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At least 19 recordsLinked to original sources
PubMed1995

The relationship between proliferating cell nuclear antigen (PCNA), nuclear DNA content and mutant p53 during genesis of cervical carcinoma.

Proliferating cell nuclear antigen (PCNA), nuclear DNA content and mutant p53 overexpression were studied by means of image cytometry and immunohistochemistry respectively in normal mucosa (n = 10), in mild (n = 16), moderate (n = 9) and severe (n = 17) atypical lesions, as well as in squamous cell carcinomas (n = 36) of the cervix uteri. The results show that increasing histopathological atypia in the cervical mucosa was correlated to an initial increase of PCNA followed by distinct aneuploidy and p53 overexpression. The data are suggested to contribute to a better understanding of the genesis of cervical carcinoma, and to indicate that the coexistence of both distinct aneuploidy and p53 immunoreactivity can be used to decide if a cell population is neoplastic, whereas the absence of p53 overexpression does not necessarily exclude neoplasia. This diagnostic procedure can be suggested to improve early detection of intraepithelial squamous neoplasia.

Aneuploidy↗
PubMed1994

End-stage cardiac failure in humans is coupled with the induction of proliferating cell nuclear antigen and nuclear mitotic division in ventricular myocytes.

Proliferating cell nuclear antigen (PCNA) is a late growth-regulated gene that is expressed at the G1-S boundary of the cell cycle and is required for DNA synthesis and cell proliferation. Since quantitative results suggest that myocyte hyperplasia occurs in the decompensated human heart, we postulated that induction of the PCNA gene may be present in the failing heart in humans. PCNA protein was detected in myocardial samples obtained from the left and right ventricles of patients with congestive heart failure. Endomyocardial biopsies collected from donor subjects were used as control tissue. The percentage of positively stained myocyte nuclei in the ventricles was established by using PCNA monoclonal antibody and the immunoperoxidase technique. The localization of PCNA in myocytes was confirmed by alpha-sarcomeric actin antibody staining. PCNA labeling was present in left ventricular myocytes of 29 of the 32 hearts examined. In the right ventricle, 24 of the 29 samples showed positive staining. In a subset of 25 patients, the percentage of PCNA-labeled myocyte nuclei was measured and found to constitute 49 +/- 22% of left ventricular myocytes. A similar analysis for the right ventricle, conducted in 21 patients, showed that 49 +/- 19% of the myocyte nuclei exhibited PCNA protein. In addition, mitotic figures in myocytes were documented. A quantitative analysis of this cellular process revealed that 11 myocyte nuclei per 1 million cells exhibited mitotic images in chronic heart failure. Immediately after myocardial infarction, two cells per million showed mitotic division, and this phenomenon was restricted to the region adjacent to the necrotic tissue. No PCNA labeling or nuclear mitotic images were detected in the ventricular myocardium of control subjects. Thus, the observation that diffuse PCNA labeling and myocyte mitotic division are present in hearts with end-stage failure strongly suggests that adult ventricular myocytes are not terminally differentiated cells and that myocyte cellular hyperplasia may constitute a growth reserve mechanism of the diseased heart.

Adult↗
PubMed1983

Antibodies to the Epstein-Barr virus nuclear antigen and to rheumatoid arthritis nuclear antigen identify the same polypeptide.

Patients with seropositive rheumatoid arthritis (RA) have elevated titers of precipitating antibody toward an antigen designated RA nuclear antigen (RANA). Anti-RANA reactivity has been associated with prior Epstein-Barr virus (EBV) infection. Using the protein blot technique, we have identified, in extracts of WI-L2, an EBV+ nonproducer B-lymphoblast line, a Mr 80,000 polypeptide that is reactive with anti-RANA-containing sera. This same polypeptide can be recovered from RANA precipitin bands. The Mr 80,000 polypeptide appears to be EBV-associated, as a homologous antigen was detected in two other EBV+ cell lines, Daudi and Raji, but was not present in three EBV- human cell lines tested, HeLa, BJAB, and Ramos. Anti-RANA antibodies and antibodies reactive with the Mr 80,000 polypeptide also appear coincidently in the sera of individuals exhibiting an EBV infection (infectious mononucleosis). Further analysis of the RANA-associated Mr 80,000 polypeptide suggested its identity with the previously recognized EBV-associated nuclear antigen designated EBNA. The Mr 80,000 antigen shares with EBNA the properties of being a heat-stable, DNA binding protein. EBNA is traditionally assayed by a complement fixation reaction and anti-Mr 80,000 antibodies were shown to be reactive when a complement fixation assay was used in the immunoblot. Finally, when the Mr 80,000 antigen was used to absorb an anti-RANA/anti-EBNA serum, both antibodies were reduced.

