Immunologic cell markers of peripheral blood and lymph node lymphocytes in chronic lymphocytic leukemia.

The immunologic phenotype of the lymphocytes of 100 patients with chronic lymphocytic leukaemia (CLL) was investigated. Peripheral blood lymphocytes (PBL) were examined in all cases; in 46 patients with lymphadenopathy, a lymph node was biopsied and the histologic and immunologic patterns were assessed: 24 had a lymphocytic-lymphoplasmocytoid histology and 22 the follicular variant of lymphocytic lymphoma (mantle zone lymphoma, MZL). For comparison, lymph node suspensions from 19 patients with non-leukemic centrocytic lymphoma (CCL) were also studied. Significant differences in the PBL immunologic features were found between stage O and stage I patients. The phenotype of the lymphocytes of patients with lymphocytic histology was similar to that of stage 0 CLL patients, whereas major differences were found between these patients and those with mantle zone histology. This enables these patients to be recognized easily on immunologic grounds.

G1 heavy chain disease: clinicopathological, ultrastructural and immunochemical study of a new case.

Clinicopathological ultrastructural, and immunohistochemical findings of a new case of IgG1 heavy chain disease are reported in detail. The abnormal protein lacks the VH and CH1 region with sequence starting at 225 residue. The main pathologic feature was plasmacytic tumor of the lymph nodes with B-cell immunoblastic sarcoma patterns. Neoplastic diffusion to other organs was also present. Plasmacytic neoplastic cells have also been found in the bone marrow. The ultrastructure of the noeplastic cells was characterized by more or less abundant plasmacytic-like endoplasmic reticulum with very frequent peculiar whorled configurations. Immunohistochemical methods revealed the abnormal protein production by neoplastic cells in different stages of differentiation. From this case and from the data of the literature it is concluded that the gamma HCD is due to a neoplastic proliferation of lymphoplasmacytic cells whereas the reticulum cells are never involved.

Argyrophilic nucleolar organizer region counts and Ki-67 scores in human renal cell carcinoma.

The proliferative activity of 21 cases of renal cell carcinoma has been investigated by means of monoclonal antibody Ki-67 and Nucleolar Organizer Regions (AgNORs) analysis. The correlation between AgNOR counts and Ki-67 scores was only slightly significant (r = 0.53, r2 = 0.28) as determined by linear regression. Positive correlation was found between Ki-67 scores and tumour histologic grade. However, no correlation was observed between Ki-67 scores and tumour pathologic stage and between AgNOR counts and tumour histologic grade and/or pathologic stage. The results suggest that AgNOR counts cannot replace Ki-67 scores in evaluating the proliferative activity of renal cell carcinoma and that such activity and both histologic grade and pathologic stage seem to be independent parametres.

Angiogenic activity of hyperplastic and neoplastic lymph nodes.

Angiogenic capacity was tested in 14 non-Hodgkin's lymphomas, 7 Hodgkin's lymphomas and 15 cervical lymph nodes nonneoplastic but draining a territory with a laryngeal carcinoma. The objective was to find out whether different groups of lymphomas showed differences in their angiogenic capacity and to compare the ability to induce neovascularization of neoplastic lymphocytes. Frequency and intensity of the angiogenic response were similar for classes of lymphomas different for morphologic and immunologic characteristics. The presence of a carcinoma was sufficient to induce in tributary, nonmetastatic lymph nodes an angiogenic activity comparable to that known to characterize antigenically stimulated lymphocytes.

Prominent plasmacytic differentiation in the recurrence of a parotid immunocytoma. A morphologic and immunohistochemical study.

We report and discuss a case consisting of 2 lesions that developed at different times in the same parotid gland. Although the first lesion showed morphologic features similar to those of the benign lymphoepithelial lesion, a monotypic IgM/K pattern was also revealed by the immunoperoxidase method in its lymphoid infiltrates. Thus, on cytologic grounds, the first lesion was classified as lymphoplasmacytic immunocytoma (according to Kiel criteria). In the second lesion, which recurred 3 years later, the prominent feature was a marked predominance of plasma cells with the same monoclonal IgM/K as the preceding lymphoma. These findings suggests that a B-lymphomatous cell monoclone may undergo morphologic maturation along the lineage of its competence. Secondly, they indicate that every case of benign lymphoepithelial lesion of the salivary glands should undergo thorough immunologic evaluation to exclude the possibility of signs of precocious lymphomatous transformation.

Cytologic localization of antigens from human saliva in pleomorphic adenomas of salivary glands.

Twenty pleomorphic adenomas of human salivary glands were studied by means of specific immunohistochemical methods. Material reacting with antihuman saliva antiserum has been detected in epithelial structures of all pleomorphic adenomas. The myxoid stroma was immunohistochemically negative. Both PAS and immunohistochemical reactions in the same section indicate that there is no close relationship between the results of these procedures. This study suggests that the pleomorphic adenomas can be considered epithelial tumors of salivary distal gland cells retaining or developing specific exocrine capability.

