Blood, Vol. 62, No. 3 (September), 1983: pp. 557-563 557

Surface Marker Analysis of Acute Myeloblastic Leukemia:Identification of Differentiation-Associated Phenotypes

By James D. Griffin, Robert J. Mayer, Howard J. Weinstein, David S. Rosenthal, Felice S. Coral, Richard P. Beveridge,

and Stuart F. Schlossman

A series of monoclonal antibodies reactive with normal

myeloid cells at different stages of differentiation (anti-

MY4. -MY7. -MY8. -Mol . -Ia) were used to characterize

the leukemic cells of 70 patients with acute myeloblastic

leukemia (AMI). Sixty-two of the leukemias expressed a

phenotype corresponding to a recognizable immature nor-

mal myeloid cell. These 62 cases could be divided into 4

phenotype groups. corresponding approximately to the

normal CFU-C (group I. 21 %). myeloblast (group II. 26%).

promyelocyte (group Ill. 8%). and promonocyte (group IV.

45%). Morphological subtyping of these leukemias tended

to agree with the immunologic phenotype. particularly with

more “differentiated” morphological subtypes. such as

acute monocytic leukemia or acute promyelocytic leuke-

mia. However. each phenotype group contained more than

A TTEMPTS TO DEFINE clinically useful classi-

fication systems for acute myeloblastic leukemia

(AML) have become particularly important because

of the recent development of highly effective chemo-

therapy. Most classification schemes of AML take

advantage of the morphological similarity of the leu-

kemic cell to a normal counterpart cell, such as a

myeloblast or promyelocyte. This type of analysis of

morphology has been particularly well suited to AML

because of the wide range of morphological variation

among the leukemic cells. For example, the French-

American-British (FAB) Cooperative Group scheme

recognizes six (M1-M6) morphological variants in

acute nonlymphocytic leukemia that reflect the degree

ofmaturation ofthe leukemic cell.”2 Although there do

not appear to be major prognostic differences among

the subtypes,2� some investigators have reported a

slightly lower response rate5 or survival rate 6 with the

monocytic (MS) variant and longer remissions in pro-

myelocytic (M3) patients.5 Cytochemical and bio-

chemical techniques have been added to morphological

analysis in an attempt to further define such subgroups

and to improve diagnostic reproducibility.2’3’5 How-

ever, many patients are difficult to classify in any

group, or have features of more than one group, and

agreement among reviewers is not always concordant.

It has recently been demonstrated that monoclonal

antibodies reactive with lineage-restricted antigens

can be reliably used to diagnose acute lymphoblastic

leukemia (ALL) and that such an immunologic analy-

sis can provide prognostically useful information.7’8 In

this article, we describe the use of a series of mono-

clonal antibodies reactive with differentiation antigens

of myeloid cells to investigate the heterogeneity of

one morphological type of AMI. indicating that the level of

differentiation of the surface membrane of AML cells may

not always be concordant with morphology. The pheno-

type groups were also analyzed with respect to cytochemi-

cal staining patterns. age. the presence of Auer rods. and

complete remission rates. Statistically significant differ-

ences among the groups were noted in the distribution of

myeloperoxidase staining. nonspecific esterase staining.

and Auer rods. The complete remission rates varied from

60% (groups Ill and IV) to 88% (group II). These results

suggest that surface marker analysis in AML may be used

as a highly reproducible classification system that will

provide additional information about the leukemic cells in

conjunction with morphological analysis.

AML. This series of antibodies can identify several

different stages of early normal myeloid and monocyte

differentiation, and expression of these antigens by

AML cells may, in part, be a reflection of the level of

maturity of those cells. The identification of different

stages of AML cell differentiation through the use of

these antibodies allows the development of a classifica-

tion system that can provide information not obtaina-

ble by morphology or cytochemistry.

