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[Carcinosarcoma of the prostate. Immunophenotype, morphologic course and clinico-pathologic differential diagnosis].

The observation of a patient with a malignant tumor of the prostate initially classified as adenocarcinoma with desmoplasia, reclassified as adenocarcinoma with a malignant spindle cell component of uncertain cell line following two large tumor recurrences over a period of 4 months, and subsequently as carcinosarcoma with heterologous mesenchymal areas has prompted us to review the 11 reported cases of prostatic carcinosarcoma and the diagnostic algorithm of the proliferative processes of the prostate with elongated cells in order to recognize (from the histological features and the inclusion and exclusion immunophenotypes) the pseudosarcomatous stromal reactions, fusicellular changes of the carcinoma, benign and malignant phyllodes tumors and carcinosarcoma.

Adenocarcinoma↗

Distinctive immunophenotypic features of t(8;21)(q22;q22) acute myeloblastic leukemia in children.

Thirty cases of newly diagnosed pediatric acute myeloblastic leukemia (AML) with French-American-British (FAB) M2 morphology were analyzed with cytogenetics and a comprehensive panel of monoclonal antibodies reactive with lymphoid-, natural killer (NK)-cell-, and myeloid-associated antigens. The t(8;21)(q22;q22), or t(8;21;V)(q22;q22;V), translocation was identified in 16 of the 30 cases. Cases with the t(8;21) did not differ significantly from the remaining M2 cases with respect to expression of CD11b, CD13, CD14, CD15, CD33, CD34, CD36, CD41a, CD42b, CDw65, TdT, or HLA-DR. Expression of the B-cell antigen CD19 was detected in 13 of the 16 t(8;21) cases (81%), but in only 1 of the 14 (7%) other M2 cases (P = .00006). Expression of the CD56 NK-cell antigen was also significantly more frequent among t(8;21) cases (63% v 14%; P = .01). Coexpression of CD19 and CD56 was found only in the t(8;21) group (9 of 16 cases, P = .0009). Furthermore, this phenotype was not found in 48 evaluable cases of de novo AML of the FAB M1, M3, M4, M5, or M7 subtypes. The 14 M2 AML cases lacking the t(8;21) commonly expressed CD2 (n = 5) or CD7 (n = 8). However, no case with the t(8;21) expressed either antigen (P = .01 and .0005, respectively). Thus, the t(8;21) biologic subgroup of pediatric M2 AML has distinct immunophenotypic characteristics that distinguish it from other types of de novo AML.

Adolescent↗

[Study of the pathology, immunophenotype, etiology and genetic marker of NK/T-cell lymphoma].

OBJECTIVE: To study the NK/T-cell lymphoma, search for a more efficacious and simpler method and establish a standard guideline for distinguishing the NK-like T-cell lymphoma from the NK-cell lymphoma. METHODS: Thirty-four NK or T-cell lymphomas from the upper aerodigestive tract (n = 22), skin (n = 2), gastrointestinal (GI) tract (n = 2), lymph nodes (n = 7), and other sites (n = 1) were studied. Immunophenotype was analyzed by immunohistochemistry. In situ hybridization with EBER 1/2 RNA probes was performed. T-cell receptor (TCR)-beta and -gamma gene rearrangement was analyzed by polymerase chain reaction (PCR). RESULTS: Eighteen cases were positive for CD(56) and 16 for TIA-1 in 34 lymphomas cases. All tumor cells in the skin cases were positive for Ki-67. Epstein-Barr virus (EBV) mRNA was detected in 12 upper aerodigestive tumors including 9 of 12 nasal and 3 extranasal tumors. EBER was also detected in 1 of 2 skin lymphomas and both of the 2 GI lymphomas. Clonal TCR-beta and -gamma gene rearrangement was detected in 2 of 22 upper aerodigestive, all of the skin and GI lymphomas, and 6 of 9 nodal and other site lymphomas. CONCLUSION: Most upper aerodigestive NK/T-cell lymphomas are genotypically NK derivation, and a few belong to T lineage. However, NK-like T-cell lymphomas more frequently seen in skin and GI tract. Nodal NK-cell lymphoma are quite rare. These two kinds of lymphomas can only be diagnosed with additional immunohistochemical markers, EBER detection by ISH, TCR gene rearrangement or NK-cell receptors (NKRs) RNA detection. Detection of TCR rearrangement remains the important standard for the diagnosis of T-cell lymphoma.

Adolescent↗

[Immunophenotyping in diagnosis of chronic lymphoproliferative diseases].

