Immunophenotyping of acute myeloid leukemia using monoclonal antibodies and the alkaline phosphatase-antialkaline phosphatase technique

Abstract The leukemic cells from 41 cases of acute myeloid leukemia (AML) and 17 cases of acute lymphocytic leukemia (ALL) were immunophenotyped by the alkaline phosphatase-antialkaline phosphatase (APAAP) immunocytochemical technique utilizing eight monoclonal antibodies (MoAb) reactive with cells of myeloid origin and seven MoAb reactive with lymphoid antigens. Ninety percent of the cases of AML reacted with one or more of the pan-myeloid MoAb, My7, My9, or 20.3. Reactivity of the myeloid panel of MoAb showed some correlation with the French- American-British (FAB) classification of AML. Five of six cases of acute promyelocytic leukemia (APL) were HLA-DR negative; the one HLA-DR- positive APL had a minor population of HLA-DR-negative promyelocytes. OKM5 and/or My4 reacted with 16 of 16 monocytic leukemias. No specific marker of early erythroid development was identified. AP3, a MoAb reactive with platelet glycoprotein (GPIIIa), was specific for acute megakaryoblastic leukemia. Immunocytochemistry was also helpful in classifying seven cases of AML with equivocal or negative routine cytochemistry. Two cases of AML had minor populations of blasts detected by the APAAP technique that were immunologically distinct from the major blast population; these minor populations emerged as the predominant cell type at relapse. Two cases of ALL expressed multiple myeloid and lymphoid antigens. Two other cases that morphologically were ALL reacted with only myeloid MoAb; one consisted entirely of immature basophils on ultrastructural examination. Immunophenotyping results using the APAAP technique were comparable with those obtained with flow cytometry. The APAAP technique is a reliable method for immunophenotyping leukemia that complements other methods of immunologic evaluation. The primary advantages of this method include its use with routinely prepared blood and bone marrow smears and the ability to correlate immunocytochemical reactions with morphology.

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Immunophenotyping of acute myeloid leukemia using monoclonal antibodies and the alkaline phosphatase-antialkaline phosphatase technique

Blood ◽

10.1182/blood.v70.1.83.bloodjournal70183 ◽

1987 ◽

Vol 70 (1) ◽

pp. 83-89 ◽

Cited By ~ 2

Author(s):

CA Hanson ◽

KJ Gajl-Peczalska ◽

JL Parkin ◽

RD Brunning

Keyword(s):

Alkaline Phosphatase ◽

Acute Myeloid Leukemia ◽

Monoclonal Antibodies ◽

Myeloid Leukemia ◽

Lymphocytic Leukemia ◽

Leukemic Cells ◽

Platelet Glycoprotein ◽

Specific Marker ◽

Hla Dr ◽

Acute Myeloid

The leukemic cells from 41 cases of acute myeloid leukemia (AML) and 17 cases of acute lymphocytic leukemia (ALL) were immunophenotyped by the alkaline phosphatase-antialkaline phosphatase (APAAP) immunocytochemical technique utilizing eight monoclonal antibodies (MoAb) reactive with cells of myeloid origin and seven MoAb reactive with lymphoid antigens. Ninety percent of the cases of AML reacted with one or more of the pan-myeloid MoAb, My7, My9, or 20.3. Reactivity of the myeloid panel of MoAb showed some correlation with the French- American-British (FAB) classification of AML. Five of six cases of acute promyelocytic leukemia (APL) were HLA-DR negative; the one HLA-DR- positive APL had a minor population of HLA-DR-negative promyelocytes. OKM5 and/or My4 reacted with 16 of 16 monocytic leukemias. No specific marker of early erythroid development was identified. AP3, a MoAb reactive with platelet glycoprotein (GPIIIa), was specific for acute megakaryoblastic leukemia. Immunocytochemistry was also helpful in classifying seven cases of AML with equivocal or negative routine cytochemistry. Two cases of AML had minor populations of blasts detected by the APAAP technique that were immunologically distinct from the major blast population; these minor populations emerged as the predominant cell type at relapse. Two cases of ALL expressed multiple myeloid and lymphoid antigens. Two other cases that morphologically were ALL reacted with only myeloid MoAb; one consisted entirely of immature basophils on ultrastructural examination. Immunophenotyping results using the APAAP technique were comparable with those obtained with flow cytometry. The APAAP technique is a reliable method for immunophenotyping leukemia that complements other methods of immunologic evaluation. The primary advantages of this method include its use with routinely prepared blood and bone marrow smears and the ability to correlate immunocytochemical reactions with morphology.

