This study was aimed to investigate the relationship between expression of CD200 antigen and clinical characteristics in AML patients and to analyse the value of CD200 in evaluation of AML prognosis. The CD200 and immunophenotypes were detected by flow cytometry, the chromosome karyotypes were determined by R banding, the FISH was used to measure the AML1/ETO, PML/RARa and inv(16), and PCR technique was used to detect the fusion genes AML1/ETO and PML/RARα. The results showed that the positive rate of CD200 antigen expression in 54 patients was 57.4% (31/54), the CD200 antigen expression between sex and age of patients was no significant different (P > 0.05). There was significant difference of CD200 expression between CD34 and CD117 (P < 0.05), but the difference of CD200 expression in chromosome karyotypes was no significant difference(P > 0.05). Moreover, there was significant difference of CD200 expression in CD34 and CD117 of CBF positive AML patients (P < 0.05). It is concluded that the CD200 antigen expression in AML may associate with a poor prognosis of patients.
Antigens, CD ; immunology ; Chromosome Banding ; Humans ; Immunophenotyping ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; immunology ; Oncogene Proteins, Fusion ; Prognosis

OBJECTIVETo analyze the immunophenotype and leukemia associated immunophenotype (LAIP) of leukemia cells from patients with acute myeloid leukemia (AML) in minimal residual disease (MRD) detection.

METHODSFour-color multiparametric flow cytometry (FCM) with CD45/SSC gating was used to determine the expression of the following antibodies of CD7, CD117, CD33, CD34, CD10, CD19, CD56, CD38, CD13, CD14, CD64, CD9, CD16, CD2, CD5, CD11b, CD123, HLA-DR in 610 AML patients and 20 normal bone marrow (NBM) samples.

RESULTSThe mean percentages of CD34+ and CD117+ side scatter low (SSC(low)) cells in NBM mononuclear cells (BMMNCs) were (0.35 +/- 0.15)% and (0.76 +/- 0.31)%, respectively. The majority (84% -95%) of CD34+ SSC(low) cells co-expressed CD13, CD33, CD38, CD117 and HLA-DR. 33% and 20% of CD34+ SSC(low) cells were CD15+ and CD9, respectively. Only a small proportion (< 10%) of CD34 SSC(low) cell co-expressed CD11b, CD56, CD19 and CD64, while co-expressed CD7 was 12%. In CD117 SSC(low) cells, the relative proportions of CD19+, CD11b+, CD56+ and CD7+ were less than 10%, while CD9+ was 14%. CD38+ and HLA-DR+ were 87% and 91%, respectively. The expressions of CD15, CD34, CD13 and CD33 on CD117 SSC(low) cells were between 30% and 50%. In AML patients, most cases were CD117+ (95.08%), CD38+ (94.74%), CD9+ (84.93%), CD33+ (84.60%), HLA-DR+ (77.23%) and CD13 (75.25%). The proportions of CD64+ and CD34+ were 64.41% and 59.51%, and that of CD15 and CD11b+ were 43.06% and 22.07%, respectively. 86.39% of AML patients were found to have at least one LAIP, the highest incidence being in AML-M1 and M3 subtypes and the lowest in AML-M4Eo subtype. The cross-lineage antigen and asynchronous antigen expression were the most frequent aberrant phenotypes. CD7, CD19 and CD56 expressing on CD34+ cells were major cross-lineage antigen. For asynchronous antigen expression, CD34+ CD64+, CD117+ CD11b+ , CD34+ CD38(-/dim) and CD34+ HLA-DR(-/dim) were seldom expressed on normal BMMNCs (about 0.01%), and the logarithm difference between AML and NBM was larger than 3.0, being the more sensitive LAIP.

CONCLUSIONMRD detection by multiparameter flow cytometry can be applied to more than 80% of AML patients.

Adult ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; immunology ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis

