BACKGROUND: Aberrant expression of myeloid antigens (MyAgs) on acute lymphoblastic leukemia (ALL) cells is a well-documented phenomenon, although its regulating mechanisms are unclear. MyAgs in ALL are interpreted e.g. as hallmarks of early differentiation stage and/or lineage indecisiveness. Granulocytic marker CD66c -- Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) is aberrantly expressed on ALL with strong correlation to genotype (negative in TEL/AML1 and MLL/AF4, positive in BCR/ABL and hyperdiploid cases). METHODS: In a cohort of 365 consecutively diagnosed Czech B-precursor ALL patients, we analyze distribution of MyAg+ cases and mutual relationship among CD13, CD15, CD33, CD65 and CD66c. The most frequent MyAg (CD66c) is studied further regarding its stability from diagnosis to relapse, prognostic significance and regulation of surface expression. For the latter, flow cytometry, Western blot and quantitative RT-PCR on sorted cells is used. RESULTS: We show CD66c is expressed in 43% patients, which is more frequent than other MyAgs studied. In addition, CD66c expression negatively correlates with CD13 (p < 0.0001), CD33 (p = 0.002) and/or CD65 (p = 0.029). Our data show that different myeloid antigens often differ in biological importance, which may be obscured by combining them into "MyAg positive ALL". We show that unlike other MyAgs, CD66c expression is not shifted from the onset of ALL to relapse (n = 39, time to relapse 0.3-5.3 years). Although opposite has previously been suggested, we show that CEACAM6 transcription is invariably followed by surface expression (by quantitative RT-PCR on sorted cells) and that malignant cells containing CD66c in cytoplasm without surface expression are not found by flow cytometry nor by Western blot in vivo. We report no prognostic significance of CD66c, globally or separately in genotype subsets of B-precursor ALL, nor an association with known risk factors (n = 254). CONCLUSION: In contrast to general notion we show that different MyAgs in lymphoblastic leukemia represent different biological circumstances. We chose the most frequent and tightly genotype-associated MyAg CD66c to show its stabile expression in patients from diagnosis to relapse, which differs from what is known on the other MyAgs. Surface expression of CD66c is regulated at the gene transcription level, in contrast to previous reports.

Department of Immunology Charles University 2nd Medical School Prague Czech Republic

Zobrazit více v PubMed

Markert CL. Neoplasia: a disease of cell differentiation. Cancer Res. 1968;28:1908–1914. PubMed

Greaves MF. Differentiation-linked leukemogenesis in lymphocytes. Science. 1986;234:697–704. PubMed

Mori T, Sugita K, Suzuki T, Okazaki T, Manabe A, Hosoya R, Mizutani S, Kinoshita A, Nakazawa S. A novel monoclonal antibody, KOR-SA3544 which reacts to Philadelphia chromosome-positive acute lymphoblastic leukemia cells with high sensitivity. Leukemia. 1995;9:1233–1239. PubMed

Hrusak O, Trka J, Zuna J, Houskova J, Bartunkova J, Stary J. Aberrant expression of KOR-SA3544 antigen in childhood acute lymphoblastic leukemia predicts TEL-AML1 negativity. The Pediatric Hematology Working Group in the Czech Republic. Leukemia. 1998;12:1064–1070. doi: 10.1038/sj.leu.2401072. PubMed DOI

Hrusak O, Porwit-MacDonald A. Antigen expression patterns reflecting genotype of acute leukemias. Leukemia. 2002;16:1233–1258. doi: 10.1038/sj.leu.2402504. PubMed DOI

Boccuni P, Di Noto R, Lo Pardo C, Villa MR, Ferrara F, Rotoli B, Del Vecchio L. CD66c antigen expression is myeloid restricted in normal bone marrow but is a common feature of CD10+ early-B-cell malignancies. Tissue Antigens. 1998;52:1–8. PubMed

Kishimoto T, Kikutani H, von dem Borne AEGK, Goyert SM, Mason DY, Miyasaka M, Moretta L, Okumura K, Shaw S, Springer TA, Sugamura K, Zola H. Leukocyte Typing VI. New York, London, Garland Publishing Inc; 1997. p. 1342.

Klein ML, McGhee SA, Baranian J, Stevens L, Hefta SA. Role of nonspecific cross-reacting antigen, a CD66 cluster antigen, in activation of human granulocytes. Infect Immun. 1996;64:4574–4579. PubMed PMC

Skubitz KM, Campbell KD, Skubitz AP. CD66a, CD66b, CD66c, and CD66d each independently stimulate neutrophils. J Leukoc Biol. 1996;60:106–117. PubMed

Jantscheff P, Terracciano L, Lowy A, Glatz-Krieger K, Grunert F, Micheel B, Brummer J, Laffer U, Metzger U, Herrmann R, Rochlitz C. Expression of CEACAM6 in resectable colorectal cancer: a factor of independent prognostic significance. J Clin Oncol. 2003;21:3638–3646. doi: 10.1200/JCO.2003.55.135. PubMed DOI

Ordonez C, Screaton RA, Ilantzis C, Stanners CP. Human carcinoembryonic antigen functions as a general inhibitor of anoikis. Cancer Res. 2000;60:3419–3424. PubMed

Duxbury MS, Ito H, Zinner MJ, Ashley SW, Whang EE. CEACAM6 gene silencing impairs anoikis resistance and in vivo metastatic ability of pancreatic adenocarcinoma cells. Oncogene. 2004;23:465–473. doi: 10.1038/sj.onc.1207036. PubMed DOI

