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Results for “Immunophenotyping”

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PubMed1990

Histopathologic, immunophenotypic, and genotypic analysis of Ki-1 anaplastic large cell lymphomas that express histiocyte-associated antigens.

CD30/Ki-1 antigen expression in 243 cases of malignant lymphomas was examined using Ber-H2 monoclonal antibody. Among them 20 cases were categorized as Ki-1 anaplastic large cell lymphoma. In two of these cases histiocyte-associated markers were also expressed. In these cases histopathologic and extensive in situ immunophenotypic analyses were used with genotypic studies in the determination of cell lineage. A sinusoid histologic pattern of involvement with partial lymph node infiltration by pleomorphic neoplastic cells was noticed in the nodes from both patients. Solid areas of node replacement resembling metastatic carcinoma were seen in Patient 1. Immunohistologically, tumor cells of both cases were positive for CD30, CD25, CD71, LN3 (HLA-DR), EMA, CD45, CD74, vimentin, alpha-1-antichymotrypsin, and CD68. Patient 1 was also CD45RO+, CD43+, whereas Patient 2 was positive for alpha-1-antitrypsin and CD4 tumor cells. Genotypic studies revealed that TCR beta and TCR gamma chain genes were clonally rearranged in Patient 1, whereas no rearrangements were detected in Patient 2. This study supports the view that some Ki-1 anaplastic large cell lymphomas may express multiple histiocyte-associated antigens and confirms that this group of neoplasms have immunophenotypic heterogeneity. The results of genotypic analyses used with immunophenotyping does not exclude that the tumor cells in these cases may be of true histiocytic origin despite the Ki-1-positive phenotype.

Adolescent↗

PubMed2000

Contribution of flow cytometric immunophenotyping to the evaluation of tissues with suspected lymphoma?

BACKGROUND: A critical analysis of the contribution of flow cytometric immunophenotyping (FCI) to the evaluation of lymph nodes and extranodal tissues with suspected lymphoma by a large, retrospective approach has not been reported previously and represents the purpose of this study. METHODS: A total of 278 lymph nodes and 95 extranodal tissue specimens submitted over a 2-year period with complete histologic, FCI, and immunohistochemical (IH) data formed the basis of the study. RESULTS: The FCI data contributed significantly to or was consistent with the final tissue diagnosis in the majority (94%) of the tissue samples. There is no well-described utility of flow cytometry markers for Hodgkin's lymphoma (HL) due to the usual scarcity of tumor cells in the final cell suspensions obtained from these tumors. However, the FCI data excluded non-Hodgkin's lymphoma (NHL) and suggested the possible usefulness of CD15 and CD30 by FCI in HL. In addition, immunophenotypic data by FCI in combination with touch imprint cytomorphology was useful in excluding a diagnosis of NHL in cases of nonhematopoietic malignancies and was particularly useful in defining the following hematopoietic tumors and malignancies: thymoma, T-cell lymphoblastic lymphoma, leukemia cutis, and plasma cell dyscrasia. Thus, IH was not essential for the diagnosis in these latter cases and was performed in only two cases (one thymoma and one plasma cell dyscrasia). Of interest, FCI supported the diagnosis in 3 cases of Ewing's sarcoma/primitive neuroectodermal tumor by detection of CD56 on the surface of the malignant cell. Only 11% of NHL were "negative" by FCI (i.e., an aberrant T-cell or monoclonal B-cell population was not identified). Reasons for these discrepancies included partial tissue involvement by the NHL with sampling differences, T-cell rich or lymphohistiocytic-rich variants with a small population of monoclonal B cells, marked tumoral sclerosis, poor tumor preservation, and T-cell NHL without an aberrant immunophenotype. Only 60% of CD30+ anaplastic large cell lymphomas (ALCL) were CD30+ by FCI. CONCLUSIONS: FCI data should always be correlated with light microscopy if no FCI abnormalities are detected; IH may need to be performed in selected cases. It is less necessary to perform microscopic examination of tissues when the FCI data are positive and indisputable. However, in selected cases in which FCI data is diagnostic, microscopic observations may provide additional information due to sampling.

