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PubMed2004

[Immunophenotyping of haematologic malignancies].

Over the last 10 years, immunophenotyping of haematologic malignancies has become an indispensable diagnostic supplement to the classical morphological approach. Immunophenotyping of haematopoietic cells is performed with the use of a number of monoclonal antibodies (MOABs), which are directed specifically against structures of blood cells that become expressed at the different stages of differentiation and maturation. Cells to which the fluorescently labelled MOABs are directed can be recognised and measured using fluorescence microscopy or fluorescence flow cytometry. Many MOABs, fluorochromes and user-friendly flow cytometers have become available in the last 15 years, as a result of which immunophenotyping is now routinely applied in clinical practice. Immunophenotyping has the potential to classify leukaemias and other malignant lymphomas according to cell type and stage of maturation. This information is important for the establishment of the right diagnosis and prognosis, and for the optimal treatment choice. In a number of cases immunophenotyping provides information which cannot be obtained by simple morphological investigation. The immunophenotyping of blood and bone-marrow cells is also a sensitive method for detecting minimal residual disease after an apparent complete remission has been achieved.

Antibodies, Monoclonal↗

PubMed2005

Rapid isolation of highly enriched and quiescent microglia from adult rat hippocampus: immunophenotypic and functional characteristics.

Isolation of microglia from CNS tissue provides a powerful tool to study basic microglia biology and examine the effects of in vivo treatments on microglia immunophenotype and function. Previous microglia isolation methodologies utilized whole brain. However, microglia immunophenotype varies across CNS anatomical loci, thus isolation of microglia from whole brain may obscure regional brain variations in microglia immunophenotype and function. In addition, it is unknown to what extent microglia isolation procedures alter the in situ immunophenotype and function of microglia. The present report details a procedure for the rapid isolation of microglia from discrete CNS anatomical loci and addresses the issue of whether the in situ microglia immunophenotype is significantly altered by the isolation procedure. The present microglia isolation method yielded highly enriched hippocampal microglia, which were devoid of other CNS macrophage subtypes and exhibited attributes reflecting a quiescent phenotype characteristic of microglia observed in situ under non-pathological conditions. Further, isolated microglia exhibited functional responsiveness to immunogenic stimuli ex vivo. The immunophenotypic and functional attributes of isolated microglia suggest that the isolation procedure preserves the in vivo phenotype of microglia, thus providing an experimental method with minimal procedural confounds for examining in vivo treatments on microglia ex vivo.

Animals↗

PubMed1986

Classifying acute leukemia by immunophenotyping: a combined FAB-immunologic classification of AML.

A panel of commercially available monoclonal antibodies and five heteroantisera were used to distinguish and subtype 138 cases of acute leukemia (AL). The immunophenotype was compared with the French-American-British (FAB) classification obtained on the cases. The immunophenotype discriminated acute myelogenous leukemia (AML) from acute lymphoblastic leukemia (ALL) and recognized cases not distinguished by cytochemistry (22% of cases), mixed lineage phenotypes (13% of cases), and cases with separate populations of lymphoblasts and myeloblasts (one case). Using the immunologic panel and derived criteria to subtype AML, correspondence of the immunophenotype to the FAB subtypes M1, M2, M4, and M5 was possible in greater than 80% of cases. A combined classification of the immunophenotype and FAB morphology/cytochemistry was devised for AML subtyping. It is recommended that immunophenotyping should be done at least in all cases with negative or inconclusive cytochemistry. At present, we suggest that until a "gold standard" for identifying leukemic subtypes is developed, the best method for typing acute leukemia is by using a combination of morphology, cytochemistry and immunophenotyping.

Antibodies, Monoclonal↗

PubMed1995

Adult acute lymphoblastic leukemia at relapse. Cytogenetic, immunophenotypic, and molecular changes.