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Coagulopathy in HLA-DR Antigen-Negative Acute Myeloid Leukemia.

Blood ◽

10.1182/blood.v104.11.4457.4457 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4457-4457

Author(s):

Hideki Uchiumi ◽

Takafumi Matsushima ◽

Arito Yamane ◽

Hiroshi Handa ◽

Hiroyuki Irisawa ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

De Novo ◽

Myeloid Cell ◽

Leukemic Cell ◽

Leukemic Cells ◽

Hla Dr ◽

Risk Of Mortality ◽

Acute Myeloid

Abstract Background: HLA-DR antigen is present on hematopoietic progenitors and granulocyte/monocyte, erythrocyte and megakaryocytic precursors but absent at the promyelocytic stage during myeloid cell maturation. In accordance with this, majority of promyelocytic leukemia (APL) cells were negative for HLA-DR. Meanwhile, some of non-APL acute myeloid leukemia (AML) cells is found to express HLA-DR. However, the clinical significance of HLA-DR antigen on AML cells is currently unclear. Purpose: We sought to determine the prevalence and clinical characteristics of negativity in HLA-DR expression by retrospectively analyzing 181 consecutive patients with de novo adult AML. Patients and Methods: AML patients examined in the current study (aged 15–86 years) had been diagnosed between August 1995 and July 2004, and categorized to M0 (8 patients), M1 (35), M2 (74), M3 (20), M4 (25), M5 (15), and M6 (4), based on the FAB classification. Median follow-up time was 19.3 months. Phenotypic analyses of leukemic cells were performed using CD45 gating methods. HLA-DR-negative AML was defined as HLA-DR expression less than 20% of cells in the CD45 leukemic cell gate. Results: Among 181 patients, HLA-DR antigens were not detected on AML cells from 46 patients; 20 with APL and 26 with non-APL (non-APL/DR(−)), the latter of which included M0 (2 patients), M1 (15), M2 (7), M4 (2). Leukemic cells from other non-APL patients were HLA-DR-positive (non-APL/DR(+)). None of non-APL/DR(−) patients had t(15;17) nor PML/RARa rearrangement on cytogenetic analysis. Twenty out of 26 patients with non-APL/DR(−) had normal chromosome, and 6 had abnormal karyotypes. In the non-APL/DR(−) group, various degrees of nuclear folding, convolution, or lobulation were observed in 9 patients. Although treatment response and overall survival rate were similar in the three groups (APL, non-APL/DR(−), and non-APL/DR(+)), both FDP levels at diagnosis (57.3 vs 13.2, p<0.05) and maximal FDP levels (232.6 vs 43.8, p<0.01) were significantly higher in non-APL/DR(−) compared with non-APL/DR(+). The maximal FDP levels in the non-APL/DR(−) patients were comparable to those in the APL patients. FDP levels greater than 40 mg/ml were significantly more prevalent in the non-APL/DR(−) than in the the non-APL/DR(−) group. Logistic regression analysis demonstrated that low HLA-DR expression was an independent risk factor for FDP > 40 mg/ml. Conclusion: Our study suggests that AML with negative HLA-DR antigen tend to be associated with abnormality in coagulation and fibrinolysis even if they are genetically non-APL. We propose that more attention should be paied for HLA-DR expression to avoid a devastating coagulopathy which carries a high risk of mortality unless specifically addressed.

