狼疮性肾炎患者血清DNA—抗—DNA免疫复合物与蛋白尿和肾脏病理…

利用单克隆抗－ＤＮＡ抗体检测了３９名狼疮性肾炎（ＬＮ）患者不同的时期内不同类型的血清ＤＮＡ－抗－ＤＮＡ免疫复合物，其中２４例做了肾活检。结果显示：活动期ＬＮ患者血清抗－ＤＮＡＩＣ的类型分布与患乾的肾脏病理类型有一定的关系，有助于推测ＬＮ肾组织病理类型，ＩｇＧ型的抗单链ＤＮＡＩＣ与２４ｈ尿蛋白定量呈正相关。进一步说明抗－ＤＮＡＩＣ在ＬＮ的发病机理中的较重要的作用。     

释放左旋十八甲基炔诺酮宫内节育器对子宫内膜DNA含量的影响

为了研究释放左旋十八甲基炔诺酮（２０μｇ／天）宫内节育器（ＬＮＧ－ＩＵＤ－２０）对子宫内膜细胞ＤＮＡ含量的影响。将置器后子宫内膜剪碎后涂片，Ｆｅｕｌｇｅｎ染色，用显微分光光度计测量内膜单细胞核ＤＮＡ含量。结果显示，放置ＩＵＤ后内膜细胞ＤＮＡ含量显著下降（Ｐ＜０．００１）。鉴于低释放量的（２μｇ／天）ＬＮＧ－ＩＵＤ（ＬＮＧ－ＩＵＤ－２）并不影响内膜细胞的ＤＮＡ含量。认为，ＬＮＧ－ＩＵＤ对内膜增殖活性的影响同释放量密切相关，而且推断对内膜活性的抑制程度与ＩＵＤ的临床表现密切相关     

体外免疫吸附对狼疮肾炎的治疗和保护作用

目的　观察体外免疫吸附（ＥＩＡ） 对狼疮肾炎（ＬＮ） 的治疗和肾脏保护作用。方法　比较树脂型ＥＩＡ、大剂量激素和ＣＴＸ联合治疗（ 简称ＥＩＡ组）与大剂量激素和ＣＴＸ治疗（ 简称对照组） 血ＩｇＧ、ＡＮＡ、ｄｓＤＮＡ、Ｃ３、２４ 小时尿蛋白、ＢＵＮ、Ｓｃｒ 及其他临床指标的变化。结果　治疗２ 周时,ＥＩＡ组血ＩｇＧ、ＡＮＡ、ｄｓＤＮＡ显著低于对照组［ 两组分别为ＩｇＧ（１０－０３ ±２－９１） 、（１３－５４ ±３－０５）ｇ／Ｌ,ＡＮＡ（１４－００ ±１３－１４）、（３１－８２ ±３０－０２）ｔｉｔｅｒ－ １,ｄｓＤＮＡ（８－３０ ±４－８３） 、（２７－００ ±１８－５１） ％］ 。Ｃ３ 显著回升［ 两组分别为（５８７－３±９７－０３） 、（４８１－５±１３２－０４）ｍｇ／Ｌ］,２４ 小时尿蛋白、ＢＵＮ、Ｓｃｒ 显著低于对照组［ 两组分别为２４ 小时尿蛋白（２－２８±２－０７） 、（５－９８ ±３－３４）ｇ／２４ｈ,ＢＵＮ（９－１７ ±２－８３）、（１５－２９ ±５－３８）ｍｍｏｌ／Ｌ、Ｓｃｒ（１５６－７５ ±８９－３３）、（２４７－１４±７４－６９）μｍｏｌ／Ｌ） 。治疗４ 周时,ＥＩＡ 组血ＩｇＧ、ｄｓＤＮＡ 仍低于对照组［ 两组分别为Ｉｇ     

充血性脾大伴脾功能亢进患者不同脾切除术前后血小板相关抗体的变化

目的 了解充血性脾肿大伴脾功能亢进（脾亢）患者血小板相关抗体（ＰＡ－ＩｇＧ）水平及不同脾切除术后的改变,探索脾肿大、血小板、ＰＡ－ＩｇＧ之间的关系。方法 采用竞争性酶联免疫吸附试验（ＥＬＩＳＡ）检测了２４例脾肿大伴脾亢患者血清ＰＡ－ＩｇＧ水平。结果 脾肿大伴脾亢患者的ＰＡ－ＩｇＧ水平明显高于正常者（Ｐ〈０．０１）,而血小板值低,ＰＡ－ＩｇＧ与血小板之间存在显著负相关（ｒ＝－０．４７４７,Ｐ〈０．０     

