31例轻链型多发性骨髓瘤实验室指标分析

目的探讨实验室检测指标在轻链型多发性骨髓瘤(MM)诊治中的应用价值。方法收集轻链型MM患者31例及对照组60例,对照组为健康者20例(健康组),以及该院诊断的IgGκ型和IgAκ型MM患者各20例。检测免疫球蛋白、血清蛋白电泳、免疫固定电泳、尿素氮(UREA)、肌酐(Cr)、β_2微球蛋白,并进行流式细胞免疫表型检测及骨髓细胞学检查。结果血清蛋白电泳检测到M蛋白的阳性率为29.03%,Durie-Salmon临床分期Ⅱ期与Ⅲ期M蛋白水平差异有统计学意义(P0.05);免疫固定电泳结果均需用琼脂糖凝胶电泳法加做IgD及IgE抗体复查;轻链型MM患者与健康者的IgG、IgA、IgM、UREA、Cr、β_2微球蛋白比较,差异有统计学意义(P0.05);MM细胞形态特征改变显著,细胞抗原表达以CD38、CD138、CD56为主。结论免疫球蛋白定量及免疫固定电泳可作为M蛋白的筛查方法,血清蛋白电泳可用于患者的临床分期和疗效观察;骨髓细胞学检查对轻链型MM起到重要的诊断作用,轻链型MM比其他类型MM有更加严重的肾功能损伤。     

Verapamil and furosemide prevent cholecystokinin-induced translocation of immunoglobulins in rat intestine

Our previous studies have shown that cholecystokinin and pilocarpine are known to be extracellular messengers promoting the release of immunoglobulins A and G antibody activity in the lumen of the rat intestine. In the present study, which was also performed in rats, we show that CCK also promotes the translocation of albumin, electrolytes, and water into the lumen of the intestine. The effect of CCK on the translocation of immunoglobulins, albumin, and electrolytes is reduced by the prior injection of the calcium-channel blocker verapamil and the chloride-channel blocker furosemide. Taken together, the above observations suggest that the translocation of immunoglobulins, albumin, and electrolytes in the intestine appears to be stimulated by identical mechanisms and to proceed simultaneously.This work was supported in part by GIF grant 1-79-063.2/88 in part by the Miriam Coven-Fish Fund and in part by the joint Research Fund of the Hebrew University and Hadassah.     

One‐year vaccination against hepatitis B virus with a MPL‐vaccine in liver transplant patients for HBV‐related cirrhosis

Conflicting results have been reported on vaccination against hepatitis B virus (HBV) as a prophylaxis against viral recurrence after liver transplantation. We investigated the efficacy of 1‐year, monthly vaccination using an adjuvant 3‐deacylated monophosphoryl‐lipid‐A (MPL) recombinant S vaccine initially administered together with hepatitis B immunoglobulins (HBIg) in 18 patients transplanted for HBV‐related cirrhosis. All received 12 vaccine doses (HBsAg, 20 mcg plus MPL, 50 mcg): the initial six doses (phase I) were administered within 7 days after intravenous HBIg (2000 IU), while the last 6 (phase II) following HBIg withdrawal. All patients received lamivudine during the study. Anti‐HBs titers were determined before each dose and then for 1 year after vaccination. After phase I anti‐HBs titers were greater than 100 IU/l in all patients and in three (16.6%) were greater than 500 IU/l. After phase II 10 patients (55.5%) achieved anti‐HBs titers greater than 100 IU/l and five (27.7%) greater than 500 IU/l. One year after vaccination eight patients (44.4%) maintained anti‐HBs titers greater than 100 IU/l, with a median titer of 234 IU/l (102–1205), and 2 (11.1%) greater than 500 IU/l. One‐year extended monthly vaccination with a MPL‐adjuvant recombinant vaccine induces a sustained protective anti‐HBs response in approximately half of transplant recipients.     