MATERIALS AND METHODS

Monoclonal Antibodies

Production and characterization of murine monoclonal antibodies

anti-MY4, -MY7 and -MY8 have been previously described.9’0

Briefly, splenocytes from mice immunized with AML cells were

fused with the P3/NSI/I-Ag4-I myeloma cellline by the method of

Kohler and Milstein.” Hybrid clones were screened for the produc-

tion of monoclonal immunoglobulin reactive with the immunizing

cells by an indirect immunofluorescence assay, recloned by limiting

dilution, and passaged as ascites tumors in pristane-primed BALB/c

mice. The anti-Mol antibody’2 was provided by Dr. Robert Todd

From the Division of Tumor Immunology. Medical and Pediatric

Oncology. Dana-Farber Cancer Institute; the Division of Hematolo-

gy, Brigham and Women’s Hospital; Children’s Hospital Medical

Center; and the Departments of Medicine and Pediatrics. Harvard

Medical School, Boston, MA.

Supported in part by NIH Grants CA 25369, CA 1 9389 Project 3.

CA 09172, CA 28740, and RR 005526. J.D.G. is supported in part

by the Medical Foundation, Boston, MA.

Submitted January 31, 1983; accepted March 24. 1983.

Address reprint requests to Dr. James D. Griffin. Division of

Tumor Immunology. Dana-Farber Cancer Institute. 44 Binney

Street, Boston, MA 02115.

© I 983 by Grune & Stratton. Inc.

0006-4971/83/6203-0006$01.00/0

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558 GRIFFIN ET AL.

(Dana-Farber Cancer Institute), and anti-I2 (anti-la)” by Dr. Lee

Nadler (Dana-Farber Cancer Institute).

Immunofluorescence Assays

Antibody reactivity with test cells was determined by indirect

immunofluorescence.9 A sample of 106 test cells was incubated (4#{176}C,

30 mm) with 100 .cl of antibody (ascites fluid diluted 1:250 in

minimal essential medium containing 2.5% human AB serum),

washed 2 times, and further incubated with fluorescein-conjugated

goat anti-mouse Ig (Tago, Burlingame, CA). After 2 additionalwash steps, fluorescent antibody-coated cells were detected on a flow

cytometer (FACS I, Becton Dickinson, Mountain View, CA; or

EPICS V. Coulter Electronics, Hialeah, FL). Background fluores-

cence, determined by using a nonreactive monoclonal IgG antibody,

was subtracted.

Isolation ofHuman Cell Fractions

Granulocytes, monocytes, T cells, B cells, erythrocytes, and

platelets were prepared from the peripheral blood of normal donors

by standard techniques, as previously described.9’0 Natural killer

(NK) cells were assayed in a 4-hr chromium release assay using

K562 cells as targets.’4 Bone marrow was obtained from healthy

volunteers by aspiration into heparin-containing syringes. A mono-

nuclear cell suspension was prepared by Ficoll-Hypaque sedimenta-

tion.’#{176}

Fluorescence-Activated Cell Sorting (FACS)

The distribution of each antigen on immature bone marrow

myeloid cells was determined by separation of marrow mononuclear

cell suspensions into antigen-positive and antigen-negative cell frac-

tions by FACS, as previously described.9’0 Cytocentrifuge smears of

each fraction were stained with Wright’s stain, and a differential cell

count of 200 cells was obtained.

