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Results for “Immunophenotyping”

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PubMed1991

[Correlation of immunophenotype with clinical features in T-cell acute lymphoblastic leukemia].

The immunophenotype of leukemicblasts from 111 patients with T-ALL or T-NHL were further examined by using a panel of standardized McAbs of CD nomenclature to human leukocyte differentiation antigens. Four major subsets of T-ALL were defined: pre T-ALL, immature T-ALL (I), common T-ALL (II) and mature T-ALL (III), with the percentages 20.7%, 20.7%, 20.7% and 37.0% respectively. In addition there was a case with M-T acute hybrid leukemia. Some of the clinical features of the patients with T-ALL and T-NHL were compared. It was found that male predominance, older age, higher leukocyte count, lower platelet level, relative higher hemoglobin level and increased incidence of extramedullary involvement, including hepatomegaly, splenomegaly and lymphadenopathy were alike for all subsets of T-ALL cases. However, the average white cell level and incidence of lymphadenopathy in the pre T-ALL subset significantly differed from those in other subsets. The correlation of immunophenotype with morphologic characterization was also discussed in this paper.

Adolescent↗

PubMedDate not supplied

Chronic B-cell lymphoproliferative disease: relationship between immunophenotype and clinical stage.

We analyzed the immunophenotype in 34 cases of B-CLD referred to our observation over a 13-month period. In 25 patients the classical phenotypic pattern of B-CLL was observed; in 9 cases clinical, morphocytochemical, and histologic findings were consistent with CLL but differed in phenotype. The results indicate that four immunophenotypic subgroups related to CD5 and FMC7 expression may be identified within B-CLL. Clinical correlations in these cases suggest that B-CLL with anomalous phenotypes are more frequently diagnosed at a later stage than classical B-CLL. Furthermore, while the percentage of CD3+ cells did not vary among the subgroups, a comparison of the CD4/CD8 ratio disclosed marked differences.

B-Lymphocytes↗

PubMed1988

Immunophenotypic characterization of primary and secondary lymphoid follicles.

The need for an immunophenotypical referential framework relative to lymphoid follicle has led us to apply a panel of monoclonal and polyclonal antibodies, by means of a sensitive immunostaining method. Lymphoid follicle is an immunophenotypically complex structure made up of three lymphoid populations (B, being its bulk, and a few T and NK cells), dendritic reticulum cells (DRCs) and Flemming's macrophages. Follicular B population is To 15 +, B1+, OKB 7 +, HLA-DR + and C3bR +. In secondary follicles there are differential characteristic reactivities for each topographic compartment: Mantle zone is positive for OKB 2 and surface IgM (sIgM) and IgD (sIgD); germinal center (GC) clear zone (with centrocytic predominance) for OKT 9, sIgM and weakly for OKB 2; and GC dark zone (with centroblastic predominance) only for OKT 9. In sections, OKT 10 allows one to see immunoblasts and plasma cells, the latter being with lymphoplasmacytoid cells the only intracytoplasmic immunoglobulin holders. 10% of GC lymphocytes are T cells, almost exclusively T-helper (Leu 3a +). Another 10% to 15% of lymphoid cells are Leu 7 (HNK-1) +. In histological sections, DRCs are specifically marked with R4/23 and Flemming's macrophages with anti-alpha1-antitrypsin and anti-alpha1-antichymotrypsin antibodies, both populations being negative to OKM 1 and OKM 5.

Antibodies, Monoclonal↗

PubMed1995

[Large-cell anaplastic lymphomas: a genetic and immunophenotypic study].