Anemia, Hemolytic, Congenital↗
PubMed1989

[Aberrant expression of major histocompatibility complex class II. (HLA-DR/DQ) antigens and proliferative nuclear antigen. (Ki-67) in renal tubular epithelial cells].

In an attempt to clarify the participations of cellular immunity in the development of tubulo-interstitial lesions, aberrant expressions of major histocompatibility complex (MHC) class II antigens and Ki-67 nuclear antigen on the renal tubular epithelial cells were studied. Ki-67 antigen was known to appear in all phases of cell cycle except for Go. Nine normal kidney specimens (4 males and 5 females) and 117 kidney specimens obtained from patients with kidney diseases (54 males and 63 females) were examined with the indirect immunofluorescence technique using murine monoclonal antibodies against HLA-DR (lal), HLA-DQ (Leu10) and Ki-67 nuclear antigen. Patients included 100 with glomerular diseases, and 16 with tubulo-interstitial diseases consisting of 4 acute tubular necrosis (ATN), 7 acute tubulo-interstitial nephritis (AIN), one renal allograft rejection and 4 sarcoidosis. In normal kidney, HLA-DR was solely noted in only two specimens (22.2%) at the basal portion of proximal tubular epithelial cells. In tubulo-interstitial diseases 11 (68.8%) out of 16 patients showed diffuse and intense expressions of HLA-DR concomitant with HLA-DQ in 6 of 13 (42.9%), and 11 of 13 (84.6%) were positive for Ki-67 nuclear antigen. Especially, in AIN and allograft rejection, intense expression of HLA-DR, DQ and Ki-67 nuclear antigen were observed in 100%, 86%, 100%, respectively. In ATN 3(75%) were positive for HLA-DR and Ki-67, but not for HLA-DQ. In contrast, only 12(15.6%). 2(2.6%) and 2(4.8%) of primary glomerular disease were weakly positive for HLA-DR, DQ and Ki-67 nuclear antigen, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1988

A monoclonal antibody to SV40 large T-antigen labels a nuclear antigen in JC virus-transformed cells and in progressive multifocal leukoencephalopathy (PML) brain infected with JC virus.

Thirty monoclonal antibodies to SV40 large T-antigen were tested for reactivity on the JC virus-transformed hamster glial cell line known as HJC-15. Two of them (PAb 416 and PAb 108) detected a nuclear antigen in both SV40-transformed CCL 75.1 cells and in HJC-15 cells, but not in control cells lacking T-antigen. These same antibodies also labeled a nuclear antigen in hamster tumor tissue derived from HJC-15 cells. In addition, the monoclonal antibody PAb 416 detected a nuclear antigen in progressive multifocal leukoencephalopathy (PML) tissue infected with JC virus, but not in normal brain tissue or tissue from other neurological diseases. Staining by PAb 416 was reduced by prior incubation with hamster anti-JCV tumor serum, suggesting that the polyclonal antiserum to JCV T-antigen may compete for an epitope at or near the PAb 416 binding site.

Animals↗
PubMed1977

Transient induction of a nuclear antigen unrelated to Epstein-Barr nuclear antigen in cells of two human B-lymphoma lines converted by Epstein-Barr virus.