With due respect to their many-year experience, the authors focused their attention on the peculiarities of implementation and registration aspects of the immunofluorescence method of immunotyping made in cell suspensions and in tissue imprints; additionally they made a system of a set of monoclonal antibodies (used for the purpose), which enable the differential diagnosis of reactive conditions related with various malignant lymphoproliferative diseases (LPD). It order to specify a nature of lymphoid cells it is suggested to undertake the immunophenotyping withing a gradually expanding set of monoclonal antibodies, which reflects different parameters of lymphoid cells like linear attributes, clonal characteristics, differential-diagnostic markers, functional status and proliferative activity. Typical marker phenotypes of lymphoid cells observed in the main B- and T-cell LPDs are described; a possibility is mentioned that there can be errors in interpreting the phenotyping results while diagnosing an LPD.

Diagnosis, Differential↗

[Comparison of clinical usefulness of immunophenotyping of leukemia using the immunofluorescence and immunoenzyme APAAP methods].

The clinical utility of the indirect immunofluorescence (IF) and the alkaline phosphatase-anti-alkaline phosphatase (APAAP) techniques was compared in 103 newly diagnosed acute leukaemia patients immunophenotyped using a panel of 19 monoclonal antibodies (MoAb). In spite of slight variations in the percentages of cells reacting with particular MoAbs when comparing the two methods we found no discrepancies in the final classification of each case. In ANLL (n = 73) the best correlation between the two methods was found for CDw65 which is a good screening marker, and for CD15 having a prognostic significance. In ALL (n = 30) the best correlation was observed for CD19 and CD10, both of great diagnostic importance. The following antigens present both in membrane and in cytoplasm displayed higher positivity with the APAAP than in IF HLA-Dr, CD71 and CD11b in ANLL, CD22 and HLA-Dr in nonT-ALL and CD3 in T-ALL. The important advantages of the APAAP technique are: 1) its use with routinely performed bone marrow or peripheral blood films, which can be stored before staining, 2) the possibility of correlating morphology with immunological characterization and documentation of the results.

Alkaline Phosphatase↗

Monoclonal gammopathy of undetermined significance: a morphologic and immunophenotypic study of the bone marrow.

A monoclonal paraprotein in the serum or urine raises the possibility of myeloma. However, in a significant proportion of individuals with serum paraproteins, particularly those with low levels of paraprotein, clinical and routine bone marrow evaluation is not diagnostic of an underlying neoplasm. The purpose of this study was to define the pathologic basis for monoclonal gammopathy in patients whose bone marrow biopsies showed no evidence of myeloma. We used immunofluorescence microscopy and flow cytometry of cell suspensions prepared from aspirated marrow, as well as immunohistochemistry of core biopsies, to perform immunopathologic evaluations of the bone marrow from 26 such patients. Eighteen patients with myeloma and seven without a serum paraprotein or evidence of myeloma were similarly studied. The data indicate that 17 of the 26 patients with monoclonal paraproteins whose routine bone marrow biopsies were normal or nondiagnostic had, in fact, a dispersed monotypic plasma cell population of concordant immunoglobulin heavy and light chain type in the bone marrow demonstrable by at least one of the three analytic methods. Among these, immunofluorescence microscopy of isolated bone marrow mononuclear cells was the most sensitive assay. Immunophenotypic evaluation of the bone marrow is useful for documenting and quantifying a monoclonal plasma cell population in patients with monoclonal gammopathy of undetermined significance.

Aged↗

Structural and immunophenotypic characterization of high endothelial venules in rat and human tissue.

OBJECTIVE: To present additional data on high endothelial venule (HEV) structure and immunophenotype. METHODS: We used the zinc iodide-osmium tetroxide technique (ZIO), which is a metallophilic fixation and staining technique to examine HEVs at light and electron microscopic levels as this technique was previously reported to be reactive with cells in HEVs. Tonsils and lymph nodes were obtained from the Surgery and Otorhinolaryngology Departments, Hacettepe University Hospital, Ankara, Turkey during 2002 and 2003. An indirect immunohistochemical technique was used to examine frozen human tissue samples. RESULTS: Organelle rich high endothelial cells, sheet-like processes of pericytes surrounding HEVs, structural relation of pericyte processes with fibroblastic reticular cells, an unusual multivesicular body-like organelle within high endothelial cells were presented. Expression of a large panel of defined and yet non-defined antigens on HEVs are also presented using an indirect immunoperoxidase technique. CONCLUSION: Presence of some of these antigens on HEVs was previously reported while no previous report is available for others. Significance of the expression of these antigens in HEVs, structural hints for trans endothelial migration of lymphocytes and their travel along the reticular cell meshwork is briefly discussed.

