Human macrophages secrete a tumoricidal activity distinct from tumour necrosis factor-alpha and reactive nitrogen intermediates

Abstract

Human macrophages, differentiated in vitro from blood monocytes, can be induced to secrete tumouricidal activity when activated by combined treatment with recombinant interferon gamma and bacterial lipopolysaccharide. We have analysed conditioned culture supernatants of activated human monocytes and in vitro differentiated macrophages cultivated under serum-free conditions for cytolytic activity against a TNF alpha-insensitive human tumour cell line and characterized this activity with respect to its relationship to TNF alpha and reactive nitrogen intermediates. Cytolytic activity was recovered in the high molecular weight fraction of culture supernatants conditioned by terminally differentiated macrophages, whereas conditioned culture supernatants of freshly isolated blood monocytes, processed under identical conditions, were devoid of significant cytolytic activity. This activity was tumour-specific, strongly affecting the human lymphoma cell line JMP, whereas freshly isolated human peripheral blood lymphocytes were not affected to a significant extent. It was inactivated by heat or trypsin treatment, but only partially inhibited by a monoclonal antibody against recombinant human TNF alpha, which completely neutralized all of the TNF alpha activity detectable in the supernatants tested. Cytolytic activity could not be reduced further even by a 1000-fold excess of anti-TNF alpha antibody, suggesting that TNF alpha has some synergistic effect on the tumouricidal activity observed, rather than being the central effector molecule. This notion was supported by enhancement of low levels of cytolytic activity by addition of recombinant human TNF alpha at concentrations not having any direct cytotoxic effect on the tumour target cells used.(ABSTRACT TRUNCATED AT 250 WORDS)