Ryuzo Ueda

Clinicopathologic study of CD56 (NCAM)-positive angiocentric lymphoma occurring in sites other than the upper and lower respiratory tract.

The expression of the neural cell adhesion molecule (NCAM) (CD56, NKH-1) is a rare phenomenon in malignant lymphoma. Recently, several authors, including our group, described the clinicopathologic, phenotypic, and genotypic features of NCAM-positive tumors as a unique subgroup within a larger category of hematolymphoid malignancies. Ten cases of CD56+ angiocentric lymphoma occurring in sites other than the upper aerodigestive tract were studied for evaluating their characteristics. The disease occurred in six men and four women varying from 24 to 85 years (mean age, 53 years) who often exhibited a striking predilection for extranodal sites of involvement, such as the skin, gastrointestinal tract, and muscle, usually in the absence of peripheral lymphadenopathy. Although the cytologic appearances and immunophenotypic profile varied from case to case, these tumors often exhibited azurophilic granules, an angiocentric growth pattern, and surface CD3−, T-cell receptor (TCR) antigens−, and CD56+ phenotype without B-cell phenotype, except for a single case of CD3+, TCRα/β+, and CD56+ phenotype. Genotypic investigation exhibited germline configuration of the TCR β and γ chain genes and the immunoglobulin heavy chain gene in all five cases of surface CD3− phenotype examined, whereas the case of CD3+ phenotype showed rearrangement of TCRβ. They seem to constitute a distinct entity of the lineage spectrum spanning from natural killer (NK) cell to NK-like T cell.

Prognostic significance of abnormal p53 accumulation in primary, resected non-small-cell lung cancers.

PURPOSE This study was conducted to evaluate the prognostic significance of p53 abnormalities in primary, resected non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS Methodologic validation of immunohistologic detection of p53 abnormalities in routine pathology sections was assessed using 31 lung cancer specimens for which p53 gene status was known from our previous molecular biologic studies. Applying the optimized cutoff value, we evaluated the prognostic significance of p53 abnormalities in an independent cohort of 208 NSCLC patients with complete follow-up data, whose resections were consecutively performed between January 1984 and December 1988. RESULTS Immunohistologic detection of p53 abnormalities appeared to be reliable and showed approximately 90% concordance with the p53 gene status. Using the selected cutoff value of 10%, 46% of 208 NSCLCs showed p53 abnormalities. There was no relationship between p53 abnormalities and clinical outcome in the entire cohort, which represented all histologic subtypes of NSCLC (P = .58). Based on the reasoning that the influence of p53 abnormalities may have been obscured by distinct biologic roles depending on histologic subtypes, we also separately analyzed subsets of patients with adenocarcinomas (n = 100) and with squamous cell carcinomas (n = 88) and found that it may be a useful prognosticator only in adenocarcinoma patients (P = .04). CONCLUSION p53 abnormalities are not a significant prognostic factor in primary, resected NSCLC when all histologic subtypes are combined, but may be a useful prognosticator for adenocarcinomas. Additional studies are warranted for further evaluation, specifically of adenocarcinomas.

Inconsistent association of Epstein-Barr virus with CD56 (NCAM)-positive angiocentric lymphoma occuring in sites other than the upper and lower respiratory tract

We previously described nine cases of angiocentric lymphoma of a possible natural killer (NK)‐cell lineage with a surface CD3− CD56+ phenotype occurring in sites other than the upper and lower respiratory tract. This study was performed to investigate the association of Epstein‐Barr virus (EBV) with these lymphomas, using the polymerase chain reaction (PCR) for the presence of EBV‐DNA, in situ hybridization (ISH) for EBV‐encoded small RNAs (EBERs) and immunohistology for EBV‐determined nuclear antigen‐2 (EBNA‐2) and latent membrane protein‐1 (LMP‐1) in paraffin sections. PCR and ISH produced almost identical results, and EBERs were identified in the nuclei of the lymphoma cells of three cases, two of which exhibited LMP‐1 in the cytoplasm of tumour cells without EBNA‐2 expression. Molecular genetic analysis revealed EBV to be incorporated into these three EBER‐positive cases either clonally or biclonally. It was revealed by re‐evaluation of their morphology with the established EBV status on each case that, in contrast to the rather variable and irregular cellular composition of the EBV‐ positive tumours, the EBV‐negative tumours stood out because of their remarkably uniform ‘blastoid’ appearance, and could be grouped as blastic NK‐cell lymphoma. The relationship of the EBV‐positive cases with nasal NK‐cell tumours has yet to be clarified.

