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PubMed2003

Immunophenotypic characterization of normal peripheral blood B lymphocyte by flow cytometry: reference for diagnosis of chronic B cell leukemia/lymphoma.

To establish reference values of various immunophenotypic markers in B lymphocyte population in healthy Chinese adults and build background information for accurate interpretation of B cell immunophenotyping data in clinical practice, peripheral blood from 41 healthy adults were collected separately into test tubes containing EDTA-K(2) and stored in room temperature no more than 24 hours before analysis. Whole blood lysis technique and multiparameter flow cytometry were applied to immunophenotype B cells gated on CD19/SSC dot-plot. The results showed that CD22, CD20, CD62L, CD40, CD24, CD79b, CD79a, and FMC-7 were almost positive in the circulating B cell population, whereas CD11a, CD80, CD103, CD10, CD40L, CD54, CD95L, CD86, and CD95 were almost negative in the peripheral blood B lymphocytes. CD18, CD44, CD23, CD5, CD11c and CD43 were positive in different B cell subpopulations. 78% of B cells were IgD positive and ratio kappa/lambda was 1.26. The significance of all these markers in the differential diagnosis of lymphoproliferative diseases was discussed. The conclusion is that it is necessary to consider the qualitative and quantitative levels of expression of various markers in normal B cell population in order to accurately interpret the pathological immunophenotypic data in clinical practice. It is also important to note the immunotypic differences of B cells between Chinese and Western populations.

Adult↗

PubMed1992

Inconsistency of the immunophenotype of Reed-Sternberg cells in simultaneous and consecutive specimens from the same patients. A paraffin section evaluation in 56 patients.

Both immunophenotypic overlaps between Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL), and evolution of one into the other have been reported. However, the underlying assumption that the antigenic expression of Reed-Sternberg (RS) cells is consistent in the same patient has not been evaluated. Such an evaluation was undertaken by immunophenotyping paraffin-embedded lymphoid tissue biopsies with HD from 56 patients in whom multiple specimens were obtained, either simultaneously from different sites or at different times. The panel of antibodies we used included: CD3 polyclonal antiserum, DAKO-M1 (CD15), L26 (CD20), BerH2 (CD30), MT1 (CD43), DAKO-LCA (CD45RB), UCHL1 (CD45R0), LN2 (CD74), and DAKO-EMA. The phenotype of RS cells was identical in simultaneous biopsies in only 11 of 39 patients (28%) and remained constant in consecutive biopsies in only 4 of 21 patients (19%). Major differences (relative to cell lineage specific antigens) were observed in 10 of 39 patients with simultaneous biopsies and in 10 of 21 patients over time; they mainly involved expression of T-cell antigens. Minor differences (relative to any other antigen) were observed in 22 of 39 patients with simultaneous biopsies and in 15 of 21 patients over time; these mainly involved CD15 or CD74. This striking variability of the immunophenotype of RS cells in the same patient may be due to aberrant marker expression, as a result of the neoplastic state, and/or to modulation of antigenic expression in relation to the host environment. This inconsistency suggests caution when interpreting the relationship between HD and NHL by paraffin immunophenotyping alone.

Antibodies↗

PubMed2004

[Immunophenotyping of acute leukemias: diagnostic and pronostic utility in Abidjan, Côte d'Ivoire].

Flow cytometry is nowadays the first-line method for immunophenotypic identification of blast cells but is not so usual in limited-resources countries. We have investigated on the usefulness of this tool in Abidjan, Côte d'Ivoire. Bone marrow sample from 13 patients with acute leukemia identified by cytology and cytochemical analysis was immunophenotyped by using monoclonal antibodies directed to: T lymphoid cells (CD3, CD5, CD7); B lymphoid cells (CD10, CD19, CD20, CD22, HLA-DR) and myeloid cells (CD13, CD33). Immunophenotyping allowed us to confirm the diagnosis of 6 de novo acute leukemias (2 acute myeloid leukaemias, 4 acute lymphoid leukemias) and 7 acute leukaemias resulting from chronic myeloid leukaemias. Immunophenotyping also characterizes the atypical/aberrant lineage essential for the prognosis: 2 biphenotypic acute leukemias (myeloid/lymphoid T) were identified. Our results suggest that flow cytometry may be a useful additional tool to identify the specific leukemic cell, to make a better classification as well as a prognosis evaluation of patients with acute leukemias.