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Immunophenotyping Features in Acute Myeloid Leukemia (AML) with NPM1+ and/or FLT3+

Blood ◽

10.1182/blood.v118.21.4908.4908 ◽

2011 ◽

Vol 118 (21) ◽

pp. 4908-4908

Author(s):

Pervin Topcuoglu ◽

Klara Dalva ◽

Sinem Civriz Bozdag ◽

Onder Arslan ◽

Muhit Ozcan ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Conflicts Of Interest ◽

Leukemic Cells ◽

Aberrant Expression ◽

Hla Dr ◽

Cd36 Expression ◽

Cd34 Cells ◽

Cd56 Expression ◽

Acute Myeloid

Abstract Abstract 4908 Immunophenotyping Features in Acute Myeloid Leukemia (AML) with NPM1 and/or FLT-3 Positive Pervin Topçuoglu, Klara Dalva, Sinem Civriz Bozdag, Önder Arslan, Muhit Özcan, Osman Ýlhan, Hamdi Akan, Meral Beksaç, Günhan Gürman Aim: We aimed to evaluate immunophenotypical (IP) features in AML pts with NPM1+ and/or FLT3+ except on acute promyelocytic leukemia. Patients&Method: Between Nov 2009 and Feb 2011, we retrospectively analyzed IP features by flow cytometry (FCM) in 51 pts (46M;17F) with new diagnosed AML. Median age was 46 years (range: 14–71 ys). The mutations of NMP1 and FLT-3 TKD&ITD were determined by the methods of RQ-PCR or RFLP, respectively in the samples of bone marrow (n=31) or periheral blood (n=20) at the diagnosis. Antigenic expression of leukemic cells was analyzed by four-color FCM (FITC, PE, PerCP&APC) based by Nomdedeu et al researh (Leuk res 2011; 35:163) (Table-1). Results: We detected NMP1+ mutation in 16 patients. Of these, three were associated with mutations of FLT3-ITD (n=2) or -TKD (n=1). Twelve patients had FLT3+ (9 ITD or 3 TKD). More than half of the patients without any mutation were CD15+/CD34+/HLA-DR+ and 11.5% for CD34 negative. Similarly, the patients with FLT-3 positive were mostly CD34+ as the pts w/o any mutations. Contrary, most of the pts with NMP1+ were CD34 negative (56.3%) (Table 1). When evaluated the complete IP in leukemic cells, the expression of CD123 was significantly marked in the patients with NPM1+ and/or FLT3+ than those w/o mutations (p=0.008). While the co-expression of CD7 and CD117 was found in 67% of the pts w/o any mutations, 30% of the pts with NMP1 and/or FLT-3 ITD (p=0.01). CD56 expression was detected in more pts with NMP1+ than those with FLT-3+ (40% vs 8%, p=0.04). Besides, CD36 expression was positive in the all pts with FLT3-ITD than TKD+ (p=0.005). More intensive CD33 expression was seen in NMP1+ pts. The expression of CD64 was similar in all three mutations. Conclusion: Though NMP1 mutation was associated more CD34+ cells, more FLT3+ pts had CD34 positivity. The expression of CD123 was especially associated with the mutations. Aberrant expression of CD56 was in more pts with NPM1+, but CD36 for FLT3-ITD. These data might be a step for a study aiming to show a correlation between the type of mutations combined with IP features of leukemic cells and clinical characteristics or disease course of AML pts. Disclosures: No relevant conflicts of interest to declare.

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Characterization of CEBPA Mutations in Acute Myeloid Leukemia in Taiwan - Most Patients with CEBPA Mutations Have Biallelic Mutations and Show a Distinct Immunophenotype of the Leukemic Cells.