^125I标记抗C—erbB—2单抗对荷人乳腺癌裸鼠的定位诊断及生？…

目的 研究＾１２５Ｉ标记抗Ｃ－ｅｒｂＢ－２单克隆抗体ＩＣＲ１２（简称＾１２５Ｉ－ＩＣＲ１２）及正常小鼠ＩｇＧ（简称＾１２５Ｉ－ｍＩｇＧ）在荷人乳腺癌裸鼠体内的分布，为放射免疫显像（ＲＩＩ）及临床应用提供依据。方法 每鼠尾静脉注抗体９２．５ＫＢｑ／０．１ｍｌ于注射后２４，４８，９６，１２０小时分批处死，测定肿瘤和血液，肝，肺等重要脏器的单位重量放射性比值（Ｔ／ＮＴ）各组织摄取百分比（％ＩＤ／ｇ）及定     

原位杂交检测乙型肝炎病毒DNA在肾脏的存在

目的：探讨乙型肝炎病毒对肾脏的致病作用。方法：应用地高辛素标记的ＨＢＶ ＤＮＡ两种探针原位杂交，检测１５例乙型肝炎病毒相关性肾炎（ＨＧＶ－ＧＮ）患者肾活检石腊包埋切片ＨＢＶ ＤＮＡ。结果：ＩＳＨ采用ＨＢＶ ＤＮＡ全长段探针９／１５例阳性，其中５例阳性和４例阴性者再用ＨＢＶ ＤＮＡ Ｘ＋Ｃ段探针检查，各获１例阳性（２０％和２５％），阳性信号主要见于肾小管上皮细胞，其次是肾小球系膜细胞，呈胞浆型或核型     

bDNA定量检测血清乙型肝炎病毒DNA在肾炎中的应用

我国是乙型肝炎病毒（ＨＢＶ）感染高发区,ＨＢＶ感染可致急性肝炎、慢性肝炎、肝硬化、肝癌或引起肝外病变如乙型肝炎病毒相关性肾小球肾炎（ＨＢＶ－ＧＮ）等。α－干扰素（ＩＦＮ－α）作为一种有效的抗病毒手段已被广泛应用于慢性乙型肝炎,并被尝试用于治疗ＨＢＶ－ＧＮ［１］。分枝ＤＮＡ技术（ｂＤＮＡ）是血清ＨＢＶ　ＤＮＡ定量先进技术［２］。现将我科应用该技术在部分血ＨＢｓＡｇ（＋）肾炎患者中的诊疗经验总结如下。 一、对象和方法 １．对象：１９９７．６～１９９８．８期间在本科住院的各类肾小球肾炎伴血清ＨＢｓＡｇ（…     

3－酮－地索高诺酮对雌、孕及雄激素受体亲和力选择性的实验研究

采用体外放射配体受体结合法测定３－酮－地索高诺酮（ＫＤＧ）与兔子宫胞浆雌激素受体（ＥｃＲ）、孕激素受体（ＰｃＲ）和大鼠腹侧前列腺胞浆雄激素受体（ＡｃＲ）结合的亲和力，并以左旋１８－甲基炔诺酮（ＬＮＧ）作对照，从受体水平探讨ＫＤＧ对当体激素受体的选择性。结果表明：ＫＤＧ对ＰｃＲ有很高的亲和力，是ＬＮＧ的１．５倍；对ＡｃＲ的亲和力很低，仅为ＬＮＧ的０．７４倍；对ＥｃＲ几乎没有亲和力，受体亲和力选择系数也明显高于ＬＮＧ。由此可见，在ＬＮＧ甾核结构中Ｃ１１位引入次甲基合成的ＫＤＧ，其受体亲和力选择性更专一。     

