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The plasmacytoid T cell or plasmacytoid monocyte--a sessile lymphoid cell with unique immunophenotype and unknown function, still awaiting lineage affiliation.

Clusters of cells with a plasmacytoid appearance have been identified in T zones of human lymphoid tissue. These cells were shown to express the CD4 antigen in the absence of B-cell antigens. Thus, they were named "plasmacytoid T cells" (PTCs). Recent studies, however, have suggested a myelomonocytic origin of this cell type, which led to the designation "plasmacytoid monocytes." In order to obtain a comprehensive immunophenotype of this cell, we used frozen serial sections of lymph nodes, an indirect immunoperoxidase technique, and a broad panel of well-defined monoclonal antibodies (mAbs). On PTCs, the antigenic density was high for CD4 and CD26 antigens and low for CD2; they furthermore reacted with the mAb OKM5 recognizing an undefined antigen associated with the myelomonocytic lineage and expressed the monocyte-associated antigen CD68. PTCs expressed HLA-A, B, C, HLA-DR, and the HLA-D associated invariant chain (Ii) while HLA-DP antigens were expressed only in low amounts and HLA-DQ antigens were lacking entirely. PTCs further expressed the adhesion molecule CD11c. The absence of detectable transferrin receptor and the proliferation antigen defined by Ki-67 indicated that PTCs were nonproliferating. Other well-defined antigens confined to the T-, B-, and myelomonocyte lineages could not be detected in the target cell population. For us, antigenic equipment is still suggestive of a sessile, terminally differentiated T cell likely to exert a secretory function as yet unknown.

Antibodies, Monoclonal↗

Morphologic and immunophenotypic heterogeneity of corneal dendritic cells.

The morphology, distribution and immunophenotype of corneal dendritic cells was investigated in frozen sections of normal human cornea using a panel of monoclonal antibodies (mAbs). Throughout the cornea, epithelial dendritic cells were labelled with mAbs LN2 and LN3, directed to HLA-DR-associated antigens. In the peripheral corneal and limbic epithelium, dendritic cells with long cytoplasmic processes in a suprabasal position were identified as being OKT6+, whereas in the central corneal epithelium, dendritic cells with short processes were located basally and corresponded to the OKT6- phenotype. Throughout the corneal stroma, LN2+ LN3+ OKT6- spindle cells were found. This study discloses that corneal dendritic cells display considerable phenotypic and morphologic heterogeneity. We suggest that this heterogeneity is partly due to local inductive effects of peripheral corneal basal cells.

Antibodies, Monoclonal↗

Species differences in the immunophenotype of osteoclasts and mononuclear phagocytes.

Human osteoclasts, in contrast to mononuclear phagocytes, are known to express a well-defined restricted range of myeloid antigens. To determine whether these antigenic differences are present in other species, we examined the immunophenotype of chicken and rabbit osteoclasts, macrophages, macrophage polykaryons, and monocytes and compared them with similarly derived and cultured human cells. Human, rabbit, and avian osteoclasts reacted with monoclonal antibodies against human beta 1 integrins (CD29, CD49b, CD49d), beta 3 integrins (CD51, CD61), as well as human macrophage-associated antigen CD68. Avian osteoclasts also reacted for CD11a/18 and CD14 which are not present on human osteoclasts. Avian and mammalian monocytes, macrophages, and macrophage polykaryons expressed all the above antigens. Both avian and human macrophage polykaryons produced by culture of peritoneal macrophages reacted with anti-CD51 antibodies indicating that expression of the vitronectin receptor alone does not distinguish between these cells in vitro.

Animals↗

Histiocytosis X arising in Hodgkin's disease: immunophenotypic characterization with a panel of monoclonal antibodies.

