Over-expression of p53 protein as an indicator of the malignant transformation in spiradenoma

Malignant spiradenomas (spiradenocarcinomas) are exceedingly rare tumours of cutaneous adnexal origin, consisting of two components: benign—the pre-existent adenoma, and malignant—developing from the former part. We studied p53 protein expression in both compartments of three cases of malignant spiradenoma and compared these results with results obtained with eight cases of spiradenoma. Nuclear staining was consistently negative in all benign tumours, whilst in the cases of malignant transformation within spiradenoma p53 protein was present in the carcinomatous component, but the immunostaining remained negative in the benign counterpart of the tumour. In the zone of transition between both components of the spiradenocarcinomas p53 expression was positive in the cells with morphological atypia, providing clear discrimination. Thus, we conclude that the accumulation of p53 protein, which results from alterations in its turnover, accompanies the process of malignant transformation within long-standing spiradenomas.     

Presence of antibodies to replication protein A in some patients with systemic lupus erythematosus (SLE)

Presence of antibodies directed against replication protein A (RPA), a DNA binding protein complex composed of three subunits (RPA-70, RPA-32 and RPA-14) was investigated among patients with SLE and other autoimmune diseases using immunoblot analysis to RPA-70 and RPA-32 recombinant proteins. Anti-RPA antibodies were found in two out of 108 sera from SLE patients, one of them showing reactivity against RPA-32 and RPA-70 and the other reacting only against RPA-32. Sera from 108 patients with other autoimmune disorders as well as from 42 healthy control individuals were negative. Thus, the frequency of these antibodies in SLE is estimated to be 2–3%. The study demonstrates that RPA is one target more of the wide array of autoantigens that elicit an immune response in SLE. The presence of anti-RPA autoantibodies seems to be circumscribed to a small number of patients with SLE.     

Suppression of haematopoiesis by IgG autoantibodies from patients with systemic lupus erythematosus (SLE).

The inhibiting activity of serum on haematopoiesis has been described in patients with SLE. To explore further the features of serum inhibitor, we first examined the suppression of granulocytic and erythroid colony formation in vitro by serum from patients with SLE using methylcellulose culture. The potent inhibiting activity was demonstrated in six of 20 patients. All of these six patients were associated with leukocytopenia and/or anaemia. Five of 10 sera from patients with active SLE suppressed the colony formation of both burst-forming units of erythrocyte (BFU-E) and colony-forming units of granulocyte/macrophage (CFU-GM), and one serum suppressed BFU-E only. IgG fraction isolated from sera with inhibiting activity suppressed colony formation without complement involvement. The elimination of monocytes and lymphocytes from target mononuclear cells did not affect the suppression by the IgG fractions. The suppressive effect was completely eliminated after incubation of the IgG fractions with progenitor-enriched mononuclear cells. Flow cytometric analysis showed these IgG bound to CD34+ haematopoietic progenitor cells, but not to CD33+ cells. These data suggest that (i) the inhibitor of colony formation in serum was observed in IgG fraction; (ii) its suppressive effect on colony formation was mediated by neither monocytes and lymphocytes nor complements; and (iii) IgG fraction could bind to primitive haematopoietic progenitor cells and suppress the growth of these cells. Thus, IgG autoantibodies to primitive haematopoietic progenitor cells are demonstrated to be present in the sera of a significant proportion of active SLE patients with anaemia and leukocytopenia and to suppress the progenitor cell growth.     

Anti-C1q receptor/calreticulin autoantibodies in patients with systemic lupus erythematosus (SLE)