孕二烯酮和左旋18甲基炔诺酮抗排卵的实验研究

本研究比较孕二烯酮（ＧＳＤ）和左旋１８甲基炔诺酮（ＬＮＧ）的抗排卵作用。灌胃（ｉｇ）或皮下注射（ｓｃ）ＧＳＤ４及３ｍｇ／ｋｇ或ＬＮＧ４０ｍｇ／ｋｇ（ｓｃ）都可以完全抑制兔的排卵作用。经ｉｇ或ｓｃ两种药物对兔的抗排卵作用ＥＤ５０：ＧＳＤ分别为０．７９和０．６７ｍｇ／ｋｇ；而ＬＮＧ分别为２２．９和１６．１ｍｇ／ｋｇ。ＧＳＤ和ＬＮＧ对大鼠的抗排卵作用：ＧＳＤ７２（ｉｇ），４８ｍｇ／ｋｇ（ｓｃ）或ＬＮＧ４８０ｍｇ／ｋｇ（ｉｇ）都可以完全抑制大鼠排卵作用，以上剂量ＧＳＤ抑制排卵的ＥＤ５０分别为２４．０（ｉｇ）及１４．４ｍｇ／ｋｇ（ｓｃ）；ＬＮＧ为１９０ｍｇ／ｋｇ（ｉｇ）。ＧＳＤ和ＬＮＧ均有明显的孕激素活性，且ＧＳＤ比ＬＮＧ较强。ＧＳＤ无雌激素活性，但具有抗雌激素活性。ＧＳＤ的抗排卵作用较ＬＮＧ为强。     

IgA肾病是否与乙型肝炎病毒有关？

探讨ＩｇＡ肾病与乙型肝炎病毒的关系。方法采用免疫组化技术检测ＩｇＡ肾病患者肾组织中ＨＢＶ抗原及Ｓｏｕｒｔｈｅｒｎ印迹分子杂交技术检测肾组织中ＨＢＶＤＮＡ。结果８５例ＩｇＡ肾病患者血清ＨＢｓＡｇ阳性１５例（１７．６５％），肾组织ＨＢＶ抗原阳性２６例（３０．５９％），其中肾小球阳性１８例（５７．１４％），ＨＢｃＡｇ在肾小管和肾间质中阳性分别为９例（３４．６７％）和２例（７．６９％）；２例测肾组织ＨＢＶＤＮＡ，１例阳性；肾组织ＨＢＶ抗原阳性组比阴性组临床和病理改变更严重。结论乙型肝炎病毒与ＩｇＡ肾病的发病密切相关     

Mesangial cell-binding anti-DNA antibodies in patients with systemic lupus erythematosus

The mechanisms by which anti-DNA antibodies contribute to the pathogenesis of lupus nephritis (LN) remain to be elucidated. This study investigates the binding of polyclonal anti-DNA immunoglobulins from patients with systemic lupus erythematosus (SLE) to human mesangial cells (HMC) in vitro. Testing of cross-sectional serum samples from 280 LN patients (108 during active disease; 172 during remission), 35 SLE patients without renal involvement, 72 patients with non-lupus primary glomerular diseases, and 37 healthy subjects with a cellular enzyme-linked immunosorbent assay showed significant IgG mesangial cell-binding activity in patients with SLE, particularly those with active LN (P < 0.0001). Significant HMC-binding activity was demonstrated in 83.9%, 42.8%, and 47.1% of patients with active LN, inactive LN, and non-renal SLE, respectively. This was predominantly attributed to binding by anti-DNA antibodies, and immune complex binding accounted for 4.6%, 3.5%, and 2.8% of seropositive samples in the respective groups. Longitudinal studies in 27 LN patients demonstrated correlation between serial levels of anti-DNA antibodies, serum HMC-binding activity, and disease activity in 18 patients (66.7%). Affinity-purified polyclonal IgG anti-DNA antibodies from sera with HMC-binding activity showed significant binding to cultured HMC, and to a lesser extent glomerular and proximal tubular epithelial cells and human umbilical vein endothelial cells, but not tumor cell lines, peritoneal mesothelial cells, bronchial epithelial cells, or fibroblasts. The binding of anti-DNA antibodies to HMC was increased 1.47-fold (P = 0.0059) after the removal of Ig-associated DNA by DNase treatment, but it was unaffected by DNase treatment of HMC membrane. Controlled trypsinization of membrane proteins in HMC resulted in a 1.26-fold (P = 0.0025) increase in their binding by anti-DNA antibodies. In conclusion, subsets of anti-DNA antibodies from patients with SLE are capable of binding to HMC. The association of such binding with renal involvement and disease activity and its modulation by DNA concentration suggest that Ig binding to HMC can be a potential marker for disease activity in selected patients and that the binding of anti-DNA antibodies to HMC may be a pathogenetic mechanism in LN.     

