This study was aimed to analyze the correlation of CD19 positive cell counts in bone marrow of multiple myeloma(MM) patients with therapeutic efficacy and investigate the characteristics of CD19 cell change in MM bone marrow. The CD19(+) and CD38(++)CD45(-), CD38(++)CD45(-)CD56(+) cells in bone marrow of 63 MM patients were detected by flow cytometry. The difference of CD19(+), CD38(++)CD45(-), CD38(++)CD45(-)CD56(+) cell counts at different stages and types, as well as their relation with results of 4 course of VADM or VD chemotherapy were analyzed. The results showed that in 63 MM patients, CD19(+) cell ratio at stage II were higher than those at stage III; CD38(++)CD45(-)CD56(+) cell ratio at stage II were lower than those at stage III; CD19(+) cell ratio in type IgA were higher than those in type IgD; the CD38(++)CD45(-), CD38(++)CD45(-)CD56(+) cell counts in type IgA were obviously lower than those in type IgG, IgD and light chain which showed a negative correlation between cell counts of CD19(+) against CD38(++)CD45(-), CD38(++)CD45(-)CD56(+). CD19(+) cell counts in effective treatment group of all 43 patients and the effective treatment group with VD were both higher than those in the ineffective treatment group; CD38(++)CD45(-) cell counts in effective treatment group with VD was obviously lower than those in ineffective treatment group, and CD38(++)CD45(-), CD38(++)CD45(-)CD56(+) in effective treatment group of all 43 patients were lower than those in ineffective treatment group. It is concluded that CD19(+) cell counts in bone marrow may be related to disease status and development stage of MM, which may be useful to predict treatment efficacy and prognosis.
Antigens, CD19 ; metabolism ; Bone Marrow Cells ; cytology ; metabolism ; Humans ; Middle Aged ; Multiple Myeloma ; diagnosis ; therapy ; Treatment Outcome

BACKGROUND: We developed a single-color multitarget flow cytometry (SM-FC) assay, a single-tube assay with graded mean fluorescence intensities (MFIs). We evaluated the repeatability of SM-FC, and its correlation with multicolor flow cytometry (MFC), to assess its application as a routine FC assay. METHODS: We selected CD19, CD3, CD4, and CD8 as antigen targets to analyze a lymphocyte subset. MFIs were graded by adjusting monoclonal antibody (mAb) volumes to detect several cell populations. Dimly labeled mAb was prepared by decreasing mAb volume and the optimum diluted volume was determined by serial dilution. SM-FC repeatability was analyzed 10 times in 2 normal controls. The correlation between SM-FC and MFC was evaluated in 20 normal and 23 patient samples. RESULTS: CV values (0.8-5.0% and 1.3-4.1% in samples 1 and 2, respectively) acquired by SM-FC with CD3-fluorescein alpha-isothyocyanate (FITC)dim+CD4-FITCbright and with CD19-FITCdim+CD3-FITCbright showed good repeatability, comparable to that acquired by MFC (1.6-3.7% and 1.0-4.8% in samples 1 and 2, respectively). Excellent correlation was observed between the 2 methods in the 20 normal samples (B cells, T cells, non-Thelper cells, and Thelper cells; r2=0.87, 0.97, 0.97, and 0.98, respectively; P<0.05). There were also linear relationships between SM-FC with CD19-FITCdim+CD3-FITCbright and CD8-PEdim+CD4-PEbright, and MFC, in the 23 patient samples (B cells, T cells, Tcytotoxic cells, and Thelper cells; r2> or =0.98, 0.99, 0.99, and 0.99, respectively; P<0.05). CONCLUSIONS: The multicolor, single-tube SM-FC technique is a potential alternative tool for identifying a lymphocyte subset.
Antibodies, Monoclonal/chemistry/*immunology ; Antigens, CD19/chemistry/metabolism ; Antigens, CD3/chemistry/metabolism ; Antigens, CD4/chemistry/metabolism ; Antigens, CD8/chemistry/metabolism ; B-Lymphocyte Subsets/immunology/metabolism ; Color ; Flow Cytometry/*methods ; Fluorescein-5-isothiocyanate/*chemistry ; Humans ; T-Lymphocyte Subsets/immunology/metabolism

IL-10 production by CD19(+)CD5(+) B cells was investigated, by determining the expression levels of CD19, a classical B cell marker. Peripheral mononuclear cells were stained with fluorescence-conjugated anti-CD5, anti-CD19, anti-IL-10, and Annexin V. Interestingly, IL-10-producing B cells were found to be localised within the CD19(low)CD5(+) B cell subset. Apoptotic changes were also observed mainly in CD19(low) cells among B cells. Thus, CD5(+) B cells should be classified as CD19(high) and CD19(low) cells, and the immunological significance of CD19 for the IL-10 production by CD5(+) B cells requires further studies.
Antigens, CD19/metabolism ; Antigens, CD5/metabolism ; Apoptosis/immunology ; B-Lymphocyte Subsets/cytology/*immunology ; Cell Separation ; Flow Cytometry ; Humans ; Interleukin-10/*biosynthesis

OBJECTIVETo investigate the application of flow cytometry in diagnosis of T-cell rich diffuse large B-cell lymphoma.