SLE is a disease characterized by the presence of multiple autoantibodies and high levels of circulating immune complexes. We studied the presence and functional relevance of autoantibodies directed against a receptor for the collagen-like stalks of the first subcomponent of complement, also known as calreticulin (cC1qR/CaR), in patients with SLE. In a cross-sectional study it was found that higher titres of antibodies against cC1qR/CaR are present in sera of SLE patients compared with normal donors. No association between anti-cC1qR/CaR titres and SLE disease activity was found. Following gel filtration of SLE serum it was found that anti-cC1qR/CaR reactivity is associated with the peak of monomeric IgG. Purified IgG from patients was able to specifically immunoprecipitate cC1qR/CaR. Since we have shown previously that cC1qR/CaR is able to inhibit the haemolytic activity of C1q, we determined a possible pathogenic role for anti-cC1qR/CaR on complement regulation. IgG derived from SLE serum reversed the inhibitory capacity of cC1qR/CaR in a dose-dependent fashion up to 63%, whereas IgG from normal donors had no significant effect. With respect to the capacity of anti-cC1qR/CaR antibodies to activate neutrophils, it was found that incubation of normal neutrophils with F(ab′)₂ anti-cC1qR/CaR resulted in a very limited oxidative burst. However, cross-linking of F(ab′)₂ anti-cC1qR/CaR on the neutrophils clearly induced neutrophil activation. Pre-incubation of the SLE-derived F(ab′)₂ with cC1qR/CaR prevented activation of neutrophils up to 81 ± 5%. These results suggest that the presence of anti-cC1qR/CaR antibodies in patients with SLE may modulate complement and neutrophil activation.     

Analysis of autoantibody specificities in selected systemic lupus erythematosus (SLE) sera

In order to design effective diagnostics for lupus, the heterogeneity in patient response must be understood. This heterogeneity in the anti-Sm and anti-U1-RNP response was examined via a frequency analysis of autoantibody fine specificities. Thus, 275 sera were studied by immunoprecipitation, immunoblotting, and immunodiffusion, and the frequency of occurrence of different autoantibodies to individual snRNP polypeptides and to other HeLa cell polypeptides was determined. The sera were found to contain autoantibodies reactive with denatured as well as native forms of HeLa-cell polypeptides. The common occurrence of several novel antibody fine specificities was noted, such as anti-p45 (different from anti-La/SS-B), anti-p105, and anti-p115. Another group of autoantibodies that is apparently not disease associated was observed in both lupus and normal sera.     

Autoimmunity to the p53 protein is a feature of systemic lupus erythematosus (SLE) related to anti-DNA antibodies

The induction of anti-DNA autoantibodies in systemic lupus erythematosus (SLE) patients is problematic because mammalian DNA is poorly immunogenic at best. Here we demonstrate a chain of connected antibodies in SLE patient sera that could account for the induction of anti-DNA antibody, and possibly for some of the pathogenic features of SLE. We now report that SLE patients, in addition to anti-DNA, produce antibodies to the carboxy-terminal domain of the tumour suppressor molecule p53; this p53 domain recognizes damaged DNA. Hence, these anti-p53 antibodies could mimic damaged DNA immunologically. Indeed, SLE sera do contain anti-idiotypic antibodies to a prototypic anti-p53 antibody. Moreover, SLE anti-DNA antibodies also recognize this type of anti-p53 antibody. Indeed, binding of affinity-purified anti-DNA both to DNA and to the anti-p53 antibody could be blocked by a p53 peptide derived from the DNA-binding domain. This mimicry of the p53 DNA-binding domain by the SLE anti-DNA antibodies is functional: activation of the p53 molecule could be inhibited by such anti-DNA antibodies. Thus, anti-DNA antibodies may arise in SLE patients by a chain of idiotypic autoimmunity centered around p53 autoimmunity. The SLE anti-DNA and anti-p53 antibodies can functionally block p53 activation, and so could affect apoptosis.     

Absence of p53 gene mutation and infrequent overexpression of p53 protein in hepatoblastoma

Ten cases of hepatoblastoma were studies for overexpression of p53 protein by immunohistochemistry and for possible p53 gene mutation by single strand conformation polymorphism (SSCP) analysis and direct DNA sequencing of the polymerase chain reaction products. Only one case of the macrotrabecular type at stage IV showed overexpression of p53 protein. No DNA mobility shift was found in any of the cases studies by SSCP analysis. DNA sequencing performed on the case showing overexpression of p53 protein revealed no mutation within exons 5 to 8. The associated adrenal cortical carcinoma of the same case also showed overexpression of p53 protein, but no mutation of the p53 gene. These results indicate that mutation of the p53 gene is infrequent in hepatoblastoma. This observation supports the view that mutation of the p53 gene is not as important in the oncogenesis of childhood neoplasms as in adult cancers.     