Hematopoietic Progenitor Cell Assays

Antibody reactivity with hematopoietic progenitor cells was deter-

mined by sterile separation of normal bone marrow cells into

antigen-positive and antigen-negative cell fractions by FACS. The

appropriate colony assays were then performed with the cells in each

fraction. In multiple experiments, recovery of progenitor cells after

sorting was similar to recovery of total cells. The committed granulo-

cyte-monocyte colony-forming cell (CFU-C) was assayed in agar by

the method of Pike and Robinson.” The erythroid burst-forming

unit (BFU-E) and colony-forming unit (CFU-E) were determined as

described by Clark and Houseman.’6

Leukemia Samples

Peripheral blood or bone marrow aspirate cells from 70 patients

with AML were collected into sterile heparin-containing syringes

prior to the initiation of antileukemic treatment. Mononuclear cells

were prepared by Ficoll-Hypaque sedimentation (1.077 g/cu cm),

and a Wright’s stained cytocentrifuge smear examined to determine

the percentage of leukemic blasts. All patients had >50% blasts in

the analyzed sample and usually >75%. Leukemic cells were cryo-

preserved in 10% dimethylsulfoxide in the vapor phase of liquid

nitrogen. Immunofluorescence assays were performed on cryopre-

served samples of all patients except those with acute promyelocytic

leukemia (APL), which were analyzed immediately because of poorviability following cryopreservation.

All patients underwent initial diagnostic testing and treatment at

the Dana-Farber Cancer Institute, the Children’s Hospital Medical

Center, or the Brigham and Women’s Hospital. All patients were

treated with one of two similar induction protocols, including

cytosine arabinoside by continuous infusion for 7 days and an

anthracycline for 3 days. Supportive care was similar at all institu-

tions. Diagnosis and morphological typing were determined by

standard techniques using Wright-Giemsa-stained smears. .2 Myelo-

peroxidase (MPO) and alpha-naphthylacetate esterase (nonspecific

esterase, NSE) staining patterns were determined on most patients’

cells. Cytochemical staining was used to confirm morphological

diagnosis. FAB types M I and M2 were grouped as “AML.” These

leukemias showed no monocytic morphological features and were

generally MPO+ and NSE- . Acute myelomonocytic leukemias

(AMML) showed morphological features of partial monocytic dif-

ferentiation (equivalent to FAB type M4). These leukemias tended

to be MPO+ and NSE+. Acute monocytic leukemias (AMoL,

equivalent to FAB MS) were MPO- and NSE+. Acute hypergran-

ular promyelocytic leukemia (APL, equivalent to FAB M3) were

MPO+ and NSE- . No patients with erythroleukemia (M6) were

tested. The presence of Auer rods was determined on a Wright’s

stained smear. A complete remission was defined as <5% blasts in

the marrow aspirate, lack of circulating leukemic cells, and a return

of normal levels of hematopoiesis.

Statistical Analysis

The influence of morphology, cytochemistry, and clinical charac-

teristics on expression of these cell surface antigens was evaluated by

chi square analysis. Antigens were studied individually, as well as

within groups noted to be expressed frequently by the study popula-

tion.

RESULTS

Immunologic Phenotype of Immature Normal

Myeloid Cells

The distribution of Ia, MY4, MY7, MY8, and Mol

antigens was determined by fluorescence-activated cell

sorting of aliquots of bone marrow stained with each

antibody and is depicted in Fig. 1 . As shown, the

differentiation of normal myeloid cells is accompanied

by an orderly, gradual acquisition (or loss) of the

surface structures defined by these antibodies.9”#{176} It is

therefore possible to determine the surface antigen

“phenotype” of myeloid cells at each level of differen-

tiation. For example, CFU-C cells are predominantly

Ia+ and MY7+, but lack MY4, MY8, and Mol

antigens. With granulocyte maturation beyond the

CFU-C stage, MY8 and Mol antigens are gradually

acquired, while the antigen density of MY7 and Ia

decreases. The promyelocyte lacks detectable Ia anti-

gen,’7’8 but MY7, MY8, and Mol antigens continue to

be expressed.

During monocyte differentiation, however, Ia anti-

gen is retained. The promonocyte is difficult to identify

morphologically, particularly after cell sorting, and

expression of antigens on this cell type cannot be

determined with certainty. Ia and MY7 antigens,

which are expressed by both the CFU-C and the

monocyte,� would presumably also be expressed by

any intermediate cells (promonocytes). Cell sorting

experiments confirm that cells with the appearance of

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cru-c

MONOCYTE

-E�,PROMONOCYTE

MV4 �IffiffiJJII�

BLAST

I�.

