We have recently reported that two EB virus-transformed human cell lines, on exposure to heat shock conditions, produce an antiviral activity. This activity has been characterized as interferon-Gamma (IFN-Gamma). One of the cell lines, GM3299, produces high levels of IFN-Alpha constitutively, whereas GM4408 produces very little (if any) IFN-Alpha. Initial characterization of the IFN-Alpha produced indicates the possibility that this is a species of interferon differing from PHA-stimulated T-lymphocyte IFN-Gamma. The very high levels of interferon activity induced in GM3299 by heat shock may reflect potentiation or synergism between IFN-Alpha and IFN-Gamma. We shall purify and characterize this 'novel' interferon. We shall characterize the induction process in detail and clone the IFN-Gamma gene. We shall attempt to define the nucleotide sequences involved in """"""""heat-induction."""""""" Using cloned EB virus DNA segments, we shall examine the relationship, if any, between EB DNA synthesis and transcription and the interferon induction process. These experiments will be paralleled with in vivo studies on the effect of fever on interferon induction. We have measured the interferon levels in genital herpes-infected patients. High levels of IFN-Alpha are detected in the serum, and IFN-Beta in herpatic vesicles. PHA-stimulated lymphocytes produce 10-100 times more IFN-Gamma than stimulated lymphocytes of control individuals. Contrary to other reports, IFN-Gamma inhibits herpes plaque formation in culture. IFN-Alpha and IFN-Beta are less active. We shall examine the molecular basis of this antiviral activity of using cloned HSV genes. Recent reports indicate that murine IFN-Gamma is identical to macrophage-activating factor. This will be examined for human IFN-Gamma using monoclonal antibodies. We shall attempt to functionally separate the three functions of IFN-Gamma: antiviral, anti-proliferative, and macrophage activation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI021898-02
Application #
3132384
Study Section
Virology Study Section (VR)
Project Start
1985-01-01
Project End
1987-12-31
Budget Start
1986-01-01
Budget End
1986-12-31
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Indiana University Bloomington
Department
Type
Schools of Arts and Sciences
DUNS #
006046700
City
Bloomington
State
IN
Country
United States
Zip Code
47402
De Stasio, P R; Taylor, M W (1990) Specific effect of interferon on the herpes simplex virus type 1 transactivation event. J Virol 64:2588-93
De Stasio, P R; Taylor, M W (1989) Trans-activation of herpes simplex virus type 1 immediate early genes is specifically inhibited by human recombinant interferons. Biochem Biophys Res Commun 159:439-44
De Stasio, P R; Etchberger, K J; Paeratakul, U et al. (1989) In situ amplification of viral signal in infected samples applied directly to nitrocellulose membranes. Biotechniques 7:442-7
Paeratakul, U; De Stasio, P R; Taylor, M W (1988) A fast and sensitive method for detecting specific viral RNA in mammalian cells. J Virol 62:1132-5
De Stasio, P R; Reichenba, K; Taylor, M W (1988) A sensitive method to titer interferons. Biotechniques 6:206, 208
Feng, G S; Gray, P W; Shepard, H M et al. (1988) Antiproliferative activity of a hybrid protein between interferon-gamma and tumor necrosis factor-beta. Science 241:1501-3
Downing, J F; Martinez-Valdez, H; Elizondo, R S et al. (1988) Hyperthermia in humans enhances interferon-gamma synthesis and alters the peripheral lymphocyte population. J Interferon Res 8:143-50
Downing, J F; Taylor, M W; Wei, K M et al. (1987) In vivo hyperthermia enhances plasma antiviral activity and stimulates peripheral lymphocytes for increased synthesis of interferon-gamma. J Interferon Res 7:185-93
Downing, J F; Taylor, M W (1987) The effect of in vivo hyperthermia on selected lymphokines in man. Lymphokine Res 6:103-9