PURPOSE: To characterize from a genetic point of view a group of non-Hodgkin's anaplastic large-cell lymphomas (ALCL) by Southern blot and PCR methods with different probes (molecular study) and with direct or post 24-78 hours cultures with GTC banding techniques (cytogenetic). To correlate the results to the immunophenotype performed with a complete panel of monoclonal antibodies (MoAb) according to avidin-biotine and alkaline phosphatase (APAAP) methods. MATERIAL AND METHODS: Sixty cases of ALCL were reviewed and only 19 selected (with frozen or fresh material) because a complete immunohistologic and genotypic correlation had been done. According to CD15 expression two groups were considered, CD15+ (the so-called Hodgkin's related or Hodgkin's like) and CD15- or classical type. RESULTS: Only 26.5% of cases showed immuno -genotypic correlation. Immunohistochemistry is an accurate method for activation, proliferation and B-cell nature, but T-cell cases were not stained because T-cell paraffin markers are not completely specific. CD15 group had only 30% rearranged cases with scarce cytogenetic abnormalities, as it occurs in Hodgkin's disease (HD). ALCL classical type showed 66% rearranged cases, and one of the T-cell cases showed an incomplete t (2:5) translocation or polyploid cell lines. CONCLUSIONS: Both groups have different genetic and immunophenotypic behaviour which resembles HD or NHL. CD15 positive cases or ALCL HD-related constitute a borderline entity which has to be recognized because of the different therapy and clinical behaviour.

Hodgkin Disease↗

PubMed1994

[Immunophenotyping in chronic B-cell lymphatic leukemia].

The authors present the results of immunophenotyping analysis of cells from 67 patients with the diagnosis of B-CLL from group of 393 adults examined by immunophenotyping because of a haematological malignity. Characteristic for B-CLL is the finding of surface molecules HLA DR, B lymphocytic superficial molecules and CD5. Aberrant membrane equipment of B-CLL cells is suggested by the rare absence of some pan B lymphocytic sign. Marked reduction of T lymphocyte ratios, their sub-populations and NK cells is associated with immunodeficiency manifestations which are common in patients with B-CLL.

Adult↗

PubMed1994

Transitional changes in immunophenotypic subpopulations of human peripheral blood CD34+ cells expanded in vitro.

We determined the appropriate incubation period to expand human peripheral blood (PB) CD34+ cells for clinical application and the role of recombinant human (rh) interleukin-3 (rhIL-3) in the expansion and differentiation of these cells. The cells were purified up to 40 +/- 16% and transitional changes in immunophenotypic subpopulations in suspension culture were examined following stimulation with three different combinations of rh colony-stimulating factors (rhCSFs): 1) rhIL-3 alone, 2) rhIL-6, rhSCF, rhG-CSF, plus rhGM-CSF, and 3) the four CSFs plus rhIL-3. With all three CSF combinations, the total cells increased continuously after day 5 until day 14, and a combination of the five CSFs always gave rise to the highest number of total cells. Immunophenotypic analysis of the expanded cells showed that with all three CSF combinations CD34+ cells reached a maximal rate on day 5 and then decreased in an inverse correlation between the logarithm of CD34 positive rate and the duration of suspension culture. The maximum expansion of CD34+ cells and PB progenitor cells (PBPC) with rhIL-3 alone, the four CSFs, or the five CSFs was observed on day 5, 10, or 7, respectively. The combination of the five CSFs was identified as the most potent stimulus for expansion of PBPC and CD34+ cells, as it increased non-erythroid PBPC 89 +/- 69-fold, with a range of 24 to 204-fold on day 7. However, differences in the expansion rate of these cells on days 5, 7, and 10 were not statistically significant. The majority of purified CD34+ cells coexpressed CD38 (91 +/- 3%) but were negative for CD33 (85 +/- 4%), and one-half coexpressed CD13. With all three combinations of CSFs, the mature CD34+ cells that coexpressed CD38, CD33, or CD13 expanded in parallel with the total CD34+ cells, while an increase in relatively immature CD34+ cells, which do not express CD38, CD33, or CD13, was only statistically significant with the five CSFs. Thus, rhIL-3 played a critical role when combined with the four CSFs by increasing both mature and immature CD34+ cells.

Adult↗

PubMed1995

Chronic myeloid leukemia: correlation between purine metabolism enzyme activities and membrane immunophenotype.

