L. Jaso-friedmann et al., Regulation of innate immunity in tilapia: activation of nonspecific cytotoxic cells by cytokine-like factors, DEV COMP IM, 24(1), 2000, pp. 25-36
Exposure of tilapia (Oreochromis niloticus) to water temperatures of 10-15
degrees C for 3-5 min produces physiological stress responses characterized
by immediate phenotypic and immunological changes. In the present study, t
his general stress response was utilized as a model system to study innate
immunity mediated by soluble factors and cytotoxic cells. Acute innate cyto
toxic responses of nonspecific cytotoxic cells (NCC) in the peripheral bloo
d (PBL), anterior kidney (AK) and spleen (SPL) were measured. Following tem
perature stress, the levels of NCC activity depended on the presence of sol
uble factors and on the cell compartments from which the NCC were obtained.
NCC from PBL of stressed tilapia had 30x or greater cytotoxic activity com
pared to nonstressed PBLs from controls. NCC activity from the AK and SPL o
f stressed tilapia was lower than controls. Flow cytometric analysis of NCC
in each tissue showed that increased cytotoxicity was not produced by incr
eased numbers of NCC. To determine the mechanism of amplification of cytoto
xicity, NCC from nonstressed tilapia were passively treated with serum from
temperature stressed tilapia. Serum containing the "stress activated serum
factor" (SASF) passively increased naive NCC cytotoxicity (from PBL) 3-4 f
old. The cytotoxic eel response was inhibited by addition of anti-NCC monoc
lonal antibody 5C6. These data indicated that NCC are (at least one of) the
target cells for SASF. SASF required only lj min pre-incubation with naive
NCC to activate cytotoxicity. Activation was nonreversible and concentrati
on dependent. Pretreatment of NCC with SASF reduced the assay time required
to amplify target cell cytotoxicity from 12-24 h to 6 h, SASF amplificatio
n of NCC cytotoxicity was not restricted by different histological types of
target cells. Determination of select physical/chemical properties of SASF
revealed: complete heat inactivation of cytotoxicity amplification followi
ng 55 degrees C and 65 degrees C pretreatment; SASF was thermostable at roo
m temperature to 45 degrees C for 15 min; and freeze-thaw treatment reduced
but did not completely remove amplification activity. The molecular weight
range of SASF activity was identified in a 50-100 kDa fraction obtained by
differential dialysis. SASF appears to be a protein sensitive to trypsin d
igestion. (C) 2000 Elsevier Science Ltd. All rights reserved.