用免疫球蛋白重/轻链配对比值发现同一泳带IgG-κ和IgM-κ双M蛋白血症1例

目的初步鉴定M蛋白峰完全重叠的双M蛋白血症。方法用血清蛋白质电泳、免疫固定电泳、免疫球蛋白定量、并根据IgG、κ、λ定量推算IgG重/轻链配对比值,对患者血清进行综合鉴定。结果血清蛋白质电泳显示在γ区有1条明显的M蛋白峰,免疫球蛋白及轻链定量初步提示为IgM-κ型M蛋白;而重/轻链配对比值IgG-κ/IgG-λ7.8,显著高于IgG-κ/IgG-λ比值正常上限2.7,提示同时存在IgG-κ型M蛋白;琼脂糖免疫固定和毛细管免疫固定均证实该患者存在IgG-κ型和IgM-κ型双M蛋白,2条M蛋白完全重叠。结论 2种单克隆M蛋白电泳条带完全重叠时,血清蛋白电泳无法区分,免疫球蛋白重/轻链配对比值可以用于免疫球蛋白克隆性的初步鉴别。     

42例多发性骨髓瘤的血清免疫电泳分析

目的应用免疫固定电泳检测多发性骨髓瘤（MM）患者血清M蛋白并进行分型。方法对42例MM患者血清进行血清蛋白电泳、免疫固定电泳及免疫球蛋白（IgG、IgA、IgM）定量分析。结果42例MM患者中：40例MM检出M带,免疫固定电泳分型：IgGκ型16例（38.1%）,IgGλ型10例（23.8%）;IgAκ型5例（11.9%）,IgAλ型3例（7.1%）;κ轻链型4例（9.5%）,λ轻链型2例（4.8%）;不分泌型2例（4.8%）。免疫球蛋白检测表明同型免疫球蛋白含量显著升高并常伴有非对应组分的偏低。结论免疫固定电泳检测MM患者的M蛋白特异性好、灵敏度高,对MM的诊断分型和临床分期及预后判断具有重要意义。     

42例多发性骨髓瘤的血清免疫电泳分析

目的应用免疫固定电泳检测多发性骨髓瘤（MM）患者血清M蛋白并进行分型。方法对42例MM患者血清进行血清蛋白电泳、免疫固定电泳及免疫球蛋白（IgG、IgA、IgM）定量分析。结果42例MM患者中：40例MM检出M带,免疫固定电泳分型：IgGκ型16例（38.1%）,IgGλ型10例（23.8%）;IgAκ型5例（11.9%）,IgAλ型3例（7.1%）;κ轻链型4例（9.5%）,λ轻链型2例（4.8%）;不分泌型2例（4.8%）。免疫球蛋白检测表明同型免疫球蛋白含量显著升高并常伴有非对应组分的偏低。结论免疫固定电泳检测MM患者的M蛋白特异性好、灵敏度高,对MM的诊断分型和临床分期及预后判断具有重要意义。     

游离轻链检测对多发性骨髓瘤诊断的临床意义

目的评价免疫固定电泳、尿液轻链定量及血清游离轻链检测在多发性骨髓瘤（MM）患者轻链检测中的价值。方法用速率散射比浊法进行重链、轻链及游离轻链定量,用蛋白电泳及免疫固定电泳法对M成份做定性分析,对多发性骨髓瘤患者轻链型21例,完整型29例（其中IgGI、gA型19例;IgD型10例）标本分别做免疫固定电泳,尿轻链定量及游离轻链检测,然后比较三种方法的检出率和灵敏度。结果对于轻链型骨髓瘤患者,免疫固定电泳、尿轻链定量及血清游离轻链检测三种方法的轻链检测阳性率分别为90.5%（19/21）;80.9%（17/21）;100%（21/21）。对于完整型骨髓瘤患者三种方法的游离轻链检测（免疫固定电泳法检测与重链结合的单克隆轻链的阳性率100%）阳性率分别为20.7%（6/29）;41.4%（12/29）;82.7%（24/29）。结论血清游离轻链检测较另外两种方法检测游离轻链更为灵敏,其与免疫固定电泳法结合检测对多发性骨髓瘤患者的明确诊断、疗效观察及改善预后起重要作用。     

