Acquired pure red cell aplasia associated with thyroid carcinoma: case report.
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Biomedical subjects
Publications and source records attributed to G Pizzolo.
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Bone marrow trephine biopsies from 17 patients with B-chronic lymphocytic leukemia (B-CLL) were studied by immunohistologic techniques in order to investigate the cellular phenotypes of both neoplastic (B-lymphoid) and reactive (T-lymphoid) infiltrates. For this purpose, several heteroantisera and monoclonal antibodies against human Ig isotypes, HLA-DR antigens, and T-cell subpopulations were used in immunofluorescence. The findings were analyzed in relationship to the histologic pattern of involvement, as well as to the immunologic data of cell suspensions from peripheral blood. In all cases, the dominant lymphoid population within the bone marrow infiltrates showed identical phenotypic characteristics of B-CLL cells from the blood (HLA-DR+, mu +, most frequently delta +, kappa +, or lambda +, and weakly RFA-1+). The infiltration by these malignant B cells was diffuse in 5 cases and nodular plus interstitial in 12. The number of T cells (UCHT1+, RFA-1+, mu) was variable (5%-25%) in the different samples, but the values were high when compared to the proportion of T cells in normal bone marrow and in the blood of most patients studied. Furthermore, a clear predominance of T cells exhibiting the inducer phenotype (Leu-3+) was observed in all bone marrow samples, which is in contrast with the findings from peripheral blood, where T cells with the suppressor/cytotoxic phenotype (Leu-2+) were dominant. These data suggest a different blood and tissue distribution of inducer and suppressor/cytotoxic cells in B-CLL, which may have important pathophysiologic significance.
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Two patients with B chronic lymphocytic leukemia having leukemic cells that bind sheep red blood cells by different mechanisms are described. In the first case, rosette formation was mediated by the anti-sheep erythrocyte activity of a monoclonal surface IgMk, related to Forssman antigen. In the second, E-rosette formation was found to be independent of both surface immunoglobulins and the classic E-rosette receptor since the leukemic cells were recognized neither by the OKT-11 monoclonal antibody nor by other markers specific to T-cell lineage. Evaluation of these rare cases emphasizes that detection of surface immunoglobulins and spontaneous rosetting are not sufficient for the characterization of leukemic clones and raises some doubts concerning the use of available surface markers in the characterization of lymphoproliferative disorders.
The enzyme glycyl-proline-naphthylamidase (dipeptidyl-amino-peptidase IV), (DAP-IV) has been histochemically analysed in normal and pathological specimens of different lymphoid tissues. In all the tissues examined, the enzyme appeared to be highly specific for T cells. Using chloroformacetone fixation and sections of proper thickness, DAP-IV could be successfully demonstrated in cortical thymocytes, which exhibited weak reactivity, as well as in medullary thymocytes which showed a more intense and variable reactivity. A similar reactivity was observed in T cells of peripheral blood, lymph node, and spleen. Various acute lymphoblastic and nonlymphoblastic leukemias and malignant lymphomas were studied using the histochemical method for DAP-IV. Malignant cells from all the acute leukemias and from the B-cell lymphomas were DAP-IV unreactive, while strong reactivity was observed in one case of T-CLL and one case of T-PLL. The possible relationship of DAP-IV positivity pattern with T-cell maturation and its potential use as a diagnostic tool in lymphoproliferative disorders are discussed.
Cryostat sections from bone marrow biopsies submerged in gum-sucrose solution and snap-frozen can be stained with various antisera to detect both the membrane and cytoplasmic antigens on normal and malignant populations in their tissue environments. This technique seems to be the method of choice to analyse the degree of BM involvement in cases of non-Hodgkin's lymphoma where conventional histology provides equivocal results. The uniform expression of only one type of light chain in B lymphoma and the analysis of the T cell infiltrate in T-CLL using monoclonal antibodies is demonstrated.
An antiserum to beta-thromboglobulin was used in immunofluorescence to detect normal and malignant megakaryocytes. In normal peripheral blood and bone marrow smears, only platelets and megakaryocytes were specifically stained by this antiserum. Among 25 cases of acute lymphoid and non-lymphoid leukaemia, only 2 exhibited a clear positivity in a % of blasts, thus proving their megakaryocytic origin. This method is proposed as a simple and useful tool to detect normal and malignant megakaryocytes on smears.
A recently developed procedure, that has been shown to be suitable for detailed immunohistological analysis, has been used to prepare cryostat sections of bone marrow to investigate whether enzyme-histochemical techniques are also feasible on such material. A selected group of enzymes, some of which are inhibited or destroyed in paraffin- or plastic-embedded samples, have been demonstrated. The morphological details obtained were satisfactory in the preparations. The enzymes were dipeptidyl(amino)peptidase IV (for T lymphocytes); tartrate-resistant acid phosphatase (for hairy cell leukaemia); acid phosphatase and non-specific esterase (for macrophages and monocytes); ATPase and 5'nucleotidase (for B lymphocytes); and peroxidase or chloroacetate esterase (for granulocytic cells). In these preparations strong enzyme activities were shown. In adjacent sections the immunological analysis of membrane markers could also be performed contributing to a comprehensive study of the normal and malignant bone marrow cells.