Blood ◽

10.1182/blood.v104.11.4321.4321 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4321-4321

Author(s):

Liang In Lin ◽

Chien Yuan Chen ◽

Dong Tsamn Lin ◽

You Chia Yeh ◽

Hwei Fang Tien

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Topoisomerase Ii ◽

Normal Karyotype ◽

Leukemic Cells ◽

Intermediate Risk ◽

Hla Dr ◽

Cebpa Mutations ◽

Biallelic Mutations ◽

Acute Myeloid

Abstract The transcription factor CCAAT/enhancer binding protein alpha (C/EBPa) encoded by the CEBPA gene, is crucial for the differentiation of immature granulocytes. Diminished or abnormal C/EBPa activity resulting from CEBPA gene mutations is widely known to contribute to the transformation of myeloid progenitors via reduction of their differentiation potential. The CEBPA mutations have been detected in approximately 7% of total acute myeloid leukemia (AML) and in 15% of those with intermediate-risk cytogenetics or those with normal karyotype. However, the age distribution of the patients with the CEBPA mutations and the immunophenotype of their leukemic cells are not known. Sequential studies of the CEBPA gene in AML patients are also limited. In this study, 104 patients with de novo acute myeloid leukemia (AML) were evaluated for the CEBPA mutation by direct sequencing. Excluding the silent mutations, 16 (15%) of the total 104 AML patients, 15 (25%) of the 61 patients with intermediate-risk cytogenetics and 11 (35%) of the 31 patients with normal karyotype showed CEBPA mutations, frequencies higher than those reported in the West. Further cloning and subsequent nucleotide sequence analysis revealed that 14 patients had heterozygous biallelic mutations: 11 had mutations involving both the N-terminal transactivation domain (TAD) and the C-terminal basic leucine zipper domain (bZIP) and three, in either the TAD region or the bZIP region. The remaining two patients had only one allele mutation in the TAD1 region. Most mutations in TAD region were repeat-number changes of simple sequence repeats and those in bZIP region were internal tandem duplications. Sequence analysis revealed that in the region spanning the bZIP mutations, there was hot spot for concensus topoisomerase II sites, which has also been shown in other AML-related mutations FLT3-ITD and MLL duplication. All but one patient with CEBPA mutations had M1 or M2 subtype of AML. The patients with CEBPA mutations had significantly higher incidences of CD7 (73%), CD15 (100%), CD34 (93%) and HLA-DR (93%) expression than others and the majority of them showed a distinct immunophenotype of the leukemic cells: HLA-DR+ CD7+ CD13+ CD14− CD15+ CD33+ CD34+. The incidence of the CEBPA mutation in children with AML was similar to that in adults. The CEBPA mutation was serially analyzed in 27 patients; the mutations disappeared at CR, but reappeared at relapse. No one developed novel mutation during the follow-up period. In conclusion, the CEBPA mutation may play an important role in the development, but not progression, of AML. The patients with the CEBPA mutations showed a distinct immunophenotype of the leukemic cells. Potential topoisomerase II cleavage sites locating in the bZIP region were first reported and we propose that this is relevant to the process of illegitimate recombination generating the internal tandem duplication pattern of bZIP mutations.

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Surface phenotype of clonogenic cells in acute myeloid leukemia defined by monoclonal antibodies

Blood ◽

10.1182/blood.v64.3.693.bloodjournal643693 ◽

1984 ◽

Vol 64 (3) ◽

pp. 693-700 ◽

Cited By ~ 1

Author(s):

B Lange ◽

D Ferrero ◽

S Pessano ◽

A Palumbo ◽

J Faust ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Monoclonal Antibodies ◽