Presence of autoantibodies to peptidyl-prolyl cis-trans isomerase (cyclosporin A-binding protein) in systemic lupus erythematosus.

Several autoantibodies against cytoplasmic or nuclear components of cells have been reported in autoimmune diseases. We report here a previously unrecognized autoantibody to peptidyl-prolyl cis-trans isomerase (PPIase) in patients with systemic lupus erythematosus (SLE). PPIase, which catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides, has recently been found to be identical to cyclophilin, a specific binding protein of a potent immunosuppressant, cyclosporin A. IgG and IgM anti-PPIase antibodies were detected in 40 and 20% of unselected patients with SLE, respectively, by ELISA. The reactivity of these sera was confirmed by immunoblotting experiments. Sera from rheumatoid arthritis patients showed no reactivity and 1 of 8 sera from systemic sclerosis patients and 1 of 25 sera from normal controls showed only weak reactivity. Unexpectedly, the anti-PPIase antibody was unable to inhibit PPIase activity, indicating that the autoantibody recognizes an epitope of PPIase which is different from the active site of PPIase. The levels of the anti-PPIase antibody in SLE patients correlated with remissions and flares of the disease. The anti-PPIase antibody was higher in patients with active SLE than those with inactive disease. The prevalence of the active stage of the disease was significantly higher in IgG anti-PPIase antibody-positive SLE patients as compared to antibody-negative SLE patients. These data define the presence of a new autoantibody against PPIase and its association with the activity and certain clinical manifestations in SLE.     

Relationship between abnormal p53 protein and failure to express p21 protein in human breast carcinomas

Alterations in the p53 protein are a common feature in most malignancies, including breast carcinomas. p53 protein alterations contribute to malignant transformation in several ways, through genomic instability and accumulation of additional genetic alterations in other genes, through alteration of the p53-dependent apoptotic pathway, and through downregulation of downstream effector proteins such as p21 (WAF1/CIP1), necessary for cell-cycle growth arrest. Cell-cycle arrest is needed to allow DNA repair after injury. This study examines the relationship between abnormalities in p53 protein and expression of p21 protein in 70 cases selected from a series of 212 sporadic human breast carcinomas. Immunohistochemistry (IHC) was used for detection of p53 and p21 protein expression. Constant denaturant gel electrophoresis (CDGE) was used for detection of mutations in exons 5–8 of the TP53 gene. A highly significant association was found between abnormalities in p53, scored as protein accumulation and/or mutations, and lack of p21 expression. p21 was also shown to be downregulated in samples without p53 alterations, indicating that other mechanisms are also involved in turning off this gene. © 1997 John Wiley & Sons, Ltd.     

Autonomic nervous dysfunction in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA): possible pathogenic role of autoantibodies to autonomic nervous structures

Autonomic nervous dysfunction has been previously reported in SLE, RA and systemic sclerosis, but the pathogenesis of such a complication is poorly understood. In the present study, four standard cardiovascular autonomic function tests were performed in 34 female patients with connective tissue diseases and in 25 healthy control subjects, and results expressed as cardiovascular (CV) test scores. Moreover, in each subject the presence of circulating complement-fixing autoantibodies directed against sympathetic and parasympathetic nervous structures, represented by superior cervical ganglia and vagus nerve, respectively, was simultaneously assessed by an indirect immunofluorescent complement-fixation technique, using rabbit tissue as substrate. None of the patients reported autonomic symptoms. However, an abnormal CV test score (≥ 5) was detected in 15% of the patients and in none of the healthy control subjects, approaching statistical significance (P = 0.07). No correlation was found between CV test results and disease duration, type of therapy or presence of conventional autoantibodies. One or two autoantibodies to autonomic nervous structures were detected in six patients (18%) and not in the control subjects (P < 0.05). Values of deep breathing test were significantly lower in autoantibody-positive patients compared with those amongst the control subjects (P < 0.05), and an abnormal CV test score was significantly associated with the presence of autoantibodies to autonomic nervous structures (P < 0.05). In conclusion, we confirm that autonomic nervous function can be impaired in patients with connective tissue diseases, and suggest that autoantibodies directed against autonomic nervous system structures may play a role in the pathogenesis of the autonomic dysfunction.     

