Nonspecific cytotoxic cells in fish (Ictalurus punctatus). I. Optimum requirements for target cell lysis

Abstract

Nonspecific cytotoxic cells (NCC) obtained from the head (anterior) kidney of fish (Ictalurus punctatus) lyse human transformed B-cell targets. Lysis depended on direct cell-cell contact. Fish size, age, environmental holding temperatures, and lytic reaction conditions such as osmolality and optimum effector:target cell ratios were optimized. Experiments to characterize optimum kinetics demonstrated highly efficient killing after two hours incubation. This rapid cytolysis was further studied by determining NCC activity against appropriately labeled target cells after 30, 60, 90 and 120 minutes of cocultivation. At 160:1 (E:T) greater than 40% of the 5 hour percent specific release value was produced after 30 minutes. After 90 minutes, more than 90% of total percent specific release was observed. At least one mechanism of regulation of NCC killing was described. In the presence of normal (homologous or heterologous) catfish serum (CFS), essentially no NCC activity was observed. This suppression was reversible by preincubation in 10% fetal bovine serum (FBS). NCC "activation" by preincubation in 10% FBS was time-dependent (at least four hours was required to generate significant lysis). NCC activation could be reversed by treating potentially lytic cells with supernatants containing dissociated CFS. In addition, reversible activation could be demonstrated by treating potentially lytic effector cells with CFS to produce suppression. Regulation occurred at the effector cell level because treated target cells did not suppress NCC activity. These data demonstrate a population of nonspecific effector cytolytic cells that potentially represent a phylogenetic precursor to mammalian natural killer cells.