IgE型多发性骨髓瘤的分型检测

目的探讨用免疫固定电泳法检测IgD和IgE单克隆免疫球蛋白，在筛选罕见IgE型多发性骨髓瘤(MM)中的应用价值。方法通过检测血清kD和IgE单克隆免疫球蛋白，从148例MM病人中，筛出只有轻链而不出现IgGAM重链的25份血清样本，用抗游离轻链的单克隆抗体及抗IgD、抗IgE单克隆抗体再进行免疫固定电泳检测。结果25例MM病人中有1例为IgE-κ单克隆免疫球蛋白阳性，24例呈游离轻链阳性(κ型7例、λ型17例)；在17例λ型游离轻链病人中有3例抗λ轻链(游离和结合)病人血清样本呈双克隆带状，用抗IgD单克隆抗体检测证实，有IgD．λ单克隆免疫球蛋白并伴有游离轻链λ。结论采用免疫固定电泳对MM病人血清进行IgG、IgA、IgM和轻链的检测后，用固定免疫技术对只有轻链而不出现IgGAM重链的样本，进行抗游离轻链、抗IgD和抗IgE单克隆抗体分型检测，成功地筛出1例罕见的IgE-κ型MM，提高了对该病的检测水平。     

免疫固定电泳检查在多发性骨髓瘤中的应用

目的探讨免疫固定电泳检查在多发性骨髓瘤患者血清单克隆免疫球蛋白（M蛋白）中的应用,并对M蛋白的特性进行分型鉴定。方法对39例血清蛋白电泳异常的患者血清进行免疫固定电泳、血清蛋白和免疫球蛋白分析。结果39例多发性骨髓瘤患者中,IgG型20例（51.3%）,其中κ轻链型9例,λ轻链型11例;IgA型11例（28.2%）,其中κ轻链型5例,λ轻链型6例;IgM型2例（5.1%）,均为λ轻链型,单纯轻链型6例（15.4%）,λ轻链型3例,κ轻链型3例。血清总蛋白升高者20例,球蛋白升高者24例。免疫球蛋白和轻链检测表明同型免疫球蛋白和轻链其含量多明显升高并常伴有其他组份的异常。结论应用免疫固定电泳对多发性骨髓瘤患者的M蛋白进行分型鉴定特异性好、灵敏度高,对多发性骨髓瘤的诊断分型和临床分期及预后具有重要意义。     

MM患者血清M蛋白轻重链的型别构成及与血清或尿液中游离轻链检出的关系

目的探讨多发性骨髓瘤患者(MM)血清单克隆免疫球蛋白(M蛋白)轻、重链型别分布的规律,以及不同重链型患者血清或尿液中游离轻链的检出状况。方法对采用免疫固定电泳对312例MM患者血清M蛋白进行分析的结果(不含非分泌型)进行回顾性分析。结果在312例MM患者中,IgG型、游离轻链型、IgA型和IgM型分别占62.82%、20.51%、15.38%和1.28%。IgG和IgM重链型MM患者均分别以κ轻链为主,κ∶λ=1.55∶1.00和3.00∶1.00,而IgA以λ轻链为主,κ∶λ=0.71∶1.00。IgG型MM患者中,血液中检出游离轻链的患者占43.88%,显著高于IgA型的20.83%和IgM型的25.00%(P=0.003、0.004)。血液中检出游离轻链的IgA型MM患者,其尿液中有70.00%亦同时出现游离轻链,这一比例显著高于IgG型的16.28%(P=0.000)。结论 MM患者仍以IgG型为主,游离轻链型居于第2位。MM患者的重链分型,与血液或尿液中检出游离轻链的概率高低有关。     

453例多发性骨髓瘤回顾性分析

目的探讨免疫球蛋白定量和κ、λ比值分析在多发性骨髓瘤实验诊断中的价值. 方法对453例经血清免疫电泳、免疫固定电泳、血、尿κ、λ轻链ELISA测定确诊为多发性骨髓瘤(MM)患者的免疫球蛋白定量和κ、λ比值及一般临床资料进行统计分析. 结果 IgG类患者212例,占46.8%,IgA类90例(19.9%),IgM类35例(7.7%),IgD类36例(7.9%),游离κ链36例(7.9%),游离λ链44例(9.7%). 结论 IgM,IgG及IgA型MM患者中以κ型轻链为主,而IgD类以λ型轻链为主.在IgG类中,IgG1亚类最多见,IgG2值最高.     