PROMYELOCVTE

MY7

-0MYELOCYTE Pot.?

MY8

Mol

-��ffl1�IfflJI

‘a �

fflBBJJJEJJIIIIIIJEIIIIIIIIIIEJEIIIIIIIIIII

Phenotype Grotip

lIIIIIIIftllhII��

AML SURFACE MARKERS 559

Fig. i . Schematic representation of the distribution of MV4. MY7, MY8, Mol .and Ia antigens on normal bone marrow myeloid cells asdetermined by fluorescence-activated cell sorting.

promonocytes (large diameter, NSE + ,high nuclear/

cytoplasmic ratio) are Ia + and MY7 + . Similarly,

MY4, MY8, and Mol are not expressed by CFU-C,

but are acquired at some point in cellular development

between CFU-C and the peripheral blood monocyte.

Cell sorting experiments suggest that some, but not all,

promonocytes express each of these antigens.

The information summarized in Fig. 1 is based on

FACS analysis for each antigen of marrow samples

from 3-6 individual normal donors. There appears to

be little variation among normal individuals with

respect to expression of these antigens on bone marrow

cells. Antigen expression on the CFU-C cell has been

confirmed by complement lysis experiments for those

antibodies that are lytic9 (anti-MY7 is not lytic) and

by a sensitive immune rosette technique for all anti-

bodies)9’2#{176}Expression of these antigens on nonmyeloid

cells has been previously reported.9”#{176}’2’ MY4, MY7,

MY8, and Mol antigens are not detectable on periph-

eral blood B cells, T cells, platelets, or erythrocytes, or

on bone marrow erythrocyte progenitors (BFU-E and

CFU-E) or megakaryocytes. Mo! antigen appears to

be associated with a receptor for a complement compo-

nent (C3bi)’2 and is also expressed on large granular

lymphocytes having natural killer cell activity.’2 Ia

antigen is also expressed by B cells, activated T cells,

and other hematopoietic stem cells, including BFU-

E.’3’8’23 MY4, MY7, MY8, and Mo! are not expressed

by acute lymphoblastic or chronic lymphocytic leuke-

mias.9 �2

Using this series of monoclonal antibodies, it can be

seen from Fig. 1 that 4 distinct stages of early myeloid

differentiation are defined by unique surface antigen

phenotypes. The most immature myeloid cell (group I,

“CFU-C”) corresponds to the phenotype of the normal

CFU-C (Ia+, MY7+ but lacking MY4, MY8, and

Mo!). Group II (“myeloblast”) is defined by the

acquisition of detectable levels of MY8 and Mo!

antigens. As the myeloblast matures to the promyelo-

cyte stage, Ia antigen is lost, defining group III (“pro-

myelocyte”). A fourth group (“promonocyte-mono-

cyte”) is defined by the acquisition of MY4 antigen,

along with expression of Ia, MY7, MY8, and Mo!

antigens. It should be noted that group I and group II

can be distinguished immunologically and functional-

ly, but are not distinguishable morphologically, as the

CFU-C cells cannot be morphologically identified with

certainty. The labels “myeloblast,” “promyelocyte,”

“promonocyte,” and “CFU-C” are used only to mdi-

cate a representative normal cell that would express

each of the immunologically defined phenotypes.

Phenotype Analysis ofAML Patients

Seventy patients with AML were analyzed for sur-

face marker expression prior to initiation of therapy.

The patients were classified as AML, AMML, AMoL,

or APL by standard criteria, using both morphology

and cytochemical stains (Table 1 ). Thirty-six patients

had AML, 25 had AMML, 6 had AMoL, and 3 APL.