Flow Cytometry↗

PubMed2000

Cell heterogeneity and subpopulations in solid tumors characterized by simultaneous immunophenotyping and DNA content analysis.

BACKGROUND: Heterogeneity in human malignant tumors is a well-described phenomenon and of interest with regard to subpopulations with differences in clonality, metastatic potential, and response to therapy under different treatment regimes. The aim of this study was the simultaneous characterization of surface markers and DNA content of solid tumors to identify tumor cell subpopulations and to study the association between the expression of antigens and DNA content. METHODS: In the present study, six different malignant tumors grown as xenografts in nude mice were characterized by five-parameter flow cytometry. Immunophenotyping was performed using a variety of direct fluorescence-conjugated antibodies. In all cases, simultaneous detection of DNA content was done after staining with 7-aminoactinomycin D. RESULTS: Tumor cells were characterized by light scatter properties, antigen expression, and DNA content. Tumor cell heterogeneity, subpopulations, and DNA content-dependent antigen expression were identified. CONCLUSIONS: This method offers the possibility of characterizing solid tumors according to their immunophenotype and DNA content. The results obtained can be used to identify changes in immunophenotypic and DNA profiles of tumor cell populations before and after therapy and might be useful to define parameters predictive for response to therapy.

Animals↗

PubMed2006

Evaluation of complete disease remission in acute myeloid leukemia: a prospective study based on cytomorphology, interphase fluorescence in situ hybridization, and immunophenotyping during follow-up in patients with acute myeloid leukemia.

BACKGROUND: Different diagnostic methods add information to define complete remission (CR) in patients with acute myeloid leukemia (AML). The detection of minimal residual disease (MRD) for predicting prognosis and for therapeutic planning still are under discussion. METHODS: The authors studied 216 patients with AML at the time of initial diagnosis and during follow-up and correlated cytomorphology, interphase fluorescence in situ hybridization (FISH), and flow cytometry results to evaluate response status. They further tested the prognostic impact of those results, especially in patients who achieved a morphologic CR. RESULTS: Interphase FISH was found to be correlated significantly with the clinical course at the time of complete cytomorphologic remission and was more reliable than morphology for defining CR. Furthermore, interphase FISH was correlated with immunophenotyping results at all times during follow-up. CONCLUSIONS: The current results indicated that interphase FISH may be used as a valid MRD parameter in patients with AML. Multiparameter immunophenotyping for MRD also was correlated strongly with the clinical course, and the authors suggest integrating such immunophenotyping into the routine diagnostic panel at the time of diagnosis and during the clinical course in patients with AML.

Acute Disease↗

PubMed2001

Fine-needle aspiration biopsy of peripheral T-cell lymphomas. A cytologic and immunophenotypic study of 33 cases.

INTRODUCTION: Peripheral T-cell lymphoma (PTCL) accounts for 10-20% of all non-Hodgkin lymphomas in the United States. In this study, the authors reviewed the cytologic and immunophenotypic findings of 33 fine-needle aspirations (FNAs) of PTCL. METHODS: Thirty-three FNAs from 26 patients (12 females and 14 males) with PTCL were identified during 1991-1999. The patients' age ranged from 19 to 96 years. Immunophenotyping was performed in 24 cases by using either flow cytometry (FC; 21 cases) or immunocytochemistry (IC; 3 cases). Follow-up included review of prior or current histology and clinical records. RESULTS: Nine cases were associated with mycosis fungoides, three cases were classified as T-cell chronic lymphocytic leukemia, and two were angioimmunoblastic adenopathy (AILD)-like T-cell lymphoma. The remaining 19 were classified as PTCL, not otherwise specified. The latter consisted of eight mixed cell variant, eight large cell variant, and three anaplastic variant. One of the mixed cell variant and one of the large cell variants contained numerous epithelioid histiocytes (Lennert lymphoma). Thirty (91%) cases had a definitive diagnosis of malignant lymphoma. Twenty-two cases (2 IC and 20 FC) showed a predominant population of T lymphocytes without a monoclonal B-cell population. In addition, FC revealed an aberrant expression of T-cell markers in 13 cases. Two cases were interpreted as "atypical lymphoid population"; one case was an AILD-like T-cell lymphoma, and the other case was PTCL, large cell type. One case initially was interpreted as granulomatous lymphadenitis; subsequent biopsy revealed PTCL, Lennert type. CONCLUSIONS: Peripheral T-cell lymphoma is a heterogeneous group of lesions with diverse cytomorphology. Cytologic analysis and immunophenotyping is an accurate method of diagnosing peripheral T-cell lymphoma.