This report describes the antigenic profile of the proliferating cells of pulmonary histiocytosis X (HX) in a patient treated with chemotherapy for Hodgkin's lymphoma; the association of pulmonary HX and Hodgkin's disease has rarely been described in the literature. The histopathological diagnosis of HX was confirmed with the aid of monoclonal antibodies (mAbs) to CD4, CD1a, and polyclonal serum anti S-100 protein. The phenotype of HX cells has been analysed using a panel of mAbs against HLA class I A, B, C monomorphic determinants, locus A and B, beta 2-microglobulin, HLA class II distinct monomorphic determinants, DP, DQ, DR, intercellular adhesion molecule-1 (ICAM-1) and vitronectin receptors. Our results indicate that HX cells express HLA class I and II, including locus A, locus B and DP, DQ, DR, like their normal counterpart (represented by Langerhans cells) and detectable levels of ICAM-1 but not vitronectin receptors. We would like to stress the possibility of the association of HX and Hodgkin's lymphoma extending the immunophenotypic profile of HX cells.

Adult↗

Large cell anaplastic lymphoma: evaluation of immunophenotype on paraffin and frozen sections in comparison with ultrastructural features.

Eleven cases of large cell anaplastic lymphoma (T type n = 5, B type n = 4, 0 type n = 2) were investigated using electron microscopy and immunophenotyping on formalin-fixed paraffin sections and frozen sections of fresh tissue, to determine whether morphological criteria exist for the discrimination of T, B, and 0 phenotypes. Tumour cell lineage could not be established from ultrastructural features. On paraffin material monoclonal B-cell markers Ki-B5 and L-26 served as reliable tools for recognizing the B phenotype of large cell anaplastic lymphomas (previously determined on fresh material). whereas monoclonal antibodies MT1 (CD43) and UCHL1 (CD45RO) were of limited value in lineage determination.

Adult↗

Morpho-immunophenotypic diversity of Castleman's disease, hyaline-vascular type: with emphasis on a stroma-rich variant and a new pathogenetic hypothesis.

A histologic review of 102 cases of Castleman's disease of the hyaline-vascular type, with a detailed paraffin immunophenotypic study of 23 of them, was undertaken to evaluate the morphologic variability of this disorder and its immuno/cyto-architectural characteristics. All cases had features in common including: abnormal follicles, with increased vascularity, poorly formed germinal centers and predominance of the mantle zone; lack of sinuses; and hypervascular interfollicular tissue containing large numbers of KP1-positive plasmacytoid monocytes. Networks of actin-positive cells [fibroblastic reticulum cells (RCs) or "myoid cells"] and KP1-positive dendritic cells (histiocytic RCs) were seen. There were differences in the proportion of follicles to interfollicular tissue, which covered a continuum from a "follicular", through a "classic", to a "stroma-rich" variant. The last-mentioned was qualitatively different as it showed loss of HECA-452 and MECA-79 reactivity in the blood vessels, decreased plasmacytoid monocytes and increased myoid cells and histiocytic RCs. In 5 cases there was formation of distinct nodular growths which varied from spindle cell foci to angio-histiocytic-RC proliferations, all of which may be confused with vascular or follicular dendritic RC neoplasms. From our findings, data from the literature and the working hypothesis that plasmacytoid monocytes are the precursors of both follicular dendritic RCs and sinus lining cells (Parwaresch et al.), a pathogenetic theory is proposed for this type of Castleman's disease which postulates that a developmental block in plasmacytoid monocytes results in their accumulation with poor formation of germinal centers and sinuses under stimulation. The lack of sinuses would lead to impaired egress of circulating lymphocytes which, however, would continue to enter the node through functional high endothelial venules and to accumulate in the mantle zone. The factors responsible for angiogenesis and for the cellular growths that characterize the stroma-rich variant remain to be determined, as do the relationships between the three variants.

Adolescent↗

Correlation between immunophenotypic diversity and clinical features in B-cell lymphoblastic lymphoma.

Lymphoblastic lymphoma (LBL) is a highly malignant subtype of non-Hodgkin's lymphoma (NHL) and generally carries a T-cell phenotype with mediastinum or central nervous system (CNS) involvement. However, only a small proportion of LBL exhibit a B-cell phenotype (B-LBL), and these frequently present at the head and neck without mediastinum or CNS involvement. Three immunological subgroups may exist. The most predominant CD10-positive pre-B-cell type, corresponding to a precursor B-cell neoplasm, frequently involves the head and neck. The second, CD10-negative or mature B-cell type, defined by the absence of CD10 or presence of surface membrane immunoglobulins combined with expression of CD19 or CD20, often involves the mediastinum. The final group is a CD5-positive B-cell type corresponding to a blastic variant of mantle cell lymphoma (MCL). Its clinical course is less aggressive, patients are often older, and nodal lesions are more frequent than extranodal involvement. Thus, B-LBL is immunologically diverse, but its biological behavior correlates with the immunophenotype.