The age, cytochemical staining, presenting white

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560 GRIFFIN ET AL.

remission.

count, and complete remission rate of these patients

are shown in Table I . Overall, 70% of patients were

adults, 91% were MPO+, 58% were NSE+, 67% had

Auer rods, 21% presented with WBC >100,000, and

71% went into complete remission. This group is

comparable to previously reported groups from our

institution’4 and other centers.’5’25

The expression of Ia, MY4, MY7, MY8, and Mol

antigens on AML patients is shown in Table 2. Overall,

89% of patients were Ia+, 41% were MY4+, 84%

were MY7+, 57% were MY8+, and 63% were

Mo! + . For a given antigen, 20% fluorescent cells was

considered “positive” and <20% was considered “neg-

ative.” However, positive antigens were usually

expressed on 50%-lOO% of the leukemic cells, while

negative antigens were detected on <5%. In most

cases, positive antigens were brightly fluorescent (high

antigen density), with two exceptions. The expression

of MY7 in AMoL was less bright in its fluorescence

that the expression of MY7 in other leukemic types.

Also, the expression of MY8 and Mol in APL was

typically less than in most AMLs (Ml and M2) and

AMMLs. It should be noted, particularly in the AML

and AMML subtypes, that there was considerable

heterogeneity of antigen expression.

In order to examine the approximate “differentia-

tion” state of AML cells, the surface antigen pheno-

types of individual leukemic patients were compared to

the phenotype groups that were previously determined

to identify stages of early normal myeloid cell matura-

tion (Fig. I and Table 3). Sixty-two of the 70 patients

expressed a phenotype compatible with one of these 4

groups, while 8 patients had phenotypes that did not fit

the pattern of a normal counterpart cell. The clinical

and laboratory parameters in the 62 leukemic patients

have been analyzed with respect to the 4 immunologi-

cally defined groups to determine the relationship of

this immunologic classification system with standard

morphological classi fication . Thirteen patients ex-

pressed a phenotype equivalent to the normal CFU-C

(Group I, Table 3). Eight-five percent ofthese patients

had leukemic cells that were considered to be morpho-

logically AML (MI or M2), while 15% had cells that

Table 2. Expression of Surface Antigens by

Morphological Diagnosis

Diagnosis Number

Per cent Positivet

Ia MY4 MY7 MY8 Mol

AML 36 92 25 81 39 50

AMML 25 92 56 84 76 72

APL 3 0 0 100 33 67

AMOL 6 100 100 iO0� 83 100

Total 70 89 41 84 57 63

. Morphological/cytochemical criteria.

tAn immunofluorescence assay was considered positive when >20%of test cells were fluorescent by FACS analysis. Generally, 50%- 100%of leukemic cells were fluorescent in a positive AML.

�The antigen density tended to be low (dim fluorescence) compared to

expression in other types of leukemia.

were morphologically classified as AMML. Group II

(“myeloblast”) included 16 patients of whom 63% had

AML and 37% had AMML. Group III (“promyelo-

cyte”) included all of the patients with APL, one

patient felt to have AML, and one patient with

AMML. The AMML patient was difficult to classify

with certainty, as the cells had features of both M3 and

M4. The largest group (group IV, “promonocyte”)

included 28 patients of whom 28% had AML, 50% had

AMML, and 22% had AMoL. All ofthe patients with

AMoL studied had the phenotype of group IV. Thus,

there was a tendency for patients considered morpho-

logically to have a less differentiated form of leukemia

to have a less differentiated phenotype as well. Of note,

unique phenotypes were observed in AMoL and APL.

However, every immunologic group contained patients

from more than one morphological group. The 8

patients whose cells did not have a phenotype that fit

the immunologic classification were also heteroge-

neous. Six patients had AML, while 2 had AMML.

For this analysis, the MY8 and Mo! antigens were

considered as one group (MY8/Mo!) because of their

very similar expression on normal myeloid cells (Fig.