Infection of cells of the Epstein-Barr virus (EBV)-negative human B-lymphoma lines BJAB and Ramos with EBV preparations from P3HR-1 or B 95-8 cells converted these cells to EBV genome carriers expressing Epstein-Barr nuclear antigen (EBNA) in almost 100% of these cells. Induction of these cells as well as of clones from P3HR-1 EBV-converted BJAB cells with iododeoxyuridine, aminopterin, and hypoxanthine resulted in the appearance of a nuclear antigen in about 1-6% of the cells 1-4 days after induction. The antigen is different from known EBV-induced antigens like EBNA, viral capsid antigen (VCA) or the D- and R-subspecificities of the early antigen (EA) complex. It is demonstrated by indirect immunofluorescence and inactivated after acetone fixation. The antigen was not detectable after induction of uninfected BJAB and Ramos cells nor has it been found in noninduced or induced P3HR-1 and Raji cells. Thus, it appears that EBV-infection mediates the expression of this antigen, for which the name TINA (transiently induced nuclear antigen) is suggested. Sera reacting against TINA generally contained high antibody titers against EBV-induced EA. Only a limited number of highly EA-reactive sera, however, were also positive for TINA. Among 200 sera tested thus far, TINA reactivity was most frequently observed in sera of patients with nasopharyngeal carcinoma (7 out of 28), in sera of the only two patients with immunoblastoma tested and occasionally in sera from patients with Hodgkin's disease and chronic lymphatic leukemia. Among 70 sera from nontumor patients, TINA reactivity was observed three times: two patients suffered from "chronic" infectious mononucleosis, the other revealed persistent splenomegaly.

Aminopterin↗
PubMed1998

Proliferating cell nuclear antigen in hypertrophied spinal ligaments. Immunohistochemical localization of proliferating cell nuclear antigen in hypertrophied posterior longitudinal ligament of the cervical spine.

STUDY DESIGN: An experimental immunohistochemical investigation using an antibody for proliferating cell nuclear antigen. Surgically-extirpated specimens of posterior longitudinal ligament tissues from patients with hypertrophy of the posterior longitudinal ligament and other disorders of the cervical spine were analyzed. OBJECTIVE: To analyze the developmental mechanism of hypertrophy of the posterior longitudinal ligament, the authors evaluated the growth activity of cells in the posterior longitudinal ligament tissues by examining the immunolocalization of the proliferating cell nuclear antigen. SUMMARY OF BACKGROUND DATA: Although a number of cases of hypertrophy of the posterior longitudinal ligament have been reported, the pathophysiology of ligament hypertrophy is still unclear. It is well established that the proliferating cell nuclear antigen is a cell proliferation marker, and immunohistochemical analysis using an anti-proliferating cell nuclear antigen antibody is of value in assessing the cell growth activity of several tissues. METHODS: During anterior decompression surgery in the cervical spine, the authors extirpated posterior longitudinal ligament tissues in one piece from patients with hypertrophy of the posterior longitudinal ligament, ossification of the posterior longitudinal ligament, cervical disc herniation, and cervical spondylotic myelopathy. Midsagittal sections of the specimens were stained with an antibody against the proliferating cell nuclear antigen. RESULTS: In cases of hypertrophy of the posterior longitudinal ligament, immunostaining with the proliferating cell nuclear antigen was detected in cells in the posterior longitudinal ligament, not only at the vertebral endplate level, but also at the midvertebral level. A similar distribution of proliferating cell nuclear antigen-positive cells was observed in cases of ossification of the posterior longitudinal ligament. In cases of cervical disc herniation, however, proliferating cell nuclear antigen-positive cells in posterior longitudinal ligament tissues were restricted to the vertebral endplate level. No immunostaining with the proliferating cell nuclear antigen was seen in posterior longitudinal ligament tissues in cases of cervical spondylotic myelopathy. CONCLUSIONS: Cell growth activity was accelerated in posterior longitudinal ligament tissues in cases of hypertrophy of the posterior longitudinal ligament; such an unusual phenotype of posterior longitudinal ligament cells was also expressed in cases of ossification of cervical disc herniation and cervical spondylotic myelopathy. Therefore, up-regulation of the growth of posterior longitudinal ligament cells may contribute to the development of hypertrophy of the posterior longitudinal ligament, and some common regulatory mechanism(s) on the proliferation of posterior longitudinal ligament cells seem to underlie the development of hypertrophy of the posterior longitudinal ligament and ossification of the posterior longitudinal ligament.

Adult↗
PubMed1979

Evidence for antigenic distinctness of the Epstein--Barr virus-determined nuclear antigen and the Herpesvirus papio-determined nuclear antigen.

