Discovering novel lung cancer associated antigens and the utilization of their autoantibodies in detection of lung cancer

ObjectiveThe identification of tumor-associated antigens (TAAs) and their corresponding autoantibodies in lung cancer (LC) may expand our vision of cancer immunity. This study aims to screen novel TAAs to distinguish LC from the healthy population.MethodsIn our previous study, 35 genes encoding LC-associated TAAs were identified from the serological analysis of recombinant cDNA expression libraries (SEREX), and Oncomine database was further used to identify potential genes in cancer progression. Autoantibody to TAAs were tested by enzyme-linked immunosorbent assay (ELISA) in sera from 1379 participants in validation set and verification set.FindingsBased on analysis of three independent microarrays in Oncomine, ten genes were consistently dysregulated in LC. The sera level and positive frequency of the anti-TOP2A, anti-ACTR3, anti-RPS6KA5 and anti-PSIP1 from LC patients were higher than normal control in validation set. The area under curve (AUC) of anti-TOP2A, anti-ACTR3, anti-RPS6KA5 and anti-PSIP1 was respectively 0.758, 0.787, 0.707, 0.668. The sensitivity of these four autoantibodies for LC detection ranged from 26.63 % to 32.07 % with the specificity over 90 %. Data from the verification set confirmed the results. Except that, the frequency of serum autoantibody against TOP2A (43.3 %) and ACTR3 (50.0 %) was significantly higher in early stage LC than late stage (23.6 % and 22.3 %, respectively).ConclusionTOP2A, ACTR3, RPS6KA5 and PSIP1 can elicit humoral immune response in LC and their autoantibodies have relationship with the tumorigenesis of LC. Anti-TOP2A and anti-ACTR3 have the potential to serve as a serological biomarkers in early stage LC.     

Naturally occuring antibodies against nerve growth factor in human and rabbit sera: comparison between control and herpes simplex virus-infected patients

Antibodies against nerve growth factor (NGF) in sera were detected by enzyme-linked immunosorbent assays (ELISA), by their isolation after passage of sera through NGF immunoadsorbent columns and by their specificity to bind and immunoprecipitate mouse NGF as well as to stain by immunohistochemical methods cellular sites of NGF synthesis. Increased levels of anti-NGF antibodies were found in sera of herpes simplex virus (HSV)-infected patients but not in HSV-inoculated rabbits. As HSV latency is known to be promoted by NGF in vitro, these results may suggest that anti-NGF antibodies modulate the cytokine function of NGF and thus might play a role in HSV infection. The biological function of circulating antibodies against NGF, in general, is now open to future investigation.     

慢性乙型肝炎及乙肝后肝硬化患者血清中自身抗体结果分析

目的 :探讨慢性乙型肝炎及乙肝后肝硬化患者血清中自身抗体的变化情况。方法 :收集了慢性乙型肝炎患者 2 4 5例 ,乙肝后肝硬化 86例血清标本测定抗核抗体 (ANA)和可提取性核抗原 (ENA)抗体。结果 :慢性乙型肝炎和乙肝后肝硬化ANA的检出率分别为 13.1% (31/ 2 4 5 )、2 3.2 % (2 0 / 86 ) ,均明显高于正常对照组的 2 .5 % (2 / 80 ) (P <0 .0 5 )。两组病例间也有显著性差异 (P <0 .0 5 )。抗ENA抗体阳性率分别为 7.34%和 12 .7%。阳性主要有nRNP/Sm、SS -A、Scl- 70、Jo - 1、dsDNA等多种类型。结论 :慢性乙型肝炎及乙肝后肝硬化患者血清自身抗体显著高于正常人。除一部分是目前已鉴定的与常见自身免疫性疾病相关的自身抗体外 ,还有许多有待进一步鉴定。     

抗人甲状腺过氧化物酶抗体与其他自身抗体的关系

甲状腺过氧化物酶（ＴＰＯ）是甲状腺微粒体的抗原成分。在纯化抗原建立测定抗ＴＰＯ抗体ＥＬＩＳＡ法的基础上，并对抗ＴＰＯ与其他自身抗体的伴随关系作了研究。发现除自身免疫性甲状腺病有较高阳性率外，ＳＬＥ等患者抗ＴＰＯ检出率也很高（１０％￣５２．１％），且抗ＴＰＯ阳性者，抗核抗体等其他自身抗体的阳性率也高达３０．８％￣９５．２％。抗ＴＰＯ抗体与其他自身抗体的相关系数ｒ＝０．７４６７（Ｐ〈０．０１）。     

The consensus workshops for the detection of autoantibodies to intracellular antigens in rheumatic diseases

In 1988 and in 1989 consensus workshops were organized in order to define the interlaboratory concordance in detecting autoantibody specificities in selected sera from patients with rheumatoid disorders and to determine the possible causes of discrepancies. In total 20 sera were tested for the presence of antibodies against nRNP, Sm, Ro (SS-A), La (SS-B), Scl-70, centromeric antigens, ribosomal RNP and Jo-1. The methods used for detection by the 28 European laboratories who participated included immunofluorescence, counter-immunoelectrophoresis, immunodiffusion, immunoblotting and ELISA.

The results showed that only a combination of two or more techniques was able to detect all specificities with an adequate efficiency. Recommendations to improve the efficiency of autoantibody detection and to standardize laboratory protocols are given.     