1). Expression of either Mol or MY8 or both was

considered as evidence of partial differentiation. 5ev-

enty-three percent of the patients considered MY8/

Mol + expressed both MY8 and Mo!, 17% expressed

only Mol, and 10% expressed only MY8. No signifi-

Table 1 . AMI Study Population

.

Morphological

Diagnosis Number

Percent With Each Characteristic

Age 0- 1 7 Age 18-60 MPO NSE Auer Rod WBC > look CR

AML

AMML

APL

AMOL

Total

36

25

3

6

70

36

24

0

33

30

64 97 5 86

76 100 96 48

100 100 0 100

67 20 100 17

70 91 58 67

14

32

33

20

21

78

68

66

50

71

MPO, myeloperox idase; NSE. alph a naphthylacetat e esterase; WBC > lOOK, white blood cell count greater than 1 00.000/cu mm; CR .complete

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AML SURFACE MARKERS 561

Table 3. D ifferentiation I evels in AML

Phenotype

Group

Approximate

Stage of

Differentiation

PhenotypeNo. of

Patients

Morphology

AML AMML AMOL APL pIa MY4 MY7 MY8/Mol t

I CFU-C + - + - 13 85 15 0 0 0.18

II Myeloblast + - + + 16 63 37 0 0 0.53

Ill Promyelocyte - - + + 5 20 20 0 60 0.001

IV Promonocyte + + + + 28 28 50 22 0 0.00 1

Chi square analysis comparing distribution of morphological types in a given phenotype group with the total population.

tMY8 and Mol antigens were considered as a group because of their similar distribution on normal cells. See text for explanation.

cant differences were detected when these patients

were analyzed separately (data not shown).

Table 4 presents an analysis of the other clinical and

laboratory parameters studied in these 62 patients.

The majority of patients in each immunologic group

were adult (range 54%-!00%). Only 6 of the 62 cases

were MPO-, but all 6 were found in group IV. NSE

was detected in 0% (group III) to 76% (group IV) of

cases. Auer rods were found in the majority of cases,

except in group IV. Similar numbers of patients with

high presenting white cell counts (> I 00,000/cu mm)

were found in all groups. The highest complete remis-

sion (CR) rate (88%) was seen in group II, and the

lowest (60%) in groups III and IV. When each group

was compared to the total population, however, the

difference in CR rates did not reach statistical signifi-

cance.

DISCUSSION

Acute myeloblastic leukemia has long been recog-

nized as a heterogeneous disease. The leukemic cells of

different patients often show morphological features

characteristic of different stages of myeloid differen-

tiation. The classification schemes of AML are largely

based on these morphological differences, which pre-

sumably reflect the maturation of the leukemic cell.”2

We have previously described the production of mono-

clonal antibodies that react with specific differentia-

tion-linked antigens of myeloid cells.9”#{176}’2’The results

presented here show that it is possible to trace the

development of early myeloid cells through the use of

anti-MY4, -MY7, -MY8, -Mo!, and -Ia to detect

differentiation-associated changes on the cell surface.

The patterns of reactivity of these monoclonal antibod-

ies identify four different stages of differentiation that

correspond approximately to the normal CFU-C, mye-

loblast, promonocyte, and promyelocyte. Since many

AML cells morphologically resemble myeloblasts, pro-

myelocytes, or promonocytes, it was of interest to

determine the relationship between the level of mor-

phological differentiation and the state of maturation

determined by the cell surface antigen phenotype.

Seventy patients with AML were analyzed with

respect to morphology, cytochemistry, and surface

antigens. Sixty-two of the 70 patients expressed a

phenotype that was identical to one of the four pheno-

types characteristic of normal immature myeloid cells.

