INTRODUCTION: The ABX Pentra DX 120 (Pentra DX 120, ABX Diagnostics, Montpellier, France) adopted new technologies to perform differential leukocyte and erythroblast counts. The double matrix can discriminate Large Immature Cell (LIC), Immature Granulocyte (IMG), Immature Monocyte (IMM), Immature Lymphocyte (IML), and Atypical lymphocyte (ALY) in addition to a routine 5-differential count. For erythroblast (ERB), a fluorescence method is employed. In this study, we evaluated the performance of the Pentra DX 120 in the performance of differential leukocyte and erythroblast counts. METHODS: Precision was evaluated using 3-level control materials. Comparison analysis was performed on 200 samples: 100 normal and 100 abnormal samples. We evaluated the 5 part differential count, LIC, IMG, IMM, IML, ALY, and ERB. These parameters were analyzed in comparison with the results from the reference method, manual differential count. RESULTS: The coefficients of variation (CVs) of precision were <5% for neutrophils and lymphocytes, <15% for eosinophils and basophils and <7% for erythroblasts. The correlation coefficients (r) were >0.9 except monocytes and basophils. IMG, ALY and erythroblasts were also well correlated with manual count (r=0.8315, 0.5602, 0.8144, respectively). The efficiency of flagging system was 84% for LIC, 80% for ALY, and 78.0% for increased ERB (>2/100WBCs). CONCLUSIONS: The Pentra DX 120 performed reliable differential leukocyte and IMG, ALY, and ERB results demonstrated comparable performance to manual count. And, the flagging system was efficient for detecting each abnormal cell population. We expect the Pentra DX 120 double matrix and erythroblast count can reduce microscopic review rate in routine laboratory and promote laboratory efficiency.
Basophils ; Eosinophils ; Erythroblasts ; Fluorescence ; Granulocytes ; Leukocytes ; Lymphocytes ; Monocytes ; Neutrophils

OBJECTIVE: To explore the relation of circulating follicular helper T cell (c Tfh) changes with B cell dysfunction in MDS patients. METHODS: 20 patients diagnosed as MDS from Auguct 2015 to October 2017 were enrolled in MDS group, and 20 healthy valuntears matching in age and sex were enrolled in healthy control (HC) group. The perepheral blood in 2 groups were collected, the mononuclear cells (PBMC) from which were isolated by densily gradient contrifugation, at the same time, the serum left in isolation process was reserved for further study. The flow cytometry was used to detect the ratio of cTfh such as CD4CXCR5 T cells and its subset CD4CXCR5ICOS T cells, CD4CXCR5PD-1 T cells in PBMC, as well as the ratio of plasmablast CD19CD20CD38 B cells. The ELISA was used to detect the concentration of IgA, IgM and IgG. The differences in ratio of cTfh cells and plasmablast B cells, as well as the concentration of IgA, IgM and IgG between MDS and HC groups were compared, at the same time, the correlation of cTfh cell ratio with the plasmablast B cell ratio and the concentration of IgA, IgM and IgG in MDS patient was analyzed. RESULTS: The ratio of CD4CXCR5T, CD4CXCR5ICOST cells and CD19CD20CD38B cells and the concentration of IgA, IgM and IgG decreased in MDS patients, while the ratio of CD4CXCR5PD-1T cells increased in MDS patients. The ratio of CD4CXCR5T cells, CD4CXCR5ICOST cells positively correlated with the ratio of CD19CD20CD38B cells, as well as with the concentration of IgA, IgM and IgG in MDS patients. However, the ratio of CD4CXCR5PD-1T cells negatively correlated with the ratio of CD19CD20CD38B cells, as well as with the concentration of IgA, IgM and IgG. CONCLUSION The ratio of circulating Tfh cells and their subsets showed significant changes, that correlate with B cell dysfunction in MDS patients.
B-Lymphocytes ; Humans ; Interleukins ; Leukocytes, Mononuclear ; Myelodysplastic Syndromes ; Plasma Cells ; Receptors, CXCR5 ; T-Lymphocytes, Helper-Inducer

