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Pathology Research International
Volume 2013 (2013), Article ID 920972, 12 pages
http://dx.doi.org/10.1155/2013/920972
Research Article

Immunoreactivity of the 14F7 Mab Raised against N-Glycolyl GM3 Ganglioside in Primary Lymphoid Tumors and Lymph Node Metastasis

1Laboratory of Recognition and Biological Activity Assays, Department of Quality Control, Center of Molecular Immunology, P.O. Box 16040, 216 Street and 15 Avenue Atabey, Playa, 11600 Havana, Cuba
2Department of Cell Biology and Tissues Banking, National Institute of Oncology and Radiobiology, 29 and F Street Vedado, Plaza de la Revolución, 10400 Havana, Cuba
3Department of Pathology, Manuel Fajardo General Hospital, Zapata and D Street Vedado, Plaza de la Revolución, 10400 Havana, Cuba
4Clinical Trials Unit, National Institute of Oncology and Radiobiology, 29 and F Street Vedado, Plaza de la Revolución, 10400 Havana, Cuba
5Department of Pathology, National Institute of Oncology and Radiobiology, 29 and F Street Vedado, Plaza de la Revolución, 10400 Havana, Cuba
6Research and Development Direction, Center of Molecular Immunology, P.O. Box 16040, 216 Street and 15 Avenue Atabey, Playa, 11600 Havana, Cuba

Received 31 July 2013; Revised 30 September 2013; Accepted 10 October 2013

Academic Editor: Shahid Pervez

Copyright © 2013 Rancés Blanco et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The reactivity of the 14F7 Mab, a highly specific IgG1 against N-glycolyl GM3 ganglioside (NeuGcGM3) in normal tissues, lymphomas, lymph node metastasis, and other metastatic sites was assessed by immunohistochemistry. In addition, the effect of chemical fixation on the 14F7 Mab staining using monolayers of P3X63Ag.653 cells was also evaluated. Moreover, the ability of 14F7 to bind NeuGcGM3 ganglioside inducing complement-independent cytotoxicity by a flow cytometry-based assay was measured. The 14F7 Mab was reactive in unfixed, 4% paraformaldehyde, 4% formaldehyde, and acetone fixed cells. Postfixation with acetone did not alter the localization of NeuGcGM3, while the staining with 14F7 Mab was significantly eliminated in both cells fixed and postfixed with methanol but only partially reduced with ethanol. The staining with 14F7 Mab was evidenced in the 89.2%, 89.4%, and 88.9% of lymphomas, lymph node metastasis, and other metastatic sites, respectively, but not in normal tissues. The treatment with 14F7 Mab affected both morphology and membrane integrity of P3X63Ag.653 cells. This cytotoxic activity was dose-dependent and ranged from 24.0 to 84.7% (10–1000 μg/mL) as compared to the negative control. Our data could support the possible use of NeuGcGM3 as target for both active and passive immunotherapy against malignancies expressing this molecule.