Cell Sorting ◽

Myeloid Leukemia ◽

Leukemic Cells ◽

Surface Phenotype ◽

Macrophage Colony ◽

Acute Myeloid ◽

Mature Colony ◽

Clonogenic Cells

Colony-forming cells in ten cases of acute myeloid leukemia (AML) were studied with six cytotoxic monoclonal antibodies that react with antigens expressed at discrete stages of differentiation of normal and leukemic hematopoietic cells. The reactivity of the whole leukemic population was measured by indirect immunofluorescence, and the reactivity of the colony-forming cells was established by complement- mediated cytotoxicity and by fluorescence activated cell sorting. Comparison of the immunofluorescent reactivity with cytotoxicity and cell sorting showed that colony-forming cells were found within a fraction of the leukemic subpopulations that expresses these antigens. This finding implies that immunofluorescence reactivity of the total leukemic population does not necessarily predict the phenotype of the clonogenic cells. When the surface phenotype of the clonogenic leukemic cells was compared to that previously established for normal marrow hemopoietic clonogenic cells, several patterns were seen: (1) in four of ten cases, the clonogenic cells expressed a phenotype like that of relatively mature normal granulocyte-macrophage colony-forming cells (late CFU-GM) or, (2) in two cases, a phenotype similar to the less mature colony-forming cells (early CFU-GM or CFU-GEMM), and (3) in four cases, a composite phenotype of early and late CFU-GM. Thus, the level of impairment of differentiation in AML may vary from case to case. In those cases phenotypically similar to the late CFU-GM, it may be possible to separate leukemic clonogenic cells from less mature normal clonogenic cells using monoclonal antibodies selectively cytotoxic for the late CFU-GM.

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Subpopulation heterogeneity in human acute myeloid leukemia determined by monoclonal antibodies

Blood ◽

10.1182/blood.v64.1.275.275 ◽

1984 ◽

Vol 64 (1) ◽

pp. 275-281 ◽

Cited By ~ 46

Author(s):

S Pessano ◽

A Palumbo ◽

D Ferrero ◽

GL Pagliardi ◽

L Bottero ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Monoclonal Antibodies ◽

Myeloid Leukemia ◽

Immunofluorescence Assay ◽

Leukemic Cells ◽

Mature Granulocyte ◽

Human Acute Myeloid Leukemia ◽

Antigenic Phenotype ◽

The Individual ◽

Acute Myeloid

Abstract The leukemic population in 63 patients with acute myeloid leukemia (AML) was studied with 15 monoclonal antibodies that detect lineage- related and stage-related antigens on normal hemopoietic cells. Indirect immunofluorescence and fluorescence-activated cell sorting showed that subpopulations of leukemic cells reacted with some or all antibodies, but the percentage of cells reacting with a single antibody varied widely among patients. The composite antigenic phenotype of the various cases, as determined by immunofluorescence assay, did not correlate with the French-American-British morphological classification. Furthermore, some cells in each case failed to express any antigen normally expressed on myelomonocytic precursors from the level of the early CFU-GM to the mature granulocyte or monocyte. In double-fluorescence experiments, the individual cells expressed none, one, or both antigens. These results demonstrate that there is considerable subpopulation heterogeneity in AML. This heterogeneity may considerably limit or complicate the use of monoclonal antibodies for diagnosis, prognosis, and treatment of acute nonlymphocytic leukemia (ANLL).

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Surface phenotype of clonogenic cells in acute myeloid leukemia defined by monoclonal antibodies

Blood ◽

10.1182/blood.v64.3.693.693 ◽

1984 ◽

Vol 64 (3) ◽

pp. 693-700 ◽

Cited By ~ 36

Author(s):

B Lange ◽

D Ferrero ◽

S Pessano ◽

A Palumbo ◽

J Faust ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Monoclonal Antibodies ◽

