FN were removed from the circulation faster than uncomplexed DNP-GL. The sites of deposition of DNP-GL were also different from those of DNP-BSA. Thus, DNP-GL demonstrated higher hepatic, splenic, and renal uptake than did DNP-BSA. Renal uptake of DNP-GL was quite high despite the fact that DNP-GL is anionic. Indeed, expressed per gram of tissue, liver and kidney deposition of DNP-GL was not significantly different. By immunofluorescence microscopy, DNP-GL could be demonstrated in hepatic sinusoids and glomerular mesangium. In vitro, DNP-GL bound to FN in the mesangium of frozen sections of kidney tissue. IC formed with DNP-GL or DNP-BSA demonstrated virtually the same size, yet the fate of DNP-GL IC was strikingly different from that of DNP-BSA IC. The removal of DNP-GL IC from the circulation was mediated by the antigen and not by Fc receptors since gelatin (an inhibitor of DNP-GL clearance) but not aggregated IgG (an inhibitor of Fc receptors) inhibited the removal of DNP-GL IC from the circulation. In summary, these studies suggest that the ability of an antigen or IC to bind to FN markedly influences the fate of that antigen or IC. Specifically, binding to FN accelerates clearance from the circulation and favors hepatic and renal (primarily mesangial) uptake of the FN binding antigen of IC.     

单细胞人白细胞抗原多基因位点检测的配型预选性研究及意义

目的　探索单细胞扩增前引物延伸法 (PEP)全基因组扩增 ,后续多重巢式聚合酶链反应 (MN PCR)同步扩增人类白细胞抗原 (HLA) A、B和DR基因位点检测技术 ,进行配型预选研究的价值。方法　采用PEP MN PCR方法测定单个淋巴细胞和单个胚胎细胞的HLA A、B和DR基因型。结果　PEP MN PCR对单个淋巴细胞的扩增率、扩增准确率、假阴性率和假阳性率分别为 91 7%、95 7%、3 0 %和 1 7% ,而单个胚胎细胞则分别为 97 7%、97 5 %、2 3%和 2 6 % ,两种细胞检测结果比较差异均无显著性意义 (P >0 0 5 )。结论　单细胞PEP MN PCR检测HLA多基因位点有较高的扩增效率和扩增准确率 ,具有应用于植入前胚胎遗传学分析 ,筛选HLA匹配胚胎 ,提供脐血造血干细胞移植治疗同胞疾病的应用价值。     

A single amino acid substitution in the human histocompatibility leukocyte antigen DR3 beta chain selectively alters antigen presentation

Activation of T lymphocytes by immunogenic peptides bound to HLA molecules is a central event in the generation of an immune response. To determine the sites on HLA molecules involved in this process, we isolated mutant EBV-transformed B cell clones that express altered HLA-DR3 molecules. One mutant has lost the ability to stimulate a T cell clone specific for a mycobacterial protein, but retains the ability to stimulate other antigen-specific T cells. The DNA sequence of the complete DR alpha and beta coding regions revealed a single nucleotide change resulting in a glutamic acid to lysine substitution at amino acid 9 in the first hypervariable region of the DR beta chain. These results are discussed in relation to a recently proposed model of class II molecule structure.     

Implication of antibodies against human leukocyte antigen in simultaneous presentation of fetal and neonatal alloimmune thrombocytopenia and neutropenia

Fetal and neonatal alloimmune thrombocytopenia (FNAIT) and neonatal alloimmune neutropenia (NAN) are two rare complications of newborns caused by antibodies against paternal inherited antigens. Human platelet (HPA) and neutrophil antigens (HNA) are the common targets. Human leukocyte antigen (HLA) class I proteins are also expressed on platelets and neutrophils and anti-HLA antibodies have occasionally been implicated in these complications. We report a premature twin infant who presented with severe thrombocytopenia and neutropenia clinically compatible with FNAIT and NAN, from a mother with no identifiable HPA or HNA antibodies, but with very high levels of complement-fixing antibodies against paternal inherited HLA. These antibodies were also detected in the infant. HLA antibodies are commonly present in multiparous women who deliver healthy infants. They can, however, be cytotoxic and cause clinical complications after blood products transfusion (TRALI and becoming refractory to platelets transfusion) and after organ transplantation (allogeneic organ rejection).     

Radiolabeled fragments of monoclonal antibodies against carcinoembryonic antigen for localization of human colon carcinoma grafted into nude mice

Four monoclonal antibodies against carcinoembryonic antigen (CEA) have been selected from 32 hybrids that produce antibodies against this antigen, by the criteria of high affinity for CEA and low cross- reactivity with granulocyte glycoprotein(s). The specificity of tumor localization in vivo of the four MAb, and their F(ab')2 and Fab fragments was compared in nude mice bearing grafts of a serially transplanted, CEA-producing, human colon carcinoma. The distribution of radiolabeled MAb and their fragments after intravenous injection was analyzed by direct measurement of radioactivity in tumor and normal organs, as well as by whole-body scanning and by autoradiography of tumor sections. Paired labeling experiments, in which 131I-labeled antibody or fragments and 125I-labeled control IgG are injected simultaneously, were undertaken to determine the relative tumor uptakes of each labeled protein. The tumor antibody uptake divided by that of control IgG defines the specificity index of localization. Tumor antibody uptakes (as compared with the whole mouse), ranging between 7 and 15, and specificity indices ranging between 3.4 and 6.8, were obtained with the four intact MAb at day 4-5 after injection. With F(ab')2 fragments of the four MAb, at day 3, the tumor antibody uptakes ranged between 12 and 24 and the specificity indices between 5.3 and 8.2. With the Fab fragments prepared from the two most promising MAb, the antibody uptakes reached values of 34 and 82 at day 2-3 and the specificity indices were as high as 12 and 19. The scanning results paralleled those obtained by direct measurement of radioactivity. With intact MAb, tumor grafts of 0.5-1 g gave very contrasted positive scans 3 d after injection. Using MAb fragments, tumors of smaller size were detectable earlier. The best results were obtained with Fab fragments of MAb 35, which gave clear detections of tumors weighing only 0.1 g as early as 48 h after injection. Autoradiographs of tumor sections from mice injected with 125I-labeled MAb demonstrated that the radioactivity was localized in the tumor tissues and not in the stromal connective tissue of mouse origin. The highest radioactivity concentration was localized in areas known to contain CEA such as the pseudolumen of glands and the apical side of carcinoma cells. The penetration of radioactivity in the central part of tumor nodules and the pseudolumen appeared to be increased with the use of MAb fragments.