Anti-DNA antibodies in the urine of lupus nephritis patients.

BACKGROUND: It has previously been reported that patients with systemic lupus erythematosus (SLE) and glomerulonephritis do not have anti- (deoxyribonucleic acid) DNA antibodies in their urine. This finding was attributed to specific entrapment of anti-DNA antibodies by the immune complexes in the glomerular capillary walls. METHODS: This phenomenon has been re-investigated as part of a study of the use of desoxyribonuclease 1 (DNase 1) to treat lupus nephritis (LN). For this purpose an ELISA was developed for the detection of anti-DNA antibodies in urine. It was found that such an assay was very susceptible to the presence of DNase in urine which destroys the antigen coating the plates and gives rise to false negative results. For this reason, it is essential that all tests for anti-DNA antibodies in the urine are carried out in the presence of EDTA to inhibit the endogenous DNase 1 activity. RESULTS: Using this assay to test the urine from 24 patients with LN and non-selective proteinurea, it was found that they all contained anti-DNA antibodies. The amount of anti-DNA antibodies detected in the urine was compared with that expected by calculations from the anti-DNA antibody titre in the serum and total immunoglobulin levels in serum and in urine. It showed that in 20 patients there was neither specific entrapment nor specific excretion of anti-DNA in urine, only the expected amount of leakage. In only three patients was any appreciable entrapment demonstrated and in only one, any excess excretion. CONCLUSIONS: It is suggested that the failure to detect anti-DNA antibodies in the urine in the previous work was due to failure to inhibit the endogenous urinary DNase. It remains to be determined whether the retention of anti-DNA antibodies or excessive secretion is correlated with clinical phases of LN.     

Heterogeneity of immune complex-derived anti-DNA antibodies associated with lupus nephritis

The mechanisms responsible for the tissue injuries associated with lupus nephritis have not yet been well explained. We have investigated the characteristics of anti-DNA antibodies in circulating immune complexes (CIC) and in the deposits of renal glomeruli in patients with active lupus nephritis. The CIC-derived antibodies expressed anti-DNA idiotypes (Id) designated as 0-81 Id and NE-1 Id, and bound mainly to single-stranded DNA but never to glomerular basement membrane (GBM) antigens. On the other hand, the immunoglobulins (Ig) eluted from renal glomeruli of lupus patients reacted not only with DNA but also with GBM, proteoglycan, and heparan sulfate. The binding of glomeruli-deposited Ig was markedly low when GBM antigens were used after treatment with heparitinase, suggesting that some anti-DNA antibodies may bind directly to GBM antigens associated with heparan sulfate, and form in situ IC in renal glomeruli. It was also revealed that the renal eluates obtained after passing through GBM antigen-coupled Sepharose lost the binding ability with GBM but still retained DNA-binding and 0-81 Id activity, showing the participation of circulating IC-derived anti-DNA antibodies in the glomerular deposits. Theoretically there may be two mechanisms in the pathogenesis of lupus nephritis through the deposition of circulating IC and through in situ formation of anti-DNA IC in renal glomeruli. The diversity of histological features in lupus kidneys may be attributed to the heterogeneity of the mechanisms.     