BACKGROUND: There have been published reports on cytogenetic, immunophenotypic, and molecular changes at relapse in childhood acute lymphoblastic leukemia (ALL) including lineage switch and secondary leukemia. There are limited data, however, on the cytogenetic, immunophenotypic, and molecular parameters of adult ALL at relapse. Because, as in children, the cytogenetic and/or immunophenotypic changes observed in adult ALL at relapse may have prognostic significance, the authors investigated the significance of such changes. METHODS: Fifty-three patients with relapsed adult ALL for whom cytogenetic, immunophenotypic, and/or molecular analyses were performed at diagnosis and at relapse were studied. Changes in any of the parameters at relapse were correlated with total survival and survival from the time of relapse. RESULTS: Of the 32 patients for whom cytogenetic studies were performed at relapse, 21 (66%) showed clonal cytogenetic changes, 40% of which were clonal evolution. None of these cases, however, showed two entirely different abnormal karyotypes at diagnosis and at relapse. The immunophenotypes showed occasional gain or loss of one or two surface markers, and the molecular genetic configurations for JH, JK, and the T-cell receptor beta were stable throughout the evolution of the disease. Patients with clonal evolution had a shorter overall survival than the rest of the group (P = 0.02). This difference, however, was not significant with respect to survival measured from the time of relapse. CONCLUSIONS: The most frequent changes in the biologic profile of adult ALL at relapse are shifts in the karyotype, with or without clonal evolution. Clonal evolution detected at relapse is associated with a higher frequency of unfavorable karyotypes at diagnosis and with a worse overall prognosis. However, survival from the time of relapse is similar in patients with and without clonal evolution.

Adolescent↗

PubMed2002

Lymphoid and myeloid neoplasms involving cerebrospinal fluid: comparison of morphologic examination and immunophenotyping by flow cytometry.

We studied 53 samples of cerebrospinal fluid (CSF) by cytologic examination and immunophenotyping by flow cytometry. The samples were taken from 43 patients; 25 had a previous diagnosis of malignant lymphoma/leukemia and the remaining 18 a variety of other diseases involving the central nervous system (CNS). Lymphoma/leukemia was detected in 21 samples: 12 by morphologic examination and immunophenotyping and nine by immunophenotyping alone. There were two cases with a suspicious morphologic examination and negative immunophenotyping in which the final diagnosis were cryptococcal and viral meningitis. In the group of 18 patients, one was diagnosed as a primary malignant lymphoma of the CNS and was positive with cytology and immunophenotyping. The other 17 were negative with both methods and follow-up showed no evidence of lymphoma/leukemia. This study shows that morphologic examination combined with flow cytometry enhances the detection rate by 75% over morphologic examination alone in CSF samples.

Adolescent↗

PubMed1993

Immunophenotyping of cytologic specimens by flow cytometry.

Immunophenotyping by flow cytometry is well established as an ancillary technique in the diagnosis of hematopoietic neoplasms. However, flow cytometry is rarely performed on cytologic specimens because most cytologist are more comfortable with direct microscopy and believe that there is inadequate cellularity for analysis. Paradoxically, cytologic material is usually cell suspensions making it ideal for flow cytometry. In order to evaluate the usefulness of immunophenotyping cytologic specimens by flow cytometry, we retrospectively reviewed all cytologic specimens submitted to our flow cytometry unit from 1988 to 1991. Thirty-one cerebrospinal fluid specimens were analyzed. There were inadequate cells for analysis in 15 cases. Five showed a monoclonal proliferation; 11 were nondiagnostic. A range (r) of one to six cell surface markers were performed. Thirty-two body cavity fluids were analyzed: 7 peritoneal, 19 pleural, 2 pericardial, and 4 bronchoalveolar lavage. There were cells to analyze in all cases. Seven had a monoclonal proliferation; 25 were nondiagnostic (r = 4-21 markers performed). One hundred eighteen fine needle aspirates (FNA) were reviewed; 58 FNA were radiologically guided, 60 were superficial lesions. There were inadequate cells for analysis in two cases. Sixty-one demonstrated a monoclonal proliferation; 55 were nondiagnostic (r = 1-22 markers performed). We conclude that immunophenotyping by flow cytometry is of limited value for cerebrospinal fluid analysis and that knowledge of previous immunophenotyping studies is essential for correct analysis; analysis of body cavity fluids is easily performed but less often demonstrates a monoclonal proliferation. Immunophenotyping by flow cytometry is a valuable adjunctive technique for FNA and yields adequate cells for analysis.