Amelioration of experimental systemic lupus erythematosus (SLE) by retrovirus infection

Experimental SLE can be induced in susceptible 129/J mice by immunization with a human anti-DNA antibody bearing a common idiotype designated 16/6 Id. Immunized mice develop autoantibodies, leukopenia, proteinuria, and immune complex deposits in renal glomeruli. Case reports have described clinical improvement in SLE in individuals becoming infected with HIV-1. Because 129/J mice are susceptible to experimental SLE and to infection with the BM5def murine leukemia virus (MuLV) mixture but do not develop the lymphoproliferative/immunodeficiency disorder known as murine AIDS (MAIDS), we superimposed this infection on immunization with the 16/6 Id. Multiple effects were observed. First, we noted an amelioration in the course of experimental SLE. Second, both in experimental SLE and in BM5def MuLV infection, immunoreactivity to HIV-1 gp120 was demonstrated, although gp120 is not present in the BM5def MuLV viruses. Third, production of autoantibodies characteristically found in SLE, e.g., anti-DNA, anti-RNP, and anti-SSA, was seen in BM5def MuLV-infected mice, demonstrating that an immune response as a consequence of infection had occurred despite the absence of MAIDS induction. We conclude that (1) retrovirus inoculation may ameliorate the course of experimental SLE; and (2) retrovirus inoculation, even in the absence of MAIDS induction, induces an immunologic response which promotes the production of potentially pathogenic autoantibodies.     

乳腺癌p53和p27蛋白的表达及意义

目的　探讨p5 3蛋白和p2 7蛋白在乳腺癌中的表达及意义。方法　应用免疫组化S -P法检测 6 7例乳腺癌中p5 3蛋白和p2 7蛋白的表达情况。结果　 6 7例乳腺癌中p5 3蛋白和p2 7蛋白的阳性表达率分别为 5 8 3%和 5 9 7% ,p5 3蛋白的表达与乳腺腋窝淋巴结转移有相关性 (P <0 0 1) ,p2 7蛋白的表达也与乳腺腋窝淋巴结转移有关 (P <0 0 5 ) ,乳腺癌p2 7和p5 3蛋白表达之间无明显相关。结论　p5 3蛋白的过表达和p2 7蛋白的失表达均与乳腺癌发生发展相关 ,二者可作为判断乳腺癌预后的有用指标     

Direct mapping of the human TATA box-binding protein (TBP) gene to 6q27 by fluorescencein situ hybridization

Summary TBP (TATA box-binding protein) participates in the expression of eukaryotic genes transcribed by RNA polymerases I, II, and III. Molecular cloning of human TBP revealed that the N-terminal region contains a polymorphic (CAG)_n repeat. We report here the direct localization of human TBP gene to chromosome 6q2705qter region by fluorescencein situ hybridization, using the cDNA clone with or without the (CAG)_n repeat as a probe.     