Acute Disease↗

PubMed2005

[Immunophenotypes in 115 patients with acute myeloid leukemia by multi-color flow cytometry].

Immunophenotyping has become common in the diagnosis and classification of leukemia. To evaluate the immunophenotype of acute myeloid leukemia (AML), multiparameter flow cytometry and CD45/SSC gating were used to analyze the surface and cytoplasmic antigen expressions in 115 cases of AML. The results were compared with the French-American-British (FAB) Cooperative Group classification to help define the best use and role of multiparameter flow cytometry in the diagnosis and proper classification of AML. The results showed that CD38, CD38 and CD13 were the most commonly expressed antigen (94.8%, 91.3% and 89.6%, respectively). CD7 was the most commonly expressed lymphoid antigen (20.2%), followed by CD19 (16.5%) and CD2 (15%). Some immunophenotypes correlated with FAB type, including increased frequency of CD2 in M(3); lack of HLA-DR, CD34 and CD56 expression in M(3); increased frequency of CD19 in M(2), CD14 and CD56 in M(5) and lack of MPO in M(0). In conclusion, multiparameter flow cytometry is a reliable technique in the diagnosis of AML, and some immunophenotypes correlate with FAB type.

Acute Disease↗

PubMed2005

[Sensitivity and specificity analysis of the lineage related antibodies in acute leukemia immunophenotyping by flow cytometry].

To evaluate the sensitivity and specificity analysis of the lineage related antibodies in acute leukemia immunophenotyping by flow cytometry (FCM), immunophenotyping in 184 patients with acute leukemia was performed by FCM analysis. The results showed that in the lineage-related antibodies of acute myelocytic leukemia (AML), the sensitivity of CD13 and CD33 was higher (95.5% and 91.2%, respectively), the specificity of them was deficient (72.5% and 62.2%, respectively); the sensitivity of MPO was low (69.1%), but the specificity was high (100%); the sensitivity and specificity of CD117 were high (88.2% and 100%, respectively); the sensitivity of CD14 and CD15 was low (18.4% and 27.2%, respectively); the specificity of CD14 with monocytes was high. As the lineage-related antibodies of B-lineage ALL were concerned, CD19 showed high sensitivity and low specificity (100% vs 83.4%); the sensitivity and specificity of CD79a (96.4% vs 100%) and CD22 (100% vs 100%) were high; the sensitivity and specificity of CD10 (53.6% vs 82.5%) and CD20 (70.4% vs 87.5%) were low. In T-lineage ALL, the specificity of CD3 was high (97.5%), but the sensitivity was below the mark (80.0%); the sensitivity of CD7 was high (100%), but the specificity was low (77.9%); while the sensitivity and specificity of CD5, CD2 and CD1a were all deficient. In conclusion, the sensitivity and specificity analysis of the lineage-related antibodies in acute leukemia immunophenotyping are coincident with St Jude immunophenotyping project. It seems only that CD117 is superior to MPO in defining AML, but the sensitivity and specificity analysis of CD22 and CD79 are similar in defining B-lineage ALL, therefore, anyone of them may be selected as your need.

Acute Disease↗

PubMed2006

[Immunophenotypic features in leukemia of NK cell series].

The aim was to investigate the immunophenotypes of NK series leukemia. Immunophenotypes of 297 cases of acute leukemia (AL) were measured by flow cytometry, and these immucopenotypic features were analyzed. The results showed that 43 out of 297 cases of AL (14.5%) were CD56 positive. 6 cases were NK series leukemia and 37 cases were acute myelogenous leukemia with CD56 expressed. One patient has been diagnosed as myeloid/NK cell precursor acute leukemia, two patients were blastic NK cell leukemia, one was supposed to be NK-like T-cell lymphoma/leukemia, while another one was large granular lymphocyte leukemia (LGLL). It is concluded that almost all of CD56 positive leukemia were acute myelogenous leukemia with CD56 expressed. The immunophenotypes of NK series leukemia were antigens from hematopoietic stem cells to T/NK progenitor cells with meyloid antigen positive, and through NK progenitors to mature NK cells. The immunophenotypes of heterogeneous NK leukemia cells are different, that should be carefully distinguished.

