Leukoagglutinins in patients with hemophilia

“…All other chemicals and biological reagents were from Sigma (St Louis, MO, USA), unless otherwise stated. 34 PBMC were separated by Ficoll-Hypaque density centrifugation 35 and suspended in RPMI 1640, supplemented with 2 mM glutamine, gentamycin and 10% heat-inactivated human AB serum (medium A). PBMC (1 × 10 5 cells/100 l of medium A) of patients with ALL (n = 87), CML (n = 7), NHL (n = 4), AML (n = 5), thalassemia (n = 5) and aplastic anemia (n = 5) and normal donors (n = 30) were incubated with ATN H (2 g) for 1 h at 37°C.…”

Section: Cell Lines Probes and Reagentsmentioning

confidence: 99%

True

1999

Leukemia

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Neoplastic transformation causes changes in cell surface architecture, most notably, aberrant sialylation. Exploiting the restricted specificity of a 9-O acetyl sialic acid (9-OAcSA) binding lectin, Achatinin H (ATN H ), we have identified two 9-O acetyl sialoglyconjugates (9-OAcSGs) on lymphoblasts of 87 children suffering from acute lymphoblastic leukemia (ALL). The preferential binding of ATN H to lymphoblasts induces their 11-fold increased agglutination (81 ± 7.8%) compared to peripheral blood mononuclear cells (PBMC) of normal donors (8 ± 4.3%) which corroborates with flow cytometry studies. Agglutination of MOLT-4 (87 ± 4.8%), a lymphoblastoid cell line and MDCK (91.25 ± 0.01%), a cell line expressing surface 9-OAcSA, confirms the preferential binding of ATN H to lymphoblasts through their surface 9-OAcSGs. Furthermore, fluorometric quantitation reveals a 4.6-fold increase in % of 9-OAcSA on lymphoblasts of ALL patients (42.1 ± 4.1%) compared to normal donors (9.2. ± 3.4%). Western blotting confirms that ATN H recognizes two membrane sialoglycoconjugates, of MW 120 kDa and 90 kDa, both having 9-OAcSA ␣2 → 6 GalNAc terminal sugar moiety as their lectinogenic epitope. We propose that these 9-OAcSGs may serve as biomarkers for detection and monitoring of lymphoblasts in ALL and accordingly merit therapeutic considerations.

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