Adolescent↗

Spontaneous splenic rupture as the initial manifestation of acute lymphoblastic leukaemia: immunophenotype and cytogenetics.

Spontaneous splenic rupture is a rare, though life-threatening complication of some hematological malignancies and is even more infrequent as the initial symptom of acute lymphoblastic leukemia. We describe the fourth case, to our knowledge, of acute lymphoblastic leukemia presenting as splenic rupture, and for the first time the immunophenotype and cytogenetic pattern observed.

Adult↗

Intrathymic lymphoid cell differentiation in myasthenia gravis: an immunophenotypic study.

Thymocytes express multiple different surface antigens according to their stage of maturation. We studied lymphocyte surface differentiation antigens using direct immunofluorescence technique in the thymus of 20 patients with myasthenia gravis (MG) and 10 controls undergoing cardiac surgery. Fluorescein isothiocyanate-conjugated monoclonal antibodies were used to stain thymic cell surface antigens. We found a decrease in the percent expression of CD1, CD5, and CD7 surface antigens and a significant increase of CD20+ cells in myasthenic thymus compared with the controls. The changes in the percent expression of CD3, CD4, and CD8 antigens were not significant. These data suggest both the decrease in the immunophenotypes corresponding to cortical thymocytes (probably reflecting the cortical atrophy of the MG thymus) and the increase of mature B Cells (CD20+), which may participate in an active immune response.

Adult↗

Immunophenotypings of malignant epithelial mesothelioma and their roles in the differential diagnosis.

To investigate the immunophenotypings of malignant epithelial mesothelioma (MEM), and to seek the valuable markers in distinguishing peritoneal MEM from peritoneal metastatic ovarian adenocarcinoma (OA) and colorectal adenocarcinoma (CA), immunohistochemical SP method was used to detect expressions of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 in paraffin-embedded tissues of 18 cases of MEM, 20 OA and 20 CA. The results showed that there was a significant difference in the expressions of E-cadherin, CA19-9 and MOC-31 between MEM and OA group (P<0.05). Similarly, the difference in the expression of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 between MEM and CA groups is significant (P<0.05). These results indicate that HBME-1 could be used as a positive marker in distinguishing MEM from CA. E-cadherin, CA19-9 and MOC-31 are considered to be useful negative markers in diagnostic distinction between MEM and metastatic adenocarcinomas, including OA and CA. CK7 is the best positive marker in distinguishing MEM from CA, but this marker appears to be valueless in discriminating MEM from OA.

Adenocarcinoma↗

Immunophenotype of thymoma-associated lymphoid cell component of T-cell type. A new analytic procedure in keeping with structural heterogeneities.

The phenotype of the lymphoid cell component of 35 thymomas was investigated by analyzing cryostat sections and lymphocyte suspensions. The morphology in each case was determined by examining multiple tissue samples from different parts of the tumor. Structural heterogeneity was shown in 14 thymomas, and a homogeneous morphology of cortical or medullary or mixed types in the others. To assess whether this heterogeneity was correlated with differences in the lymphoid phenotype, we analyzed both lymphocyte suspensions and frozen sections from the same samples. Phenotypical differences in the suspensions of each thymoma in the heterogeneous group were noted and similar differences were also observed in the cryostat sections. Phenotypical abnormalities were found in some thymomas. They consisted of the simultaneous expression of cortical and medullary markers, which was most marked in the heterogenous mixed-type thymomas invading the lung. Furthermore, the global phenotype was tested on a pool of lymphocyte suspensions in all thymomas. This procedure distinguished cortical, medullary and intermediate cortico-medullary immunophenotype models which closely correlated with the tumor histology. It was concluded that, due to the frequent structural and immunological heterogeneity of thymomas, correct assessment of their lymphoid component requires a two-step analysis. This comprises: 1) individual suspensions from samples taken from different areas of the same thymoma, and 2) a pool of these suspensions. The first step will reveal the different immunological characteristics. In the second, the lymphocyte phenotype, which may vary widely throughout the tumor, will be represented in its totality. These findings may be of great help in predicting clinical patterns, especially possible malignant evolution.