Peripheral blood or bone marrow of 24 patients with chronic myeloid leukemia (CML) were characterized for their surface membrane marker profiles using flow cytometry and fluorescence microscopy. Purine metabolism enzyme activities were compared with membrane immunophenotype and cytochemical stains. CML subtypes were correlated with the expression of surface membrane antigens detected by the monoclonal antibodies. On the basis of immunophenotyping we found the following characteristic marker profiles: In stable phase of CML (CML-SP)-CD15, CD11b, CDw65, CD13, in accelerated phase of CML (CML-AP)-CD15, CDw65, CD11b, CD13 and CD33, in myeloid blastic phase of CML(CML-BP-M)-CD13, CD33, HLA-DR, CD11b, CD15, CDw65, in myeloid and lymphoid (mixed) blastic phase of CML (CML-BP-M+L)-CD13, CD33, CD34, HLA-DR, CD11b, CD10 and in chronic myelomonocytic leukemia (CMML)-CD14, CDw65, CD11b, CD33 and HLA-DR. Analysis of purine metabolism enzyme activities showed that there was a correlation between the values of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) and various types of CML. ADA levels in CML-SP, CML-AP and CMML were comparable with those in normal cells. In CML-BP-M, which represents proliferation of less mature myeloid cells (similar to less mature AML subtypes), ADA activity increased and PNP activity decreased. ADA activity was significantly different between control group and CML-BP-M (p < 0.01), between CML-SP and CML-BP-M (p < 0.05). The values of PNP activity were the highest in stable phase of CML (125 pkat. 10(-6) cells) and the lowest (23 pkat.10(-6) cells) in CML-BP-M+L. PNP activity in the other groups corresponded to control values. High ADA/PNP ratio was found in CML-BP-M and CML-BP-M+L (0.7 and 2.0, respectively) in comparison to CML-SP (0.2). It follows from our results that ADA/PNP ratio enables to discriminate between stable and blast phases of CML (p < 0.01). The level of the cytochemical enzymes (CHAE, MPO, SBB, ANAE and 5' NT) varied and reflected the degree of cell differentiation and maturation. CHAE and MPO were characteristic enzymes for CML, ANBE for CMML and 5' NT for CML-BP-lymphoid.

Adenosine Deaminase↗

PubMed1995

Detection of EBV mRNA in nodular sclerosis type of Hodgkin's disease using in situ hybridization with EBER 1 and EBER 2 probes with regard to the immunophenotype of Reed-Sternberg cells.

Hodgkin's disease (HD) is a heterogenous disorder in terms of morphology, immunophenotype, genotype and association with viruses. Seroepidemiologic, immunohistochemical and molecular studies suggest that Epstein-Barr Virus (EBV) plays an important role in the pathogenesis of HD. We used in situ hybridization (ISH) with digoxigenin-labeled EBER 1 and EBER2 probes to examine a series of 28 lymph node specimens from untreated patients from Poland with the nodular sclerosis NS 1 and NS 2 subtypes of HD to detect EBV mRNA in Reed-Sternberg cells (RSC), or in small non-neoplastic lymphocytes called "bystander cell" (ByC). EBV mRNA was detected in 12 of 28 cases (32.1%). Furthermore, the expression of Latent Membrane Protein (LMP) of EBV strongly correlated with expression of EBER 1 and EBER 2 in RSC but not in "bystander cells", which were LMP negative. The correlation between EBV status and immunophenotype of RSC are discussed.

Herpesvirus 4, Human↗

PubMed1995

Acute leukemia and the transient myeloproliferative disorder associated with Down syndrome: morphologic, immunophenotypic and cytogenetic manifestations.

Individuals with Down syndrome have an increased incidence of leukemia compared to the general population. In addition, Down syndrome children may acquire a myeloproliferation that resembles acute leukemia that undergoes a spontaneous, durable remission. To clarify the relationship between these two disorders, the morphologic, immunophenotypic and cytogenetic characteristics of 28 patients with Down syndrome and the morphologic manifestations of acute leukemia were examined. Three cytomorphological groups were discerned. The first two groups consisted of five patients with acute lymphoblastic leukemia (group I) and three patients with acute myeloid leukemia (group II). These leukemias resembled those of non-Down individuals. The third and largest group (group III) consisted of 20 cases of acute myeloid leukemia that showed prominent megakaryocytic and/or erythroid differentiation and occurred in children under 6 years of age. The blasts in this group were non-reactive for myeloperoxidase or non-specific esterase and expressed CD7, CD34 and CD36 with variable expression of CD61, CD13 and CD33. Four patients in this group had an acquired trisomy 8. Four group III leukemias underwent a durable, spontaneous remission within 2 months of diagnosis. There were no morphologic differences between those leukemias in this group that progressed and those that remitted; however, all remissions occurred in newborns. It is concluded that Down syndrome children acquire a characteristic acute myeloid leukemia that has prominent megakaryocytic and/or erythroid differentiation and an unusual immunophenotype. This group of leukemias may undergo a durable, spontaneous remission in the newborn period.