METHODSHistopathologic features, immunohistochemical findings and flow cytometry results of three cases of T-cell rich diffuse large B-cell lymphoma were reviewed retrospectively.

RESULTSIn CD45-side scatter (SSC) dot plot of the first patient, two different CD45-positive lymphoid cell populations were identified. The bright population consisted of both T and B cells, with a T-cell predominance. The dim population consisted mainly of B cells which showed lambda light chain restriction. In the second patient, CD45-positive cells were subdivided into two groups according to CD45-SSC dot plot. The small population consisted of both T and B cells, with a T-cell predominance. The large population consisted mainly of B cells which showed kappa light chain restriction. In the third patient, CD19-positive cells were subdivided into two groups according to the expression of CD20 in CD19-CD20 dot plot. The CD20-positive population expressed both kappa and lambda light chains, while the CD20-negative population demonstrated kappa light chain restriction.

CONCLUSIONSNeoplastic B cells can be distinguished from reactive lymphoid cells in T-cell rich diffuse large B-cell lymphoma by flow cytometry, according to a number of parameters which include intensity of antigen expression, loss of antigens, expression of non-B-cell lineage antigens, patterns of forward scatter (FSC) and/or SSC, and expression of immature B-cell antigens.

Aged ; Antigens, CD19 ; metabolism ; Antigens, CD20 ; metabolism ; Diagnosis, Differential ; Female ; Flow Cytometry ; methods ; Humans ; Immunoglobulin lambda-Chains ; metabolism ; Immunohistochemistry ; Leukocyte Common Antigens ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; metabolism ; Male ; Middle Aged ; T-Lymphocytes ; metabolism ; pathology

BACKGROUND: HLA-DR negativity is known to be useful for distinguishing acute promyelocytic leukemia (APL) from other subtypes of AML, but non-APL cases without HLA-DR antigen expression have been reported. The purpose of this study was to evaluate and compare the characteristics of APL, HLA-DR negative non-APL, and HLA-DR positive non-APL cases. METHODS: A total of 114 cases of AML admitted at Ewha Womans University, Mokdong Hospital between March 1997 and June 2006 were included in this study. A diagnosis of AML was made based on the results of morphology, cytochemistry, immunophenotype, cytogenetics, and/or fluorescence in situ hybridization. RESULTS: Among the 114 AML patients, HLA-DR antigen was not expressed in 39 (34%), including 24 non-APL (62%) and 15 APL patients (38%). The HLA-DR negative non-APL group showed higher leukocyte counts and positive rate of CD19 expression than did APL group (P<0.05). The remaining laboratory findings were not statistically different between the HLA-DR negative non-APL and APL groups. CD34 expression was more frequent in the HLA-DR positive non-APL group than in the HLA-DR negative non-APL group and APL group. Of the 24 patients with HLA-DR negative non-APL, 7 patients had disseminated intravascular coagulation and 2 patients showed morphologic features similar to those of APL. CONCLUSIONS: CD19 expression and leukocyte count may be helpful for differentiating HLA-DR negative non-APL from APL. However, the final diagnosis and classification should be confirmed by cytogenetic or molecular studies.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD19/metabolism ; Antigens, CD34/metabolism ; Child ; Child, Preschool ; Female ; HLA-DR Antigens/*metabolism ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute/*diagnosis/pathology ; Leukocyte Count ; Male ; Middle Aged ; Retrospective Studies