Sugita K, Mori T, Yokota S, Kuroki M, Koyama TO, Inukai T, Iijima K, Goi K, Tezuka T, Kojika S, Shiraishi K, Nakamura M, Miyamoto N, Karakida N, Kagami K, Nakazawa S. The KOR-SA3544 antigen predominantly expressed on the surface of Philadelphia chromosome-positive acute lymphoblastic leukemia cells is nonspecific cross-reacting antigen-50/90 (CD66c) and invariably expressed in cytoplasm of human leukemia cells. Leukemia. 1999;13:779–785. doi: 10.1038/sj/leu/2401408. PubMed DOI

Campana D, Coustan-Smith E. Detection of minimal residual disease in acute leukemia by flow cytometry. Cytometry. 1999;38:139–152. doi: 10.1002/(SICI)1097-0320(19990815)38:4<139::AID-CYTO1>3.0.CO;2-H. PubMed DOI

Campana D, Coustan-Smith E. Advances in the immunological monitoring of childhood acute lymphoblastic leukaemia. Best Pract Res Clin Haematol. 2002;15:1–19. doi: 10.1053/beha.2002.0182. PubMed DOI

Bene MC, Castoldi G, Knapp W, Ludwig WD, Matutes E, Orfao A, van't Veer MB. Proposals for the immunological classification of acute leukemias. European Group for the Immunological Characterization of Leukemias (EGIL) Leukemia. 1995;9:1783–1786. PubMed

Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–159. doi: 10.1016/0003-2697(87)90021-2. PubMed DOI

Baranov V, Yeung MM, Hammarstrom S. Expression of carcinoembryonic antigen and nonspecific cross-reacting 50-kDa antigen in human normal and cancerous colon mucosa: comparative ultrastructural study with monoclonal antibodies. Cancer Res. 1994;54:3305–3314. PubMed

Madzo J, Zuna J, Muzikova K, Kalinova M, Krejci O, Hrusak O, Otova B, Stary J, Trka J. Slower molecular response to treatment predicts poor outcome in patients with TEL/AML1 positive acute lymphoblastic leukemia: prospective real-time quantitative reverse transcriptase-polymerase chain reaction study. Cancer. 2003;97:105–113. doi: 10.1002/cncr.11043. PubMed DOI

Muller HJ, Beier R, Loning L, Blutters-Sawatzki R, Dorffel W, Maass E, Muller-Weihrich S, Scheel-Walter HG, Scherer F, Stahnke K, Schrappe M, Horn A, Lumkemann K, Boos J. Pharmacokinetics of native Escherichia coli asparaginase (Asparaginase medac) and hypersensitivity reactions in ALL-BFM 95 reinduction treatment. Br J Haematol. 2001;114:794–799. doi: 10.1046/j.1365-2141.2001.03009.x. PubMed DOI

Dworzak MN, Froschl G, Printz D, Mann G, Potschger U, Muhlegger N, Fritsch G, Gadner H. Prognostic significance and modalities of flow cytometric minimal residual disease detection in childhood acute lymphoblastic leukemia. Blood. 2002;99:1952–1958. doi: 10.1182/blood.V99.6.1952. PubMed DOI

Hanenberg H, Baumann M, Quentin I, Nagel G, Grosse-Wilde H, von Kleist S, Gobel U, Burdach S, Grunert F. Expression of the CEA gene family members NCA-50/90 and NCA-160 (CD66) in childhood acute lymphoblastic leukemias (ALLs) and in cell lines of B-cell origin. Leukemia. 1994;8:2127–2133. PubMed

Scholzel S, Zimmermann W, Schwarzkopf G, Grunert F, Rogaczewski B, Thompson J. Carcinoembryonic Antigen Family Members CEACAM6 and CEACAM7 Are Differentially Expressed in Normal Tissues and Oppositely Deregulated in Hyperplastic Colorectal Polyps and Early Adenomas. Am J Pathol. 2000;156:595–605. PubMed PMC

Duxbury MS, Ito H, Benoit E, Zinner MJ, Ashley SW, Whang EE. Overexpression of CEACAM6 promotes insulin-like growth factor I-induced pancreatic adenocarcinoma cellular invasiveness. Oncogene. 2004;23:5834–5842. doi: 10.1038/sj.onc.1207775. PubMed DOI

Duxbury MS, Ito H, Ashley SW, Whang EE. CEACAM6 as a novel target for indirect type 1 immunotoxin-based therapy in pancreatic adenocarcinoma. Biochem Biophys Res Commun. 2004;317:837–843. doi: 10.1016/j.bbrc.2004.03.128. PubMed DOI

Putti MC, Rondelli R, Cocito MG, Arico M, Sainati L, Conter V, Guglielmi C, Cantu-Rajnoldi A, Consolini R, Pession A, Zanesco L, Masera G, Biondi A, Basso G. Expression of Myeloid Markers Lacks Prognostic Impact in Children Treated for Acute Lymphoblastic Leukemia: Italian Experience in AIEOP-ALL 88-91 Studies. Blood. 1998;92:795–801. PubMed

San Miguel JF, Ciudad J, Vidriales MB, Orfao A, Lucio P, Porwit-MacDonald A, Gaipa G, van Wering E, van Dongen JJ. Immunophenotypical detection of minimal residual disease in acute leukemia. Crit Rev Oncol Hematol. 1999;32:175–185. PubMed

Mejstrikova E, Kalina T, Trka J, Stary J, Hrusak O. Correlation of CD33 with poorer prognosis in childhood ALL implicates a potential of anti-CD33 frontline therapy. Leukemia. 2005;in press doi: 10.1038/sj.leu.2403737. PubMed DOI

Standardized flow cytometry for highly sensitive MRD measurements in B-cell acute lymphoblastic leukemia

Prognosis of children with mixed phenotype acute leukemia treated on the basis of consistent immunophenotypic criteria