Acute Disease↗

PubMed1994

Flow cytometric analysis of low grade B-cell non-Hodgkin's lymphoma: correlation between histology, immunophenotyping and cell cycle analysis.

In the present study the immunophenotyping and the DNA parameters in the group of 38 patients with B-cell non-Hodgkin's lymphoma of low grade malignancy were evaluated in relation to the Kiel Classification. The study was performed with flow cytometry. The evaluation of the FSC/SSC light scatters enables selective immunophenotyping and also corresponds to tumor histology. The lymphoma cells reactivity with selected anti-lymphocyte antibodies depended on their histological type, however no characteristic phenotype, except for lymphocytic lymphoma, for each lymphoma type could be distinguished. Low grade lymphomas were characterized, in general, by low proliferative activity. The differences in percentage of cells in S-phase fraction within selected groups of lymphoma could be a valuable indicator of the clinical outcome.

Adult↗

PubMed1994

Flow cytometric analysis of high grade B-cell non-Hodgkin's lymphoma: correlation between histology, immunophenotyping and cell cycle analysis.

The immunophenotype and the DNA parameters of 21 high grade B-cell non-Hodgkin's lymphomas considered in terms of the Kiel Classification were studied by flow cytometry. The centroblastic and immunoblastic lymphomas differed in the expression of lymphocyte markers, however no characteristic immunophenotype for each lymphoma type could be distinguished. Assessment of the FSC/SSC dot plots with histology enables to distinguish subpopulations of the lymphoma cells. The immunoblastic and centroblastic lymphomas were characterized by high proliferative activity although no significant differences in S-phase values between both groups could be found.

Adult↗

PubMed1995

The effect of nickel compounds on immunophenotype and natural killer cell function of normal human lymphocytes.

In order to elucidate effects of nickel on human lymphocytes in vitro, peripheral blood mononuclear cells from normal donors were initially tested for viability in the presence of increasing concentrations of two selected nickel salts, sparingly-soluble nickel subsulfide (Ni3S2), and promptly-soluble nickel sulfate (NiSO4). After establishing the toxicity profile, the cells were cultured for 24 h with each compound at three nontoxic concentrations, 0.01 mM, 0.02 mM, and 0.04 mM, to determine its effect on lymphocyte immunophenotype and function. Cells were also cultured in the presence of 0.01-0.04 mM magnesium acetate, Mg(CH3COO)2, while still other cell samples were subjected to a mixture of Mg(CH3COO)2 plus either Ni3S2 or NiSO4 at equimolar concentration. Following the culture, the immunophenotype of the cells was determined by indirect immunofluorescence, using monoclonal antibodies to major differentiation antigens of peripheral blood mononuclear cells, and their natural killer activity toward K562 target cells was measured. Both nickel salts were found to exert distinct effects on lymphocyte phenotype. Exposure of cells to Ni3S2 resulted in the decline of CD4 and natural killer cell populations. NiSO4 diminished the abundance of natural killer cells and, to a limited extent, also of CD4 cells. The nickel salts tested suppressed natural cytotoxicity of peripheral blood mononuclear cells, with Ni3S2 acting more strongly than NiSO4. The addition of Mg(CH3COO)2 to a nickel salt during in vitro culture abolished the above inhibitory effects. Nickel and magnesium salts did not affect CD3, CD8, CD20, and CD11a cell populations. The results indicate that nickel salts have deleterious effects on human peripheral blood mononuclear cells in short-term in vitro culture, but the magnitude of these effects varies, depending on the cell subsets.

Cell Survival↗

PubMed1995

Ultrastructural and immunophenotypic differentiation of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor (GM-CSF).