Immunophenotype is critical for diagnosing common B-cell acute lymphoblastic leukemia (common ALL) and detecting minimal residual disease. We developed a protocol to explore the immunophenotypic profiles of common ALL based on the expression levels of the antigens associated with B lymphoid development, including IL-7Rα (CD127), cytoplasmic CD79a (cCD79a), CD19, VpreB (CD179a), and sIgM, which are successive and essential for progression of B cells along their developmental pathway. Analysis of the immunophenotypes of 48 common ALL cases showed that the immunophenotypic patterns were highly heterogeneous, with the leukemic cell population differing from case to case. Through the comprehensive analysis of immunophenotypic patterns, the profiles of patient-specific composite leukemia cell populations could provide detailed information helpful for the diagnosis, therapeutic monitoring, and individualized therapies for common ALL.
Adult ; Antigens, CD19 ; metabolism ; B-Lymphocytes ; immunology ; metabolism ; CD79 Antigens ; metabolism ; Female ; Humans ; Immunoglobulin Light Chains, Surrogate ; metabolism ; Immunophenotyping ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; pathology ; Receptors, Interleukin-7 ; metabolism

OBJECTIVE: To explore the expression of CD23, CD19 on peripheral blood lymphocytes as well as its association with serum total IgE levels and nasal allergic symptoms in patients with allergic rhinitis (AR). METHOD: Symptom scores were evaluated in 46 AR patients, expression of CD23, CD19 on peripheral blood lymphocytes were measured by flow cytometry, and serum total IgE levels were determined by immune chemiluminescence. Thirty two healthy individuals were enrolled as controls. RESULT: The percentage of CD23+, CD19+ and CD23+/ CD19+,on peripheral blood lymphocytes in AR patients were 11.6 +/- 1. , 22.8 +/- 3.3,10.2 +/- 1.7, respectively. Higher frequencies of CD23+, CD19+, and CD23+/CD19+ were found in AR patients compared with controls (P < 0.05). There were positive correlations between expression rates of CD23+, CD19+, CD23+/CD19+ and levels of serum total IgE, nasal allergic symptom scores, respectively. CD23+/CD19+ demonstrated greater correlations with serum total IgE and nasal allergic symptom (r = 0.65 and 0.49, P < 0.05) than CD23+ and CD19+ did. Correlation between CD23+/CD19+ and nasal allergic symptom scores was greater than the corresponding correlations of serum total IgE (r = 0.33, P < 0.05). CONCLUSION CD23 and CD19 are important factors that associated with serum total IgE in the pathogenesis of AR, Analysis on the expression of CD23+/CD19+ on peripheral blood lymphocytes is helpful for evaluating the severity of AR.
Adolescent ; Adult ; Antigens, CD19 ; blood ; Case-Control Studies ; Female ; Humans ; Immunoglobulin E ; blood ; Lymphocyte Count ; Lymphocytes ; metabolism ; Middle Aged ; Receptors, IgE ; blood ; Rhinitis, Allergic, Perennial ; blood ; Young Adult

BACKGROUND: Bone marrow biopsies are routinely performed for staging patients with B-cell non-Hodgkin lymphoma (NHL). In addition to histomorphological studies, ancillary tools may be needed for accurate diagnosis. We investigated the clinical utility of multiparameter flow cytometric examination of bone marrow aspirates. METHODS: A total of 248 bone marrow specimens from 232 patients diagnosed with B-cell NHL were examined. Monoclonal antibodies directed against CD19, CD20, CD10 (or CD5), and kappa and lambda immunoglobulins were used. Multi-stage sequential gating was performed to select specific cells of interest, and the results were compared with bone marrow histology. RESULTS: The concordance rate between histomorphology and flow cytometry was 91.5% (n=227). Eight cases (3.2%) were detected by flow cytometry alone and were missed by histomorphology analysis, and 6 of these 8 cases showed minimal bone marrow involvement (0.09-2.2%). The diagnosis in these cases included large cell lymphoma (n=3), mantle cell lymphoma (n=3), and mucosa-associated lymphoid tissue (MALT) lymphoma (n=2). Thirteen cases were histopathologically positive and immunophenotypically negative, and the diagnoses in these cases included diffuse large cell lymphoma (n=7), T-cell/histiocyte-rich large B-cell lymphoma (n=2), anaplastic lymphoma kinase (ALK)-positive large B-cell lymphoma (n=1), follicular lymphoma (n=1), MALT lymphoma (n=1), and unclassifiable lymphoma (n=1). CONCLUSIONS: Multi-color flow cytometry can be a useful method for assessing bone marrow in staging NHL and also plays a complementary role, especially in detecting small numbers of lymphoma cells.
Antibodies, Monoclonal/immunology ; Antigens, CD19/immunology/metabolism ; Antigens, CD20/immunology/metabolism ; Bone Marrow/*pathology ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; Lymphoma, B-Cell/*pathology ; Male ; Neoplasm Staging ; Neprilysin/immunology/metabolism

BACKGROUNDThe cause of late-onset hemorrhagic cystitis (LOHC) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains obscure. In clinical practice, some LOHC patients respond to immunosuppression. The aim of this study was to determine the immune pathogenesis of LOHC post allo-HSCT.