α重链病临床特征和实验室检查特征

目的探讨α重链病的临床特征和实验室检查特征。方法收集华中科技大学同济医学院附属同济医院收治的1例α重链病患者及文献报道的3例α重链病患者的临床资料,分析α重链病临床表现及实验室特点。结果 4例患者中男性3例,女性1例;年龄11~58岁,平均年龄38.5岁;临床表现为腹痛3例;3例通过治疗后恢复健康,1例手术治疗后死亡。α重链病患者血清免疫球蛋白定量检测中IgA水平大多数偏高,蛋白电泳中在β和α2区间常会出现一宽峰,免疫固定电泳上通常显示为在IgA泳道上形成特异性反应沉淀带,与IgG、IgM和K、L无特异性反应沉淀带。结论重链病非常少见,早期α重链病患者临床表现不明显,分型和诊断依赖血清免疫固定电泳,早期对抗菌药物敏感,中晚期重链病患者倾向发展为淋巴瘤。     

肾淀粉样变性病患者临床病理分析（ 附 15 例报告）

目的探讨淀粉样变性肾脏损害的临床及病理特征，提高对肾淀粉样变性病的认识。方法回顾性分析徐州医学院附属淮海医院10年来收治的15例肾淀粉样变性患者的临床及病理资料。结果本研究15例肾淀粉样变性肾脏损害均表现为蛋白尿，其中表现为肾病综合征者13例，4例伴镜下血尿，4例伴肾功能衰竭。肾外表现：本研究14例患者中4例表现为高血压，4例血压〈90／60mmHg（1mmHg=0．133kPa）。15例中14例有不同程度的心脏损害，代谢性脑病1例，腹水3例，胸腔积液2例，舌体增大1例，肋骨病理性骨折2例；肝脏肿大4例；脾脏肿大1例，所有患者肾脏无明显缩小。24h尿蛋白定量（6．78±2．92）g，尿免疫球蛋白入轻链（629．14±1139．90）mg／L，尿免疫球蛋白κ轻链（349．83±565．94）mg／L，血κ轻链（4．62±2．18）g/L，血入轻链（3．93±4．36）g／L，血清白蛋白（24．02±6．45）g／L；血尿素氮（8．35±5．11）mmol／L，肌酐（94．51±67．94）μmol／L。病理改变：光镜下15例患者中14例以肾小球损害为主，伴不同程度的小管间质及血管病变。光镜下可见肾小球体积明显增大，系膜区中重度增宽。15例患者中10例患者PASM-Masson染色可见上皮侧较多的毛刺状嗜银物，全部15例患者肾间质小动脉壁亦见PAS及HE淡染的均质无结构物质沉积；免疫荧光示免疫复合物弥漫分布，呈颗粒状及团块状沉积于系膜区及血管袢；肾小管沉积以IgG、C3为主。肾组织轻链染色均阳性，其中7例患者肾小球K轻链染色呈阳性，11例患者肾小球入轻链染色呈阳性，l例患者肾小球K、入轻链染色均阴性，仅肾小管κ、λ轻链染色阳性。15例患者肾组织刚果红染色均阳性，其中14例经高锰酸钾处理后再行刚果红染色仍然阳性，1例经高锰酸钾处理后再行刚果红染色为阴性。结论淀粉样变性肾脏易受累，临床表现多样，确诊主要依靠病理活检刚果红染色。     