Adolescent↗

PubMed2006

Immunophenotyping of Hodgkin's disease--an aid to its classification.

Based on morphological, phenotypic, genotypic and clinical findings, Hodgkin's disease has recently been classified into two subtypes: Nodular lymphocyte predominant and classical Hodgkin's disease. Forty-two cases of Hodgkin's disease were subjected to detailed morphological assessment and immunophenotyping. The commonest subtype was Nodular Sclerosis seen in 27 cases. The panel of antibodies used was CD 15, CD 30, CD 3, CD 20, CD 45 and Epithelial Membrane Antigen. Immunophenotyping was done by Streptavidin Biotin Peroxidase complex technique. CD 30 was expressed in 86% and CD 15 in 76% cases. Immunophenotype helped reclassify two cases, according to the WHO classification scheme. Although morphology remains the gold standard in the diagnosis of Hodgkin's disease, immunophenotype is a useful adjunct in differentiating prognostically distinct subtypes.

Adolescent↗

PubMed2006

Comparison of immunophenotype and clinical manifestations between patients with M5a and M5b of acute monocytic leukemia.

Acute monocytic leukemia is a distinct subtype of acute myeloid leukemia (AML) with characteristic biology and clinical features. This study was designed to compare the immunophenotypical features and clinical manifestations of the patients with AML-M(5a) to that of patients with AML-M(5b), and to identify differences between M(5a) and M(5b) and to explore their relations. A total of 58 cases of de novo adult patients with AML M(5) were investigated. Immunofluorescence analysis by flow cytometry was performed to determine the immunophenotype of the leukemic cells in all cases. Meanwhile, clinical data of these cases were studied retrospectively. The results showed that the immunophenotypes of monocytic leukemic cells in patients with AML M(5) were heterogeneous, and CD68 and CD11b were expressed higher in patients with AML M(5a), compared with that in patients with AML M(5b) (P < 0.01). The significant differences in sex, extramedullary infiltration, WBC counts of peripheral blood, complete remission rate and disease-free survival (DFS > 300 days) between the patients with AML M(5a) and M(5b) did not exist (P > 0.05). It is concluded that the special individual immunophenotype features can be detected in patients with either of AML M(5a) or M(5b), and that expressions of CD68 and CD11b were much higher in M(5a). It seems that the complete remission rate and disease-free survival of patients with M(5a) and M(5b) are not different from that of currently available therapy.

Adolescent↗

PubMed1991

Small lymphocytic lymphomas with predominant splenomegaly: a comparison of immunophenotypes with cases of predominant lymphadenopathy.

In this study, we compared small lymphocytic lymphomas with predominant lymphadenopathy with those with predominant splenomegaly and found differences in morphology and immunophenotype as well as clinical features. Cases with lymphadenopathy were characterized by widespread disease, CLL type morphology with proliferation centers, and a CD5, CD11c, CD23, CD43 positive, CD45Ro negative immunophenotype. Cases with predominant splenomegaly had more localized disease, a mantle zone pattern or a diffuse growth pattern without proliferation centers, and a CD5, CD11c, CD23, CD43 negative, and sometimes CD45Ro positive immunophenotype. CD45Ro (UCHL1) positivity and alkaline phosphatase staining were associated with a mantle zone growth pattern. Comparison with other small lymphocytic lymphoma subtypes indicated that each has its own specific immunophenotype.

Aged↗

PubMed1995

True T-cell chronic lymphocytic leukemia: a morphologic and immunophenotypic study of 25 cases.