Animals↗

[Immunophenotypic analysis in acute leukemia].

Acute leukemias have been classified on French-American-British (FAB) criteria depending on the morphocytochemical features of blasts. Immunophenotyping and clonal rearrangement analysis of lineage-associated genes can decide a frozen stage of the hematopoietic differentiation process in blasts from acute leukemias. B-lineage acute lymphoblastic leukemia (ALL) and T-lineage ALL are systematically classified according to the sequential expression of differentiation-associated antigens. In acute myelogenous leukemia (AML), several new entities are proposed: AML-M0 is an AML without cytologic maturation, in which the myeloid commitment should be demonstrated by myeloperoxidase-positive microgranule on immunohistochemical staining or electron-microscopy, or by positive reaction for CD13 or CD33 antigens. CD7-positive AML is considered to be one of immature subtypes of AML, rather than hybrid leukemia. Thus, immunological studies on blasts enable us to discriminate a subgroup of leukemias, which will perhaps contribute to the improvement of treatment approach.

Acute Disease↗

[Immunophenotype of chronic lymphocytic leukemia and prolymphocytic leukemia].

The immunophenotype of chronic lymphocytic leukemia (CLL) and prolymphocytic leukemia (PLL) is described, mainly according to the FAB classification. CLL and related diseases have been recognized as highly variable diseases and their classification is still confusing. Is there true T-CLL? How should we deal with non-T, non-B-CLL such as NK-GLL? What about the patient with borderline manifestations? Is the concept "MLUS" appropriate? Further examination and more detailed analysis is required in order to clarify the disease entity of CLL and related disorders.

Antigens, CD↗

Kinetics and immunophenotypic characterization of circulating hematopoietic progenitor cells after peripheral blood stem cell transplantation.

BACKGROUND AND OBJECTIVES: Hematopoietic progenitor cells (HPC) circulate in the peripheral blood (PB) before and after engraftment following autologous or allogeneic peripheral blood stem cell transplantation (PBSCT), although the characteristics of these cells are not known. CD34 protein is a reliable marker for identifying the fraction of hematopoietic cells in which HPC are contained. The CD34(+) cells represent a heterogeneous cell population consisting of both primitive uncommitted as well as pluripotent committed progenitors. The aim of this study was to investigate the kinetics and immunophenotypic characteristics of these post-transplant circulating progenitor cells. DESIGN AND METHODS: Forty-seven auto-PBSCT and nine allo-PBSCT recipients were selected for this study. Samples of PB were taken from each patient 4, 9, 11, 14, 16 and 18 days after the transplant. Cells were incubated with the following combinations of monoclonal antibodies: CD34-FITC/CD90-PE/CD38-CyCrome; CD34-FITC/CD117-PE/HLA-DR-PerCP; CD34-FITC/CD13-PE/CD33-CyCrome and the cells were then analyzed by flow cytometry. RESULTS: CD34(+) cells were undetectable on day +4; they reappeared from day +9 to day +18 along with neutrophil and platelet recovery. Subsets of CD34(+) HPC enriched in pluripotent stem cells (CD90(+)/CD38(low) or HLADR-) were hardly detected during the very early post-transplant period. HPC that expressed myeloid associated antigens (CD33, CD13, and CD117) increased after engraftment and constituted the largest proportion of the hematopoietic progenitor cells. INTERPRETATION AND CONCLUSIONS: Circulating HPC could be detected in the early period after PBSCT. The qualitative and quantitative composition of these cells is similar to that found among HPC from mobilized PB.

Adolescent↗

[Morphological characteristics and immunophenotype of dendritic cells generated from monocytes of human peripheral blood].

Morphological features and immunophenotype of mature dendritic cells (DC) generated from the monocytes of healthy donor blood after culture with GM-CSF and IL-4 with the addition of TNF, were studied using light and electron microscopy, as well as flow-cytometry. It was shown that DC were characterized by a number of morphological features such as: large size, eccentrically located nucleus, highly developed system of extensions, large vacuolated cytoplasm, prominent and activated Golgi complex and ribosomal apparatus. Mature DC are characterized by active surface expression of MHC and co-stimulatory molecules (MHC I, MHC II, CD40, CD80, CD86).

Antigens, CD↗

[Adult T-cell leukemia with cervical bone tumor showing immunophenotypic change].