p53、p21~(WAF1)蛋白在非小细胞肺癌中的表达及其临床意义

目的　探讨原发性非小细胞肺癌中p5 3、p2 1WAF1蛋白表达与临床病理及预后的关系。方法　应用免疫组织化学 (SP法 )方法。共检测非小细胞肺癌 147例 ,其中腺癌 6 6例 ,鳞癌 6 3例 ,腺鳞癌 14例 ,大细胞癌 4例。结果　p5 3蛋白总阳性率为 6 1.2 % (90 / 147) ,腺癌为 5 7.6 % (38/ 6 6 ) ,鳞癌阳性率为 6 3.5 % (4 0 / 6 3) ,腺鳞癌为 71.4% (10 / 14) ,大细胞癌 2例阳性。p2 1WAF1蛋白总阳性率为40 1% (5 9/ 147) ,腺癌为 42 .4% (2 8/ 6 6 ) ,鳞癌为 41.3% (2 6 / 6 3) ,腺鳞癌 2 8.6 % (4 / 14) ,大细胞癌 1例阳性。肺腺癌p5 3蛋白阳性表达与其预后相关 ,6 6例腺癌中 ,生存率低于 3年组和高于 3年组的p5 3蛋白阳性率分别为 75 % (2 1/ 2 8)和 44 .7% (17/ 38) ,差异有显著性意义 (P <0 .0 2 5 )。p2 1WAF1阳性表达与肺癌预后有关 ,p2 1WAF1阳性表达者 3年生存率 (6 4.4% )高于阴性表达者 (4 6 .6 % ) (P <0 .0 5 )。p5 3阳性而p2 1WAF1阴性的非小细胞肺癌患者的预后比p5 3阴性而p2 1WAF1阳性者差 (P <0 .0 1)。结论　检测p5 3蛋白表达可作为判断肺腺癌预后的指标之一 ;检测p2 1WAF1蛋白表达有利于对非小细胞肺癌预后的判断 ;联合检测p5 3、p2 1WAF1蛋白对判断非小细胞肺癌的预后有重要的意义 ,似可作     

Increased binding of circulating systemic lupus erythematosus autoantibodies to recombinant interferon alpha 2b

Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease characterized by various types of immunological abnormalities including circulating and tissue‐fixed autoantibodies reactive with autoantigens. The mechanism that can explain the production of these antibodies is unclear. Here we address the binding specificity of SLE autoantibodies with recombinant alpha interferon 2b (hrIFN α‐2b), commercially available interferon (IFN α‐2b), and the gene (cIFN α‐2b) encoding this interferon. hrIFN α‐2b showed higher binding with naturally occurring SLE autoantibodies as compared to IFN α‐2b (p < 0.05) or cIFN α‐2b gene (p < 0.001) as assessed by direct binding, inhibition ELISA, and quantitative precipitin titration. The relative affinity of SLE autoantibodies for hrIFN α‐2b, IFN α‐2b, and cIFN α‐2b gene was in the order of 1.13 × 10^?7, 1.38 × 10^?6, and 1.22 × 10^?6, respectively. hrIFN α‐2b is shown to have unique epitopes that would explain the possible antigenic role of hrIFN α‐2b in the generation of SLE autoantibodies. Anti‐hrIFN α‐2b antibodies have been shown to represent an alternative immunological probe for the estimation of interferon alpha 2b in the serum of SLE patients.     

错配修复蛋白和p53蛋白表达与结直肠癌的临床病理关系及其相关性

目的探讨错配修复蛋白(mismatch repair protein,MMRP)及p53蛋白在结直肠癌(colorectal cancer,CRC)中的表达,进而分析微卫星不稳定(microsatellite instability,MSI)、p53与CRC临床病理特征的关系及其相关性。方法采用组织芯片及免疫组化法对980例CRC中4种MMRP及p53进行检测,将4种MMMP中的1种及以上表达缺失定为MSI组,全部阳性表达定为微卫星稳定(microsatellite stable,MSS)组。结果 (1)MMRP表达缺失率为11%,MLH1、PMS2、MSH2及MSH6的表达缺失率分别为7.3%、7.1%、2.0%及1.9%;其中共同缺失表达类型为MLH1-PMS2、MSH2-MSH6者分别为52例、14例,统计分析结果显示二者均呈正相关(rs=0.712),4种蛋白均缺失者3例。(2)p53阳性率为59.1%。(3)MSI与患者年龄、肿瘤部位、大小、组织学类型、淋巴结转移、临床分期及Ki-67有关(P0.05)。(4)p53与组织学类型、大体分型、浸润深度、远处转移及Ki-67有关(P0.05)。(5)MSI与p53呈负相关(rs=-0.118)。结论 MLH1、PMS2缺失表达较MSH2和MSH6多见。MLH1与PMS2、MSH2与MSH6常常协同表达或缺失。MSI及p53与CRC临床病理特征关系密切,对预测CRC的风险、恶性程度的评估等具有指导意义。MSI与p53表达呈负相关,提示二者可能参与CRC的不同发生、发展过程。