Lymphocyte populations of non-scleronodular Hodgkin's disease subtypes in different stages of lymphocyte depletion. An immunophenotypic and quantitative study.

Twenty-one cases of non-scleronodular Hodgkin's disease with variable lymphocyte contents were studied immunophenotypically and quantitatively to analyse the distribution of different lymphocyte populations and to determine whether selective loss of lymphocyte subpopulations accompanies overall lymphocyte depletion. In Hodgkin's tissue B-cells were scanty and unevenly distributed in samples with both many and few lymphocytes. Several large B, LN1-positive (possibly activated) cells were observed in a few cases. CD3-positive T-lymphocytes predominated in all cases; the same cells were also UCHL1-positive, thus expressing characteristics of mature T-memory cells. CD4-positive lymphocytes were usually more numerous than CD8-positive lymphocytes, but quantitative evaluation of the latter showed that they did not decrease in proportion to any diminution of the whole lymphocyte population. This finding suggests that in the process of lymphocyte depletion more CD4-positive lymphocytes than CD8-positive lymphocytes are lost, and this might account for the impairment of cell-mediated immunity in Hodgkin's disease.

Identification of rhinopharyngeal plasmacytic tumours by immunofluorescence on paraffin-embedded sections.

Solitary myeloma is an uncommon lesion which may occur in many organs, mainly in the rhinopharynx. This lesion usually appears as non-secretory, and it is constituted by either typical plasma cells or large and atypical cells resembling epithelial anaplastic cells. Therefore, and immunofluorescent method on paraffin-embedded sections may be applied to reveal the presence and the monoclonality of the intracytoplasmic Ig in the pathological cells. In this paper we report a study on 22 patients suspected to bear a rhinopharyngeal tumor investigated by immunofluorescence. Moreover, in 4 cases peripheral blood lymphocytes were also studied. The results are discussed on the basis of the modern immunological trends.

Signet ring cell lymphoma in salivary gland. An immunohistochemical and ultrastructural study.

A case of recurrent signet ring cell lymphoma involving a salivary gland is reported. The tumour was centroblastic-centrocytic nodular and diffuse. The cells were vacuolated, oil red, Sudan III, PAS and Alcian blue negative. They displayed the Ia-like antigen but were negative for SIg. Ultrastructurally the vacuoles appeared empty or containing residual membrane and/or myelinoid bodies. Sometimes microvesicles were found in them. The vacuoles were mainly located in the Golgi area but they were also found in mitochondria. Cytoplasmic bodies containing microvesicles were also seen. The case is compared with those of the literature and the significance of the vacuoles is discussed.

Histochemical study on human germinal centre, mantle-zone and extra-follicular area lymphoid cell subpopulations. Immunological and cytochemical correlations with lymphomatous cells, peripheral normal and leukemic lymphocytes.

Tissue sections of lymph nodes, appendices and tonsils, together with smears of immunologically separated peripheral lymphoid cells from a B-CLL and lymphomatous cells from an immunocytoma were submitted to combined enzyme cytochemical investigations with acid alpha-naphthyl acetate esterase (ANAE), beta-glucuronidase (B-G), acid phosphatase (AcPh), adenosine triphosphatase (ATPase), a,d 5'nucleotidase (5'N). T-cells were Acph+, ATPase- and 5'N-. The vast majority of T- and B-cells displayed ANAE and B-G activities with two distinct staining patterns (T-like and B-like pattern). A high proportion of lymphoid cells in the germinal centre (G.C.) and the vast majority of lymphoid cells in the mantle-zone (M.Z.) were shown to belong to B-cell system because of the expression of ATPase and 5'N in their membranes. Some lymphoid cells positive for ANAE and B-G with a B-like pattern and for AcPh were recognizable in the G.C. In the M.Z. only a few lymphoid cells being ANAE+, with a T-like pattern, and AcPh+ were shown to belong to the T-cell system. In contrast, in this zone a high proportion of small lymphoid cells (64% +/- 10%) showed ANAE activity, mostly with B-like pattern. Therefore, these findings indicate that in the M.Z. a high proportion of B-cells ATPase+ and 5'N+ also display ANAE activity. By comparison of the results obtained from lymphoid tissue sections, B-CLL and immunocytoma cell suspensions and normal circulating lymphocytes we can conclude that B-ANAE-positive cells of the M.Z. do not usually appear in the peripheral blood. They circulate in large numbers only in some pathological conditions (like our reported B-CLL). Therefore, B-ANAE-positive lymphoid cells of the mantle, with a B-like staining pattern, include a wide range of subsets which exclude large lymphoid cells and plasma cells.

Selective binding of peanut lectin by T lineage lymphocytes in paraffin sections.

On sections of formalin fixed paraffin embedded lymph nodes, thymuses and spleens, binding of the lectin from peanut (Arachis hypogaea) was detected on some histiocytes, while the lymphoid cells were negative. After treatment of the sections with neuraminidase, sialic acid covered beta-D-galactosyl (1-3)-N-acetyl-D-galactosamine residues were revealed in a more extensive number of histiocytic cells and on a sub-population of lymphoid cells with the typical distribution of T lineage lymphocytes. The technique here described allowing the identification of subclasses of lymphocytes on paraffin sections might provide new data for the study and the diagnosis of lymph node diseases.