Cryopreservation↗

Immunophenotyping of insulitis in control and essential fatty acid deficient mice treated with multiple low-dose streptozotocin.

Multiple injections of low-dose streptozotocin induce lymphocytic insulitis and autoimmune diabetes in male CD-1 mice. Prior to the onset of insulitis, macrophages infiltrate the islets (single cell insulitis) and presumably help initiate the lymphocytic response directed at streptozotocin-induced neoantigens on islet beta cells. Essential fatty acid deficiency ameliorates the lymphocytic insulitis and prevents diabetes in this model. We hypothesize that essential fatty acid deficiency, which perturbs eicosanoid pathways and blocks the production of inflammatory mediators such as leukotriene B4, might prevent or diminish the single cell insulitis and, thus, abrogate the lymphocytic response. The purpose of the study was to determine whether essential fatty acid deficiency causes any differences in the immunophenotype or the time course of single cell insulitis or insulitis after low-dose streptozotocin. Three to five essential fatty acid deficient and 3-5 control mice were treated with low-dose streptozotocin and killed on days 0, 3, 5, 8, 10, 12 and 15. Frozen sections of the pancreata were stained using an immunoperoxidase method with antibodies against mouse macrophages, CD4T-lymphocytes and CD8 T-lymphocytes. Sections were assessed for the presence and severity of single cell insulitis and insulitis. Based on cell counts in the most severely involved islet from each pancreas, there was no difference in the single cell insulitis in control and essential fatty acid deficient mice. Islets from control pancreata had a higher mean grade of lymphocytic insulitis. These findings suggest that autoimmune diabetes following low-dose streptozotocin in control mice is the result of both lymphocytic and histiocytic infiltrates with subsequent beta-cell destruction. Our results do not support the hypothesis that the protective effect of essential fatty acid deficiency is due to diminished influx of macrophages into the islets. It is, however, possible that essential fatty acid deficiency deleteriously affects macrophage function and, thus, blunts the lymphocytic response.

Animals↗

Association between trisomy 8 and the immunophenotype of blast cells from acute leukemias secondary to a myelodysplastic syndrome or chronic myeloproliferative disorders.

In the present study we have used FISH to analyze the incidence of trisomy 8 in acute leukemias following either a primary myeloproliferative disorder (MPD) or a myelodysplastic syndrome (MDS) and correlated it with both the immunophenotype and the cell-cycle distribution of the leukemic blast cells. Six of the 21 (28%) acute leukemias studied displayed trisomy 8 by FISH. The number of trisomic cells in these cases ranged from 20 to 84%, with a mean of 46 +/- 24%. Trisomy 8 was associated with a homogeneous population of leukemic cells, phenotypically characterized by CD34+/HLADR+/CD13+/CD33+/CD11b-/ CD15-/CD14-. No significant differences were observed on the proliferative rate of cases with trisomy 8, as compared with blast cells from the remaining patients. Overall, our findings suggest that in acute leukemias secondary to MPD or MDS, trisomy 8 is associated with a blockade of myeloid maturation at an early step of the differentiation process.

Acute Disease↗

[Primary brain lymphoma in acquired immunodeficiency syndrome. Immunophenotype and molecular pathologic characterization in stereotactic biopsy, autopsy and cerebrospinal fluid cytology].