Background: The diagnosis of acute leukemia is multidisciplinary with histology, immunology, and cytogenetics. Among these, cytogenetics is important for diagnosis and analysis of prognosis and some of the chromosomal abnormalities are specific for the particular subtypes of acute leukemia. However, cytogenetic analysis is laborious and sometimes does not provide sufficient metaphases. To detect the common 7 gene rearrangements in acute leukemia, a reverse transcription-polymerase chain reaction (RT-PCR) was performed simultaneously under the same conditions. Methods: The author analyzed 38 cases of acute myeloid leukemia (AML) and 20 cases of acute lymphocytic leukemia (ALL) for the evaluation and treatment of acute leukemia. The simultaneous RT-PCR assays were performed under the same conditions to detect 7 chromosomal abnormalities of t(1:19), t(8; 21), t(9; 22), dupMLL (11q23), t(12; 21), t(15; 17), and inv(16) in acute leukemia. Results: The simultaneous RT-PCR assay detected the expression of 7 fusion genes generated by chromosomal rearrangement. The gene rearrangements were found in 53% of AML and 40% of ALL. In AML, there were 7 cases of PML/RARA, 6 cases of AML1/ETO, 4 cases of dupMLL, and 3 cases of CBF/MYH11. In ALL, 4 cases of dupMLL, 2 cases of BCR/ABL, 1 case of E2A/PBX1, and 1 case of TEL/AML1 were detected. The discrepant results between simultaneous RT-PCR and cytogenetic analysis were found in 11 cases. Nine cases were positive by simultaneous RT-PCR but negative in the cytogenetic analysis and each case of variant t(9; 22) and t(15; 17) was negative in simultaneous RT-PCR. Conclusions: It suggests that the simultaneous RT-PCR assay is an efficient and fast procedure for the detection of genetic changes in acute leukemia and it appears to be an useful method for rapid diagnosis of acute leukemia.
Allergy and Immunology ; Chromosome Aberrations ; Cytogenetic Analysis ; Cytogenetics ; Diagnosis ; Gene Rearrangement* ; Leukemia* ; Leukemia, Myeloid, Acute ; Metaphase ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Prognosis

OBJECTIVETo study the clinicopathologic features, diagnosis and differential diagnosis of extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma.

METHODSFive cases of extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma were selected from 102 cases of myeloid sarcoma diagnosed during the period from 1990 to 2006. The clinicopathologic findings and followup data were retrospectively analyzed. Immunohistochemical study was also carried out with SP method.

RESULTSAmong the 5 cases studied, 3 were males and 2 were females, including 2 children and 3 adults. Generalized lymphadenopathy was found in 4 patients and skin lesions were observed in 2 patients. The tumor cells in all cases were positive for CD68 (KP1), CD68 (PGM1), lysozyme and CD45. They were negative for MPO, CD15, CD163, TdT, CD117, T and B cell markers. The Ki-67 index ranged from 40% to 80%. Follow-up data were available in all the 5 patients. Four of the 5 patients died of the disease, with the average survival time being 6.25 months.

CONCLUSIONSMonoblastic sarcoma is a rare disease with poor prognosis. It is almost impossible to distinguish monoblastic sarcoma from granulocytic sarcoma and other types of small round cell tumors on the basis of morphologic examination alone. Immunohistochemistry is mandatory for a correct diagnosis.

Adult ; Antigens, CD ; immunology ; Antigens, Differentiation, Myelomonocytic ; immunology ; Child ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; methods ; Immunophenotyping ; Leukemia, Monocytic, Acute ; immunology ; pathology ; Leukocyte Common Antigens ; Lewis X Antigen ; immunology ; Male ; Receptors, Cell Surface ; immunology ; Sarcoma ; immunology ; pathology ; Sarcoma, Myeloid ; immunology ; pathology

In order to analyze the clinical characteristics and biological features of acute leukemia in elderly, 104 acute leukemia patients in elderly were retrospectively analyzed and compared with 71 acute leukemia patients below 60 years old. The results showed that: (1) the proportion of AML in the aged group (73%) was higher than that in the young group (54.9%), and the difference was statistically significant (P < 0.05), but AML (M3) was absent in the aged group; (2) the median of bone marrow blast cell in the aged group was significantly lower than that in the young group (P < 0.05); (3) in AML, the frequently of CD14 expression was higher in the aged group (18.8%) than that in the young group (2.6%), while the expression frequencies of CD15 (37.5%), CD117 (62.5%), and CD38 (59.4%) were respectively lower in the aged group than that in the young group which were (69.2%) for CD15, (89.7%) for CD17, and (84.6%) for CD38 respectively, and the difference was also statistically significant (P < 0.05). (4) CD19 was most frequently expressed in ALL of the aged group and the positive rate was 100%; (5) there was no significant difference in expression of special lineage antigens and overlapping lineage antigens between the aged group and the young group (P > 0.05); (6) the expression frequency of unfavorable karyotypes in the aged group was higher than that in the young group, and the difference was statistically very significant (P < 0.01); (7) the complete remission rate (CR rate) in the aged group was 42.9%, 2-year survival rate in the aged group was 5.4%, and treatment-related mortality rate in the aged group was 26.8%, while the CR rate in the young group was 76.6%, the difference was statistically significant (P < 0.05). It is concluded that the expression frequency of CD14 associated with unfavorable prognosis is higher in the aged group than that in the young group, while the expression frequency of CD15 associated with favorable prognosis is lower in the aged group than that in the young group. The expression frequency of unfavorable karyotypes in the aged group is higher than that in the young group. The CR rate of acute leukemia in elderly is low, thus the patients in elderly often have unfavorable prognosis.
Adult ; Age Factors ; Aged ; Aged, 80 and over ; Female ; Humans ; Immunophenotyping ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; immunology ; Lipopolysaccharide Receptors ; analysis ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; immunology ; Prognosis ; Receptor, Macrophage Colony-Stimulating Factor ; analysis ; Remission Induction ; Retrospective Studies