益元活血汤对恶性肿瘤患者免疫功能的影响

目的:观察益元活血汤对恶性肿瘤患者的治疗作用和化疗期间对免疫功能的影响。方法:采用完全随机设计方法,将恶性肿瘤患者分为益元活血汤配合化疗组(治疗组)29例和单纯化疗组(对照组)27例,在化疗前及化疗4周期后,以外周血分别测定患者CD3,CD4,CD8,淋巴细胞非组蛋白(Ag-Nors),免疫球蛋白,同时观察本时期内患者的中医证候表现。对比两组之间的差异及治疗前后的差异。结果:恶性肿瘤患者化疗期间的免疫紊乱主要表现为细胞免疫下降,其中CD3,CD4,CD4/CD8,Ag-Nors水平均有所减低,免疫球蛋白变化不明显。通过益元活血汤治疗,能够延缓化疗导致的细胞免疫功能下降,但对体液免疫无明显影响。结论:益元活血汤对与化疗导致的细胞免疫紊乱有调节作用。     

体外循环围术期检测超敏C反应蛋白、免疫球蛋白及补体的临床意义

目的探讨风湿性心脏病(RHD)患者在体外循环(CPB)围手术期监测外周血超敏C反应蛋白(hs-CRP)、免疫球蛋白以及补体的临床价值。方法选取28例施行CPB手术的RHD患者分别检测其术前、术毕以及术后第1天、3天和7天外周血hs-CRP、C3、C4、IgG、IgM及IgA的浓度。结果 RHD患者术前hs-CRP、免疫球蛋白和补体的检测结果均在正常范围内。hs-CRP在术后第1天显著升高,第3天仍处于较高水平,术后第7天恢复术前水平;术毕时C3、C4均降低,术后第1天低至谷值,第3天升高至术前水平;IgG、IgM和IgA于术毕至术后第1天均有不同程度的降低,IgM下降速度最快,下降幅度以IgG最为明显,手术第7天后恢复至术前。结论联合监测hs-CRP、免疫球蛋白及补体指标能反映RHD患者在CPB围术期机体免疫功能病理性损害的程度,对疾病的监护和判定预后转归具有重要的意义。     

2种不同试剂检测血清免疫球蛋白结果的可比性及偏倚评估

目的进行2种不同免疫球蛋白诊断试剂对血清免疫球蛋白测定结果的可比性及偏倚评估研究,为血清免疫球蛋白测定的标准化和临床实验室认可提供实验数据。方法参考美国临床和实验室标准协会(CLSI)的EP9A2文件,分别用执诚试剂和优利特试剂在HITACHI7600-010全自动生化分析仪上测定免疫球蛋白IgG、IgA和IgM,以执诚试剂为对照组,优利特试剂为实验组。用线性回归统计法分析对照组(Y)和实验组(X)测定结果决定水平处的方法间误差,以美国临床实验室修正法规(CLIA′88)规定的室间质量评价允许误差范围的1/2为标准,判断不同试剂的临床可接受性。结果两种试剂对患者血清免疫球蛋白测定结果显示,方法内重复性良好,无离群点,除IgA在决定水平处的系统误差不能被接受外,其余项目测定结果的偏差临床可以接受。结论当同一实验室同一检验项目存在2个或以上的试剂时,应进行方法比对和偏差评估,判断其临床可接受性,以保证检验结果的可比性。     

Supportive care in multiple myeloma

In myeloma bone disease, bisphosphonates have been shown to delay the progression of osteolytic lesions and prevent fractures. In the case of painful vertebral fractures, vertebroplasty and kyphoplasty have become standard procedures to control symptoms and restore the original height of the vertebrae. Adequate pain control is of crucial importance for the quality of life of myeloma patients, as is maintaining adequate hemoglobin levels with the use of erythropoietic growth factors. Infections should be treated aggressively in myeloma patients, as these contribute significantly to morbidity and mortality. In patients with repeated infectious complications, prophylactic measures such as long-term application of antibiotic or antiviral medication or use of intravenous immunoglobulins is recommended. The concerted action of these supportive therapies can significantly improve the wellbeing of myeloma patients in phases of disease progression as well as during phases of remission. In progressive disease, certain measures such as adequate pain control and radiotherapy can ameliorate symptoms until the therapeutic effect of systemic anti-neoplastic therapy becomes evident.     

