Melanoma cells constitutively release an anchor-positive soluble form of protectin (sCD59) that retains functional activities in homologous complement-mediated cytotoxicity

Lorelei I. Brasoveanu, Ester Fonsatti, Alberto Visintin, Mirjana Pavlovic, Ilaria Cattarossi, Francesca Colizzi, Aldo Gasparollo, Sandra Coral, Vaclav Horejsi, Maresa Altomonte, Michele Maio

Research output: Contribution to journalArticle

Abstract

Protectin (CD59), a glycosylphosphatidylinositol-anchored cell membrane glycoprotein, is differentially expressed on melanocytic cells and represents the main restriction factor of C-mediated lysis of melanoma cells. In this study, we report that CD59-positive melanoma cells constitutively release a soluble form of CD59 (sCD59), and that its levels directly correlate (r = 0.926; P <0.05) with the amount of membrane-bound CD59. SDS-PAGE analysis showed that the molecular components of sCD59 are similar to those of cellular CD59 expressed by melanoma cells. Melanoma-released sCD59 is anchor positive since it inserts into cell membranes of homologous cells that transiently increase their expression of CD59. Moreover, sCD59 is functional: it blocks the binding of the anti-CD59 mAb YTH53.1 to melanoma cells and reverses its effects on C-mediated lysis. In fact, preincubation of mAb YTH53.1 with scalar doses of conditioned media of CD59-positive but not of CD59-negative melanoma cells reduced significantly (P <0.05), and in a dose- dependent fashion, the enhancement of C-mediated lysis of anti-GD3- sensitized melanoma cells induced by the masking of cellular CD59 by mAb YTH53.1. Altogether, these data demonstrate that CD59-positive human melanoma cells release a soluble form of CD59 that is structurally similar to cellular CD59, retains its anchoring ability, is functional, and may impair the effectiveness of clinical approaches to humoral immunotherapy for human melanoma.

Original languageEnglish
Pages (from-to)1248-1255
Number of pages8
JournalJournal of Clinical Investigation
Volume100
Issue number5
Publication statusPublished - Sep 1 1997

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CD59 Antigens
Melanoma
Cell Membrane
Glycosylphosphatidylinositols
Membrane Glycoproteins
Conditioned Culture Medium
Immunotherapy
Polyacrylamide Gel Electrophoresis

Keywords

  • GPI anchor
  • Immunotherapy
  • Malignant disease
  • PI-PLC
  • Shedding

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Melanoma cells constitutively release an anchor-positive soluble form of protectin (sCD59) that retains functional activities in homologous complement-mediated cytotoxicity. / Brasoveanu, Lorelei I.; Fonsatti, Ester; Visintin, Alberto; Pavlovic, Mirjana; Cattarossi, Ilaria; Colizzi, Francesca; Gasparollo, Aldo; Coral, Sandra; Horejsi, Vaclav; Altomonte, Maresa; Maio, Michele.

In: Journal of Clinical Investigation, Vol. 100, No. 5, 01.09.1997, p. 1248-1255.

Research output: Contribution to journalArticle

Brasoveanu, LI, Fonsatti, E, Visintin, A, Pavlovic, M, Cattarossi, I, Colizzi, F, Gasparollo, A, Coral, S, Horejsi, V, Altomonte, M & Maio, M 1997, 'Melanoma cells constitutively release an anchor-positive soluble form of protectin (sCD59) that retains functional activities in homologous complement-mediated cytotoxicity', Journal of Clinical Investigation, vol. 100, no. 5, pp. 1248-1255.
Brasoveanu, Lorelei I. ; Fonsatti, Ester ; Visintin, Alberto ; Pavlovic, Mirjana ; Cattarossi, Ilaria ; Colizzi, Francesca ; Gasparollo, Aldo ; Coral, Sandra ; Horejsi, Vaclav ; Altomonte, Maresa ; Maio, Michele. / Melanoma cells constitutively release an anchor-positive soluble form of protectin (sCD59) that retains functional activities in homologous complement-mediated cytotoxicity. In: Journal of Clinical Investigation. 1997 ; Vol. 100, No. 5. pp. 1248-1255.
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AU - Brasoveanu, Lorelei I.

AU - Fonsatti, Ester

AU - Visintin, Alberto

AU - Pavlovic, Mirjana

AU - Cattarossi, Ilaria

AU - Colizzi, Francesca

AU - Gasparollo, Aldo

AU - Coral, Sandra

AU - Horejsi, Vaclav

AU - Altomonte, Maresa

AU - Maio, Michele

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AB - Protectin (CD59), a glycosylphosphatidylinositol-anchored cell membrane glycoprotein, is differentially expressed on melanocytic cells and represents the main restriction factor of C-mediated lysis of melanoma cells. In this study, we report that CD59-positive melanoma cells constitutively release a soluble form of CD59 (sCD59), and that its levels directly correlate (r = 0.926; P <0.05) with the amount of membrane-bound CD59. SDS-PAGE analysis showed that the molecular components of sCD59 are similar to those of cellular CD59 expressed by melanoma cells. Melanoma-released sCD59 is anchor positive since it inserts into cell membranes of homologous cells that transiently increase their expression of CD59. Moreover, sCD59 is functional: it blocks the binding of the anti-CD59 mAb YTH53.1 to melanoma cells and reverses its effects on C-mediated lysis. In fact, preincubation of mAb YTH53.1 with scalar doses of conditioned media of CD59-positive but not of CD59-negative melanoma cells reduced significantly (P <0.05), and in a dose- dependent fashion, the enhancement of C-mediated lysis of anti-GD3- sensitized melanoma cells induced by the masking of cellular CD59 by mAb YTH53.1. Altogether, these data demonstrate that CD59-positive human melanoma cells release a soluble form of CD59 that is structurally similar to cellular CD59, retains its anchoring ability, is functional, and may impair the effectiveness of clinical approaches to humoral immunotherapy for human melanoma.

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