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Cell–cell adherence as a selection method for the generation of anti-melanoma monoclonal antibodies
Linköping University, Department of Clinical and Experimental Medicine, Cellbiology. Linköping University, Faculty of Health Sciences.
Molecular Immunology Center (CIMAB), Havana, Cuba.
National Institute of Oncology and Radiobiology, Havana, Cuba.
National Institute of Oncology and Radiobiology, Havana, Cuba.
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1997 (English)In: Journal of Immunological Methods, ISSN 0022-1759, Vol. 203, no 1, 103-109 p.Article in journal (Refereed) Published
Abstract [en]

The aim of the present study was to obtain monoclonal antibodies (mAbs) recognising human melanoma-associated antigens after immunisation of BALB/c mice with a 70–150 kDa membrane fraction from melanoma tumour tissues. Screening of specific antibody- producing hybridomas was performed using a novel cell–cell adherence method with the melanoma cell line M-14. Three mAbs of IgG1 isotype were selected: Mel-1, Mel-2 and Mel-3 which recognised the immunogen by ELISA and stained several melanoma cell lines positive in immunofluorescence. The molecular weight of the antigen was studied by different methods; a 170-kDa band was identified following immunoblotting of tumour lysate and a 72-kDa band was observed following immunoaffinity purification. Cell–cell adherence appears to be a reliable procedure for the generation of mAbs against native cellular antigens.

Place, publisher, year, edition, pages
1997. Vol. 203, no 1, 103-109 p.
Malignant melanoma; Hybridoma selection; Monoclonal antibody
National Category
Medical and Health Sciences
URN: urn:nbn:se:liu:diva-13713DOI: 10.1016/S0022-1759(97)00025-2OAI: diva2:21195
Available from: 2002-01-11 Created: 2002-01-11
In thesis
1. Immunological Studies in Malignant Melanoma: Importance of TNF and the Thioredoxin System
Open this publication in new window or tab >>Immunological Studies in Malignant Melanoma: Importance of TNF and the Thioredoxin System
2001 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Malignant melanoma is a tumor whose incidence is dramatically increasing in persons with light-coloured skin in all parts of the world. Due to its resistance against traditional chemo- and radiotherapy, melanoma has been a favourite target of alternative therapies, in particular those involving immunological mechanisms. Cytokines and particularly tumor necrosis factor (TNF) have been studied as possible antitumoral agents, but also as endogenous growth or differentiation factors. Previous studies showed that melanomas could express TNF in situ and that this expression correlated to decreased lymphocyte infiltration. On the other hand, redox reagents can modulate expression of cytokines, and the thioredoxin (Trx) system is particularly known to influence expression and secretion of TNF in vitro.

The overall aim of this research was to explore immunological aspects of melanoma, particularly the role of TNF both in vitro and in vivo, as well as its possible modulation by Trx.

In the in vitro studies first we developed a novel method for obtention of monoclonal antibodies against melanoma antigens, and generated and characterized specific monoclonal antibodies against both full-length and truncated Trx. We studied the cytokine expression of a panel of normal and transformed melanocytic cells by immunofluorescence, all of which presented TNF and Trx at levels comparable to monocytic cells, and TNF-receptors (TNFR) at low but detectable levels. Melanoma cells did not secrete TNF upon stimulation in spite of its presence in the Golgi apparatus. However, melanoma cells expressed the TNF-processing enzyme TACE and were capable of cleaving transfected GFP-tagged TNF. Imaging studies point to a possible cell-cell tranfer of endogenous TNF in melanoma cells.

On the other hand, TNF and Trx expression in melanoma cell lines correlated to resistance against exogenous TNF. We studied then the in situ expression of TNF and Trx by immunohistochemistry in a group of 44 cutaneous melanoma patients. Trx expression did not correlated to survival or other clinicalpathological parameters. TNF expression significantly correlated to better survival in tumors thicker than 0,8 mm, and constituted an independent prognostic factor.

These results point to a biological role of endogenous TNF in malignant melanoma, either by constituting an autocrine/paracrine differentiation factor or by modulating communication with other cell types, particularly of the host’s immune system.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2001. 79 p.
Linköping University Medical Dissertations, ISSN 0345-0082 ; 703
National Category
Medical and Health Sciences
urn:nbn:se:liu:diva-5226 (URN)91-7373-142-0 (ISBN)
Public defence
2001-12-06, Berzeliussalen, Campus US, Linköpings universitet, Linköping, 09:30 (English)
Article 4 changed significantly during the publication process.Available from: 2002-01-11 Created: 2002-01-11 Last updated: 2012-01-24Bibliographically approved

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Barral, Anna-MariaRosén, A.
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