Stimulation of TNF-alpha production by 2-(1-adamantylamino)-6-methylpyridine (AdAMP) - a novel immunomodulator with potential application in tumour immunotherapy.
The immunomodulatory effects of a recently synthesized adamantane derivative of aminopyridine - 2-(1-adamantylamino)-6-methylpyridine (AdAMP) - were tested on normal and neoplastic cells in vitro. When incubated with TNF-alpha gene-transduced mouse melanoma cells (B78/TNF), AdAMP significantly enhanced basal production of TNF-alpha by these cells, both by "high" and "moderate" TNF-alpha-producer cells. A similar TNF-alpha production-enhancing effect was observed in cultures of human ovarian carcinoma cells (CAOV1) which spontaneously produce TNF-alpha but not in cultures of tumour cells incapable of TNF-alpha secretion. RT-PCR analysis showed that the enhancement of TNF-alpha production by AdAMP was associated with an increase in TNF-alpha mRNA expression in the treated cells. The results of an electrophoretic mobility shift assay (EMSA) showed that AdAMP significantly activated nuclear factor kappaB (NF-kappaB) in both CAOV1 and B78/TNF cells. The role of NF-kappaB in enhancement of TNF-alpha production was confirmed in experiments in which MG132, an inhibitor of NF-kappaB activation, reversed the effect of AdAMP. Unexpectedly, dexamethasone, a potent antiinflammatory agent and a strong inhibitor of TNF-alpha production in vivo, increased both spontaneous and AdAMP-augmented production of TNF-alpha in in vitro cultures of ovarian carcinoma cells and B78/TNF cells. AdAMP also enhanced TNF-alpha secretion by LPS-induced monocytes. AdAMP-induced augmentation of TNF-alpha production by B78/TNF cells was accompanied by morphological changes in the treated cells and a decrease in their adherence to fibrinogen and collagen IV. In view of these properties, AdAMP seems to be a therapeutically promising compound with potential application as an adjuvant augmenting the efficacy of cancer vaccine-based therapies or in the local treatment of certain tumours.
Lasek
Witold
W
Department of Immunology, Centre of Biostructure, Medical University, Chalubinskiego 5, 02-004 Warsaw, Poland. wlasek@ib.amwaw.edu.pl
Switaj
Tomasz
T
Sieńko
Jacek
J
Kasprzycka
Monika
M
Basak
Grzegorz
G
Mikłaszewicz
Piotr
P
Maj
Małgorzata
M
Nowis
Dominika
D
Grzela
Tomasz
T
Gołab
Jakub
J
Młynarczuk
Izabela
I
Jalili
Ahmad
A
Kamińska
Bozena
B
Dziembowska
Magdalena
M
Czajkowski
Krzysztof
K
Nowaczyk
Maria
M
Górska
Agata
A
Kazimierczuk
Zygmunt
Z
D000218:Adamantane / Q000031:analogs & derivatives / Q000494:pharmacology*; D000631:Aminopyridines / Q000494:pharmacology*; D000818:Animals; D005260:Female; D006801:Humans; D008545:Melanoma / Q000473:pathology*; D051379:Mice; D016328:NF-kappa B / Q000096:biosynthesis; D010051:Ovarian Neoplasms / Q000473:pathology; D012333:RNA, Messenger / Q000032:analysis; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
C466498:2-(1-adamantylamino)-6-methylpyridine; D000631:Aminopyridines; D016328:NF-kappa B; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D000218:Adamantane