METHODSWith the diagnosis of LOHC, patients were given initial treatment consisting of fluid hydration, alkalization and forced diuresis, and empirical anti-viral therapy for 10 - 14 days or until a week after the virus became negative. The nonresponders were applied corticosteroid. Seven to ten days later, patients' response was evaluated. Along with treatment, CD19(+) B lymphocyte subsets were measured at various study points.

RESULTSFrom October 2009 to March 2010, we found 28 cases of LOHC occurred in 25 patients who underwent allo-HSCT in our hospital. Except that three cases were not treated according to the protocol, the other 25 cases were divided into three groups: anti-virus responders (Group A, n = 6), corticosteroid responders (Group B1, n = 16), corticosteroid and anti-virus nonresponders (Group C, n = 3) according to their clinical response. Percentages of CD19(+)CD5(+) B lymphocytes were not significantly different among three groups at onset of LOCH. However, in Group B1 after the first anti-virus phase, percentages of CD19(+)CD5(+) lymphocytes significantly increased comparing with those at onset (P = 0.022), and then significantly decreased at PR (P = 0.003) and CR (P = 0.002) with corticosteroid treatment. But significant change was not observed in Groups A and C.

CONCLUSIONThe immune etiology seems to be involved in the development of LOHC and the proportion of CD19(+)CD5(+) lymphocytes may serve as a cellular biomarker to predict the response to corticosteroid in LOHC.

Adolescent ; Adrenal Cortex Hormones ; therapeutic use ; Adult ; Antigens, CD19 ; metabolism ; B-Lymphocytes ; metabolism ; CD5 Antigens ; metabolism ; Child ; Child, Preschool ; Cystitis ; drug therapy ; immunology ; therapy ; Female ; Flow Cytometry ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Young Adult

BACKGROUND: The aberrant, leukemia-associated antigen expression patterns allow us to discriminate leukemic blasts from normal precursor cells. Our major goal was to determine a guideline for the detection of minimal residual disease using CD20+/CD34+ and myeloid Ag+/CD19+ combination in the bone marrow of acute leukemia in complete remission (CR) after chemotherapy. METHODS: Bone marrow samples from 117 patients with acute leukemia in complete remission after chemotherapy and from 22 healthy controls were immunophenotyped by triple staining and measured by flow cytometry. RESULTS: The CD20+/CD34+ cells in the large lymphocyte gate (R1) ranged from 0% to 3.24% (0.8+/-0.82%, P=0.000) in CD20+/CD34+ B-lineage ALL CR (N=31), from 0.03% to 4.2% (0.7+/-0.83%, P=0.000) in CD20-/CD34- B-lineage ALL CR (N=66), from 0.1% to 0.96% (0.45+/-0.32%, P=0.016) in T-ALL CR (N=10), and from 0.02% to 0.48% (0.18+/-0.15%, P=0.776) in AML CR (N=10). The CD13,33+/CD19+ cells in R1 gate ranged from 0% to 2.69% (0.37+/-0.48%, P<0.001) in CD13,33+/CD19+ B-lineage ALL CR (N=31), from 0% to 1.8% (0.31+/-0.28%, P<0.001) in CD13,33-/CD19+B-lineage ALL CR (N=65), from 0.02% to 0.64% (0.29+/-0.22%, P=0.071) in T-ALL CR (N=9), and from 0% to 0.17% (0.07+/-0.09%, P=0.341) in AML CR (N=3). CONCLUSIONS: Using an immunophenotypic method for the detection of early relapse or minimal residual disease of B-lineage ALL bone marrow in CR after chemotherapy, different cutoff values should be applied according to antigen combination and gating. When the proportion of aberrant antigen combination was less than 5% in large lymphocyte gate, the results should be interpreted with caution.
Acute Disease ; Antigens, CD/*metabolism ; Antigens, CD19/metabolism ; Antigens, CD20/metabolism ; Antigens, CD34/metabolism ; Antigens, Differentiation, Myelomonocytic/analysis/metabolism ; Bone Marrow Cells/*classification/metabolism ; Flow Cytometry ; Hematopoietic Stem Cells/classification/metabolism ; Humans ; Immunophenotyping ; Leukemia/*diagnosis/drug therapy ; Leukemia, Myeloid, Acute/diagnosis/drug therapy ; Neoplasm, Residual ; Remission Induction ; Tumor Markers, Biological/immunology