Antigens, Surface↗

PubMed1996

Immunophenotypic characterization of acute leukemias and chronic lymphoproliferative disorders: practical recommendations and classifications.

Immunophenotypic characterization of leukemic cells has become essential for the diagnosis of acute leukemias (AL) and chronic lymphoproliferative disorders (CLPD). Immunophenotyping allows to classify AL according to (i) lineage assignment of the leukemic clone based on the degree of specificity (or "score") of expressed markers, (ii) the differentiation level of the clone and (iii) the presence of irrelevant markers. In addition, some rare AL subtypes may be identified, such as M0, M6 "variant" and M7 FAB types, as well as "biphenotypic" (hybrid) AL. Finally particular immunophenotypic profiles are of pronostic value or associated with specific cytogenetic abnormalities. In leukemic phase CLPD setting, some circulating lymphoid cell immunophenotypic profiles are strongly correlated with morphology as defined by the FAB and REAL classifications. In addition, some marker expressions are of pronostic value. However, proper choices of sample nature, monoclonal antibodies and immunophenotyping methods are essential to improve quality, reliability and reproducibility of the results and must be carefully controlled in and between laboratories.

Acute Disease↗

PubMed1993

Immunophenotypic and genotypic analysis in cutaneous lymphoid hyperplasias.

BACKGROUND: The clinical utility of immunophenotyping and Southern blot analysis in the evaluation of patients with cutaneous lymphoid hyperplasia (CLH) is controversial. OBJECTIVE: Our purpose was to determine whether adjunctive immunophenotyping and Southern blot analysis are of diagnostic and prognostic value in patients with CLH. METHODS: Immunophenotyping was performed on skin biopsy specimens from 26 patients with a routine histologic diagnosis of CLH. Southern blot analysis for immunoglobulin (Ig) gene rearrangements was done on 13 of 26 cases. RESULTS: Twenty-four of 26 patients had polyclonal CLH on immunophenotyping: 2 of 26 had monoclonal lymphoma. Two of 11 patients with polyclonal CLH studied by Southern blot analysis had clonal Ig gene rearrangements. In both, lymphoma developed within 1 to 6 years; comparison of CLH and malignant lymphoma demonstrated overlapping and different clonal bands. Two additional patients with polyclonal CLH developed lymphoma. No clonal gene rearrangements were detected in the CLH or lymphoma from one; the other was not studied. CONCLUSION: Immunophenotyping will identify some patients with lymphoma with nondiagnostic histologic features. Southern blot analysis will predict some patients with polyclonal CLH in whom malignant lymphoma will develop and who may benefit from definitive therapy.

Adult↗

PubMedDate not supplied

Immunophenotype and TCR-Vbeta repertoire of peripheral blood T-cells in acute infectious mononucleosis.