PURPOSE: To evaluate the changes of differential counts and lymphocyte subsets in cancer patients' leukocyte before and after radiotherapy. METHODS AND MATERIALS: From Dec. 1994 to May 1995, the changes of leukocyte and its subsets in 16 patients who received radiotherapy in the Dept. of Radiation Oncology of Dong-A University Hospiatal were investigated. Radiation was delivered from 2700 cGy to 6660 cGy with median dose of 5400 cGy. The results of pre- and post-radiotherapy were analyzed by paired T-test. The results of patients who received < 50 Gy and > or = 50 Gy were analyzed by wilcoxon test. RESULTS: Before and after radiotherapy, there was not any significant differences in the counts of leukocyte, granulocyte and monocyte. A remarkable decrease was noted in lymphocyte counts after radiotherapy(p=0.015). T cells, B cells and natural killer cells were also decreased in number after radiotherapy but it was not significant statistically. T helper cells and T suppressor cells were also decreased in number(p>0.05). The ratio of T helper/suppressor cell was decreased from 1.52 to 1.11 and it was significant statistically(p=0.016). The portion of T suppressor cell among all T cells was increased after radiotherapy (p=0.0195). No significant difference was observed in the analysis of leukocyte and its subsets between patients who reveived < 50 Gy and > or = 50 Gy. CONCLUSION: Radiotherapy caused remarkable decrease in lymphocyte count and its subsets. Among all lymphocyte subsets, T helper cell might be the most vulnerable to radiation, considering decreased ratio of T helper/surppressor cell count after radiotherapy.
B-Lymphocytes ; Cell Count ; Granulocytes ; Humans ; Killer Cells, Natural ; Leukocytes ; Lymphocyte Count ; Lymphocyte Subsets* ; Lymphocytes* ; Monocytes ; Radiation Oncology ; Radiotherapy* ; T-Lymphocytes ; T-Lymphocytes, Helper-Inducer

BACKGROUND: We evaluated the performance of leukocyte differential counting and clinical usefulness of the morphologic flags of the SE-000, and set optimal criteria for selecting and reviewing the specimens with increased abnormal cells. METHODS: From the results of SE-000 and manual leukocyte differential counting in 100 healthy control and 520 patient specimens we evaluated the correlations on the leukocyte fractions as well as the frequency, sensitivity and false positivity of the flags. After determination of the review criteria we calculated total review rate from the 3,403 consecutive CBC specimens. RESULTS: In both control and patient groups the correlation between two methods was high with the exception of monocytes and basophils. Regarding the morphologic flags, Blast was sensitive (86.9%) however could not detect mature looking lymphoblasts. Immature granulocyte showed high sensitivity (93.7%). Left shift showed the highest frequency (34.6%) and false positive rate (82.8%). Atypical lymphocytes and NRBC showed relatively low sensitivity (63.6%, and 50.5%, respectively). We determined to review the slide when 1) All morphologic flags except Left shift are marked, 2) WBC <3,000/microliter or >20,000/microliter, Hb <8.0 g/dL or 18.0 g/dL, Platelet <100,000/microliter or >600,000/microliter, 3) Severe deviation of leukocyte fractions or 4) Specially requested by physician. As a result, total review rate was 25.0% while 14 abnormal cases with no flags could be additionally detected. CONCLUSIONS: A new review criteria determined from the results of CBC and leukocyte differential together with morphologic flags could reduce the review rate without skipping the abnormal cases.
Basophils ; Blood Platelets ; Granulocytes ; Humans ; Leukocytes ; Lymphocytes ; Monocytes

BACKGROUND: We evaluated the performance of leukocyte differential counting and clinical usefulness of the morphologic flags of the SE-000, and set optimal criteria for selecting and reviewing the specimens with increased abnormal cells. METHODS: From the results of SE-000 and manual leukocyte differential counting in 100 healthy control and 520 patient specimens we evaluated the correlations on the leukocyte fractions as well as the frequency, sensitivity and false positivity of the flags. After determination of the review criteria we calculated total review rate from the 3,403 consecutive CBC specimens. RESULTS: In both control and patient groups the correlation between two methods was high with the exception of monocytes and basophils. Regarding the morphologic flags, Blast was sensitive (86.9%) however could not detect mature looking lymphoblasts. Immature granulocyte showed high sensitivity (93.7%). Left shift showed the highest frequency (34.6%) and false positive rate (82.8%). Atypical lymphocytes and NRBC showed relatively low sensitivity (63.6%, and 50.5%, respectively). We determined to review the slide when 1) All morphologic flags except Left shift are marked, 2) WBC <3,000/microliter or >20,000/microliter, Hb <8.0 g/dL or 18.0 g/dL, Platelet <100,000/microliter or >600,000/microliter, 3) Severe deviation of leukocyte fractions or 4) Specially requested by physician. As a result, total review rate was 25.0% while 14 abnormal cases with no flags could be additionally detected. CONCLUSIONS: A new review criteria determined from the results of CBC and leukocyte differential together with morphologic flags could reduce the review rate without skipping the abnormal cases.
Basophils ; Blood Platelets ; Granulocytes ; Humans ; Leukocytes ; Lymphocytes ; Monocytes