A 60-year-old man born in Miyazaki prefecture was admitted to our hospital complaining of skin rash in December 1989. On hematological examinations, leukocyte count was 14,200/microliters with 49% of abnormal lymphocytes showing lobulated nuclei. The surface marker study revealed their phenotype as CD4+8-. Anti human T cell leukemia virus type I (HTLV-I) antibody and monoclonal integration of proviral DNA were positive. From the above results, he was diagnosed as adult T-cell leukemia (ATL). Abnormal lymphocytes gradually decreased without treatment after the first admission. In January, 1990, he began to complain of neck pain. Two months later he was readmitted because of paresis of extremities and disturbance of urination. Vertebral bone mass and a compressed spinal cord in the 4th cervic level were confirmed by MR imaging. He received a resection of tumor and an anterior fusion of vertebrae. The bone tumor was histologically diagnosed as malignant lymphoma, diffuse medium-size cell type and the infiltrating cells had their phenotype as CD4+8+. He was postoperatively treated with combination chemotherapies, but neurological abnormalities did not improve. He died of pneumonia on 35 days after the operation. A postmortem examination revealed extradural tumor formation with ATL cells. This case is considered to be rare in respect of both the disappearance of most peripheral abnormal lymphocytes without any treatments and the cervical bone tumor showing immunophenotypic change.

CD4 Antigens↗

[Immunophenotype of lymphoblastic leukaemia in children in relation to clinical symptoms and laboratory tests, preceding its diagnosis].

The aim of study was to compare the clinical picture and results of laboratory tests according to the acute lymphoblastic leukaemia (ALL) immunophenotype. The observation was carried out on a group of 67 patients treated in the IIIrd Department of Paediatrics and Department of Children Oncology in the Medical Academy of Białystok from January 1994 to April 2001. This group consists of 4 children with pro-B acute lymphoblastic leukaemia, 52 children with pre-B cell ALL, 1 child with B-cell acute lymphoblastic leukaemia and 9 children with T-cell acute lymphoblastic leukaemia. Haemorrhagic diathesis. splenomegaly, enlargement of peripheral lymph nodes as well as higher values of white blood cells count, blasts count, haemoglobin concentration, haematocrit and LDH activity were observed more frequently in patients with T-cell leukaemia than in others.

Blood Cell Count↗

Morphologic and immunophenotypic diversity in Ewing family tumors: a study of 66 genetically confirmed cases.

More than 85% of Ewing sarcoma/primitive neuroectodermal tumor (ES/PNET), or "Ewing family of tumors" (EFTs), have the translocation, t (11;22) (q24;q12), with others having variant translocations. Identification of these by cytogenetic and/or molecular genetic techniques is specific for EFT and is increasingly recognized as the "gold standard" for diagnosis. However, these techniques are not universally available. We therefore studied a large group of genetically confirmed EFTs to more completely understand the morphologic and immunophenotypic spectrum of this rare sarcoma. Sixty-six cytogenetically, FISH or RT-PCR proven-EFTs were retrieved. In 56 cases, immunohistochemistry (IHC) was performed for pan-cytokeratins (PanCK), high molecular weight cytokeratins (HMWCK), desmin (DES), CD99, CD117, and FLI1 protein using heat-induced epitope retrieval and the Dako Envision system. The cases arose chiefly in children and young adults (median 18 years; range, 3-65 years) of both sexes (male, 32; female, 31; unknown, 3) in a variety of bone (N = 39) and soft tissue (N = 27) sites. Histologically, 46 cases (73%) showed only typical features of ES, 9 cases (16%) showed features of PNET, 3 cases (5%) showed "adamantinoma-like" features, 3 cases (5%) corresponded to "atypical Ewing sarcoma," 3 cases (5%) showed principally intersecting fascicles of spindled cells, and 2 cases had abundant hyalinized matrix. IHC results were as follows: PanCK (18 of 56, 32%), HMWCK (3 of 55, 5%), DES (1 of 56, 2%), CD99 (52 of 52, 100%), CD117 (13 of 54, 24%), and FLI1 (44 of 47, 94%). HMWCK was expressed only in "adamantinoma-like" EFTs, none of which expressed DES. In conclusion, most, but not all, EFTs can be accurately diagnosed using time-honored morphologic criteria and ancillary immunohistochemistry. However, genetic confirmation remains essential for the diagnosis of unusual morphologic variants of EFT, including "adamantinoma-like," spindled, sclerosing, and clear cell/anaplastic variants. Therefore, to exclude or confirm the diagnosis of Ewing's sarcoma in round cell sarcomas having a variety of patterns but not specifically conforming to a tumor of known lineage (eg, rhabdomyosarcoma), cytogenetics, and/or molecular analysis is required.