The present study examined electron microscopical, immunohistochemical and morphometric changes in the development, differentiation, and maturation of dendritic cells in the culture of mouse bone marrow cells supplemented with granulocyte/macrophage colony-stimulating factor (GM-CSF). A large number of large dendritic cells were released from the aggregates of small round cells. These released cells showed morphological, immunophenotypic, and functional characteristics of typical dendritic cells. These dendritic cells possessed irregularly-shaped nuclei in abundant cytoplasm with rough endoplasmic reticulum (rER), vesicles, multivesicular bodies, and tubulovesicular system, and projected long dendritic cytoplasmic processes. The tubulovesicular system and dendritic surface projections were characteristic of interdigitating cells usually residing in the paracortical area of lymph nodes. Small cells in aggregates were round in shape, had oval nuclei in narrow cytoplasm with poorly developed intracellular organelles, and projected a few short processes. These cells were a proliferating population distinct from monocytes in both ultrastructure and immunophenotype. Morphometrical analysis of cultured cells provided evidence that the small cells differentiate into typical dendritic cells via intermediate cells. These results imply that dendritic cells differentiate from small round dendritic precursor cells and that GM-CSF is a major cytokine capable of inducing the development, differentiation, and proliferation of dendritic cells.

Animals↗

PubMed1995

Expression of the E2A-PBX1 fusion transcripts in t(1;19)(q23;p13) and der(19)t(1;19) at diagnosis and in remission of acute lymphoblastic leukemia with different B lineage immunophenotypes.

The chromosomal translocation t(1;19)(q23;p13) and its variant form der(19)t(1;19) found in 3-5% of acute lymphoblastic leukemia (ALL) results in the expression of the E2A-PBX1 fusion transcript. Although strongly associated with a pre-B immunophenotype, we report the occurrence of t(1;19) in bone marrow or peripheral blood in nine patients with ALL with the following immunophenotypes: pre-B ALL (four), c-ALL (two), c-ALL clg not tested (one), null-ALL (one) and mature B-ALL (one). The E2A-PBX1 fusion transcript investigated by reverse-transcriptase polymerase chain reaction (RT-PCR) was seen in all patients at diagnosis and/or on follow-up samples. Six patients are alive in first clinical remission. Of these patients, three were PCR+ve from between 2 and 38 months from diagnosis, and three were PCR-ve when examined at 5, 26 and 51 months from diagnosis. Two patients are in second remission. One was PCR+ve at 18 months, suffered a CNS relapse at 21 months but was PCR-ve 1 month later. The other was PCR+ve in remission at 2 and 11 months from diagnosis and in testicular relapse at 31 months, but was PCR-ve 5 months later. The remaining patient died 2 months from diagnosis and was not investigated in remission. The prognostic significance of these findings remains to be investigated.

Adolescent↗

PubMedDate not supplied

Fine needle aspiration cytology of non-Hodgkin's lymphomas. A morphologic and immunophenotypic study.

To assess the value of fine needle aspiration cytology (FNAC) in the diagnosis of non-Hodgkin's lymphomas (NHL), we retrospectively studied all the cases diagnosed cytologically as NHL in our laboratory during a five-year period (1987-1991). We also traced cases in which FNAC failed to diagnose NHL and where the diagnosis was made subsequently by histopathology. Fine needle aspiration (FNA) was performed on both peripheral/palpable and deeply situated lesions. A total of 164 specimens were studied cytologically, and for 130 of them a histologic report was available. In 83 of the cases, FNA was carried out as part of the initial evaluation, and in 81 the diagnosis of NHL was known and FNA was performed to confirm or exclude a relapse. In 76 cases for which morphology was inconclusive the immunophenotype was assessed by immunocytochemistry. There were three false-negative and one false-positive result; in none of them was immunophenotyping performed. No discrepancy was observed in the distinction between low and high grade lymphomas, but this was feasible in only 115 of the 164 specimens studied. We conclude that the method is a feasible, rapid and inexpensive first approach in the evaluation of patients with NHL. FNAC may be substituted for histology in the occasional patient for whom the surgical risk outweighs the inaccuracies of the procedure.

Adolescent↗

PubMed1994

The immunophenotype of acute promyelocytic leukemia (APL): an ECOG study.