The most immature cell phenotype (group 1,“CFU-C”) was characterized by the expression of Ia and

MY7 antigens, but lacked MY4, MY8, or Mo!. This

group included 1 3 AML patients, of whom 85% were

considered by morphological criteria as having AML

(Ml or M2). Group II (“myeloblast”) was character-

ized by the expression of Ia, MY7, and Mo! or MY8

antigens, but lacked MY4. The majority of the 16

patients in this group had leukemic cells that were

thought morphologically to be AML, but 37% were

considered AMML. The smallest group (III, “pro-

Table 4. Ana lysi s of Clin ical and La boratory Parameters

Phenotype

Group

Differentiation

Level No.

Percent

< 1 7

Age

> 1 8 MPO NSE Auer Rod WBC > 1 00K CR

I

II

III

IV

CFU-C

Myeloblast

Promyelocyte

Promonocyte

13

16

5

28

46

31

0

21

54

69

100

79

100

100

100

78t

43

(7)#{149}

64

(11)

0

(4)

76t

(21)

85

76

100

46t

31

25

20

21

69

(p-0.88)

88

(p-0.15)

60

( p - 0.68)

60

(p-0.18)

Number of patients tested if less than all patients in a group.

tP < 0.0 1 ; chi square analysis comparing frequency of a given characteristic in each phenotype group with the frequency of that characteristic in the

total population.

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562 GRIFFIN ET AL.

myelocyte”) included all of the patients with APL and

was characterized by loss of Ia antigen with continued

expression of MY7. The largest group (IV, “promono-

cytic”) included 28 patients and was characterized by

the expression of Ia, MY4, MY7, MY8, and Mol

antigens. Seventy-eight percent of patients were con-

sidered to have monocytic morphology (AMML or

AMoL), while 22% were considered to have AML.

This analysis shows that although there is a tendency

for the morphology to correlate with the surface anti-

gen phenotype, each phenotypic group contains

patients having different morphological types. This

suggests that the surface antigen phenotype reflects a

related, but somewhat different, view of the state of

leukemic cell differentiation than does morphology,

thereby providing useful supplemental information to

the standard morphological classification . Another

advantage of immunodiagnosis is the high degree of

reproducibility and objectivity of the technique, not

always encountered in morphological classification

systems in AML.3’5

In order to further evaluate the utility of an immu-

nologic classification scheme in AML, the phenotype

groups were analyzed to determine the distribution of

patients by age, presence of marked leukocytosis,

myeloperoxidase activity, nonspecific esterase activity,

presence of Auer rods, and complete remission rate.

The majority of patients in each group were adults,

and the fraction of patients presenting with high WBC

was similar in each group. NSE positivity was lowest in

groups I and III (CFU-C and promyelocyte) and

highest in group IV (promonocyte). Group IV had the

lowest level of myeloperoxidase activity and also Auer

rods. The latter finding is of interest because of the

previous reports of the presence of Auer rods being

prognostically favorable.3 The complete remission rate

varied from 60% (groups III and IV) to a high of 88%

(group II). However, this trend was not statistically

significant. With larger numbers of patients, pheno-

type analysis in AML may prove useful in the identifi-

cation of patient groups with particularly good or bad

response to treatment. It will also be important to

determine in the future if there are any significant

differences in duration of remission among any of the

phenotype groups. Finally, certain chromosome abnor-

malities have been described in association with spe-

cific morphological types in AML. For example,

t(15;!7) is associated with APL,26 t(8;21) with FAB

type M2,27’28 and rearrangement of the long arm of

chromosome 1 1 with AMoL.29 It is likely that specific

surface antigen phenotypes will similarly be associated

with certain chromosomal abnormalities.

ACKNOWLEDGMENT

The authors would like to thank Dr. Lee Nadler for the use of

anti-I2 monoclonal antibody, Dr. Robert Todd for the use of

anti-Mol antibody, and John Daley and Herbert Levine for assis-

tance in FACS analysis.

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1983 62: 557-563  

JD Griffin, RJ Mayer, HJ Weinstein, DS Rosenthal, FS Coral, RP Beveridge and SF Schlossman differentiation-associated phenotypesSurface marker analysis of acute myeloblastic leukemia: identification of 

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