Viable head and neck tumor spheroids stimulate in vitro autologous monocyte MCP-1 secretion through soluble substances and CD14/lectin-like receptors.
Biopsies from carcinoma tissue and benign control mucosa from head and neck squamous cell carcinoma (HNSCC) patients were used to establish fragment (F)-spheroids in vitro. We have previously shown that autologous monocytes co-cultured with F-spheroids in vitro augment their secretion of monocyte chemotactic protein-1 (MCP-1). Presently, the aims of the present work were to study whether the metabolic activity, secreted products and/or specific receptor/ligand on the surface of the F-spheroids and monocytes are necessary for stimulation of the monocyte MCP-1 secretion upon F-spheroid co-culture. Actinomycin D (1 mug/ml for 24 h) pre-treatment of the F-spheroids abolished the monocyte MCP-1 co-culture response. Co-culture of monocytes and F-spheroids separated by a semi-permeable membrane showed a decreased, but still present, monocyte MCP-1 co-culture response. Conditioned medium from F-spheroids stimulated allogenous monocytes to secrete MCP-1. The addition of glucose or galactose, but not mannose, to co-cultures partially inhibited the monocyte MCP-1 co-culture response. The addition of anti-CD14 antibody diminished the MCP-1 co-culture response. In conclusion, the monocyte MCP-1 co-culture response is dependent on metabolically active spheroids, secreted stimuli, and is augmented by direct contact with F-spheroids, possibly via lectin-like receptors and the CD14 receptor.
Olsnes
Carla
C
Department of Otolaryngology and Head and Neck Surgery, Haukeland University Hospital, Bergen, Norway.
Heimdal
John-Helge
JH
Kross
Kenneth W
KW
Olofsson
Jan
J
Aarstad
Hans Jørgen
HJ
D001323:Autoantibodies / Q000276:immunology; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000473:pathology; D018932:Chemokine CCL2 / Q000096:biosynthesis* / Q000378:metabolism; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D004797:Enzyme-Linked Immunosorbent Assay; D005690:Galactose / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D006258:Head and Neck Neoplasms / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008024:Ligands; D018950:Lipopolysaccharide Receptors / Q000276:immunology / Q000494:pharmacology*; D008358:Mannose / Q000494:pharmacology; D009000:Monocytes / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000378:metabolism*; D018874:Spheroids, Cellular / Q000378:metabolism* / Q000473:pathology; D014407:Tumor Cells, Cultured
D001323:Autoantibodies; C111835:CCL2 protein, human; D018932:Chemokine CCL2; D017077:Culture Media, Conditioned; D008024:Ligands; D018950:Lipopolysaccharide Receptors; D011975:Receptors, Mitogen; D005947:Glucose; D008358:Mannose; D005690:Galactose