Severe steroid-resistant post-infectious encephalomyelitis

Based on their presumed immuno-mediated etiology, post-infectious CNS disorders are commonly treated with high-dose steroids. Factors influencing treatment effectiveness, possible alternative options for steroid-resistant cases, and their outcome profiles, remain unclear. We here describe the clinical features, the prognosis and the efficacy of i. v. immunoglobulins (IVIg) in a series of severe ADEM refractory to steroids. We performed an inception cohort study on inpatients of the Neurologic and Infectious Disease Clinics, consecutively admitted over eight years, with a minimum two-year follow-up. Nineteen patients affected by classic and site-restricted ADEM were treated with IVIg after steroid failure. Five other patients received IVIg as first-line treatment due to steroids contraindications: although not included in the analysis, they were monitored for anecdotal comparison. Steroids were administered as IV 6-methylprednisolone (6-MP) 500/1000 mg daily until a maximum dose of 6-8 g; IVIg were administered at 0.4 g/kg/day for 5 days. The outcome was assessed by the Scripps Neurological Rating Scale (SNRS) score with determined periodicity. We observed that steroid-resistant patients showed high prevalence of PNS damage (89%) and myelitis (95 %). Other features were old age, severe disability at onset, and moderate to severe blood-brain-barrier (BBB) damage on CSF. In 10/19 patients (53 %) IVIg were effective, the clinical improvement beginning within the end of the five-day cycle,without relapses. Prominent effects of IVIg were detectable on motor dysfunction. Milder onset disability (p = 0.013) and lower CSF albumin (p = 0.006) were the predictors of IVIg response. Among steroid-free patients, 3/5 were responsive to IVIg. We conclude that IVIg can be useful in a portion of patients with severe steroid-resistant ADEM and prominent motor dysfunction. Unsolved issues regard the usefulness of IVIg in less selected groups, and the spectrum of their clinical effects.     

Human combinatorial libraries yield rare antibodies that broadly neutralize hepatitis C virus

One way to dissect the antibody response to an invading microorganism is to clone the antibody repertoire from immune donors and subsequently characterize the specific antibodies. Recently, methodological advances have allowed investigations of neutralizing antibodies against hepatitis C virus (HCV) in vitro. We have investigated three human mAbs, previously isolated from an individual infected with HCV of genotype 2b, that are known to cross-react in a binding assay to the envelope E2 protein of genotypes 1a and 1b. We now report that two of them have a neutralizing activity with a breadth not previously observed. Indeed, mAbs 1:7 and A8 recognized E2 from all of the six major genotypes, and they neutralized retroviral pseudoparticles [HCV pseudoparticles (HCVpp)] carrying genetically equally diverse HCV envelope glycoproteins. Importantly, these antibodies were also able to neutralize the cell culture infectious HCV clone JFH-1 in vitro, with IC(50) values of 60 ng/ml and 560 ng/ml, respectively. The conformational epitopes of these two broadly reactive antibodies were overlapping yet distinct and involved amino acid residues in the 523-535 region of E2, known to be important for the E2-CD81 interaction. The third antibody clone, representing a dominant population in the initial screen for these antibodies, was less broadly reactive and was unable to neutralize the genotype 2a infectious clone JFH-1. Our results confirm at the clonal level that broadly neutralizing human anti-HCV antibodies can be elicited and that the region amino acids 523-535 of the HCV envelope glycoprotein E2 carries neutralizing epitopes conserved across all genotypes.     

Molecular and Immuno-Characteristics of Immunoglobulin-like Glycoproteins in Cancer Cell-expressed Biomarker,CA215

RP215 monoclonal antibody (Mab) was shown to recognize a specific carbohydrate-associated epitope found in cancer cell-expressed glycoproteins, known as CA215. The membrane-bound and soluble forms of CA215 were detected in almost all of the cancer cells in humans, but rarely found in normal tissues. Through MALDI-TOF MS analysis, it has been reported previously that as much as 40% of the detected tryptic peptides of CA215 showed high degrees of sequence homology to those found in immunoglobulin heavy chains. The cancer cell-derived immunoglobulins were further purified from CA215 by affinity column-linked with goat anti-human IgG for molecular characterizations. Semi-quantitative RT-PCR was used to determine the mRNA levels of various immunoglobulin genes expressed by cancer cells of single or multi-cell origins and compared with those found in normal human serum. The stability of CA215 was investigated under different experimental conditions. It was observed that the RP215-specific epitope in CA215 is stable at neutral pH, in human serum or in mice (half life of 5–18 days), but unstable at extreme pH’s (pH ≤ 2.0; pH ≥ 12.0) or high temperatures. Enzyme immunoassays were performed with several secondary antibody probes related to human IgG. It was demonstrated that cancer cell-expressed immunoglobulins with RP215-specific epitope have much lower immunoactivity than that of normal human IgG (≤ 5%), despite the fact that both showed almost identical amino acid sequence in the respective Fc region reported previously. This could be the result of aberrant glycosylation of CA215 in cancer cells. Aberrant glycosylation of glycoproteins may have important biological implications on the proliferation of cancer cells in vitro or in vivo.