Although a number of studies on the phenotypic changes that occur after T-cell activation have already been published, the specific immunophenotypic features of T-lymphocytes and the frequency at which TCR-variable region (TCR-V) restricted T-cell expansions occur "in vivo" during acute viral infection still remains to be established. We report on the immunophenotype and TCR-V repertoire of peripheral blood T-cells from 28 patients with acute infectious mononucleosis. Immunophenotypic studies were performed by flow cytometry using direct immunofluorescence techniques and stain-and-then-lyse sample preparation protocols with three- and four-colour combinations of monoclonal antibodies directed against a large panel of T- and NK-cell associated markers, activation- and adhesion-related molecules and TCR-Vbeta, -Vgamma and -Vdelta families. Nearly all patients (27/28) showed a massive expansion of CD8(+)/TCRalphabeta(+) T cells, the majority (>90%) of which displayed an immunophenotype compatible with T-cell activation: CD2(+high), CD7(+low), CD11a(+high), CD38(+high), HLA-DR(+high), CD28(+/-low), CD45RO(+high), CD45RA(-/+low), CD11b(-/+low), CD11c(+/-low), CD16(-), CD56(-), CD57(-), CD62L(-), CD94(-), CD158a(-), CD161(-), NKB1(-). Additionally, the levels of both CD3 and CD5 were slightly decreased compared to those found in normal individuals. Late-activation antigens, such as CD57, were found in small proportions of CD8(+)/TCRalphabeta(+) T-cells. Increased numbers of CD4(+)/TCRalphabeta(+) T-cells, TCRgammadelta(+) T-cells and NK-cells were also noticed in 17, 16 and 13 of the 28 cases studied, respectively. Evidence for activation of CD4(+)/TCRalphabeta(+) and TCRgammadelta(+) T-cells relied on changes similar to those described for CD8(+)/TCRalphabeta(+) although less pronounced, except for higher levels of both CD5 and CD28 in the absence of reactivity for CD11c on CD4(+)/TCRalphabeta(+) T-cells and higher levels of CD161 and CD94 on TCRgammadelta(+) T-cells. Small expansions of one or more TCR-Vbeta families accounting for 12 +/- 7% of either the CD8(+)/TCRalphabeta(+) or the CD4(+)/TCRalphabeta(+) T-cell compartment were found in 12 of 14 patients studied, whereas the distribution of the TCR-Vgamma and -Vdelta repertoires tested in 2 of the individuals with expanded TCRgammadelta(+) T-cells was similar to that observed in control individuals. The results presented here provide evidence for an extensive T-cell activation during acute viral infection and establish the immunophenotype patterns associated with this condition.

Acute Disease↗

PubMed1998

Immunophenotypic patterns and cytogenetic anomalies in acute non-lymphoblastic leukemia subtypes: a prospective study of 432 patients.

This study prospectively analysed the relationships between immunophenotypic and cytogenetic features of blast cells in 432 acute non-lymphoblastic leukemias (ANLL) at presentation. An abnormal karyotype was detected in 232 cases (54%). These abnormalities were related to immunophenotypic markers as detected using a consensual panel of monoclonal antibodies allowing lineage assignment and investigation of myeloid marker expression on blast cells. In univariate analysis, CD9, CD10, CD15, CD34 and TdT expression appeared significantly associated with chromosomal anomalies. Multivariate analysis identified CD34 and CD9 expression as independently predictive of the presence of at least one cytogenetic abnormality (P < 10(-4) and P < 0.03, respectively). Significant associations between immunophenotypic and karyotypic features were observed both within individual FAB subgroups and independently from morphological criteria. Specific features were seen in five ANLL entities: M0 or M1/B lineage antigen positivity/t(9;22) or del(11)(q23); M2/CD13-/t(8;21); M4/CD13+, CD34+, CD36+/inv(16); M4 or M5/lack of B lineage antigen/del(11)(q23) or t(9;11). More practically, and although the relationships demonstrated only represent a fraction of homogeneous immunophenotypic subgroups, identification of such immunophenotypic features should prompt careful karyotypic examination, eventually using molecular biology analysis on non-growing cells.

Antigens, CD↗

PubMed1994

Immunophenotypic and genotypic analysis of acquired immunodeficiency syndrome-related non-Hodgkin's lymphomas. Correlation with histologic features in 36 cases. French Study Group of Pathology for HIV-Associated Tumors.