BACKGROUND: Recent advances of hematology analyzers have improved performance of leukocyte differential counts and have reduced work load of clinical hematology laboratories. We evaluated CELL-DYN Sapphire (Abbott Diagnostics, Santa Clara, CA, USA) performance on leukocyte differential counts according to Clinical and Laboratory Standards Institute (CLSI) document H20-A. METHODS: We evaluated imprecision (short term imprecision from duplication of 147 patients' sample and long term imprecision from three level commercial controls) and accuracy (n=462) of leukocyte differential counts of CELL-DYN Sapphire and compared with those of Sysmex XE-2100 (TOA Medical Electronics Co., Kobe, Japan), ADVIA 120 (Bayer Diagnostics, Tarrytown, NY, USA) and Beckman Coulter LH 750 (Beckman Coulter, Miami, FL, USA). RESULTS: The imprecision of CELL-DYN Sapphire for neutrophils and lymphocytes differentials was low with coefficients of variation (CV) from 1.4 to 6.2%, but the imprecision for basophils was high with CV from 34.7 to 79.6%. The correlation with manual count was good in samples without flags (n=314), with the exception of basophils (r: neutrophils, 0.921; lymphocytes, 0.921; monocytes, 0.653; eosinophils, 0.869; basophils 0.272). The correlation with other hematology analyzers was high except basophils (r: neutrophils, 0.969-0.986; lymphocytes, 0.986-0.990; monocytes, 0.787-0.887; eosinophils, 0.881-0.962; basophils 0.086-0.327). CONCLUSION: The performance on leukocyte differential counts of CELL-DYN Sapphire is comparable to Sysmex XE-2100, ADVIA 120 and Beckman Coulter LH 750. In regards of enumeration of basophils, the comparison with manual counts and other hematology analyzers shows poor agreement.
Aluminum Oxide* ; Basophils ; Electronics, Medical ; Eosinophils ; Hematology* ; Leukocytes* ; Lymphocytes ; Monocytes ; Neutrophils

PURPOSE: Radiation-induced alteration in the immune function is well known phenomenon in cancer patients. Our purpose is to evaluate the extent of immune suppression immediately after mediastinal or pelvic irradiation, which include significant volume of active bone marrow in adults. METHODS AND MATERIALS: 48 cancer patients with mediastinal(N=29) and pelvic irradiation(N=19) were the basis of this analysis. Age ranged from 36 to 76 and mean and median value was 57 years, respectively. Sex ratio was 1.3(M:F = 27/21). The immunological parameters were the complete blood cell(CBC) with differenial cell(D/C) count, T cel subset(CD3, CD4, CD8, CD19), NK cell test(CD16,CD56), and serum immunoglobulin (lgG,lgA,lgM) level. RESULTS: The mean value of white blood cell(WBC) was reduced from 7017 to 4470 after irradiation (p=0.0000). In the differential count, the number of lymphocyte, neutrophil, and basophil was markedly reduced with statistical significance(p<0.01) and the number of monocyte was not changed and, on the contrary, that of eosinophil was increased by irradiation.In the lymphocyte subpopulation analysis, the number of all subpopulations, CD3(T cell), CD4(helper T cell), CD8(suppressor T cell), CD16(NK cell), CD19(B cell) was reduced with statistical significance. The mean ratio of CD4 to CD8 in all patients was 1.09 initially and reduced to 0.99 after radiotherapy(p = 0.34), but the proportional percentage of all subpopulations was not changed except CD19(B cell) after irradiation.In the immunoglobulin study, initial values of lg G, lg A, and lg M were relatively above the normal range and the only lg M was statistically significantly reduced after radiotherapy(p=0.02) CONCLUSION: Mediastinal and pelvic irradiation resulted in remarkable suppression of lymphocyte count in contrast to the relatively good preservation of other components of white blood cells. But the further study on the functional changes of lymphocyte after radiotherapy may be necessary to conclude the effects of the radiation on the immunity of the cancer patients.
Adult ; Basophils ; Bone Marrow ; Eosinophils ; Humans ; Immunoglobulins ; Killer Cells, Natural ; Leukocytes ; Lymphocyte Count ; Lymphocyte Subsets* ; Lymphocytes* ; Monocytes ; Neutrophils ; Radiotherapy ; Reference Values ; Sex Ratio