Loss of Heterozygosity at the bcl‐2 Gene Locus and Expression of bcl‐2 in Human Gastric and Colorectal Carcinomas

The loss of heterozygosity (LOH) at the bcl‐2 gene locus and the expression of the bcl‐2 gene were examined in gastric and colorectal carcinoma cell lines and carcinoma tissues. LOH at the bcl‐2 locus was detected in 24% (4/17) of gastric and 60% (6/10) of colonic carcinomas, all of which were well differentiated adenocarcinomas, whereas LOH was not seen in poorly differentiated ones. On the other hand, 24% (5/21) of poorly differentiated stomach cancers overexpressed bcl‐2 gene, whereas no overexpression was detected in well differentiated stomach cancer. Three gastric and three colorectal carcinoma cell lines, all of which were derived from poorly differentiated adenocarcinomas, expressed considerable levels of bcl‐2 mRNA and protein. These results suggest that LOH at the bcl‐2 locus is frequently associated with well differentiated adenocarcinomas of the stomach and colon, and bcl‐2 overexpression has implications for the development of poorly differentiated adenocarcinomas of the gastrointestinal tract.

MONOCLONAL ANTIBODY AGAINST PRAD1/CYCLIN D1 STAINS NUCLEI OF TUMOR CELLS WITH TRANSLOCATION OR AMPLIFICATION AT BCL-1 LOCUS

Mouse monoclonal antibodies were produced against the bacterial product encoded by human PRAD1/ cyclin Dl gene, which is known to be involved in tumors with translocation or amplification at BCL‐1 locus of 11ql3. The immunizing antigens used were GST‐PRAD1 and T7 gene 10‐PRAD1 fusion products. Four antibodies were reactive with both PRAD1 fusion products and cell lysates of B‐cell tumor cell lines with t(ll;14)(ql3;q32) and a breast cancer cell line with 11ql3 amplification, on immunoblotting. An immunofluorescence study showed that only one of them stained nuclei of cells with 11q13 abnormalities. Since this antibody proved applicable for conventional paraffin‐embedded tissue sections, immunohistologic staining of various lymphoma tissues was performed. Eight of 11 mantle cell lymphomas showed intermediate to strong positivity and 6 of the positive cases demonstrated characteristic staining patterns that were either predominantly nuclear or both nuclear and cytoplasmic. The nuclear staining pattern was not observed with other types of lymphoma and thus may correlate with PRAD1 mRNA overexpression.

Phenotypic analysis of peripheral T cell lymphoma among the Japanese

From 1980 to 1990, 174 peripheral T cell lymphomas were studied morphologically and immunophenotypically with a panel of monoclonal antibodies which were reactive with T cell differentiation antigens in cryostat sections and/or cell suspensions. Histologically, 57% of the lymphomas were categorized into low‐grade tumors according to the updated Kiel classification, while 41% were high‐grade tumors. By immunologic studies, 50% of the lymphomas were of helper/inducer (CD4) phenotype, 6% were of cytotoxic/suppressor (CD8) phenotype, 3% expressed both CD4 and CD8, 3% lacked both CD4 and CD8, and 36% were phenotypically undetermined because of an admixture of a fairly even number of CD4 and CD8‐positive cells. The phenotypically undetermined cases were more frequently noted in the low‐grade groups than in the high‐grade group, and the latter often showed a loss of pan‐T antigens, although there was no definite correlation between the histologic category and the immunophenotype. CD25, which is strongly manifested in anti‐HTLV‐1 antibody‐positive cases, was negative or only weakly expressed in anti‐HTLV‐1 antibody‐negative cases. Anaplastic large cell lymphomas (LC‐Ana) strongly expressed CD30, which was also detectable in only large blast‐like cells in the low‐grade tumors. Seventy‐one per cent of the lymphomas expressed la antigens. In this series, the clinical data were available on 154 patients. For individual markers, the expression of CD30 and HLA‐DR were associated with a longer actuarial survival (P< 0.01 and P < 0.05 by the generalized Wilcoxon test). The absence of CD25 or the presence of CD3 on tumor cells correlated with a relatively favorable prognosis, but not significantly. The detection of CD4 and CD8 had relatively little prognostic value. In the cases excluding LC‐Ana, a significant difference was also recognized between the groups with and without CD25, CD30 and HLA‐DR (P < 0.05 by the generalized Wilcoxon test). These results suggest that the immunophenotypic analysis of peripheral T cell lymphoma provided its use as an adjunct to a histopathologic diagnosis and was related to prognostic prediction.