Previous serological evidence indicated that Epstein--Barr virus-determined nuclear antigen (EBNA) has cross reactive components with the herpesvirus papio-determined nuclear antigen (HUPNA) and, in addition, it has distinct components of its own, not present in HUPNA. In the present study, this hypothesis was studied by absorption experiments. Absorption of anti-EBNA positive human sera with EBNA abolished both anti-EBNA and anti-HUPNA reactivity. Absorption with HUPNA removed all anti-HUPNA reactivity but did not reduce anti-EBNA to any detectable degree, thus confirming the hypothesis.

Animals↗
PubMed1993

Prognostic value of the combined assessment of proliferating cell nuclear antigen immunostaining and nuclear DNA content in invasive human mammary carcinomas.

The expression of the S-phase associated, nuclear protein proliferating cell nuclear antigen (PCNA) was investigated in routinely paraffin-embedded surgical specimens from 209 breast cancer patients. Cytometric DNA assessments were performed on fine-needle aspirates, upon which the primary diagnosis of breast cancer had been based. The mean clinical follow-up was 16 years (range 13-20 years). The percentage of PCNA immunoreactive tumour cells ranged between less than 5 to 60% (mean value 13.34%). There was a direct association between PCNA expression, high histological tumour grade (p < 0.01), and DNA aneuploidy (p = 0.009). In a subgroup of 22 patients with near-diploid DNA distribution patterns the PCNA expression yielded additional prognostic information. Patients with tumours of near-diploid DNA histograms and more than 20% of PCNA immunoreactive neoplastic cells had a significantly worse clinical course, than patients with near-diploid tumours containing less than 20% PCNA immunoreactive cells (p = 0.0001). In contrast, the PCNA immunoreactivity did not yield additional prognostic information for patients with distinctly diploid or highly aneuploid tumour variants. In a multivariate analysis comprising all 209 patients, nodal status (p < 0.01), tumour size (p < 0.01), and DNA ploidy (p < 0.01) were found to have significant prognostic effect. The findings indicate that carcinomas characterised by high proliferative activity and near-diploid DNA distribution patterns can show rapid tumour progression. The combined assessment of the PCNA immunoreactivity and of the nuclear DNA content in routinely processed surgical specimens of breast cancer patients appears to be of prognostic value.

Breast Neoplasms↗
PubMed1983

Functional mapping of the Epstein-Barr virus genome: identification of sites coding for the restricted early antigen, the diffuse early antigen, and the nuclear antigen.

Attempts were made to functionally map antigenic expression of the Epstein-Barr virus (EBV) to specific regions on the EBV genome, using the B95-8 strain. Experiments were performed to map the expression of early antigen (EA), both restricted and diffuse (R and D, respectively), and the EBV nuclear antigen (EBNA), using intact B95-8 DNA, cloned BamHI fragments or Charon 4A fragments. DNA preparations were microinjected into two EBV genome-negative epithelial tumor cell lines. Expression of EBV antigens was monitored using precharacterized human sera, as well as monoclonal antibodies to EA-R and EA-D. The data suggest that EA-R maps to the BamHI H fragment, and EA-D maps to the Charon 4A fragment 7. A previous report that BamHI K is associated with the expression of a nuclear neoantigen tentatively identified as EBNA (W.P. Summers, E.A. Grogan, D. Shedd, M. Robert, C.R. Liu, and G. Miller, Proc. Nat. Acad. Sci. USA 79, 5688-5692, 1982) was also confirmed.

Antibodies, Monoclonal↗
PubMed1986

Correlation between the presence of antibodies to the Epstein-Barr virus nuclear antigen type 2 and antibodies to the rheumatoid arthritis nuclear antigen in patients with rheumatoid arthritis.

The incidence of antibodies to Epstein-Barr nuclear antigen type 2 (EBNA-2) was determined in sera from rheumatoid arthritis (RA) patients and control subjects, by protein immunoblotting. Sixty-eight percent of the RA patients and 48% of the controls possessed anti-EBNA-2 antibodies. The titer of anti-rheumatoid arthritis nuclear antigen (RANA) in RA patient sera showed a stronger correlation with serum reactions to EBNA-2 than with reactions to EBNA-1. Our results indicate that the presence of EBNA-2 may make a major contribution to the RANA reaction.