In 452 adult patients with de novo acute myeloid leukemia (AML), a series of 22 monoclonal antibodies was used to identify immunophenotypic characteristics of acute promyelocytic leukemia (APL) as compared to other AMLs (groups FAB M1/M2 and M4/M5). Only those patients with FAB M3 cytology were included in the analysis for which APL was confirmed by the presence of the t(15;17) cytogenetic aberration and the detection of the PML/RAR alpha gene fusion transcript by PCR amplification (35 cases). Significantly fewer APL blast cells were positive for the stem cell antigen, CD34 (p = 0.0001) as well as for HLA-DR (p < 0.0001). With respect to myeloid antigens, APLs less frequently expressed the myelomonocytic antigens, CD11b (p = 0.0001) and CD14 (p = 0.0013), whereas expression of CD33, a pan-myeloid marker, was more frequent in APL (p = 0.0001). CD15, the X-hapten carbohydrate structure (lacto-N-fucopentaose-III), typically expressed at the maturation stage of normal promyelocytes, was found to be sialylated on APL blasts as recognized by differential binding of the anti-CD15 antibodies, VIM-D5 (non-sialylated CD15) and VEP-9 (sialylated CD15). Expression of the T-cell associated CD7 antigen was rarer on APL than non-APL cells (p = 0.0001), as was that of the multidrug resistance P-glycoprotein (p = 0.0038). Marginal correlations existed between antigen profile (particularly CD2) and the type of PML/RAR alpha transcripts. In addition to its unique genotypic features, these data establish APL as a distinct immunophenotypic entity.

Adult↗

PubMedDate not supplied

Single step immunophenotyping of acute leukemias not classifiable by standard morphology and cytochemistry: a practical approach.

BACKGROUND: Immunophenotyping is at present a useful aid and often an essential step for correct diagnosis of acute leukemia and for this purpose some investigators have proposed several immunomarker panels. This approach is particularly important in the differential diagnosis of acute leukemias not classifiable by standard morphology and cytochemistry. METHODS: We have tested peripheral blood and/or bone marrow samples from 125 patients not classifiable by FAB criteria. In all the cases, the reactivities of the same panel of 17 monoclonal antibodies were analyzed by flow cytometry, using both single and double fluorescent labeling. RESULTS: Of the 125 patients investigated, 75 (60%) were classifiable as ALL, 58 as B-lineage ALL and 17 as T-lineage ALL; 33 (26.4%) as AML, of which 2 M7; 6 (4.8%) as biphenotypic and 11 (8.8%) as immunophenotypically undifferentiated. CONCLUSIONS: From a critical analysis of our cases and a review of the literature, we suggest that a panel of 9 monoclonal antibodies (CD2, CD5, CD7, CD10, CD19, CD20, CD13, CD33, CD41), is sufficient for reliable, rapid and reasonably low cost typing of acute leukemia, useful for an immediate therapeutic decision.

Acute Disease↗

PubMed1994

The immunophenotype of splenic lymphoma with villous lymphocytes and its relevance to the differential diagnosis with other B-cell disorders.

Splenic lymphoma with villous lymphocytes (SLVL) is a low-grade disorder that regularly presents with peripheral blood involvement. We describe the immunophenotype of the circulating cells from 100 SLVL patients whose disease has been characterized on clinical, morphologic, and histologic grounds. Cells from all cases expressed B-cell antigens (CD19 and CD37) and/or HLA-Dr and showed light chain restriction (kappa/lambda: 1.5/1) with moderate to strong intensity of membrane Ig staining. Cells from most cases (> 80%) were CD24+, FMC7+, and expressed strongly membrane CD22. The monoclonal antibodies CD10, CD23, and CD38 were positive in one-third of the cases; CD11c in 47%; and CD25 in 25% of cases. A minority of cases (< 20%) were positive with HC2, B-ly-7, and CD5. However, none of the 19 CD5+ cases had the phenotype characteristic of chronic lymphocytic leukemia (CD5+, CD23+, FMC7-, weak surface Ig and membrane CD22). None of the 17 CD25+ cases had the immunophenotype typical of hairy cell leukemia (CD25+, CD11c+, HC2+, B-ly-7+). HC2 and B-ly-7 were the most useful reagents to distinguish SLVL from hairy cell leukemia. Our findings demonstrate that SLVL has a distinct immunologic profile and that monoclonal antibodies are important for the differential diagnosis between this disease and other B-lymphoproliferative disorders with which SLVL can be confused.

Adult↗