Glomerular Autoimmune Multicomponents of Human Lupus Nephritis In Vivo (2): Planted Antigens

Glomerular planted antigens (histones, DNA, and C1q) are potential targets of autoimmunity in lupus nephritis (LN). However, the characterization of these antigens in human glomeruli in vivo remains inconsistent. We eluted glomerular autoantibodies recognizing planted antigens from laser-microdissected renal biopsy samples of 20 patients with LN. Prevalent antibody isotypes were defined, levels were determined, and glomerular colocalization was investigated. Renal and circulating antibodies were matched, and serum levels were compared in 104 patients with LN, 84 patients with SLE without LN, and 50 patients with rheumatoid arthritis (RA). Autoantibodies against podocyte antigens (anti–α-enolase/antiannexin AI) were also investigated. IgG2 autoantibodies against DNA, histones (H2A, H3, and H4), and C1q were detected in 50%, 55%, and 70% of biopsy samples, respectively. Anti-DNA IgG3 was the unique non-IgG2 anti-DNA deposit, and anti-C1q IgG4 was mainly detected in subepithelial membranous deposits. Anti-H3, anti-DNA, and anti-C1q IgG2 autoantibodies were also prevalent in LN serum, which also contained IgG3 against the antigen panel and anti-C1q IgG4. Serum and glomerular levels of autoantibodies were not strictly associated. High serum levels of all autoantibodies detected, including anti–α-enolase and antiannexin AI, identified LN versus SLE and RA. Anti-H3 and anti–α-enolase IgG2 levels had the most remarkable increase in LN serum and represented a discriminating feature of LN in principal component analysis. The highest levels of these two autoantibodies were also associated with proteinuria>3.5 g/24 hours and creatinine>1.2 mg/dl. Our findings suggest that timely autoantibody characterization might allow outcome prediction and targeted therapies for patients with nephritis.     

正常人与原发性肾小球肾炎及狼疮性肾炎患者血尿IL-18水平比较

目的:探讨IL-18在原发性肾小球肾炎(PGN)及狼疮性肾炎(LN)发生、发展中的作用,以及寻找有助于两类疾病的鉴别诊断和对肾组织炎症活动程度进行评估的指标。方法:应用酶联免疫吸附检测(enzyme-linkedimmunosorbent assay,ELISA)法测定16例正常人、21例原发性肾小球肾炎(PGN)患者和18例LN患者血浆和尿液IL-18水平的变化。结果:LN患者血浆及尿液IL-18水平显著高于正常对照组(P均<0.001)和PGN组(P均<0.05),而且WHOⅣ型LN患者血浆及尿IL-18水平均明显高于非Ⅳ型LN患者(P<0.05);PGN患者尿液IL-18水平也高于正常人(P<0.05),但血浆IL-18水平与正常人比较无统计学差异(P>0.05);LN患者血浆IL-18水平与SLEDAI呈正相关(P<0.01),而尿IL-18水平与狼疮性肾炎RHSAI是密切正相关(P<0.001),但尿IL-18水平与血浆IL-18水平相关性没有统计学意义(P>0.05)。结论:IL-18参与LN的全身免疫病理过程,但可能仅参与PGN肾组织局部炎症过程;血浆IL-18检测可能有助于区分LN和PGN,尿IL-18的检测可望作为一项估计LN肾组织病变活动程度的有用指标。     

Clinical significance of the detection of circulating immune complexes in lupus nephritis

32 patients (22 biopsed) with lupus nephritis (LN) were observed for circulating immune complexes (IC). Solid phase C1q (SPC1q) and polyethylene glycol (PEG) precipitation tests were used. The patients were studied during the clinical follow-up in different phases of disease activity. Comparative studies between each histological class of LN and corresponding forms of idiopathic glomerulonephritis (IGN) were made: no significant differences were found between either mesangial LN and stalk mesangial IGN, or between focal proliferative LN an focal proliferative IGN. However, a significant difference was found for SPC1q data between diffuse proliferative LN and mesangiocapillary IGN, and between membranous LN and membranous IGN. LN, with an acute nephritic syndrome and hypocomplementemia, displayed SPC1q data significantly above the levels of IC found in IGN with similar clinical features. IC serum data would seem an important element for the diagnosis and the clinical management of patients affected by LN.     