We studied 25 T-cell chronic lymphocytic leukemia (T-CLL) cases collected over a 15-year period. Immunophenotypic analysis was performed in each case; 12 cases were evaluated by cytogenetics, and gene rearrangement studies were performed in 14 cases. The median age was 57 years with a male predominance (M:F, 15:10). The median presenting lymphocyte count was 36.3 x 10(9)/L (range, 3.9 to 438 x 10(9)/L). Fourteen patients (56%) had shotty adenopathy and ten (40%) had mild-to-moderate splenomegaly at presentation; four (16%) had erythematous skin lesions. The lymphocytes were predominantly small; some cases had a minor component of medium-sized cells (< 10%). The nuclear: cytoplasmic ratios were uniformly high with round to oval nuclei; however, a wide spectrum of nuclear outlines could be found, ranging from minimally to markedly convoluted. Nucleoli were either absent or small and inconspicuous. These lymphocytes did not have the morphology of prolymphocytes and did not contain cytoplasmic granules. Bone marrow infiltration was generally in an interstitial pattern; the degree of involvement ranged from 15% to 90%. Immunophenotyping showed that the lymphocytes were mature T-cells with a predominant CD4+ immunophenotype. Three cases displayed a CD8+ immunophenotype. The patients were treated with a variety of chemotherapeutic regimens with only a minimal response observed in two of 20 patients. We conclude that T-CLL is an uncommon chronic lymphoproliferative disorder (CLPD) that can be morphologically similar to B-CLL, is distinct from T-prolymphocytic leukemia, and has an aggressive clinical course that is refractory to therapy. It may also be difficult to distinguish T-CLL from other T-CLPD, especially the leukemic phase of peripheral T-cell lymphoma and some cases of Sézary syndrome.

Adult↗

PubMed1995

Immunophenotypic characteristics of acute leukaemia after myelodysplastic syndromes.

PURPOSE: To analyse the immunophenotype of acute leukaemia (AL) after myelodysplastic syndromes (MDS) (MDS-AL) and to compare the immunophenotypic profile of acute myeloblastic leukaemia (AML) secondary to MDS (MDS-AML) with that of "de novo"-AML. PATIENTS AND METHODS: Twenty patients with MDS-AL and 29 patients with "de novo"-AML were studied. Morphocytochemical and flow cytometric studies were done in each case. RESULTS: All the MDS-AL studied displayed a myeloid phenotype (MDS-AML). The main difference between MDS-AML and "de novo"-AML was a significantly higher frequency of CD34 expression in the first group. Differences concerning the expression of other non-lineage related or myeloid-associated markers were not statistically significant, although the percentage of cases CD15(+) was lower in MDS-AML. The overall frequency of expression of lymphoid-associated markers was similar in both groups, T-cell markers being more frequently detected. CONCLUSIONS: Our findings support the usefulness of immunophenotyping studies to characterize MDS-AL and suggest some immunophenotyping differences between MDS-AML and "de novo"-AML which might have biological and prognostic significance.

Adult↗

PubMed1995

Primary lymphomatous effusions in AIDS: a morphological, immunophenotypic, and molecular study.

Lymphomas were documented in pleural effusions or ascites in 18 human immunodeficiency virus-positive (HIV+) patients. Eleven of 12 with clinical data had acquired immunodeficiency syndrome before the diagnosis of lymphoma. In 13 of 15 with data available, a body cavity was the site of initial presentation of lymphoma. Cytological subtypes were large cell immunoblastic, n = 7; large cell anaplastic, n = 6; and large cell NOS, n = 5. The high incidence of anaplastic large cell lymphoma and the conspicuous absence of Burkitt's lymphoma differ strikingly from HIV-associated lymphomas generally. Immunophenotypically, two cases were B-cell (CD19/20+, sIg+, CD/5-), one was T-cell (CD3+, CD5+, CD4+, CD8-, CD19/20-, sIg-), and 15 were null (CD45+, HLA-DR+ CD19/20-, sIg-, CD3/5-). This 83% incidence of null immunophenotype contrasts sharply with a 9% incidence among 35 tissue-based lymphomas in HIV+ patients that were similarly studied and a 0% null immunophenotype among 11 lymphomatous effusions in patients without HIV risk factors. Seven of the 18 HIV-associated lymphomas expressed CD30. Four of five cases with null immunophenotype showed Ig heavy-chain gene rearrangement, two had clonal Epstein-Barr virus integration, and none had MYC protooncogene rearrangement. These cases belong to a subgroup of high-grade HIV-associated lymphomas that occur in the setting of profound immunosuppression in which immunoblastic morphology predominates and MYC rearrangement is encountered only infrequently.