10例轻链型多发性骨髓瘤的实验室结果分析

目的探讨轻链型多发性骨髓瘤(MM)实验室检查的特点。方法对10例轻链型MM病例的实验室检查结果进行回顾性分析。结果轻链型MM的RBC、HB、UREA、CRE、IgG与IgG型MM比较,差异有显著性意义(P<0.05);轻链型MM的WBC、PLT、IgA、IgM、骨髓浆细胞比例、骨髓原幼浆细胞比例、骨髓成熟浆细胞比例以及溶骨性损害比例与IgG型MM比较,差异无显著性意义(P>0.05)。轻链κ型MM和IgG、κ型MM血清、尿液游离轻链大部分指标比较,差异有显著性意义(P<0.05)。轻链λ型MM和IgG、λ型MM血清、尿液游离轻链各指标比较,差异无显著性意义(P>0.05)。结论轻链型MM的贫血、肾功能损害较突出,完整的免疫球蛋白水平明显低于相应类型的MM。血清、尿液游离轻链定量及血清免疫固定电泳在MM的诊断及分型中起到非常重要的作用。     

208例多发性骨髓瘤细胞形态学与血清免疫球蛋白比对分析

目的回顾分析208例多发性骨髓瘤(MM)病例的骨髓细胞学、临床分型以及血清免疫球蛋白、免疫固定电泳结果 ,探讨其在MM诊断中的作用。方法回顾分析208例MM患者骨髓细胞学特点,按照欧洲血液学会议形态学分型标准将骨髓瘤细胞分型,计数骨髓瘤细胞比例;按临床特点和分期标准将病人分期;记录病人血清免疫球蛋白、免疫固定电泳结果。结果 208例MM病例中:骨髓涂片中骨髓瘤细胞比例≥10%的204例(98.1%),骨髓瘤细胞比例在6.0%-10.0%的3例(1.4%),骨髓瘤细胞比例不增高的1例(0.5%);血清免疫固定电泳:IgG型128例(61%),IgA型47例(22.7%),轻链型28例(13.7%),未分泌型5例(2.8%);临床表现主要有骨破坏168例(80.1%)、贫血163例(79.5%)、骨痛146例(70.1%)、肾功能损害102例(49%)、出血8例(3.8%)、粘滞血症2例(0.9%)。结论骨髓细胞形态学在MM的诊断中尤为重要。血清学特点及明显的临床表现也对MM的诊断有提示作用,三者需紧密结合。     

血清κ/λ轻链比值及蛋白电泳在诊断多发性骨髓瘤中的临床应用

目的 探讨多发性骨髓瘤(MM)患者血清轻链(sLC)κ/λ比值及血清蛋白电泳(SPE)在MM诊断中的临床价值.方法 采用速率散射免疫比浊法测定282例MM患者及521例健康对照人群sLC水平并计算κ/λ比值,同时做SPE和免疫固定电泳(IFE).以临床已确诊为MM的患者分型结果为标准,评价sLC比值联合SPE检测对MM...     

Localization of the membrane attack complex (MAC) in experimental immune complex glomerulonephritis

The role of the membrane attack complex (MAC) as a mediator of renal tissue injury was evaluated in rats affected by bovine serum albumin (BSA)-induced immune complex glomerulonephritis. Immunofluorescence studies revealed concurrent deposits of IgG, BSA, C3, and the MAC along glomerular capillary walls, although the MAC manifested a more restricted distribution than that observed for immune complexes. Immunoelectron microscopic techniques were utilized to demonstrate immune complexes, C3, and the MAC within dense deposits in the subepithelial aspect of the basement membrane. Visceral epithelial foot processes were fused in areas overlying large dense deposits and exhibited intense staining for the MAC, lesser reactivity for C3 but IgG was absent from the foot process membranes. Smaller granular deposits of immune complexes, C3, and the MAC were observed in the subendothelial region of the lamina rara interna and the lamina densa. Immune complexes may activate the classical complement pathway causing diffuse injury to the glomerular basement membrane (GBM), allowing subepithelial accumulation of complexes. These observations implicate the MAC as a mediator of GBM and juxtaposed podocyte membrane injury, thereby contributing to disruption of the glomerular filtration barrier. IgG and C3 were demonstrated within tubulointerstitial regions on the surface of collagen fibers in close proximity to the tubular basement membrane (TBM) of proximal convoluted tubules. Within the TBM, C3 localization was prominent with diminished reactivity for the MAC, but IgG was not detectable. The demonstration of C3 and scant MAC deposits in the TBM of nonimmunized control rats without evidence of interstitial IgG and C3 deposits suggests that both nonimmune and immune processes play a role in the pathogenesis of extraglomerular lesions. Evidence derived from these morphologic studies indicates that the MAC is associated with injury to the GBM, foot process membranes of visceral epithelium, and the TBM. Further experiments designed to selectively enhance or inhibit the deposition of MAC and assess consequent renal dysfunction are required to substantiate hypotheses concerning the in vivo membranolytic potential of the MAC in experimental immune complex glomerulonephritis.     