High-grade B-cell-type non-Hodgkin's lymphomas are observed in 5% to 8% of patients positive for the human immunodeficiency virus. Nearly all cases belong to one of the three major histologic types: centroblastic or large noncleaved cell, immunoblastic and Burkitt's lymphoma, or small noncleaved cell. Some cases that are polymorphic are termed high-grade B-cell, not otherwise specified (NOS). The authors determined the immunophenotype of each histologic category of acquired immunodeficiency syndrome (AIDS)-related non-Hodgkins' lymphoma and sought a relationship with the presence of the Epstein-Barr virus (EBV). B-cell differentiation antigens, activation marker expression (human leukocyte antigen-DR, CD10, CD19, CD20, CD21, CD22, CD23, CD25, CD30, CD38), and epithelial membrane antigen were analyzed. The clonality was determined by the detection of cytoplasmic immunoglobulin, surface immunoglobulin, and the analysis of joining region (JH) immunoglobulin gene configuration by Southern blot. Epstein-Barr virus was detected either by Southern blot analysis using BamHI W probe fragment or by in situ hybridization with EBV-encoded RNA transcripts-1 specific probe. The immunophenotypic and genotypic results were compared with the morphology results and with the presence or absence of EBV. Burkitt's lymphomas were associated with EBV in 50% of cases, were monoclonal, and expressed mostly immunoglobulin (Ig) MK, CD10, CD19, CD20, CD22, and CD38. This immunophenotypic profile closely resembled those of the centroblastic cases (large noncleaved cell), in which EBV was absent. Epstein-Barr virus was associated with 90% of immunoblastic cases, and only CD10, CD20, and CD38 were expressed. CD71 was expressed in all categories of non-Hodgkin's lymphoma, and CD21 and CD23 were rarely expressed. Two cases of immunoblastic lymphoma and one case of high-grade B-NOS were polyclonal regarding JH rearrangement, but EBV tested with 1.9-Kb Xhol fragment was clonal. No significant immunophenotypic changes were noted in relation to the presence of EBV. Such studies comparing morphology, immunophenotype, and genotype could help classify and better understand the pathogenesis of AIDS-related non-Hodgkin's lymphoma.

Adult↗

PubMed1992

Histopathologic, immunophenotypic and genotypic analyses in ocular adnexal lymphoproliferative disorders.

This study reports on fourteen biopsies from patients presenting to our orbital and oncology service with ocular adnexal lymphoid proliferations between November 1988 to September 1991. The biopsies were studied using histologic, immunophenotypic and genotypic analyses. By histologic criteria, there were two reactive, five indeterminate and seven lymphomatous lesions. On immunophenotypic analysis, there were two monoclonal and 10 polyclonal lesions in the 12 specimens analysed. Genotypic analysis confirmed the histopathologic diagnoses for the reactive lesions by showing them to be germline. It also confirmed that both the histopathologic lymphomas and immunophenotypically monoclonal lesions were clonally rearranged. Genotypic analysis was able to separate the histologically indeterminate group into two subsets: clonally rearranged, of which there was one, and germline, of which there were four. In addition, it demonstrated that immunophenotypic polyclonality cannot always be equated with genotypic polyclonality as was the situation in four out of 10 lesions in our series. The significance of clonal arrangements in the histologically indeterminate and immunophenotypically polyclonal groups can only be determined by prospective study.

Adult↗

PubMed1996

Multiparametric immunophenotyping of B cells in peripheral blood of healthy adults by flow cytometry.

The investigation of patients suffering from malignant lymphomas of the B-cell type requires flow cytometric immunophenotyping. Several reports described the expression of almost all B lineage antigens on normal and abnormal B lymphocytes. Thus, immunophenotyping of lymphomas must be interpreted in the context of the reference values obtained for healthy control individuals. For this purpose multiparametric flow cytometric analysis offers the unique feature for lymphocyte subset analysis. In the present study B lymphocytes in the peripheral blood of healthy adults were investigated by multiparametric flow cytometric immunophenotyping for the detection of the frequency (in percent) of antigens provided by the revised European-American classification of lymphoid neoplasms (REAL) classification. Thus, 84 healthy adults were investigated and grouped by age (average ages were as follows: group 1, 25.38 years; group 2, 33.86 years; group 3, 44.17 years; group 4, 55.67 years; group 5, 66.67 years). Analysis was done for surface immunoglobulins (kappa and lambda chains of immunoglobulin M [IgM] and IgD) as well as CD10, CD11c, CD23, CD38, CD103, FMC-7, and B-B4. Three-color immunophenotyping was performed for kappa/CD19/CD5, lambda/CD19/CD5, surface IgM/surface IgD/CD19, FMC-7/CD19/CD5, CD103/CD11c/CD19, CD10/CD23/CD19, and CD38/B-B4/CD19 by live gating of CD19+ events (n = 2,000). Although some numerical differences could be obtained for the different groups, statistical differences (P < 0.005) could only be obtained for the CD19+/CD5+ B-cell subset, which was decreased in the elderly patients (group 5). The established two-color and three-color stainings will serve as a basis for future multiparametric immunophenotyping of abnormal lymphocytes (e.g., for patients suffering from non-Hodgkin's lymphoma of the B-cell type).