12E7 Antigen↗

Immunophenotyping characteristic of the major lymphocyte populations and subpopulations in patients during salmonella infection.

AIM: To study the quantitative changes in the major lymphocyte populations and subpopulations in the peripheral blood of patients during salmonellosis and find correlations of these changes with disease severity and bacterial clearance. MATERIAL AND METHODS: The study included 24 adult patients with culture-proven gastrointestinal salmonellosis. Flow-cytometry was used to identify CD19+ (B lymphocytes), CD2+ (total T lymphocytes), CD3(+)CD4+ (helper T cells), CD3(+)CD8+ (suppressor/cytotoxic T lymphocytes), CD4(+)CD29+ and CD4(+)CD45(-)RA+ (helper/ inducer subpopulation and naive Tlymphocytes) in the acute and the convalescent phases of disease. The absolute number and percentage of cells in 1 microl of peripheral blood were also determined. Immunophenotype analysis was conducted on an EPICS XL-MCL flow cytometer, Coulter, USA using monoclonal antibodies produced by the same firm. RESULTS: T and B lymphocytes and the immunocompetent T cells were slightly decreased transiently in the acute phase of the disease. Helper T lymphocytes were slightly increased with a significant increase observed of helper/inducer cells and decrease of the naive T lymphocytes. CONCLUSION: The increase of B lymphocytes at the height of salmonellosis bears additionally a diagnostic significance in determining the severity of the disease while the increase of the helper T lymphocytes can be a prognostic marker of early bacterial clearance.

Adolescent↗

[The virogenic status of cultured continuous simian lymphoid cells and their immunophenotypical characteristics].

Immunophenotyping of cultured continuous simian lymphoid cultures using a panel containing monoclonal antibodies to human B- and T-cell antigens has been carried out, by employing enzyme immunoassay and flow cytofluorometry. The test cultures showed a wide variety of cells containing B- and T-cell antigens with their varying expression. The cultures were also found to comprise lymphoblasts simultaneously containing B- and T-cell markers. Simian lymphotropic viruses, such as EBV-like and STLV-1 retrovirus, detected apart or simultaneously, have been verified by polymerase chain reaction. Whether there is a possible relationship between the type of cells in the culture and the type of their replicating virus(es) is discussed in the paper.

Animals↗

Immunophenotypic heterogeneity of bone marrow-derived mesenchymal stromal cells from patients with hematologic disorders: correlation with bone marrow microenvironment.

The immunophenotypic analysis of ex vivo-expanded mesenchymal stromal cells (MSC) has so far been confined to single or dual staining analysis in normal subjects. In this study, using a four-color cytofluorimetric protocol, we demonstrated that cultured MSC derived from the bone marrow of patients with hematologic malignancies showed alterations in the expression of CD105, CD90, CD184, and HLA-DR molecules. The decrease in the percentage of CD105+ and CD90+ MSC correlated with an increased bone marrow angiogenesis. This paper provides evidence that multiparametric flow cytometry is essential for the establishment of a standardized protocol to identify various MSCs subsets and aberrant phenotypes.

Adult↗

Prognostic significance of immunophenotype in aggressive non-Hodgkin's lymphoma.

The purpose of the study was to evaluate prognostic value of immunophenotype in aggressive Non-Hodgkin's lymphoma. 87 patients with immunohistologically confirmed diagnosis of aggressive Non-Hodgkin's lymphoma according to the WHO classification (2001) were under observation. Patients were treated at the Department of Hematology and Transfusiology, State Medical Academy, Tbilisi, Georgia. The IPI was calculated for each case. Overall survival was calculated from the date of diagnosis to the last follow-up or death regardless of the cause. 17 patients out of 87 were diagnosed as T-cell lymphomas. Overall survival in patients with IPI greater than 2 (poor prognosis) was shorter for T-cell lymphomas (7,6 months) in comparison with B-cell lymphomas (17,3 months) (p<0,001). T-cell phenotype should be considered as an independent factor that strongly influences the survival for patients with diagnosis of aggressive non-Hodgkin's lymphomas. Besides petipherial T-cell lymphomas occurs more frequently in the elderly, with advanced stage, frequent extranodal site involvement, and often detected high level of LDH compared with B-cell lymphomas.

Adult↗