Cell Sorting ◽

Myeloid Leukemia ◽

Leukemic Cells ◽

Surface Phenotype ◽

Macrophage Colony ◽

Acute Myeloid ◽

Mature Colony ◽

Clonogenic Cells

Abstract Colony-forming cells in ten cases of acute myeloid leukemia (AML) were studied with six cytotoxic monoclonal antibodies that react with antigens expressed at discrete stages of differentiation of normal and leukemic hematopoietic cells. The reactivity of the whole leukemic population was measured by indirect immunofluorescence, and the reactivity of the colony-forming cells was established by complement- mediated cytotoxicity and by fluorescence activated cell sorting. Comparison of the immunofluorescent reactivity with cytotoxicity and cell sorting showed that colony-forming cells were found within a fraction of the leukemic subpopulations that expresses these antigens. This finding implies that immunofluorescence reactivity of the total leukemic population does not necessarily predict the phenotype of the clonogenic cells. When the surface phenotype of the clonogenic leukemic cells was compared to that previously established for normal marrow hemopoietic clonogenic cells, several patterns were seen: (1) in four of ten cases, the clonogenic cells expressed a phenotype like that of relatively mature normal granulocyte-macrophage colony-forming cells (late CFU-GM) or, (2) in two cases, a phenotype similar to the less mature colony-forming cells (early CFU-GM or CFU-GEMM), and (3) in four cases, a composite phenotype of early and late CFU-GM. Thus, the level of impairment of differentiation in AML may vary from case to case. In those cases phenotypically similar to the late CFU-GM, it may be possible to separate leukemic clonogenic cells from less mature normal clonogenic cells using monoclonal antibodies selectively cytotoxic for the late CFU-GM.

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Cytotoxicity Assessment and Apoptosis-related Gene Profiling of Antibody Treated Acute Myeloid Leukemia (AML) and Acute Lymphocytic Leukemia (ALL) Cancerous Cell Lines

Iranian Journal of Allergy Asthma and Immunology ◽

10.18502/ijaai.v18i6.2181 ◽

2020 ◽

Author(s):

Mahdi Habibi-Anbouhi ◽

Zahra Kafi ◽

Leila Ghazizadeh ◽

Shabnam Kharazi ◽

Mahdi Behdani ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Monoclonal Antibodies ◽

Cell Lines ◽

Myeloid Leukemia ◽

Acute Lymphocytic Leukemia ◽

Lymphocytic Leukemia ◽

Surface Markers ◽

Hematopoietic Stem ◽

Cancerous Cell ◽

Acute Myeloid

Acute myeloid leukemia (AML) and acute lymphocytic leukemia (ALL) are common acute leukemia in adults and children, respectively. In these malignancies, chemotherapy is the main treatment strategy that fails in many cases and is usually associated with adverse effects on healthy cells. In this regard, the development of new therapies is essential. Monoclonal antibodies directed to the cell surface markers of leukemic blasts may have promising consequences with minimal toxic effects on normal cells. Since cluster of differentiation 45Ra (CD45Ra) and CD123 antigens, two considered surface markers of leukemic blasts in AML and ALL respectively, are overexpressed on AML and ALL blasts, CD34+ leukemic progenitors, and AML-LSCs in comparison with normal hematopoietic stem cells (HSCs), they were selected to be targeted; using specific monoclonal antibodies. In this project, CD45Ra+ cells and CD123+ cells were targeted by anti-CD45Ra and/or anti-CD123 monoclonal antibodies. Cytotoxicity effect and cell death induction was determined by 3-(4,5-dimethylthiazol-2-yl)-2–5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Changes in the expression profile of MCL1, cMyc, Survivin, Id1, and PIM1 genes were assessed by real-time PCR. Statistical analysis of the results showed effective antibody-mediated cytotoxicity and induction of apoptosis in KG1α (CD45Ra+) and Nalm6 (CD123+) cell lines. Also, a significant change in the expression level of some of the apoptosis-related genes was observed. According to the results of this study, it can be concluded that an effective targeting of AML and ALL cancerous cell lines can be performed by anti-CD45Ra and anti-CD123 monoclonal antibodies through their effector functions and apoptosis induction.

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Subpopulation heterogeneity in human acute myeloid leukemia determined by monoclonal antibodies

Blood ◽

10.1182/blood.v64.1.275.bloodjournal641275 ◽

1984 ◽

Vol 64 (1) ◽

pp. 275-281

Author(s):

S Pessano ◽

A Palumbo ◽

D Ferrero ◽

GL Pagliardi ◽

L Bottero ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Monoclonal Antibodies ◽

