Treatment of Phalloides intoxication by hemoperfusion with aktivcarbon

FLORIN, I. and THELESTAM, M. Intoxication of cultured human lung fibroblasts with Clostridium djfJcile toxin. Inject. Immunity 33, 67 (1981). THE crroP.+THOCENIC effect of partially purified toxin from Clostridium djffrcile on cultured human lung fibroblasts was studied. Conditions for dettrmination of SO% tissue culture dose were standardized. The cytopathogrnic effect of the toxin was dependent on toxin concentration, exposure time and density of the cells. Transfer of the cells to 0°C did not inhibit binding of toxin to the fibroblast surface, but prevented the development of the cytopathogenic effect . Both binding of toxin and some intracellular steps) were prevrnted by 2,4dinitrophenol . These preventative effects were reversible . Before and concomitantly with the appearance of thecytopathogrnic effect, the cellular uptake of uridine and of amino acids was markedly stimulated . Protein synthesis was depressed whrn 1000 of the cells showed the cytopathogenic effect, but the synthesis of nucleic acids was inhibited only several hours later. The primary cellular target for the toxin is still unknown. Reprints : I. FLORIN Departmrnt Of BBCteI7010$y Karolinska Institutet S-104 Ol Stockholm Sweden

G. HAHERMEHL

THELFSTAM, M. Snd BRONNEGARD, M. Interaction of cytopathogenic toxin from Clostridium djjfici(e with cells in tissue culture . Scnnd. J. Inject. Dis. Suppl . 22, 16 (1980) . PARTIALLY purified cytopathogrnic toxin from Clostridium d(jfrcile induced morphological changes in five cell lines in tissue culture . The relative sensitivity scale of the all lines was human lung and intestinal fibroblasts > Chinese hamster ovary cells D mouse adrrnal cells > mouse neuroblastoma cells. The cytopathogenic effect did not occur in toxin-treated lung fibroblasts incubated at 0°C. Pre-incubation of lung fibroblasts with 2,4-dinitrophrnol prevrnted the cytopathogrnic effect . The toxin bound to as yet unidentified receptors at the surface of human lung and intestinal fibroblasts. The toxin-induced morphological (actinomorphic) changes in lung and intestinal fibroblasts closely resembled the effects induced by the fungal metabolite cytochalasin B (CB), which is known to disrupt microfilamrnts reversibly . Indirect immunofluorescence with anti-actin antiserum demonstrated that the C. d(fficile toxin disrupted the straight actin t-tlament bundles seen in normal fibroblasts . The cytopathogenic effect became apparent 3 - S hr after exposure to toxin. However, irreversible intoxication had already occurred within 20 min of exposure, as toxinueated fibroblasts which were trypsinized and reseeded wen not able to attach to the solid substratum and regenerate their typical shape, a process requiring reorganization of actin into microfilamrnt bundles. Two possible differrnt modes of action of the toxin, leading to microfilament disruption, are suggested: (1) uansmembrane signal by surface-bound toxin via microfilamrnt-linked integral membrane protein(s) ; (2) penetration of surface-bound whole toxin or an active fragmrnt, followed by its intracellular action. The experimrntal evidence so far is consistent with either of these mechanisms . Reprints : M. THELESTAM Department of Bacteriology KaroGnska Institutes 5-104 O1 Stockholm Swedrn

G. HAHERMEHt

BARTELS, C. and SEEGER, R. Treatmrnt of Phalloides intoxication by hemoperfusion with aktivcarbon . In: Champignons Toriques, p . 106 . Paris: Mason (1978) . THIRTEEN Phalloides intoxications were treated with hemoperfusion. The severity of the intoxication was different in every case, and the 6 hr hemoperfusions were more or less early. Ten patients survived . These clinical results are compared with experimrntal studies which proved that charcoal hemoperfusion eliminated the main phalloidin and amanitine toxins. Early and repeated hemodialyses are recotnmrnded. Reprints: C. BARTEIS Medizinische Universitâts-Klinik Krankenhausstrasse 12 -852 Erlangen FRG

G. HABERMEHL