Antibodies↗
PubMed1988

Reaction of antibodies to rheumatoid arthritis nuclear antigen with a synthetic peptide corresponding to part of Epstein-Barr nuclear antigen 1.

Antibodies to rheumatoid arthritis nuclear antigen (RANA) are detected by immunodiffusion (ID) and immunofluorescence (IF), though reports of the identity of the antigen(s) have been conflicting. In this study it is shown conclusively that ID and IF anti-RANA react with epitopes on Epstein-Barr nuclear antigen 1 (EBNA-1) and that the major epitope detected by immunofluorescence is represented by a synthetic peptide, P62, corresponding to part of EBNA-1. In an enzyme linked immunosorbent assay (ELISA) anti-P62 antibodies in 35 rheumatoid arthritis sera were threefold higher than those of 35 age and sex matched controls, with the highest levels occurring in young patients with active joint disease.

Adult↗
PubMed1982

Epstein-Barr virus and rheumatoid arthritis: are rheumatoid arthritis associated nuclear antigen and Epstein-Barr virus nuclear antigen different?

In order to investigate the relationship between antibodies to RANA and other Epstein-Barr virus induced antigens, we have tested 50 sera from subjects without rheumatoid arthritis and with various EBV serology patterns for aRANA, aEA, aVCA, aEBNA. Patients were either suffering from Burkitt's lymphoma, infectious mononucleosis, nasopharyngeal carcinoma, Hodgkin's disease or immunodeficiencies, or were healthy controls. RANA was detected by indirect immunofluorescence on G1 synchronized Raji cells. The correlation was very strong between aEBNA and aRANA (r = 0.86) without any correlation between aRANA and aVCA. These data not only strongly support the opinion that aRANA is frequently found in non-rheumatoid diseases but cast doubt on the distinction between RANA and EBNA.

Antigens, Viral↗
PubMedDate not supplied

Combined expression of HLA-DR antigen and proliferating cell nuclear antigen correlate with colorectal cancer prognosis.

HLA-DR antigen expression and the proliferating cell nuclear antigen labeling index (PCNA-LI) were examined immmunohistochemically in 59 surgically resected advanced colorectal cancers to clarify the clinicopathologic significance of the HLA-DR antigen. HLA-DR antigen expression was detected in 33 (56%) of the 59 lesions, which showed less frequent liver metastasis than lesions without HLA-DR antigen expression (40 vs. 68%; p < 0.05). Five-year survival rates of patients with and without HLA-DR antigen expression were 50 and 19%, respectively. Lesions combining HLA-DR antigen expression and a relatively low PCNA-LI had the best prognosis. These results indicate that HLA-DR antigen expression, particularly combined with a low PCNA-LI, is an important outcome predictor in colorectal cancer.

Aged↗
PubMed1985

Localization of nuclear antigens during preparation of nuclear matrices in situ.

Nuclear matrix structure closely resembles the organization of nonchromatin components of nuclei in situ. However, reports on the extent to which nuclear components are reorganized during matrix isolation have produced conflicting results, and the reality of an in situ nuclear matrix is still in question. We have prepared nuclear matrices by processing cells still attached to the growth substrate through the extraction steps, thus avoiding mechanical disruption due to homogenization and centrifugation. Furthermore, the extensive residual cytoskeleton seems to keep the residual nuclei "stretched out" so that they retain many features of intact nuclei. Indirect immunofluorescence staining was used to compare the distribution of nuclear antigens in intact nuclei with their organization in nuclear matrices, as well as at each stage of nuclear matrix preparation. We have applied monoclonal antibodies P1, I1, PI1, and PI2, which had been generated against isolated matrices, as well as autoimmune sera detecting lamins, perichromin, and centromere antigens. Chromatin and RNA extraction was monitored with Hoechst 33258, ethidium bromide, and antihistone. The lamins, PI1, and, to a great extent, PI2 and centromere antigens were little affected by the extraction. The data suggest furthermore that PI1 is a fundamental nuclear matrix component and may serve in integrating peripheral and internal nuclear functions. P1 and perichromin were extensively redistributed after chromatin extraction, supporting a role for these antigens in spatial ordering of chromatin. I1 was progressively extracted at each stage of nuclear matrix preparation and was artifactually associated with matrices which had not been digested with RNase. This study demonstrates unequivocally that the organization of many nuclear matrix components in final preparations reflects their organization in situ. It does indicate, however, that some components retained in matrices are extensively redistributed during nuclear matrix preparation and that their role in nuclear organization must be evaluated in consequence.

