Induction of gamma-interferon activity by elevated temperatures in human B-lymphoblastoid cell lines.

Abstract

When the human B-lymphoblastoid cell lines GM3299 and GM4408 were heat shocked, elevated levels of antiviral activity were detected. This antiviral activity has been characterized as the result of de novo synthesis of interferon (IFN) y. Levels of antiviral activity as high as 20,000 units/ml per 106 cells were found after heat treatment. By using anti-IFN-a and anti-IFN-y sera, it was shown that the high antiviral activity in GM3299 after heat shock is due to a synergistic effect of constitutively produced IFN-a and induced IFN-'y.During the last few years, attention has been focused on the effects of fever on the immune response (1-3). Febrile temperatures have been associated with increased resistance to bacterial challenge (4-7). In addition, the fever response seems to have a beneficial impact on the outcome of many viral infections: e.g., canine herpes virus (8), porcine gastroenteritis (9), and murine rabies virus (10). Elevated temperature is also currently being investigated as an anti-neoplastic treatment. High temperatures may either have a direct lethal effect on tumor growth or act indirectly via stimulation of some immune function (11).Among the responses of organisms as diverse as Escherichia coli, Drosophila melanogaster, and man (HeLa cells) to elevated temperature is the synthesis of a specific class of proteins, the heat shock proteins (12). Although a biological explanation of their function is still not forthcoming, we raise the possibility that some heat shock proteins may be involved in the fever response. Specifically, this study shows that interferon activity is increased in certain human lymphoblastoid cell lines transformed by Epstein-Barr virus when they are incubated at temperatures between 39°C and 42°C. This increased activity is due to de novo synthesis of interferon (IFN) y.We suggest that EB-transformed human lymphocytes are primed to produce IFN as a result of the infection process and that elevated temperature activates the primed interferon activity, resulting in potent antiviral activity. MATERIALS AND METHODSCell Lines. GM3299 is a normal EB-transformed B-cell line, negative for the T-cell Mr 65,000 glycoprotein. It was obtained from the National Institute of General Medical Sciences Human Genetic Mutant Cell Repository (Camden, NJ). Bloom syndrome lymphoblast (GM4408) is an EBtransformed lymphoblast cell line also negative for the T-cell Mr 65,000 glycoprotein and positive for surface immunoglobulin sIg and Ia (B-cell) antigens. It was originally isolated from a 4-year-old Caucasian male diagnosed as suffering from Bloom syndrome. Each cell line was grown in RPMI medium 1640 supplemented with 20% fetal bovine serum, streptomycin (100 ,ug/ml), and penicillin (60 ,tg/ml).IFN Assay. IFN was assayed as described by Familletti et al. (13) with minor modifications. Test cells (human Hep-2 and bovine MDBK) were plated in 96-well tissue culture plates at 5 x 104 cells per well in Hank's minimal essential medium 10% heat-inactivated fetal calf serum. Aliquots of medium cont...

Interferon gamma upregulates its own gene expression in mouse peritoneal macrophages.

“…Moreover, IFN-3' can modulate the proliferation and function ofT lymphocytes (5,6) and enhance the cytolytic activity of NK cells (7,8) The expression of IFN-'y is generally considered to be restricted to T cells and LGL (9). There are a limited number of reports on the expression of IFN-y in other cell types (10)(11)(12)(13), but the consistency and biological relevance of these observations are still unclear (9). The production of IFN-'y is generally considered to be strictly controlled, and significant amounts of this cytokine are produced only after specific stimulation (9) or in the course of certain pathologic conditions (12).…”

Interferon pretreatment lowers the threshold for maximal heat‐shock response in mouse cells

“…IFNs and stress might have related and complementary effects: viral infection has been reported to induce HSP synthesis (Collins and Hightower, 1982;Notarianni and Preston, 1982;Kao and Nevins, 1983;Khandjian and Tiirler, 1983) and IFN stimulates the transcription of the MT-2 gene coding for metallothionein (Friedman et al, 1984;Friedman and Stark, 19851, a stress protein induced by toxic levels of transition metal ions. It should also be mentioned that heat shock induces the production of 7-D" in a human 0-lymphoblastoid cell line (Taylor et al, 1984).…”

Burkitt's cells can be triggered by teleocidin to secrete interferon‐γ

“…Taylor et a1 [16] recently demonstrated the induction of IFN-7 in two EBVtransformed cell lines following incubation at 42°C. They suggest that the transformation by EBV primes the cell lines for IFN-y secretion by the induction of IFN-(r and that heat shock triggers the synthesis of IFN-7.…”