Adult↗

PubMed2002

Near-infrared dyes for six-color immunophenotyping by laser scanning cytometry.

BACKGROUND: To adequately analyze the complexity of the immune system and reduce the required sample volume for immunophenotyping in general, more measurable colors for the discrimination of leukocyte subsets are necessary. Immunophenotyping by the laser scanning cytometer (LSC), a slide-based cytometric technology, combines cell detection based on multiple colors with their subsequent visualization without the need for physical cell sorting. In the present study, the filter setting of the LSC was adapted for the measurement of the far-red emitting dye cyanine 7 (Cy7), thereby increasing the number of measurable commercially available fluorochromes. METHODS: The optical filters of the LSC were replaced-photomultiplier (PMT) 3/allophycocyanin (APC): 740-nm dichroic long pass, and 670-/55-nm bandpass; PMT 4/Cy7: 810-/90-nm bandpass. Peripheral blood leukocytes were stained directly by fluorochrome-labeled antibodies or by indirect staining. The tandem dyes of Cy7 (phycoerythrin [PE]-Cy7, APC-Cy7) and the fluorochromes fluorescein isothiocyanate (FITC), PE, PE-Cy5, and APC were tested alone and in different combinations. RESULTS: With the new filter combination and tandem fluorochromes, Cy7 was measurable at 488-nm (argon laser) or 633-nm (helium-neon laser) excitation. Resolution was in the range of FITC for PE-Cy7 but approximately 30% lower for APC-Cy7; spillover into the respective donor fluorochrome channel for both tandem dyes was prominent. A six-color panel for leukocyte subtyping was designed. CONCLUSIONS: With this adaptation, it is possible to measure the tandem conjugates PE-Cy7 and APC-Cy7. This new setup opens the way for six-color immunophenotyping by LSC.

Benzothiazoles↗

PubMed1994

Immunophenotyping specimens from HIV-infected persons: laboratory guidelines from the Centers for Disease Control and Prevention.

The Centers for Disease Control and Prevention has published laboratory guidelines for enumerating CD4+ T-cells from human immunodeficiency virus (HIV)-infected persons to help improve the quality of lymphocyte immunophenotyping. New data that clarify some of the variables in the process of immunophenotyping have been obtained. These include information relevant to both hematology and immunophenotyping procedures as well as quality control parameters that are important in producing reliable results.

CD4-Positive T-Lymphocytes↗

PubMed2004

Fluorescence-intensity multiplexing: simultaneous seven-marker, two-color immunophenotyping using flow cytometry.