Acquired Immunodeficiency Syndrome↗

PubMed1995

Discordant immunophenotype of chronic B-cell lymphoproliferative disorders in simultaneous specimens from bone marrow and peripheral sites.

This study consisted of 10 cases of chronic B-cell lymphoproliferative disorders that had simultaneous specimens obtained from both bone marrow and peripheral sites for flow cytometric immunophenotyping. The immunophenotyping results of peripheral sites from all 10 cases showed a monoclonal B-cell proliferation expressing monoclonal surface immunoglobulin, CD19, CD20, HLA-DR, and CD5 (except 1 case). Eight (80%) of the 10 cases, however, demonstrated discordant immunophenotypes with myeloid-associated marker expression (CD13, CD11b, and/or CD15) found only in the bone marrow. Patients with CD13 or CD11b marker expression in the bone marrow followed an aggressive clinical course with advanced Rai's stage and a diffuse or mixed bone marrow infiltration pattern or disease transformation. These results indicate that discordant immunophenotypes of malignant cells from different body sites occur in chronic B-cell lymphoproliferative disorders and are not uncommon. Additionally; myeloid-associated markers, which some investigators have described as being associated with an unfavorable clinical course, may be expressed only in bone marrow specimens in these disorders. Thus, bone marrow specimens may be preferential in determining myeloid-associated marker expression in chronic B-cell lymphoproliferative disorders.

Aged↗

PubMed1993

Immunophenotypic and immunogenotypic approaches useful in distinguishing benign and malignant lymphoid proliferations.

Distinguishing benign and malignant lymphoproliferative disorders by purely morphologic criteria can be difficult. Immunophenotypic analysis is a useful adjunct to the morphologic evaluation of lymphoproliferative disorders and is of considerable assistance in resolving difficult diagnostic dilemmas. Immunophenotypic analysis can be performed on virtually any pathologic specimen using a variety of methodologic approaches. The analysis should be tailored to the individual pathologic specimen and the specific diagnostic problem. The results should be correlated with the clinical and morphologic findings. Southern blot hybridization analysis of the antigen receptor genes has proven to be an objective, accurate, and sensitive method by which to determine the lineage and clonality of lymphoid proliferations and to detect clonal B- and T-cell populations that are not recognizable morphologically or immunophenotypically. Polymerase chain reaction-based strategies extraordinarily more sensitive than Southern blotting are currently being developed. However, the demonstration of clonal antigen receptor gene rearrangements is an indication of clonality and not necessarily of malignancy, although the two are closely related. This and all the other biologic and technical limitations of immunogenotypic analysis must be considered when evaluating the results of such studies. Ideally, the results of immunogenotypic analysis should be interpreted only in conjunction with the results of morphologic evaluation and immunophenotypic analysis.

Antibodies, Monoclonal↗

PubMed1995

Fine needle aspiration for flow cytometry immunophenotyping of non Hodgkin lymphoma.

The aim was to evaluate the usefulness of lymph node biopsies obtained by fine needle aspiration (FNA) for immunophenotyping of non Hodgkin lymphoma (NHL). Seventeen superficial and deep lymph node samples were fractioned for conventional cytological examination and immunophenotyping studies. Out of ten NHL, nine were readily detected by flow cytometry (FC), while failure on the remaining case was due to selective loss of large cell population, which is liable to occur with this procedure. A single case, which proved negative for all markers employed, was finally diagnosed by immunohistochemistry as germ cell tumor. The other six cases, presenting lymphoid population without phenotypic abnormalities, were diagnosed by cytology and/or histology as Hodgkin disease or hyperplasic disorders. To conclude, FC immunophenotyping seems to improve the efficacy of FNA in NHL diagnosis, whereas for Hodgkin disease and hyperplasic disorders, classic morphological criteria are more useful for differential diagnosis. Although FNA for FC immunophenotyping cannot replace histopathological examination for NHL diagnosis, it proves to be a useful tool for staging and follow up, making surgical procedures for sample collection unnecessary.

Biopsy, Needle↗

PubMed1996

[Immunophenotype. Clinical and laboratory features of acute lymphoblastic leukemia in Chile. Study of 500 children and 131 adults].