Overexpression of PRAD1 in a mantle zone lymphoma patient with a t(11;22)(q13;q11) translocation

Summary The PRAD1 gene identified from the chromosome band 11q13 region was previously demonstrated to be overexpressed in cell lines with t(11:14)(q13;q32) trans‐location and was suggested to be a candidate BCL‐1 gene. We report here one case of mantle zone lymphoma with a t(11;22)(q13:q11), a variant translocation at the BCL‐1 locus, having the PRAD1 overexpression. By analogy with the c‐myc gene in Burkitts lymphoma and the BCL‐2 gene in follicular lymphoma, this case supports strongly the idea that the PRAD1 is the candidate BCL‐1 gene.

Clinicopathologic study of 212 cases of peripheral T-cell lymphoma among the Japanese

Background. Postthymic/peripheral T‐cell malignancy shows significant histopathologic and clinical diversity, even in its prognosis, and the correlations remain to be debated.

The positive nuclear staining observed with monoclonal antibody against PRAD1/cyclin D1 correlates with mRNA expression in mantle cell lymphoma

Recently, we produced a monoclonal antibody, 5D4, against the PRAD1/cyclin D1 product and suggested positive nuclear staining to be associated with mantle cell lymphoma (MCL). Now we have further characterized the specificity of this antibody and studied the relation of immunohistochemical detection to PRAD1/cyclin D1 mRNA expression and DNA rearrangement. Immunofluorescence and immunoblotting studies demonstrated the 5D4 antibody to be crossreactive with cyclin D2, but not cyclin D3. On immunostaining, 15 of 19 MCL cases (79%) presented the nuclear staining pattern and PRAD1/cyclin D1 mRNA expression was detected by Northern blot analysis in 12 of 15 MCL cases studied (80%); all cases with the mRNA expression showed the nuclear staining pattern. Southern blot analysis with 11q13 BCL‐1 probes detected DNA rearrangements in 8 of 19 MCL cases (42%), all 8 exhibiting PRAD1/cyclin D1 mRNA expression. In 21 lymphoma cases of types other than MCL, neither the mRNA expression nor the nuclear staining were observed, although cytoplasmic staining was often apparent. These results indicated that positive nuclear staining of lymphoma cells by 5D4 antibody reflects PRAD1/cyclin D1 mRNA expression, and showed that this monoclonal antibody has diagnostic value for differentiating MCL from other types of lymphomas.

Ki-1 (CD30) Positive anaplastic large cell lymphoma of T-cell phenotype developing in association with long-standing tuberculous pyothorax: Report of a case with detection of Epstein-Barr virus genome in the tumor cells

We report a case of CD30 positive anaplastic large cell lymphoma of T-cell phenotype developing in association with long-standing tuberculous pyothorax. Phenotypic analysis showed CD1a-, CD2+, CD3+, CD4+, CD5-, CD8-, CD10-, CD19-, CD20 +/-, CD21-, CD25-, CD56-, T-cell receptor (TCR) alpha/beta antigens-, and HLA-DR+ phenotype. Neither rearrangement of TCR beta and gamma chain genes or of immunoglobulin heavy chain gene was detected in DNA extract from fresh material. The lymphoma cells were also shown to express the latent membrane protein-1 and the Epstein-Barr virus (EBV)-encoded nuclear antigen-2 by immunohistochemistry and EBV-encoded small RNAs by in situ hybridization.