Efficacy and selectivity of monoclonal-antibody-targeted drugs and free methotrexate in fluorescence-labelled mixed tumour-cell monolayer cultures and multicellular spheroids.
Free methotrexate (MTX) and 2 monoclonal antibody (MAb)-MTX conjugates were tested against mixed human tumour-cell cultures, in which 2 cell lines of differing antigenicity or drug sensitivity, pre-labelled with fluorescent dyes, were added together in microtitre wells. Conjugates were selectively cytotoxic for cells bearing high concentrations of the relevant antigen, and MTX was preferentially cytotoxic for wild-type cells rather than MTX-resistant variants when tested on separate cultures. In mixed cultures these selectivities were substantially retained, although there was a varying tendency towards intermediate cytotoxicity for each cell line of the pair. MTX was cytotoxic for cell line 79 IT grown as multicellular spheroids, but a MAb-MTX conjugate and a MAb-RTA immunotoxin showed little cytotoxicity against spheroids at the highest concentrations tested, although they were highly effective against monolayer cells. Mixed spheroids could be formed efficiently from most cell lines, although in some cases cell distribution was non-random. In mixed spheroids prepared between wild-type and MTX-resistant 79 IT variants, relative MTX sensitivities conformed broadly to those seen in separate monolayer cultures. Fluorescence-labelling was a reliable method for determining cell behaviour under the above conditions. We conclude that (a) selectivity of therapeutic agents in mixed cultures was partially, but not completely, impaired compared to that observed in separate cultures, and (b) that low Mr drugs are effective against 3-dimensional tumour-cell structures but antibody-targeted conjugates are not.
Cancer Research Campaign Laboratory, University of Nottingham, University Park, UK.
M J
MJ
Embleton
A
A
Charleston
K
K
Affleck
D000911:Antibodies, Monoclonal; D002470:Cell Survival / Q000187:drug effects*; D003110:Colonic Neoplasms; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D005260:Female; D006801:Humans; D000922:Immunotoxins / Q000633:toxicity*; D008727:Methotrexate / Q000494:pharmacology*; D012516:Osteosarcoma; D010051:Ovarian Neoplasms; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms
D000911:Antibodies, Monoclonal; D000922:Immunotoxins; D008727:Methotrexate