系统性红斑狼疮患者免疫球蛋白轻链及补体B因子测定的临床意义

目的 探讨血清中免疫球蛋白轻链κ和λ含量、κ/λ比值以及补体B因子(PFB)含量的检测在系统性红斑狼疮(SLE)诊疗中的应用价值.方法 在特定蛋白仪上,采用免疫速率散射比浊法测定SLE患者及对照组血清中轻链κ和轻链λ、PFB含量,并计算κ/λ比值.结果与30例对照组比较,51例SLE患者血清中轻链κ[(16.51±6.27)g/L,P<0.01]和轻链λ[(10.30±4.35)g/L,P<0.01]均升高;κ/λ比值下降(1.60±0.32,P<0.01),PFB含量下降[(0.214±0.063)g/L,P<0.01].结论 血清中免疫球蛋白轻链κ和轻链λ含量、κ/λ比值及PFB含量检测对SLE的临床诊疗具有重要的意义.     

Chemical compositions of glomerular and tubular basement membranes of human kidney.

Glomerular basement membranes (GBM) and tubular basement membranes (TBM) were prepared from human kidney and their chemical compositions were studied. Electron microscopic figures showed no contamination of the cellular components components or collagen fibers, indicating a high degree of purity of the preparations. Low contents of phospholipid supported this indication. Amino acid compositions of the human GBM and TBM resembled that of the bovine GBM, containing hydroxyproline, hydroxylsine, half-cystine, methionine and a large amount of glycine as the characteristic amino acids. The human GBM and TBM had similar carbohydrate compositions consisting of glucose and galactose, as the major sugars, together with mannose, L-fucose, hexosamine and sialic acid. Glucosylgalactosylhydroxylysine and a small amount of galactosylhydroxylysine were detected in alkaline hydrolyzates of both membranes. The human GBM and TBM separated into more than ten subunits having molecular weights ranging from 2.5 times 10-4 to more than 2.5 times 10-5 on polyacrylamide gel electrophoresis in the presence of sodium laurylsulfate and 2-mercaptoethanol. The electrophoretograms of both membranes resembled that of the bovine GBM. These observations indicate that the human TBM has a similar chemical structure to those of the human GBM and the bovine GBM.     

Immunoglobulin free light chain assay using latex agglutination

Summary Under normal biological conditions, immunoglobulin light chains are generally formed in conjunction with heavy chains; however, small quantities of free light chains are also produced. Thus, two types of immunoglobulin light chains may be identified; immunoglobulin-bound light chains and free light chains. In mature neoplastic B cells, immunoglobulin synthesis, including that of the free light chain, is generally increased. Furthermore, elevated free light chain levels also occur in the presence of renal glomerular and tubular impairment. Thus, the quantitative and qualitative measurement of free light chains is of considerable clinical benefit. Although methods for measurement of free light chains do exist, they are technically complicated and time consuming. Previously, we have reported a quantitative nephelometric immunoassay for measuring free light chain levels. We report here a refinement of this method using latex agglutination techniques. This new method enables quantitative measurement of free light chains in serum at levels as lows as 1 mg/l and may allow the clinical application of serum free light chain estimation.     

Structure of a monoclonal kappa chain of the V kappa IV subgroup in the kidney and plasma cells in light chain deposition disease.