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Tissue sections of frozen biopsy specimens obtained from normal and hyperplastic human lymphoid tissues as well as 40 cases of non-Hodgkin's lymphomas were analysed in immunofluorescence tests (using red TRITC and green FITC double-labeling). A panel of antisera including well-characterized conventional reagents to immunoglobulin classes, T-lymphoid and Ia-like antigens and monoclonal antibodies, including the OKT range made by Ortho Laboratories, was used. The findings show that the immunological methods can give a very accurate analysis of the normal and malignant lymphoid cells and can provide complementary information to conventional histology. Furthermore, the monoclonal antibodies to well defined lymphocyte subsets can provide data which suggests (although does not prove) functional relationships between different types of cells in normal and malignant lymph nodes. Thus, the technology described fills the gap between conventional histology and cellular immunology.
So far, enzyme histochemical examination has been applied, with few exceptions, either to tissue imprints or to cryostat sections of freshly collected samples. This procedure is not easily applicable in routine histopathologic examination. In this study, a simplified tissue embedding procedure is presented which can be performed using an automatic tissue changer. The paraffin embedded samples can be used for both conventional histopathologic examination and for demonstrating enzymes in sections. The enzymes studied (alkaline phosphatase, alpha-naphthyl acetate esterase, acid phosphatase, tartrate resistant acid phosphatase, ATPase, peroxidase, and chloroacetate esterase) gave comparable results in formalin-fixed cryostat sections and paraffin sections in both normal and pathologic lymphoid samples. The only exception was ATPase, which could not be demonstrated on paraffin-embedded material. The technic described has broad application in the analysis of lymphoid diseases.
Indirect immunofluorescence for terminal transferase enzyme (TdT) was used to study the blasts of 64 patients with acute non-lymphoid leukaemia (ANLL). In 32 patients no TdT positive cells were seen. In 19 cases a small subpopulation of cells expressing TdT was detected; these constituted up to 5% of total nucleated cells, and it was not clear whether these TdT positive cells were part of the leukaemic process or represented residual normal bone marrow lymphoid cells. The remaining 13 patients had TdT positive cells accounting for 7-90% of the total. In two of these cases TdT was expressed on blasts with myeloid features, representing an aberrant expression of TdT by myeloid cells; in contrast, in three cases mixed populations of TdT positive lymphoid blasts and TdT negative myeloid blasts were observed. In the remaining cases it was not possible to determine whether the TdT positive cells had definite lymphoid or myeloid features. Cytogenetic analysis showed no evidence of the Philadelphia chromosome. Response to treatment was assessed in 11 of the 13 patients. Only one patient remitted with the initial choice of therapy (DAT); four failed to respond to initial regimes of vincristine and prednisone (V & P) while the other five patients did not respond to myelotoxic combinations (DAT). Only one patient subsequently entered complete remission on second line therapy (V & P). This group of patients with TdT+ ANLL had a particularly bad prognosis, and appeared to differ from cases of TdT positive acute undifferentiated leukaemia, which often respond to V & P therapy.
In three cases of acute myeloid leukaemia marked increases in expression of the nuclear enzyme, terminal deoxynucleotidyl transferase (TdT), were observed during disease relapse. The first case was heterogeneous at diagnosis, consisting of subpopulations of large TdT- myeloblasts and small TDT+ blasts; however, at relapse there was complete replacement by TdT+ lymphoblasts. The other cases at diagnosis were both typical acute myeloid leukaemia, TdT-; at relapse, one showed a mixture of TdT- myeloblasts and TdT+ lymphoblasts, while in the other, TdT was demonstrated on a subpopulation of myeloblasts. No chromosomal abnormalities were found. It is suggested that in the first two cases these phenomena may have been due to leukaemic involvement of a pluripotential stem cell, or that sub-clones with different properties may have coexisted. In the third case at relapse, TdT was expressed aberrantly by malignant myeloid cells.
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Alpha-Naphthyl acetate esterase (ANAE) and fluoride resistant alpha-naphthyl acetate esterase (FRANAE) have been compared as histochemical methods to identify T lymphocytes in sections of normal and pathological human lymphoid tissues. In addition, the FRANAE method was combined with alkaline phosphatase (ALP) in order to simultaneously evaluate the relationship between T lymphocytes and fibroblastic reticular cells (ALP) positive). The "dot like" esterase positivity of T lymphocyte was better evaluated by using FRANAE when compared to ANAE because of fluoride inhibitor of the strong esterase activity of dendritic cells and most macrophages. The combined ALP-FRANAE method clearly demonstrated a large number of fibroblastic reticular cells within the T-areas in various normal and pathological tissues such as hyperplastic lymph nodes and especially in the lymph nodes and spleens from patients with Hodgkin's disease.