CD5-positive B-cells of the fetal and adult spleen lymphoid tissue: an immunophenotypical study.

The lymphoid tissue of the human fetal spleen at various stages of gestation was studied on frozen and paraffin sections and with two-colour flow cytometry. On the sections scattered lymphoid cells and perivascular lymphoid aggregates were found starting from the 15th week of gestation. CD3, CD5, CD19, CD20, CD21, CD22, CD24, CD35, CD38-positive cells were observed. No CD10- and CD23-positive cells were detected. Flow cytometry showed a prevalence of B-lymphocytes. The majority of them expressed CD5 antigen. These cells were also IgM/D, CD19, CD20, CD21, CD22, CD24 and CD35-positive. Only few of them expressed CD10 and CD23. A similar phenotype was found in human cord blood. By contrast, in adult spleens CD5 B-cells never exceeded 8% of the B-cells. The comparison between CD5 B-cells of fetal spleen and CD5 B neoplastic cells of 72 cases of small B-cell lymphomas showed that CD23, which was usually expressed by a high percentage of the neoplastic cells, particularly in B-CLL, was not displayed by the majority of the CD5 non neoplastic cells. The reverse was shown by CD35. These findings suggest that different states of activation distinguish the normal CD5 B-cells from their malignant counterpart.

B-cell chronic lymphocytic leukaemia stage 0. An immunophenotypic study of 66 cases and comparison with B small cell lymphomas.

The cells of 66 B-CLL stage 0 patients were analyzed using a large panel of monoclonal antibodies in order to better define the immunophenotype of B-CLL. The data were compared with the immunophenotypes of lymph node cells from 51 patients with diffuse B small cell lymphoma or leukaemia with lymph node enlargement. The most frequent immunophenotype of B-CLL stage 0 was SIgM(D)+ EmR+ CD5+ CD9+ CD21+ CD23+ CD35- CD38-. Among the lymphomas, EmR-positive and EmR-negative cases were identified. The vast majority of the EmR-positive cases usually showed the leukaemic pattern, immunophenotype and lymph node histology of B-CLL. The EmR-negative cases usually had immunophenotypes quite different from those of B-CLL and the histology of indented cell lymphoma (centrocytic or intermediate) or features of lymphoplasmacytoid/cytic lymphoma. More than 20% of EmR-positive cells proved to be the most important marker to distinguish B-CLL from other lymphocytic lymphomas. Indeed this was a sign of better prognosis. Lymphoplasmacytoid/cytic lymphomas were EmR-positive with the immunophenotype of B-CLL or EmR-negative with a definitely different immunological profile. This suggests two morphologically similar but biologically different subgroups of these diseases.

A new case of gamma heavy chain disease: clinical, immunochemical and structural characterization.

A new case of gamma heavy chain disease (gamma HCD) is described in a 77-year-old woman. The serum and urine contain an M-component and electrophoretic, antigenic and ultracentrifugal properties resembling those of the Fc fragment of IgG globulin. Analysis on SDS-polyacrylamide gel electrophoresis and ultracentrifugal studies show that gamma HCD portein is present in the serum as a dimer with a molecular weight of 58,000 daltons. Analysis of isotypic and allotypic markers along with the structural studies show that this HCD protein belongs to the IgG subclass and that deletion includes the total VH and CH1 regions with sequence starting at residue 225 in the middle of the hinge region.

Heterogeneous subgroups among malignant diffuse small B cell lymphomas. A combined nucleometric and immunocytologic study.

The degree of heterogeneity among subtypes was evaluated in 39 of the most frequent malignant, diffusely growing small B cell lymphomas by a combination of morphometry, automated image analysis, and immunocytologic techniques. Cluster analysis of nuclear profile parameters, including nuclear area, circularity factor, and chromatin distribution pattern, distinguished 3 groups. Each group was characterized by the preponderance of certain nuclear profile types, i.e. (a) small, rather regular (roundish) and dark staining (typical small lymphocytic lymphomas and immunocytomas); (b) small to intermediate size, rather regular (roundish) and pale staining (small to intermediate size variant); or (c) small to intermediate size, irregular, rather pale staining (diffuse follicular small cleaved cell (centrocytic) lymphomas and, probably, polymorphous immunocytomas). Group 3 was clearly distinct from the rest, but groups 1 and 2 also differed significantly. Every case displayed a mixture of the 13 registered nuclear profile types. Lymphoid cells with similar nuclear profile features occurred in B cell zones of non-neoplastic lymph nodes. Frequency distribution of nuclear profile parameters significantly differed from one lymphoma group to another. However, there were considerable overlaps. No clear correlation was found between nuclear profile types and immunophenotypes. Cellular surface antigen patterns showed an extensive intra- and inter-case variability. The findings support the notion of a certain individuality of malignant B cell lymphomas and of a marked heterogeneity among their subtypes.