This study was purposed to compare the immunophenotypic and clinical characteristics of NPM1 mutated acute myeloid leukemia with a normal karyotype under the similar constituent ratio of FAB subtypes. Immunophenotyping and NPM1 gene mutation type-A,B and D and other leukemic related fusion genes were detected by multiparameter flow cytometry and real time RT-PCR or PCR, respectively. 77 AML patients with a normal karyotype (NK) and mutated NPM1 gene (NPM1m(+)AML) detected by immunophenotyping assay were included in this study. 55 cases without NPM1 mutation (NPM1m(-)AML) and with normal karyotype were served as negative control. The results showed that there was significant difference between NPM1m(+)AML and NPM1m(-)AML in terms of sex, white blood count, platelet counts, blast, WT1 expression level, FLT3-ITD mutation positive rate and response to treatment. The characteristic immunophenotype is lower expression of early differentiation-associated antigens (CD34, HLA-DR, CD117, CD38), lymphocytic antigens (CD7, CD4, CD19, CD2) and higher expression of CD33 and CD123 (P < 0.05). When above features was further analyzed between the M1/2 and M4/5 subgroups in NPM1m(+)AML patients, the M1/2 cases retained a higher frequency in women and a higher WT1 expression level (P < 0.05) . Monocytic differentiation-associated antigens including HLA-DR and lymphocytic antigens CD7 were higher expressed and CD117 was lower expressed in M4/5 subgroup (P < 0.05). It is concluded that under condition of similar constituent ratio of M1/2 and M4/5 subtype and normal karyotype, NPM1m(+)AML patients have higher WT1 expression level and better response to treatment. The major immunophenotype features of NPM1m(+)AML patients are lower expression of early differentiation antigens and lymphoid lineage antigens and higher expression of CD33 and CD123. Monocytic differentiation-associated antigens only higher are expressed in M4/5 cases when compared with M1/2 cases within NPM1m(+) AML patients.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; Karyotype ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; immunology ; Male ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; Young Adult

This study was purposed to investigate the acute myeloid leukemia with complex karyotype t(2;21;8)(p12;q22;q22) (AML-M(2)) by using morphologic, immunologic, cytogenetic and molecular biologic classification technique (MICM) and to analyze the MICM characteristics of AML-M(2) and their diagnostic significance. The FAB typing of bone marrow cells (BMCs) was performed by Wright-Giemsa staining and histochemical staining of BM smears; the immunophenotype of leukemic cells was detected by flow cytometry; the karyotypes of chromosome samples prepared by short-term (48 hours) conventional culture of fresh BMCs were analyzed by RHG banding technique; the FISH signaling in mitotic metaphase was determined by dual color and dual fusion AML/ETO probe and chromosome painting probe, and was compared with results of conventional cytogenetic assay; the AML/ETO fusion transcripts were detected by nested RT-PCR. The results indicated that the bone marrow smears of case 1 showed extremely hyperplasia with myeloblasts in which a ratio of eosinophilic granulocytes and monocytes increased. Case 2 accorded with AML-M(2b) in which abnormal increase of myelocytes mainly appeared. The complex karyotype t(2;21;8)(p12;q22;q22) was detected by cytogenetic analysis combined with FISH in both two cases and AML1/ETO fusion transcripts were found by RT-PCR as well. The immunophenotype assay showed high co-expression of CD34 and HLA-DR accompanied with CD19 and CD56 expressions. It is concluded that application of MICM has an important significance for correct diagnostic typing of AML-M2 with complex karyotype variant of t(8; 21)(p12;q22;q22).
Adult ; Chromosomes, Human, Pair 2 ; Chromosomes, Human, Pair 21 ; Chromosomes, Human, Pair 8 ; Female ; Humans ; Immunophenotyping ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; immunology ; Male ; Middle Aged

OBJECTIVETo analyse the correlation between the cellular immunity and TCM Syndrome typing of patients with minimal residual leukemia (MRI).

METHODSThe analysis was performed by detecting cellular immunological parameters in 30 MRI patients, 55 healthy persons and 36 patients with acute leukemia (AL) using three fluoresceine-conjugated monoclonal antibodies and flow cytometer.

RESULTSAs compared with those in the healthy persons, universal reductions of various parameters were shown in the MRI patients, including WBC count, absolute value of total T-lymphocytes, T-helper lymphocyte (P < 0.05) and total lymphocytes; the percentage and absolute value of NK cells (P < 0.01); and the percentages of total T-lymphocytes, CD4+ CD29+, T-suppressor cells and T-memory cells (P < 0.05 or P < 0.01), but without any rising of absolute value. As compared with those in the patients with AL, parameters were similar in the two groups with insignificant difference. The disturbances, including the lowering on ratio of T-helper/T-suppressor lymphocytes, in MRI patients of Qi-blood insufficiency type was the severest, that in the patients of Qi-Yin deficiency type was the mildest, and that in patients of Yin-deficiency with excess Fire type located between them.