Adult↗

PubMedDate not supplied

Combined cytomorphologic and immunophenotypic analysis in the diagnostic workup of lymphomatous effusions.

OBJECTIVE: To analyze the results of cytomorphology and immunophenotyping in 54 patients with lymphomatous effusions. STUDY DESIGN: We report the results of cytomorphology and immunophenotyping in 54 patients with lymphomatous effusions. Twenty-three of the 54 had a previous diagnosis of NHL. In the remaining 31 patients, lymphomatous involvement was clinically suspected. RESULTS: Thirty-three lymphomatous effusions were positive for involvement by NHL. Twenty-one of these 33 patients (64%) had a previous diagnosis of NHL. Of the remaining 12 patients with newly diagnosed NHL, 11 had high grade lymphoma, and one had follicular center lymphoma. Twenty effusions were considered to be reactive; only two of these patients had NHL. One effusion revealed involvement by a previously unknown carcinoma. We observed seven false negative results if only one of both methods was considered. A high grade NHL was not diagnosed by immunophenotyping in one case, and six cases of low grade NHL could not be detected by cytomorphology. The combined strategy of cytomorphology and immunophenotyping had a sensitivity of 100% and specificity of 100% in our study, confirmed by follow-up studies. CONCLUSION: Both methods have shown difficulties in the examination of lymphomatous effusions. Cytomorphology has problems distinguishing reactive effusions from low grade NHL. The detection of high grade NHL by immunophenotyping is difficult. However, both methods together offer the advantage of dual staining ability and are most helpful in distinguishing clonal lymphomatous from reactive effusions.

Ascitic Fluid↗

PubMed2003

Minimal residual disease in adolescent (older than 14 years) and adult acute lymphoblastic leukemias: early immunophenotypic evaluation has high clinical value.

Investigation of minimal residual disease (MRD) in acute leukemias by immunophenotyping and/or molecular techniques is proving to be increasingly valuable for disease monitoring. In acute lymphoblastic leukemia (ALL), most MRD studies have focused on children, whereas in contrast, information on the value of MRD on adult ALL is scanty, and almost exclusively restricted to polymerase chain reaction (PCR) studies. Early response to therapy is one of the most important prognostic factors in acute leukemia, which prompted us to investigate whether or not early immunophenotypic assessment of MRD could also be a valuable tool for predicting relapse in adult patients with ALL. For that purpose we have analyzed the level of MRD during the initial phase of treatment (induction phase) by multiparameter flow cytometry in a series of 102 adolescent (older than 14 years) and adult patients with ALL. Immunophenotypic evaluation of the bone marrow (BM) at day +35 showed that patients with low MRD levels (< 0.05% leukemia-associated phenotype [LAP+] cells) had a significantly longer relapse-free survival (RFS) than patients with high MRD levels, and this prognostic influence was retained when only those patients in morphologic complete remission (mCR) at day +35 were considered (median RFS: 42 months vs 16 months; P =.001). Moreover, immunophenotyping helped to identify a small subset of patients (n = 12) with negative or low MRD levels (< 0.03% LAP+ cells) by day +14, with an excellent prognosis (projected RFS of 90% at 5 years). The contrary is true of patients who achieved late mCR (after day +35), since immunophenotypic investigation of MRD showed that, in spite of the mCR, none of the cases with more than 0.1% LAP+ cells would be relapse-free after 2 years. Multivariate analysis showed that the immunologic evaluation of MRD at day +35 was the most relevant independent prognostic parameter for adult patients with ALL, and together with age, white blood cell (WBC) count at diagnosis, and presence of the Philadelphia (Ph) chromosome, represented the most informative combination of variables for predicting relapse-free survival.

