Expansions of clonal and oligoclonal T cells in B-cell chronic lymphocytic leukemia are primarily restricted to the CD3(+)CD8(+) T-cell population.

B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation of mature-appearing clonal B cells exhibiting coexpression of CD5 and CD23. In addition to the accumulation of neoplastic B cells, numerous T-cell abnormalities also occur in B-CLL patients. In this study, the presence, and distribution within the T-cell subsets, of clonal/oligoclonal T cells was studied. Multicolor flow cytometric techniques were employed using combinations of anti-CD3, anti-CD4, and anti-CD8 antibodies coupled with antibodies specific for V(alpha) and V(beta) T-cell receptor (TCR) epitopes. Molecular studies of TCR gene sequences were done to confirm the presence of clonal/oligoclonal T-cell populations. In the flow cytometric studies, examination of V(alpha)/V(beta)expression found evidence of clonal/oligoclonal expansion in 9 of 19 patients studied. In eight of the nine patients, the expansions were restricted to the CD3(+)CD8(+) cell population. Molecular analyses were performed in 16 patients, 12 of whom showed a clonal or oligoclonal pattern. Of the four patients who were negative in the molecular analyses, all demonstrated flow cytometric evidence of clonal/oligoclonal expansions. Thus, when the flow cytometric and molecular analyses were considered together, all 16 patients for whom parallel analyses were done showed evidence of clonal/oligoclonal expansions. These results confirm previous work demonstrating that the majority of B-CLL patients harbor clonal/oligoclonal expansions within the T-cell population. Additionally, based on the relative numbers of cells expressing specific V(alpha) or V(beta)epitopes, these results show that these expansions occur primarily within the CD3(+)CD8(+) T-cell population.

Extramedullary myeloid cell tumors. An immunohistochemical study of 29 cases using routinely fixed and processed paraffin-embedded tissue sections.

OBJECTIVE: Extramedullary myeloid cell tumors (EMCTs) may be unsuspected clinically and difficult to recognize histologically. Fresh or frozen tissue is often not available for analysis. We studied 29 cases of EMCT using routinely fixed and processed paraffin-embedded tissue, an immunohistochemical method, and a panel of antibodies. PATIENTS: We studied 29 patients with EMCTs: 22 males and 7 females, with a median age of 48 years (range, 5 to 80 years). Histologically, 9 tumors were well differentiated, 16 were poorly differentiated, and 4 were blastic. RESULTS: The Leder stain (napthol-ASD-chloroacetate esterase) was positive in 21 (77.7%) of 27 tumors. Immunohistochemically, the following antibodies reacted with the greatest number of cases: Leu-22 or MT1 (CD43) in 28 (96.6%) of 29, antilysozyme in 27 (96.4%) of 28, and antimyeloperoxidase (MP07) in 21 (91.3%) of 23 cases. Other myeloid lineage-associated antibodies were positive in a subset of cases: antineutrophil elastase (NP57) in 10 (62.5%) of 16, Leu-M1 (CD15) in 7 (46.6%) of 15, and Mac-387 in 6 (40.0%) of 15 cases. The well-differentiated EMCTs reacted with most myeloid-associated antibodies; poorly differentiated and blastic tumors were more often negative. The pan-leukocyte antibody LCA (CD45RB) reacted with 15 (60%) of 25 neoplasms. Three (16.6%) of 18 tumors contained numerous p53-positive cells, ranging from 10% to 50% of the tumor cell population. In 10 cases, exons 5 through 8 of the p53 gene were analyzed using the polymerase chain reaction and single-stranded conformational polymorphism analysis. Gel shifts consistent with mutations were identified in exon 8 of one tumor (10%) that exhibited abundant p53 immunostaining. CONCLUSIONS: Immunohistochemical studies using fixed, paraffin-embedded sections are very useful in the diagnosis of EMCTs. The most sensitive antibodies are anti-CD43, antilysozyme, and antimyeloperoxidase. Immunohistochemical methods are more sensitive than the Leder stain. We found p53 staining in a small subset of cases, in which we were able to confirm evidence of p53 gene mutation using the polymerase chain reaction and single-stranded conformational polymorphism analysis in one case; p53 gene mutations appear to be uncommon in EMCTs.

Flow cytometric detection of neoplastic T cells in patients with mycosis fungoides based on levels of T-cell receptor expression.