BACKGROUND: The complete blood cell count (CBC) and leukocyte differential counts are useful tools for making a diagnosis, treating and monitoring a disease. This study evaluated the perfor-mance of Beckman-Coulter's new model, the Coulter GEN-S system (Beckman Coulter Corpora-tion, Miami, USA; GEN-S), and compared it with the Sysmex NE-8000 (Sysmex Corporation, Kobe, Japan; NE-8000) and Sysmex R-3000 (Sysmex Corporation, Kobe, Japan; R-3000). METHODS: One hundred and four blood samples and 120 blood samples were randomly chosen for a comparison analysis from various in-patients and healthy persons, who visited Samsung med-ical center, respectively. The GEN-S system was evaluated according to the linearity and how well it compared with the NE-8000 in terms of the CBC, and the efficiency of the leukocyte suspect flags. RESULTS: The GEN-S showed that the determination coefficients (R(2)) of the WBC, RBC, platelet count and hemoglobin level were more than 0.94 (P < 0.05) and the correlation coefficients (r) of the WBC, RBC, platelet count, reticulocyte count, hemoglobin and MCV were > 0.95 (P < 0.001) when compared with the NE-8000. In addition, the correlations of the leukocyte differential counts for neutrophils, eosinophils, and lymphocytes were good, but those for monocytes and basophils were poor. The sensitivity, specificity and positive predictive value of the leukocyte suspect flags of the GEN-S system were 89%, 73%, and 43%, respectively. CONCLUSIONS: These results demonstrate the comparable performance of the GEN-S system in clinical laboratories. However, a separate microscopic differential count is required due to the low positive predictive value for the leukocyte suspect flags.
Basophils ; Blood Cell Count ; Diagnosis ; Eosinophils ; Hematology* ; Humans ; Japan ; Leukocytes ; Lymphocytes ; Monocytes ; Neutrophils ; Platelet Count ; Reticulocyte Count ; Sensitivity and Specificity

BACKGROUND: The present study was designed to assess the effects of preemptive epidural analgesia on postoperative peripheral WBC response and pain in patients undergoing laparoscopic hysterectomy under general anesthesia. METHODS: Patients were randomly assigned to one of two groups; a preemptive epidural group (Pre-E group, n = 25) or a postoperative epidural group (Post-E group, n = 25). In the Pre-E group, 10 ml of 1% lidocaine and 2 mg morphine were given to achieve T6-level sensory block via an epidural catheter before the induction of anesthesia, but this was not done in the Post-E group. Postoperative pain was assessed by patients using VAS, and venous blood samples were collected four times throughout the study period (30 minutes before induction, immediately after surgery, and on postoperative days 1, and 3). RESULTS: Pain scores 1, 3, and 6 hours after surgery were significantly lower in the Pre-E group than in the Post-E group, but became similar 12 hours after surgery. Significantly more patients requested additional analgesics in the Post-E group (24%) than in the Pre-E group (0%). Monocyte percentages differed significantly in the two groups at 1 day after surgery. However, total WBC counts and percentages of neutrophils, lymphocytes, eosinophils, and basophils were similar in the two groups. CONCLUSIONS: Preemptive epidural analgesia provides more effective postoperative pain control, but shows no significant beneficial effect with respect to postoperative WBC response in patients undergoing general anesthesia for laparoscopic hysterectomy.
Analgesia, Epidural* ; Analgesics ; Anesthesia ; Anesthesia, General ; Basophils ; Catheters ; Eosinophils ; Humans ; Hysterectomy* ; Laparoscopy ; Leukocytes* ; Lidocaine ; Lymphocytes ; Monocytes ; Morphine ; Neutrophils ; Pain, Postoperative*

Flow cytometry was used to identify and characterize monoclonal antibodies (mAbs) that react with rabbit leukocyte differentiation molecules (LDM). Screening sets of mAbs, developed against LDM in other species, for reactivity with rabbit LDM yielded 11 mAbs that recognize conserved epitopes on rabbit LDM orthologues and multiple mAbs that recognize epitopes expressed on the major histocompatibility class I or class II molecules. Screening of mAbs submitted to the Animal Homologues Section of the Eighth Human Leukocyte Differentiation Workshop yielded 7 additional mAbs. Screening of mAbs generated from mice immunized with leukocytes from rabbit thymus or spleen or concanavalin A activated peripheral blood and/or spleen lymphocytes has yielded 42 mAbs that recognize species restricted epitopes expressed on one or more lineages of leukocytes. Screening of the anti-rabbit mAbs against leukocytes from other species yielded one additional mAb. The studies show that screening of existing sets of mAbs for reactivity with rabbit LDM will not be productive and that a direct approach will be needed to develop mAbs for research in rabbits. The flow cytometric approach we developed to screen for mAbs of interest offers a way for individual laboratories to identify and characterize mAbs to LDM in rabbits and other species. A web-based program we developed provides a source of information that will facilitate analysis. It contains a searchable data base on known CD molecules and a data base on mAbs, known to react with LDM in one or more species of artiodactyla, equidae, carnivora, and or lagomorpha.
Animals ; Antibodies, Monoclonal/*immunology ; Antigens, Differentiation/*metabolism ; B-Lymphocytes/cytology/metabolism ; Basophils/cytology/metabolism ; Epitopes/genetics/metabolism ; *Flow Cytometry ; Gene Expression Regulation ; Granulocytes/cytology/metabolism ; Leukocytes/immunology/*metabolism ; Mice ; Monocytes/cytology/metabolism ; Rabbits ; T-Lymphocytes/cytology/metabolism