Adolescent↗

PubMed2002

Comparison of immunophenotypes of small B-cell neoplasms in primary lymph node and concurrent blood or marrow samples.

Immunophenotyping of small B-cell neoplasms (SBCNs) may have a critical role in diagnosis. However, there are few data addressing whether the immunophenotypes of SBCNs in bone marrow (BM) and peripheral blood (PB) are representative of those in other tissue sites. We compared the immunophenotypic features of concurrently analyzed lymph node (LN) and BM/PB specimens using multiparameter flow cytometry. Fifty-five SBCNs were identified: 27 follicular lymphomas (FLs), 16 chronic lymphocytic leukemia/small lymphocytic lymphomas (CLL/SLLs), and 12 mantle cell lymphomas (MCLs). Major (presence vs absence) or minor (alteration of intensity) variations in expression of individual antigens between LN and BM/PB were observed in up to 25% of cases within a particular SBCN category. All FLs and CLL/SLLs maintained characteristic immunophenotypes in BM/PB. Potentially misleading variations included 1 case of MCL that failed to express CD5 in BM and likely would have been immunophenotypically misclassified as a marginal zone lymphoma and another MCL that expressed moderate CD23 in PB and would have required additional studies for precise classification. The remaining major and minor variations would not have affected interpretation.

Antigens, Neoplasm↗

PubMed2001

Critical analysis and diagnostic usefulness of limited immunophenotyping of B-cell non-Hodgkin lymphomas by flow cytometry.

We report the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of various immunophenotypes characteristic of each class of B-cell non-Hodgkin lymphoma (NHL) based on analysis of 352 morphologically well-characterized B-cell NHLs and 175 benign lymph nodes (LNs) using 2-color flow cytometry. All B-cell NHLs that exhibited a characteristic immunophenotype (except diffuse large B-cell lymphoma) had a high NPV. The immunophenotypes of small lymphocytic lymphoma and mantle cell lymphoma showed high specificity, but only small lymphocytic lymphoma also showed a high PPV. One third of follicular lymphomas coexpressed CD23 and CD10. Diffuse large B-cell NHL showed no consistent immunophenotype. About 90% of all benign LNs expressed no substantial amounts of CD5, CD10, or CD23. Most benign LNs also failed to express substantial amounts of immunoglobulin heavy chains. In contrast, about 90% of NHLs showed expression of 1 or 2 heavy chains. The expression pattern of immunoglobulin light chains was not found helpful in favoring one lymphoma type over another. The usefulness of each immunophenotype for each lymphoma group is of particular diagnostic importance in limited specimens, such as fine-needle aspiration biopsies, small core biopsies, body effusions, extranodal sites, and nodal tissues with various artifacts.

CD5 Antigens↗

PubMed2001

Transient myeloproliferative disorder and acute myeloid leukemia in Down syndrome. An immunophenotypic analysis.

Immunophenotypic analysis of transient myeloproliferative disorder (TMD) and acute myeloid leukemia (AML) using multiparameter flow cytometry might provide insight into their relationship. We retrospectively analyzed the expression of multiple lymphoid, myelomonocytic, and megakaryocytic antigens on blast proliferations in 18 patients with Down syndrome (DS; AML, 9; TMD, 9). The AMLs and TMDs shared several immunophenotypic characteristics. Blasts in all expressed CD45, CD38, and CD33; most AMLs and all TMDs were CD36+; and the majority expressed CD41 and CD61, suggesting megakaryocytic differentiation. The majority of cases were CD34+, CD14-, and CD64-. There was aberrant expression of the T-cell-associated antigen CD7 in most AMLs and TMDs. CD56 was expressed aberrantly in 5 AMLs and 7 TMDs. The major difference between the disorders was the pattern of expression of myeloid markers CD11b and CD13; each was expressed in 8 AMLs but only 2 TMDs. Blasts were HLA-DR-positive in 3 AMLs vs 7 TMDs. Blasts in TMD and AML in DS have a characteristic immunophenotype distinct from AML in other settings. The immunophenotypic similarities suggest a biologic relationship between the disorders; however, distinct immunophenotypic differences also were observed.

Adolescent↗