BACKGROUND: Conventional immuno-based multiparameter flow cytometric analysis has been limited by the requirement of a dedicated detection channel for each antibody-fluorophore set. To address the need to resolve multiple biological targets simultaneously, flow cytometers with as many as 10-15 detection channels have been developed. In this study, a new Zenon immunolabeling technology is developed that allows for multiple antigen detection per detection channel using a single fluorophore, through a unique method of fluorescence-intensity multiplexing. By varying the Zenon labeling reagent-to-antibody molar ratio, the fluorescence intensity of the antibody-labeled cellular targets can be used as a unique identifier. Although demonstrated in the present study with lymphocyte immunophenotyping, this approach is broadly applicable for any immuno-based multiplexed flow cytomety assay. METHODS: Lymphocyte immunophenotyping of 38 clinical blood specimens using CD3, CD4, CD8, CD16, CD56, CD19, and CD20 antibodies was performed using conventional flow cytometric analysis and fluorescence-intensity multiplexing analysis. Conventional analysis measures a single antibody-fluorophore per photomultiplier tube (PMT). Fluorescence-intensity multiplex analysis simultaneously measures seven markers with two PMTs, using Zenon labeling reagent-antibody complexes in a single tube: CD19, CD4, CD8, and CD16 antibodies labeled with Zenon Alexa Fluor 488 Mouse IgG(1) labeling reagent and CD56, CD3, and CD20 antibodies labeled with Zenon R-Phycoerythrin (R-PE) Mouse IgG(1) or IgG(2b) labeling reagents. RESULTS: The lymphocyte immunophenotyping results from fluorescence-intensity multiplexing using Zenon labeling reagents in a single tube were comparable to results from conventional flow cytometric analysis. CONCLUSIONS: Simultaneous evaluation of multiple antigens using a single fluorophore can be performed using antibodies labeled with varying ratios of a Zenon labeling reagent. Labeling two sets of antibodies with different Zenon labeling reagents can generate characteristic and distinguishable multivariate patterns. Combining multiple antibodies and fluorescent labels with fluorescence intensity multiplexing enables the resolution of more cellular targets than detection-channels, allowing sophisticated multiparameter flow cytometric studies to be performed on less complex 2- or 3-detection-channel flow cytometers. For typical biological samples, approximately 2-4 cellular targets per detection channel can be resolved using this technique.

Antigens, CD19↗

PubMed2004

Immunophenotypic analysis of mast cells in mastocytosis: When and how to do it. Proposals of the Spanish Network on Mastocytosis (REMA).

BACKGROUND: Mastocytosis is a term used for a heterogeneous group of disorders characterized by an abnormal proliferation and accumulation of mast cells (MCs) in one or multiple tissues including skin, bone marrow, liver, spleen, and lymph nodes, among others. METHODS: In recent years, multiparameter flow cytometric studies have shown that pathologic MCs from patients with mastocytosis display unique aberrant immunophenotypic characteristics as compared with normal MCs. RESULTS: Among other features, pathologic MCs show aberrant expression of CD25 and CD2 antigens and abnormally high levels of the CD11c and CD35 complement receptors, the CD59 complement regulatory molecule, the CD63 lysosomal membrane antigen, and the CD69 early-activation antigen. In addition, MCs from mastocytosis express abnormally low levels of CD117 and unexpectedly high light scatter and autofluorescence characteristics. CONCLUSIONS: These aberrant immunophenotypic features are of great relevance for the assessment of tissue involvement in mastocytosis with consequences in the diagnosis, classification, and follow-up of the disease and in its differential diagnosis with other entities. In this paper we provide the reader with information for the objective and reproducible identification of pathologic MCs by using quantitative multiparametric flow cytometry, information for their phenotypic characterization, and the criteria currently used for a correct interpretation of the immunophenotypic results obtained.

Antibodies, Monoclonal↗

PubMed2005

Immunophenotypic differences between diagnosis and relapse in childhood AML: Implications for MRD monitoring.

BACKGROUND: Determination of antigen expression patterns is, in addition to morphologic analysis, essential to the diagnosis of acute myeloid leukemia (AML). The present study was performed to determine (a) the degree of changes in immunophenotype and their consequences on the monitoring of minimal residual disease (MRD) in childhood AML and (b) whether certain clusters of changes in antigen expression patterns exist between diagnosis and relapse. METHODS: Bone marrow specimens of 48 children enrolled in the German AML-BFM-93/98 (Acute Myeloid Leukemia-Berlin-Frankfurt-Munster) studies were analyzed immunologically, morphologically, and genetically at diagnosis and at first relapse. RESULTS: The immunophenotypes by flow cytometry differed by at least one antigen between samples at presentation and relapse in 42 of 48 children (88%). More children displayed an immature phenotype at relapse (43 of 47, 91.5%, vs. 37 of 48, 77%; P = 0.05) with expression of CD34 and/or CD117. This was reflected by a gain of markers that are associated with lineage immaturity in 18 of 25 (72%) of cases, whereas the loss of such antigens was observed in 6 of 25 (24%) patients. We did not observe significant changes for lineage specific markers, with comparable occurrences of loss or gain of myeloid and lymphoid antigens in the sample pairs. Only minimal changes were seen for morphologic and genetic features. CONCLUSION: An antigenic shift was observed in 88% of cases in this study. The antibody panels used for MRD monitoring in childhood AML should therefore not be restricted to the immunophenotype detected at presentation but should include in particular markers of lineage immaturity. The clinical observation of a shift toward a more immature phenotype of the myeloblasts is consistent with the model of a clonal evolution of a leukemic stem cell.