The authors report the flow cytometric detection of neoplastic T cells in the peripheral blood of four out of five (80%) patients with peripheral blood involvement with mycosis fungoides (Sezary syndrome) based on the levels of T-cell receptor expression as measured by CD3 and TCR-alpha beta staining. Antigen receptor expression was abnormal in terms of increased density of surface CD3 or TCR-alpha beta per cell. Other immunophenotypic abnormalities were present in three of these patients. However, in one patient abnormal T-cell receptor expression was the only immunophenotypic evidence of neoplasia, although morphologically abnormal lymphocytes were present and a T-cell clone was detected by polymerase chain reaction (PCR). In another patient, the authors were able to detect development of a new, more aggressive neoplastic T-cell population based on levels of T-cell receptor expression. Levels of T-cell receptor expression may be of diagnostic utility in the evaluation of peripheral blood for the presence of neoplastic T-cell populations.

Flow cytometry and Zollinger-Ellison syndrome: relationship to clinical course.

BACKGROUND: With successful means of controlling gastric acid secretion in patients with Zollinger-Ellison syndrome, the gastrinoma itself is becoming the major determinant of long-term survival. No methods have yet been described to predict which tumors will have more malignant courses thereby indicating which patients should undergo aggressive surgery or antitumor therapy. Because DNA analysis, using flow cytometry, has proved helpful in this regard in other tumors, the current study was designed to evaluate its utility in gastrinoma patients. METHODS: Flow cytometry was performed on 81 paraffin-embedded gastrinoma specimens from 59 patients. Results were compared with preoperative patient characteristics, findings at surgery, and postoperative follow up. RESULTS: Tumors were diploid in 54% of patients, near diploid in 15%, pure tetraploid in 0%, nontetraploid aneuploid in 25%, and multiple stem line aneuploid in 5%. All patients with multiple stem line aneuploid tumors had wide-spread metastases whereas all patients with nontetraploid aneuploid tumors had localized or regional disease. Median S phase percentage was 3.6. S phase percentages were higher in patients with widespread metastatic disease than in patients with localized or regional disease. Disease extent also correlated closely with fasting serum gastrin level. After removing this variable with logistic regression analysis, the significant correlation between disease extent and DNA analysis persisted. CONCLUSIONS: DNA analysis of gastrinoma tissue specimens correlates independently with the extent of disease and may be useful in planning therapeutic strategies for patients with Zollinger-Ellison syndrome.

[DNA--ploidy in non hodgkin's lymphomas of low malignancy]. / DNA-ploidia w chloniakach nieziarniczych o niskiej zlosliwosci.

Lymphoma cell DNA cytometry in imprints of dissected lymph nodes was performed in 98 patients with low grade non-Hodgkin lymphomas and in 15 patients with reactive lymphadenopathy. The percentage of resting and proliferating cells was also calculated on the basis of computer analysis of DNA amount and cell image. It has been shown that, in contrast to the control group characterized by diploidic DNA amount, DNA aneuploidy was observed in 63.3% of lymphoma patients (DNA hyperdiploidy in 58.2% and DNA aneuploidio-polyploidy in 5.1% of the studied group). The proliferative activity of lymphoma cells was higher in patients with DNA aneuploidy than in DNA diploidic patients and in the control group.

Evaluation of spermatozoa of the rat in hyperprolactinaemia induced by metoclopramide.

Metoclopramide (MCP) in doses of 2.2 mg kg-1 body weight, being administered intraperitoneally to rats for 14 d, provokes a premature release of germ cells in the testes. The number of spermatozoa is reduced. In addition the spermatozoa exhibit defects of head and tail structures. After the administration of MCP for 14 d, followed by a 14-d break in applying the drug, the number of spermatozoa in the lumen of the epididymal duct continues to be low. Many spermatozoa are damaged. Only after a 66-d-break in MCP administration is the morphology of the seminiferous epithelium restored, and a large number of spermatozoa appear in the lumen of the epididymal duct.

DNA cytometric and histologic findings in mouse tumors (BP and S 180) with different response to treatment with tumor necrosis factor.

DNA cytometric and histopathologic investigations were performed in two tumor models (BP and S 180) which differed in their sensitivity to tumor necrosis factor (TNF). TNF induced strong necrosis in both tumors, but only the sarcoma 180 showed total regression. After TNF administration DNA cytometry revealed in the BP tumor an increase of cells in the S-phase, and in the S 180 tumor a loss of aneuploid cell populations. Histologic examination revealed a more obvious effect of TNF on tumor blood vessels in BP tumor, whereas infiltration of inflammatory cells was observed only in the S 180 tumor. We concluded that cell infiltration may be of importance for tumor regression and that aneuploid cell populations are more sensitive to TNF treatment than eudiploid cells.

[Periarteritis nodosa with clinical manifestations of acute cholecystitis]. / Guzkowe zapalenie tetnic z klinicznymi objawami ostrego zapalenia pecherzyka zólciowego.

A 69-year-old male patient is reported with clinical symptoms and signs of acute cholecystitis in whom microscopic examination of the surgically removed gallbladder showed evidence of periarteritis nodosa.