In this study we present morphological, cytological, immunophenotypical, and molecular genetic features of ten cases of AIDS-related primary brain lymphomas obtained as sterotactic biopsies, autopsy specimens, or cerebrospinal fluid samples. Histologically, a very characteristic perivascular and diffuse lymphomatous spread in the brain parenchyma was observed. By applying a highly sensitive in situ hybridization technique (ISH) using digoxigenin-labeled antisense riboprobes, abundant Epstein-Barr virus (EBV)-encoded small nuclear RNAs (EBER) transcripts could be demonstrated in each case studied. A combination of ISH with immunohistochemical staining for latent membrane protein (LMP-1) and EBV nuclear antigen-2 (EBNA-2) showed different patterns of EBV latency with a predominance of the broad spectrum (EBER+/EBNA-2+/LMP-1+). Clonal rearrangements of immunoglobulin heavy chain (IgH) genes were demonstrated in four cases using a sensitive polymerase chain reaction. In one patient the rearrangement pattern suggested biclonal lymphoproliferation. Our observations confirm the implication of the EBV in the etiology of AIDS-related primary malignant lymphomas of the brain. The demonstration of EBV gene products and clonal IgH rearrangements even in small biopsy samples is a useful tool for distinguishing between reactive and neoplastic lymphocytic infiltrates, which are frequently observed in brain and cerebrospinal fluid specimens obtained from AIDS patients.

Adult↗

Secondary glioblastoma with advanced neuronal immunophenotype.

We describe an unusual progression of astrocytoma into secondary glioblastoma exhibiting advanced neuronal immunophenotype. A tumor of the left frontal lobe of a 35-year-old man was diagnosed as astrocytoma. The tumor was treated by partial removal with postoperative chemoradiotherapy, followed by extensive removal of the residual regrowing tumor 5 month later. A secondary tumor was discovered and partially resected 8 years later, but the patient died 11 months following the operation due to extensive tumor progression showing subarachnoidal and intraventricular dissemination. The secondary tumor was small cell-predominant, highly proliferative tumor with an extremely high MIB-1 labeling index (80%). Unexpectedly, most of the tumor cells were positive for neuronal markers (synaptophysin and NeuN), but not for glial fibrillary acidic protein (GFAP). Retrospective examination of the original tumor revealed not only diffuse GFAP expression, but also neuronal marker expressions in small numbers of tumor cells that were hard to discriminate from the other cells on hematoxylin-eosin (HE) stain. This way of malignant progression of astrocytoma was quite unusual. Although the secondary tumor was classified as glioblastoma according to World Health Organization (WHO) classification (2000), it might be categorized into new variants of malignant glioneuronal tumors proposed recently.

Adult↗

Utility of immunophenotypic and immunogenotypic analysis in the study of necrotic lymph nodes.

We report a case of complete lymph node necrosis. No specific aetiology could be determined by morphology, but a B lymphoid population and clonal rearrangement of the immunoglobulin heavy chain gene were demonstrated in immunophenotypic and immunogenotypic studies performed using DNA extracted from paraffin embedded necrotic tissue. In the setting of lymph node necrosis, we suggest that immunohistochemical and gene rearrangement studies may provide additional diagnostic information.

Female↗

Isolation of human periosteum-derived progenitor cells using immunophenotypes for chondrogenesis.

Periosteum-derived progenitor cells (PDPCs) were isolated by characteristic surface markers. Reproducibility of immunophenotypes of the PDPCs was characterized by flow cytometric analysis using fluorescence-activated cell sorter (FACS). SH2(+), SH3(+), SH4(+), CD9(+), CD90(+) and CD105(+) were important eternal characteristic cell surface markers for the PDPCs. The characterized PDPCs maintained their chondrogenic potential in pellet cultures until the 15th passage from primary cell culture.

Cell Membrane↗

Urticaria pigmentosa and mastocytosis: the role of immunophenotyping in diagnosis and determining response to treatment.

Recent advances in our understanding of mast cell biology and disease resulted in identification of important differences in expression of mast cell surface antigens in normal and neoplastic mast cells. Most notably, detection of aberrant expression of CD25 and CD2 on the surface of neoplastic mast cells but not on their normal counterparts leads to the inclusion of this immunophenotypic abnormality in the World Health Organization's diagnostic criteria for systemic mastocytosis. Aberrant mast cell surface marker expression can be detected in the bone marrow aspirate by flow cytometry, even in patients with limited disease that lacks histopathologically detectable aggregates of mast cells in bone marrow biopsy sections. Flow cytometric analysis of bone marrow mast cells is therefore a sensitive method of diagnosis of mast cell disease and is expected to find increasing use in determining response to emerging mast cell cytoreductive therapies.

Biomarkers, Tumor↗