Acute Disease↗

PubMed1991

In situ immunophenotyping study of endothelial cells of the human hepatic sinusoid: results and functional implications.

Hepatic sinusoids are highly specialized capillary vessels characterized by the presence of resident macrophages adhering to the endothelial lining. Although it is likely that sinusoidal endothelial cells have specific adaptations, little is known about the roles that they actually play in vivo. We therefore designed an in situ immunophenotyping study of sinusoidal endothelial cells in normal human liver to compare this population with those of the other vascular compartments of the liver and to determine whether it expresses molecules involved in the following physiological processes: scavenging and nonspecific immune functions, antigen presentation and endothelial-leukocyte adhesion. Our study showed that sinusoidal endothelial cells displayed a highly distinctive immunophenotype characterized by the expression of several molecules not found in the other vascular compartments of the liver. A first group of molecules restricted to sinusoidal endothelial cells were: the receptors II and III for the Fc fragment of IgG; the CD14 molecule, serving as a receptor for the lipopolysaccharide-binding protein; and aminopeptidase N. The presence of those molecules suggests that sinusoidal endothelial cells contribute to the scavenger and nonspecific immune functions of hepatic sinusoids. No direct evidence for an antigen-presenting function of this cell population was obtained. Another group of proteins restricted to sinusoidal endothelial cells comprised the leukocyte adhesion molecules CD4 and intercellular adhesion molecule-1, which may be involved in the adhesion of Kupffer cells to the sinusoidal wall. Therefore our immunophenotyping study makes it possible to provide an in situ demonstration of the phenotypical and functional diversity of endothelial cells in normal human liver.(ABSTRACT TRUNCATED AT 250 WORDS)

Arteries↗

PubMed2001

Immunophenotyping of lymphocyte T and B in the peripheral blood of systemic lupus erythematosus.

The immunophenotyping expression levels of lymphocyte in the peripheral blood from 21 patients with active systemic lupus erythematosus (SLE) were analyzed by using the immunofluorescence labeling-flow cytometry technique to investigate the immunophenotyping expression of lymphocytes T and B in the peripheral blood of active SLE patients and its clinical value. It was showed that, compared with normal controls, the expression of CD3+, CD4+ and the ratio of CD4+/CD8+ in the peripheral blood of these patients were decreased (P < 0.01), while the expression of CD8+, CD20+ was significantly increased (P < 0.01). It was suggested that both T and B cells in patients with active SLE involved in immunoregulation, were activated. The abnormal expression of lymphocyte immunophenotyping could influence the immune reaction in SLE patients, which might be one of the important pathogenesis factors in SLE.

Adolescent↗

PubMed2001

Early prognosis in severe sepsis via analyzing the monocyte immunophenotype.

OBJECTIVE: To analyze the early discriminative predictive information regarding the immunophenotype components of patients with sepsis, and its potential use as a prognosis tool. DESIGN: Observational prospective clinical study. SETTING: Intensive care unit (ICU) in a University Hospital. PATIENTS: Thirty-five patients admitted with severe sepsis. MEASUREMENTS: Analysis of peripheral blood on admission and 48 h later of the absolute white cell count and the immunophenotype of lymphocyte (CD3, CD3-HLADR, CD4, CD8, CD4/CD8 ratio, CD19, and CD25) and monocyte (CD13, CD13-HLADR, CD14, CD14-HLADR, CD13-CD14, and CD4) subpopulations. RESULTS: Due to its high correlation, the immunophenotypic profile studied at admission and 48 h later showed the same prognosis power regardless of the time of performance. The univariate analysis between groups (survival versus death) confirmed the prognostic significance of the total monocyte count and its subpopulations; significant differences were observed from the beginning only in the CD19 lymphocyte subpopulation. Multivariate analysis was performed using logistic regression with survival as the dependent variable. The final model comprised monocytes beta = 0.002 (P = 0.025) and CD13-HLADR beta = 0.016 (P = 0.029). The monocytes receiver operating characteristic (ROC) area obtained was 0.819 (confidence interval 0.663-0.976 at 95 %), the CD13-HLADR ROC area was 0.810 (confidence interval 0.658-0.963), and the monocytes + CD13-HLADR ROC area was 0.918 (confidence interval 0.807-1.000). CONCLUSIONS: A single blood sample test obtaining the absolute monocyte and CD13-HLADR subpopulation count in the first days of admission could contribute to simplifying the classification of patients with severe sepsis into high- and low-mortality risk.