Association analyses of DNA methyltransferase-1 (<Emphasis Type="Italic">DNMT1</Emphasis>) polymorphisms with systemic lupus erythematosus

The etiology of systemic lupus erythematosus (SLE) is very complex, and genetic factors appear to play a significant role in susceptibility to SLE, in determining the disease expression, and in the autoantibody profiles of individuals with SLE. DNA methyltransferase-1 (DNMT1) is a major enzyme that determines genomic methylation patterns and both maintains methyltransferase and exhibits de novo DNA methylation activity in vivo. In order to clarify the association of DNMT1 polymorphisms with SLE, we scrutinized the genetic polymorphisms in exons and their boundaries of DNMT1, including the –1,500 bp promoter region, by direct sequencing in 24 Korean individuals. Twenty-nine sequence variants were identified: two in 5UTR, six in exons, and 21 in introns. Eight of these polymorphisms were selected for a larger-scale genotyping (n=680) by considering their allele frequencies, haplotype-tagging status, and linkage disequilibrium coefficiencies (LDs) among polymorphisms. The associations between DNMT1 polymorphisms and the clinical profiles of SLE were analyzed. No significant associations with the risk of SLE were detected. However, further analyses of association with autoantibody production among SLE patients revealed that one nonsynonymous SNP, +14463G>C (V120L) in exon 4, was weakly associated with an increased risk of anti-La antibody production (P=0.04), although the significance could not be retained after correction of multiple tests. The DNMT1 variations and haplotypes clarified in this study would provide valuable information for future genetic studies of other autoimmune diseases.     

特发性右室心律失常患者血清CMV、CVB病毒抗体分析及其临床意义探讨

目的：通过检测病毒血清抗体,探讨相关病毒感染与特发性右室心律失常（IRVA）发生的相关性.方法：病例对照研究分为3组：IRVA组、其他心脏病平行对照组（Heart-Disease-Control）和健康对照组（Healthy-Control）,每组30例,性别年龄匹配.接受常规检查后进行血清学检查,随访6～12个月.结果：IRVA组与其他2组的X线心胸比值、超声心脏测值比较,无显著性差异（P＞0.05）.3组的柯萨奇B组病毒（CVB）血清IgM阳性率组间差异无显著性;而IRVA组的巨细胞病毒（CMV）血清IgM阳性率（73.3%）显著高于其他2组（P＜0.01）,随访6～12个月后,该组CMV IgM阳性率仍然持续增高（66.7%）.相关性分析发现,CVB感染与IRVA发生的联系强度低（P＞0.05）,而CMV感染与IRVA发生的联系强度高（P＜0.001）.4种常见的病毒血清抗心肌自身抗体检测中发现IRVA组抗β1受体抗体阳性率（60.0%）显著高于其他2组（P＜0.01）.结论：IRVA患者血清CMV IgM阳性率高,该抗体的出现与IRVA的发生高度相关;CMV感染引起IRVA的发生可能与免疫机制（抗β1受体抗体介导）有关.     

丙型肝炎病毒感染者血清中自身抗体结果分析

目的 :观察丙型肝炎病毒 (HCV)感染者血清中自身抗体的变化情况。方法 :对85例HCV感染者血清标本测定抗核抗体(ANA)和可提取性核抗原 (ENA)抗体。结果 :HCV感染者ANA的检出率为24 7 % (21/85) ,明显高于正常对照组的2 5% (2/80)(x2=16.8,P<0 01)。抗ENA抗体阳性率为15 3% (13/85)与对照组比较差异有显著性 (x2=8.16,P<0 01)12 7 %。阳性主要有nRNP/Sm、SS-A、Scl-70、Jo-1、dsDNA等多种类型。结论 :自身免疫可能是HCV感染后肝组织损伤的重要因素     

Low prevalence of autoantibodies to CENP-H, -I, -K, -L, -M, -N, -T and -U in a Japanese cohort of anti-centromere positive samples

Objective: The constituents of the centromere region, centromere protein (CENP)-A, -B, and -C, are mainly targeted by anticentromere antibodies (ACA). Many other proteins also assemble around CENP-A nucleosomes in interphase nuclei to form the interphase centromere complex (ICEN). CENP-H, -I, -K, -L, -M, -N, -T, and -U have been reported as the constitutive components of ICEN. In this study, we examined the reactivities of ACA to the 8 CENPs for the purpose of investigating their autoantigenicity.

Methods: Sera from 95 patients with ACA were tested by western blotting (WB) and enzyme-linked immunosorbent assay (ELISA) with the recombinant C-terminal of CENP-B (Ct-CENP-B). Next, the sera were examined for autoantibodies against the 8 CENPs by WB with each recombinant protein. Furthermore, the coiled-coil motifs and granzyme B (GB) cleavage for various CENPs were analyzed with computer tools.

Results: Out of 95 ACA-positive sera, 85 and 93 sera were positive for anti-Ct-CENP-B antibodies in WB and in ELISA, respectively. In WB using the 8 CENPs, no sera reacted to any other 7 CENPs, except 1 serum, which reacted weakly to CENP-T. We were unable to find any obvious relationships between the autoantigenicity of CENPs and coiled-coil-forming probabilities or potential substrates for GB.

Conclusion: This study demonstrates that ACA rarely target the 8 CENPs, in contrast to CENP-B. Protein structures might not contribute in a major way to the autoantigenicity of CENPs.