Animals↗
PubMed1996

Expression of Ki-67 antigen and proliferative cell nuclear antigen in benign and malignant epithelial lesions of the larynx.

In an attempt to analyse the proliferative activity in benign and malignant laryngeal epithelial lesions, and to determine the relationship to their histologic grade, we studied the expression of proliferative cell nuclear antigen (PCNA) and Ki-67 antigen on 20 squamous carcinomas, and on 30 biopsies of epithelial hyperplasia categorized according to the Kambic-Lenart classification into simple, abnormal, and atypical hyperplasias. In simple hyperplasia, both antibodies stained the nuclei of the occasional cells in the basal layer. In abnormal hyperplasia (mild dyplasia), positive cells occupied up to a third, and in atypical hyperplasia (moderate and severe dysplasia) they occupied from two-thirds to the entire epithelial thickness. In squamous carcinoma, we have found a statistically significant correlation between its grade and the percentage of Ki-67-(p < 0.01) and PCNA-(p < 0.00001) positive cells. Our results suggest that the proliferative fraction progressively increases with the degree of epithelial hyperplasia and the grade of carcinoma. We conclude that the patterns of immunoreactivity to PCNA and Ki-67 antigen correspond to the histologic grade of both benign and malignant epithelial lesions of the larynx. This method should be regarded as a useful adjunct to traditional histological techniques allowing more objective grading of benign and malignant epithelial lesions.

Adult↗
PubMed1994

Developmental expression of Ki-67 antigen and proliferating cell nuclear antigen in macaque placentas.

Placental growth is largely determined by the proliferation of cytotrophoblast cells. However, the distribution of cytotrophoblast cells engaged in the cell cycle during placental development is poorly understood. Recently, antibodies have been developed that identify two proteins directly involved with DNA synthesis: Ki-67 protein and proliferating cell nuclear antigen (PCNA). Immunolocalization of Ki-67 and PCNA provides a measure of the proliferating cells in tissues. We examined, in macaque placentas, the spatiotemporal pattern of expression of these proteins during gestation. Tissues from 24 macaque placentas collected from 22-153 days of pregnancy were prepared for paraffin sections. Standard immunoperoxidase techniques were used to identify Ki-67 and PCNA. The proteins generally co-localized, although PCNA was usually represented in more cells than Ki-67. Early in gestation the cell columns contained many labeled cells. The cytotrophoblastic shell was occupied by numerous cells with PCNA positive nuclei, but few were reactive for Ki-67. By 45 days of pregnancy the immunolabeled cells in the cell columns were concentrated in the proximal regions, adjacent to the anchoring villus tips. The number of positive cells decreased by 100 days when the cell columns were diminished, leaving the anchoring villus tips buried in the shell. Labeled cells were rarely present in the shell at late pregnancy. The single layer of cytotrophoblast cells in the chorionic plate contained numerous reactive cells throughout early and mid-gestation. After approximately 100 days the cytotrophoblast layer of the chorionic plate was stratified over large areas. Soon thereafter few cells of the chorionic plate were labeled. The chorionic villi contained reactive cytotrophoblastic cells throughout gestation. Extravillous cytotrophoblast cells invading spiral arteries were sometimes labeled for PCNA but not Ki-67. We conclude that compartments of the placenta are distinguished by specific patterns of cytotrophoblast cell proliferation. Moreover, these patterns correspond to macroscopic growth parameters of the placenta. Evidence suggests that the macaque placenta slows its rate of diametrical growth at approximately 100 days of gestation. It is at about this time that the cell columns are absorbed into the trophoblastic shell and this pool of proliferating cells is diminished. The growth in diameter of the chorionic plate matches that of the shell. In this compartment also the architecture changes at about 100 days as the cytotrophoblast layer stratifies. This stratification may result from continued proliferation of cytotrophoblast cells when the diametrical rate of growth is decreasing. Soon thereafter, proliferation decreases in this compartment also. By contrast, labeled cells were found in chorionic villi throughout gestation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