APACHE↗

PubMed2000

Feasibility of simultaneous fluorescence immunophenotyping and fluorescence in situ hybridization study for the detection of estrogen receptor expression and deletions of the estrogen receptor gene in breast carcinoma cell lines.

For the first time, combined immunophenotyping and fluorescence in situ hybridization (FISH) technique according to the "fluorescence immunophenotyping and interphase cytogenetics as a tool for investigation of neoplasms" (FICTION) technique have been successfully applied in solid tumors. Thus, we were able to visualize the antigen expression of cells with chromosomal deletions of a tumor suppressor region directly. In six breast carcinoma cell lines, we investigated the correlation between estrogen receptor (ER) expression status and deletions of the estrogen receptor gene (ESR). To screen for deletions of the ESR gene, dual-color FISH was performed with a YAC (yeast artificial chromosome) probe containing the ESR gene and, as internal control, with a centromeric probe of chromosome 6. Deletions of the ESR gene were detected in four of six cell lines. For direct comparison of ER expression with the copy number of the ESR gene at the single cell level, immunophenotyping with mouse anti-human ER antibody was combined with FISH with the YAC probe containing the ESR gene according to the FICTION technique. There was no correlation between lack of or reduced ER expression and deletions of the ESR gene. One cell line with deletions of the ESR gene did express ER on the protein level, while another cell line without a deletion did not. Cells with deletions of the ESR gene were either ER expression positive or negative. The staining intensity of ER expression was not associated with the copy number of the ESR gene. Thus, this FICTION study unequivocally shows that deletions of the ESR gene are not the major cause of absent or reduced ER expression in breast carcinoma cell lines.

Animals↗

PubMed1991

Flow cytometric immunophenotyping of leukemia cells in clotted blood and bone marrow.

Immunophenotyping of leukemia cells is generally performed on cells in suspension. These suspensions are usually prepared from anticoagulated peripheral blood or bone marrow samples but when anticoagulation is suboptimal clotted samples may reach the laboratory. In this study cell suspensions were prepared from clotted blood and bone marrow samples of leukemia patients by lysis of the clots with streptokinase. The cell suspensions were then labeled with monoclonal or polyclonal antibodies and examined by flow cytometry or fluorescence microscopy. Enough cells could be isolated from small volumes of clotted blood or bone marrow aspirate to determine the immunophenotype of the cells. The morphology of the cells was well preserved and accurate identification of the cells was possible. The immunophenotypes determined on clotted samples from four patients with acute myeloblastic leukemia, five with acute lymphoblastic leukemia and two with chronic lymphocytic leukemia were identical to those established using EDTA anticoagulated samples of the same patients. When no unclotted samples are available this approach may avoid the necessity for a new sample and the concomitant delay in treatment of the patient.

Antibodies, Monoclonal↗

PubMed1995

Immunophenotyping of canine bronchoalveolar and peripheral blood lymphocytes.