Class V lupus nephritis: a clinicopathologic study in 152 patients

BACKGROUND: Class V lupus nephritis (LN) can be divided into two subgroups according to the 1995 WHO modified classification, but the difference in clinical characteristics between these subgroups is not well known. METHODS: We classified 152 patients with Class V LN, confirmed by renal biopsy, into two subgroups (61 Class Va, 91 Class Vb), and enrolled 488 patients with Class IV as controls. The clinical manifestations, serologic results and prognosis were compared for Classes Va and Vb. RESULTS: The incidence of hypertension and anemia in Class Vb patients was significantly higher than in Class Va (38.5% vs 21.3%, 72.5% vs 52.5%, p<0.05). The incidence of hematuria and renal insufficiency in Class Vb was 64.8%and 15.4%, which was higher than Class Va (44.2% and 3.3%), but lower than Class IV (89.1% and 35%), p<0.05. The percentage of patients with positive anti-dsDNA antibody and hypocomplementemia in Class Vb tended to be higher than Class Va (35.2% vs 26.2%, 50.6% vs 31.2%). Repeated renal biopsies in 24 patients (11 Class Va, 13 Class Vb) showed that eight Class Vb patients had "transformed" to Class IV LN, while only two Class Va patients did (p<0.05). In three Class Va patients serum creatinine doubled during follow-up, but none of them progressed to end-stage renal disease (ESRD). In Class Vb serum creatinine doubled in ten patients, and three progressed to ESRD. CONCLUSIONS: The renal injury and extrarenal manifestations of Class Vb patients were severer than Class Va. Class Vb patients were more likely to shift to Class IV LN, and the prognosis was poorer than for Class Va.     

Clinical significance of renal biopsies showing mixed mesangial and global proliferative lupus nephritis

Renal biopsies of 70 children with systemic lupus erythematosus were categorized, according to the World Health Organization classification, as normal (five, 7%), mesangial (23, 33%), focal segmental proliferative (11, 16%), diffuse global proliferative (20, 29%) (ie, greater than or equal to 80% of glomeruli showing mesangioendothelial cell proliferation and/or deposition of immune complexes along the subendothelial margin of glomerular capillaries), and membranous (six, 8%) lupus nephritis (LN). In addition, five (7%) biopsies showed global proliferative LN in less than 80% of glomeruli and mesangial LN in the others. We assessed the renal status of these five patients at the time of renal biopsy and at outcome following prednisone treatment, with or without azathioprine. Four patients improved and later had either a normal urinalysis or only trace proteinuria. A low chronicity index was calculated on the biopsies of these patients. The fifth patient, whose condition did not improve, demonstrated a high chronicity index on renal biopsy. Overall, the renal status at outcome in the patients showing mixed mesangial and global proliferative LN more closely resembled that of patients with mesangial than diffuse global proliferative LN.     

Association between polymorphisms of the renin-angiotensin system and more severe histological forms of lupus nephritis

AIMS: The pathogenesis of lupus nephritis (LN) has not been fully understood. The renin-angiotensin system (RAS) is implicated in various immunological and non-immunological phenomena, and the polymorphism of the RAS genes has been associated with cardiovascular and renal disease onset and outcome. Therefore, we evaluated the possible association between the polymorphism of the renin-angiotensin system genes and the development of the different types of histological lesions of lupus nephritis in Brazilian patients. METHODS: 72 LN patients and 65 healthy subjects (sex-and ethnic-matched) were enrolled and compared in this study. Following the extraction of genomic DNA from the leukocytes of the peripheral blood, the genotypes of the angiotensin converting enzyme (ACE I/D), of the angiotensinogen (AGT M235T) and of the angiotensin II type 1 receptor (AGTR1 A1166C) were determined by the polymerase chain reaction. The renal lesions of the patients with LN were classified by the histological findings according to the WHO criteria. In addition, the activity and chronicity indices were used to assess the severity of renal involvement. RESULTS: Among the 72 patients with LN, there were 17 class II, 8 class III, 40 class IV and 7 class V, according to the WHO criteria. Individuals with the III and IV classes of LN (WHO) showed a significantly increased DD genotype frequency of ACE I/D genes when compared to the control group (48% vs. 27.7%, chi2 = 4.885, df = 1, p = 0.0442). No difference was found in the distribution of the AGT M235T and AGTR1 A1166C genotype frequencies among the LN of the different histological classes (WHO) and healthy controls. There was no association between genetic polymorphism of ACE, AGT M235T and AGTR1 A1166C and susceptibility to lupus nephritis, nor histological activity and chronicity indices in renal biopsy among the patients studied. CONCLUSIONS: This study suggests that the DD genotype of the ACE may be associated with the development of the more severe histological forms of lupus nephritis.