CONCLUSIONThe immune function of MRI patients is low, belonging to the TCM Syndrome of vital energy deficiency with evil-lingering. Since the degree of cellular immune disturbance is different in various TCM Syndrome types, therefore, they should be treated with different dosages of different drugs.

Adolescent ; Adult ; Aged ; CD4-CD8 Ratio ; Child ; Diagnosis, Differential ; Female ; Humans ; Immunity, Cellular ; Killer Cells, Natural ; immunology ; Leukemia, Myeloid, Acute ; immunology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Neoplasm, Residual ; immunology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; T-Lymphocyte Subsets ; immunology ; Yang Deficiency ; immunology ; Yin Deficiency ; immunology

OBJECTIVETo compare the immunophenotypic and clinical characteristics between NPM1 mutated acute myeloid leukemia (AML) (NPM1m(+)AML) and unmutated AML(NPM1m(-)AML) not otherwise characterized (NOS) under similar FAB subtypes constituent ratio.

METHODSImmunophenotyping and NPM1 gene mutation type-A, B and D and other leukemic related fusion genes were detected by multiparameter flow cytometry and real time RT-PCR or PCR, respectively. 104 AML patients with NPM1m(+)AML and performed immunophenotyping assay were included, 97 with NPM1m(-)AML.

RESULTSThere were significant difference between the two groups at presentation in terms of sex, white blood count(WBC), platelet counts (PLT), blast ratio, normal karyotype ratio, WT1 expression level, FLT3-ITD mutation positive rate and remission rate of first course of induction therapy (P < 0.05). On the immunophenotype, the expression of early differentiation antigens (CD34, HLA-DR, CD117, CD38), lymphocytic antigens (CD7, CD4, CD19, CD2), myeloid and monocytic differentiation-associated antigens (CD13, CD14, CD15) were lower, and that of CD33 as well as CD123 were higher in NPM1m(+)AML patients. Among them, only CD34, HLA-DR, CD7, and CD4 positive cases were significantly lower in NPM1m(+)AML group than in NPM1m(-)AML group (P < 0.05), the rest of them had significant difference in the number of positive cells (P < 0.05). Above features were further analyzed between the M1/M2 and M4/M5 subgroups. M1/M2 cases retained the women prominent and had a higher WT1 expression level (P < 0.05). The expression of monocytic differentiation-associated antigens including HLA-DR and lymphocytic antigens were higher and that of CD117 were lower in M4/M5 subtype (P < 0.05). Among them, the positive rates of HLA-DR, CD64, CD11b, CD10, CD15, and CD4 were significantly higher in M4/M5 than in M1/M2 in NPM1m(+)AML group (P < 0.05).

CONCLUSIONThe most clinical characteristics in NPM1m(+)AML patients are consistent with reports, but some immunophenotype are different to the previous reports under similar FAB subtypes constituent ratio. The major immunophenotypic features of NPM1m(+)AML patients are lower expression of progenitor, myeloid and lymphoid lineage antigens. Monocytic differentiation-associated antigens are only higher expression in M4/M5 cases when comparison with M1/M2 cases within NPM1m(+)AML group.

Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Child ; Child, Preschool ; Female ; HLA-DR Antigens ; immunology ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; immunology ; Male ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; Young Adult

We report five cases of cytomegalovirus infection in immunocompromised patients which were detected by either cytomegalovirus antigenemia assay or in situ hybridization. Four cases had leukemia and the other had chronic renal failure. All the three BMT recipients suffered from GvHD. Interestingly, there was an unique case of CMV disease without a history of BMT, which reminded us that CMV could attack immunocompromised patients who had not undergone transplantation, too. Four out of five cases died. We think that cytomegalovirus infection or disease should not be regarded as a minor problem in post-transplantation infection in Korea.
Adolescent ; Adult ; Antigens, Viral/*blood ; *Bone Marrow Transplantation ; Case Report ; Cytomegalovirus/*immunology ; Cytomegalovirus Infections/complications/*diagnosis ; Fatal Outcome ; Graft vs Host Disease/complications ; Human ; Immunocompromised Host ; In Situ Hybridization ; Kidney Failure, Chronic/complications ; Kidney Transplantation ; Leukemia/*complications/therapy ; Leukemia, Lymphocytic, Acute, L2/complications/therapy ; Leukemia, Myelocytic, Acute/complications/therapy ; Leukemia, Myeloid, Chronic/complications/therapy ; Male ; Viremia/*diagnosis