That structural abnormalities may be responsible for nonamyloid immunoglobulin (Ig) light chain deposition disease (LCDD) is suggested by previous results of Ig biosynthesis studies, but this hypothesis was not documented at the molecular level. We report on the first complete primary sequence deduced from cDNA analysis of a kappa light chain responsible for LCDD associated with an apparently nonsecretory myeloma. Bone marrow myeloma cells contained intracellular kappa chains and no heavy chains by immunofluorescence. Kidney biopsy showed typical nonamyloid PAS-positive kappa chain deposits. SDS-PAGE analysis of material extracted from a kidney biopsy specimen and of Ig produced by the myeloma cells revealed kappa chains of abnormally high apparent molecular mass (30,000). Comparison of the NH2-terminal aminoacid sequence of the kappa chain deposited in the kidney and of the complete sequence of several identical kappa cDNA clones from bone marrow cells showed the identity of the tissue deposited and plasma cell kappa chain. The kappa mRNA had an overall normal structure and corresponded to the V kappa IV gene rearranged to J kappa 1 and followed by a normal constant exon of the Km(3) allotype. The variable sequence differed from the V kappa IV germline gene by nine point mutations, including an Asp----Asn substitution at position +70 resulting in a potential N-glycosylation site. In vitro biosynthesis experiments and treatment with N-glycosidase provided evidence for the intracellular glycosylation of the monoclonal kappa chain. The peculiar sequence and the glycosylation of a kappa chain of the rare V kappa IV subgroup might be responsible for structural abnormalities leading to tissue deposition.     

Clinical features and outcome of patients with thin and ultrathin glomerular membranes

There is considerable disagreement regarding the natural historyof renal disease associated with thin glomerular basement membranes(TGBM). We followed 43 patients (19 male), mean age41.6 years(range 19–73) for a mean of 88 months (48–140).TGBM was recognized in adults when glomerular basement membranethickness, measured from multiple sites in electronmicrographsof renal biopsy tissue as the harmonic mean, was <320 nm.At presentation, 95% had microscopic haematuria, 12% macroscopichaematuria, 14% loin pain, 28% proteinuria, and 14% hypertension.There was no difference in GBM width between the sexes (male258 nm vs. female 251 nm) but there was a significant negativecorrelation between age and GBM width (r=–0.53, p<0.001),with older patients having the thinnest membranes. Twenty sixpatients had ultrathin GBM (<270 nm), of whom 54% had 3$haematuria vs. 12% of the group with BM >270 nm (p<0.01).In the ultrathin group, 71% had loss of anionic charge fromthe GBM, vs. 17% in those with membranes which were thin but>270 nm (p<0.05). Proteinuria occurred more frequentlyin those with GBM >270 nm, 65% vs. 8% in the ultrathin group(p<0.01). Thin GBM were associated with a benign prognosis,as after a mean follow-up of 85 months (48–140), therewas no significant change in either serum creatinine or meanarterial blood pressure. Patients with ultrathin GBM had greaterloss ofGBM anionic charge, which might result in both an alterationof flow characteristics within the glomerular capillaries andalso increased fragility of the glomerular basement membranewithlikelihood of rupture and resultant macroscopic haematuria.     

Isolation and characterization of the nephritogenic antigen producing anti-tubular basement membrane disease

Using monoclonal antibody affinity chromatography, we isolated a 48,000 mol wt, glucose-rich glycoprotein (3M-1) from collagenase-solubilized rabbit renal tubular basement membrane (SRTA). The purified 3M-1 protein is noncollagenous, and is capable of inducing anti-TBM (tubular basement membrane) antibodies and interstitial nephritis in susceptible hosts. Further, when SRTA, at a normally nephritogenic dose, was selectively depleted of 3M-1, it lost its ability to induce disease. As shown by immunofluorescent techniques, 3M-1 appears to be localized on rodent TBM to the exclusion of the glomerular basement membrane, but was lacking in the TBM of the LEW rat, a strain devoid of the relevant antigen of anti-TBM disease. Immunoelectron microscopy revealed that 3M-1 was associated with the most lateral aspect of the TBM, which borders, and lies in the interstitium. These results indicate that 3M-1 is the nephritogenic antigen producing experimental anti-TBM disease.