The immunophenotype of canine lymphocytes obtained by bronchoalveolar lavage (BAL) was investigated and compared with that of peripheral blood leukocytes (PBL). Indirect immunofluorescence, generated by monoclonal antibodies (mAb) specific for canine CD5, CD4, CD8, CD45pan, CD45RA, MHCII and THY-1, was detected by flow cytometry. In comparison with PBL, in BAL there are fewer lymphocytes positive for CD45RA (75.4 +/- 12.6% vs. 42.3 +/- 9.4%; P < 0.05) and MHCI I (97.0 +/- 2.9% vs. 74.0 +/- 7.6%; P < 0.01), while there are more cells positive for CD8 (19.0 +/- 3.6% vs. 29.5 +/- 12.0%; P < 0.05). Thus there is a lower CD4/CD8 ratio in the cell compartment accessible by BAL (2.2 +/- 0.3 vs. 1.3 +/- 0.6; P < 0.005). The immunophenotype may be stable over time, as indicated by reexamination of cells obtained from one dog at four times over 1 year. Investigating the phenotype of lymphocytes from three different locations of the right lung, the cranial lobe lymphocytes show a lower CD4/CD8 ratio in comparison with PBL (1.81 +/- 0.35 vs. 1.12 +/- 0.31, n = 5; P < 0.02). In general, less MHCII positive lymphocytes (P < 0.001) and greater immunophenotype variability of results were found in these separate samples compared with pooled samples from these locations.

Animals↗

PubMed2004

Histopathologic classification of lung cancer: Relevance of cytokeratin and TTF-1 immunophenotyping.

Histopathologic classification of lung carcinoma is important, as a prognostic factor and in the evaluation of treatment modalities. Although the World Health Organization classification of lung cancer is based on routine microscopy, immunohistochemistry is an important additional aspect in modern pathologic practice. This study examines whether the main histologic types of lung carcinomas are more reliably diagnosed with immunohistochemical technique using antibodies for the lung tissue-specific antigen thyroid transcription factor-1 (TTF-1) and a panel of cytokeratin (CK) antibodies. Forty-five cases of lung cancer (12 squamous cell carcinoma, 13 small cell carcinoma, 11 adenocarcinoma, 9 large cell carcinoma [LCC]/pleomorphic carcinoma) were stained with antibodies to CK CAM5.2, CK5, CK7, CK20, and TTF-1. All 45 cases were positive with CAM5.2, 16 of 45 cases with CK5, 34 of 45 cases with CK7, 4 of 45 cases with CK20, and 29 of 45 with TTF-1. Squamous cell carcinoma (epidermoid carcinoma) had the immunophenotype CK5+/TTF-1-, and at least 20% were also positive with CK7. Most nonepidermoid tumors had the "lung-specific" phenotype CK5-/TTF-1+; all small cell carcinomas had the phenotype CK5-/CK8+/TTF-1+, all adenocarcinomas CK5-/CK7+/TTF-1+ and (more than 50%) of LCC CK5-/CK7+/TTF-1+. Thus, more than 50% of LCCs were of the same phenotype as adenocarcinomas. The immunophenotypes of the main histologic types of lung carcinoma are stable and highly reproducible. However, because of considerable overlapping, immunophenotyping should not be used alone for histopathologic classification of lung cancer, but only as an adjunct to light microscopy. It is also suggested that CK5+ lung carcinomas with basaloid features should be regarded as variants of squamous cell carcinoma and not as LCC.

Biomarkers, Tumor↗

PubMed2005

Hematological, immunophenotypic, and cytogenetic characteristics of acute myeloblastic leukemia with trisomy 11.

We evaluated the incidence of trisomy 11 in acute myeloblastic leukemia (AML) and its correlation with the most relevant clinical, biological, and immunophenotypic disease characteristics in a total of 399 consecutive AML patients. Trisomy 11 was found in 15 patients (3.8%), in 3 of them as the sole abnormality. Median age was 68 years (range 48-87); 87% of patients were older than 60 years. Seven patients displayed multilineage dysplasia. Cytogenetics showed an association with trisomy 8 in six patients, and in five cases with a del(5q); nine patients had complex karyotypes. In all cases, in situ hybridization studies revealed three copies of the MLL gene, but no rearrangements or tandem duplications of MLL. Immunophenotypic analysis of blast cells showed a constant immature immunophenotypic profile with CD34 or CD117 expression (or both) in all cases analyzed. The complete remission rate was 43%; median survival time was only 2 months.

Aged↗