We describe the clinical features and immunophenotype of 500 children and 131 adults with acute lymphoblastic leukemia (ALL), diagnosed between 1984 and 1993. Cases were classified, according to immunophenotype in B-cell ALL with three subtypes (pro-B or null, common and B) and T-cell ALL. Among children, common ALL accounted for 74% of cases and pro-B all was more common in children of less than one year (14%). B ALL was observed in 2% of children. Ten percent of children, mostly males, had T-cell ALL. The third part of these children had high leukocyte counts and a mediastinal mass. Children from Mapuche origin, compared with Caucasian children had a lower proportion of common ALL (36 and 74% respectively) and a higher proportions of T-cell ALL (41 and 10% respectively). Among adults common ALL was the most common phenotype (72%) followed by T-cell ALL (15%), pro-B ALL (11%) and B-cell ALL (2%). There was a lower incidence of children with common ALL with positive cytoplasmic immunoglobulin compared to North American or European studies (2 and 15-33% respectively) and a higher proportion of adults with common ALL compared with pro-B cell ALL, in contrast to European studies that show a higher proportion of patients with pro-B cell ALL. No other immunophenotypic, clinical or laboratory differences were observed with ALL from developed countries. It is concluded that the immunophenotyping of ALL allows a more precise diagnosis of this disease.

Adolescent↗

PubMed1996

Lymphocyte immunophenotype reference ranges in healthy Thai adults: implications for management of HIV/AIDS in Thailand.

Lymphocyte immunophenotype reference ranges for T, B, and NK subsets were determined for healthy adult Thais in a multi-center study in Bangkok. Immunophenotyping was by flow cytometry using lysed whole blood. A standard protocol for flow cytometry instrumentation, reagents and quality control was used to minimize site differences and to facilitate comparison of the Thai reference values to those found for Caucasians in previous studies. Major differences were determined for CD3(T), CD4 (T helper/inducer) and CD16+56 (NK) lymphocyte percentages and CD4 lymphocyte absolute counts. Age trends and sex differences were also observed. Compared to Caucasians, Thais, particularly Thai males, had lower CD3 and CD4 T lymphocyte percentages and absolute numbers whereas the percentage of NK lymphocytes was higher. Heterogeneity attributed to biological variation of CD4 T lymphocyte but not other immunophenotype subset distributions was also observed in a well defined geographic population. This study demonstrates the importance of ethnicity, age, sex and possibly environment as factors that influence distribution characteristics of normal lymphocyte immunophenotype reference values. These observations have important implications for the use of lymphocyte subsets-particularly CD3+ CD4+ T lymphocyte measurements as applied to HIV disease staging, AIDS definition and the overall clinical management of HIV/AIDS in Thailand.

Adult↗

PubMedDate not supplied

Immunophenotypic characteristics of the blast crisis in chronic myeloid leukemia.

OBJECTIVE: To characterize the immunophenotype of blast crisis (BC) in Mexican patients with chronic myeloid leukemia (CML). MATERIAL AND METHODS: Mononuclear cells of 17 patients with CML in BC were immunophenotyped employing a panel of 18 monoclonal antibodies: CD5, CD10, CD14, CD22, and anti-HLA-DR used in all patients; CD2, CD15, CD19, CD34, and CD41 in 13 to 16 patients; and CD3, CD7, CD13, CD20, CD21, CD33, CD42b, and CD61 in less than 10 patients. RESULTS: Myeloid was the most frequent type (9/17 cases) followed by lymphoid (6/17) and hybrid or mixed lineage (2/17). Four of the myeloid BC expressed megakaryocyte/platelet associated antigens; 5 of 6 cases with lymphoid BC showed an early precursor B cell immunophenotype (HLA-DR+, CD10+), and the other was an uncommon case of lymphoid B/T transformation (CD19+, CD5+). The CD34 antigen was present in 6 out of 15 cases: 4 patients with lymphoid BC, 1 with myeloid transformation, and 1 with megakaryoblastic BC. CONCLUSIONS: Our findings are comparable to those found in the literature comprising 192 patients. The present study confirms the lineage heterogeneity of CML BC and suggests that extensive immunophenotyping may allow insight for a more precise recognition of normal and leukemic ontogenesis.

Antigens, CD↗