Myeloid Leukemia ◽

Immunofluorescence Assay ◽

Leukemic Cells ◽

Mature Granulocyte ◽

Human Acute Myeloid Leukemia ◽

Antigenic Phenotype ◽

The Individual ◽

Acute Myeloid

The leukemic population in 63 patients with acute myeloid leukemia (AML) was studied with 15 monoclonal antibodies that detect lineage- related and stage-related antigens on normal hemopoietic cells. Indirect immunofluorescence and fluorescence-activated cell sorting showed that subpopulations of leukemic cells reacted with some or all antibodies, but the percentage of cells reacting with a single antibody varied widely among patients. The composite antigenic phenotype of the various cases, as determined by immunofluorescence assay, did not correlate with the French-American-British morphological classification. Furthermore, some cells in each case failed to express any antigen normally expressed on myelomonocytic precursors from the level of the early CFU-GM to the mature granulocyte or monocyte. In double-fluorescence experiments, the individual cells expressed none, one, or both antigens. These results demonstrate that there is considerable subpopulation heterogeneity in AML. This heterogeneity may considerably limit or complicate the use of monoclonal antibodies for diagnosis, prognosis, and treatment of acute nonlymphocytic leukemia (ANLL).

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Down-Regulation of Cellular Vascular Endothelial Growth Factor (VEGF) Levels Induces Differentiation of Leukemic Cells into Leukemic-Dendritic Cells in Patients with Acute Myeloid Leukemia.

Blood ◽

10.1182/blood.v106.11.2226.2226 ◽

2005 ◽

Vol 106 (11) ◽

pp. 2226-2226

Author(s):

Hyun-Kyu Kang ◽

Jung-Sun Park ◽

Bo-Hwa Choi ◽

Thanh-Nhan Nguyen ◽

Xiao-Wei Zhu ◽

...

Keyword(s):

Vascular Endothelial Growth Factor ◽

Acute Myeloid Leukemia ◽

Dendritic Cells ◽

Myeloid Leukemia ◽

Leukemic Cells ◽

Vascular Endothelial ◽

Hla Dr ◽

Endothelial Growth Factor ◽

Vegf Level ◽

Acute Myeloid

Abstract Vascular endothelial growth factor (VEGF), produced by most tumors, not only plays an important role in tumor angiogenesis but also can inhibit the maturation of dendritic cells (DCs) from hematopoietic progenitors. In this study, we investigated the relationship between the clinical characteristics of patients with acute myeloid leukemia (AML) and the generation of DCs from the leukemic cells, and found that cellular VEGF level in leukemic cells affected the generation of leukemic-DCs in vitro. Leukemic-DCs were successfully generated from 16 of 21 AML patients, and generated leukemic DCs exhibited high level of HLA-DR and CD54 expression, and intermediate level of CD80, CD86, CD83, and CD1a expression. Leukemic-DCs also were able to stimulate allogeneic T cell proliferation and cytotoxic activities against autologous leukemic cells. The failure of generation of leukemic-DCs from leukemic cells in some AML patients was related with its cellular VEGF levels. The leukemic cells that failed to generate leukemic-DCs had significantly higher cellular VEGF levels than that of generated leukimic-DCs (p = 0.005), and neutralizing of VEGF with anti-VEGF Ab inhibited the cellular VEGF level at the transcription level as determined by RT-PCR analysis, and at the protein level determined by West blotting and ELISA. Inhibition of cellular VEGF levels induced the generation of leukemic-DCs determined by increased levels of surface markers, namely, CD40, CD80, CD86, and HLA-DR. Moreover, inhibition of cellular VEGF levels also increased IL-12 production and allosimulatory capacity of leukemic-DCs, suggesting functional differentiation of leukemic-DCs. Interestingly, cellular VEGF levels also are inversely related to the achievement of complete remission for induction therapy (p = 0.02). Taken together, these results suggest that cellular VEGF levels during the differentiation of AML cells to leukemic-DCs are a predictive marker of the positive response of the AML patients to the remission-inducing therapy, and down-regulation of cellular VEGF levels can be one target of cancer immunotherapy by generating of sufficient number of DCs in AML patients.

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