16-16-dimethylprostaglandin-e2 and Hepatitis--Viral--Animal

The development of hepatitis secondary to murine hepatitis virus strain 3 (MHV-3) infection correlates with the induction of macrophage procoagulant activity (PCA). 16,16 dimethyl prostaglandin E2 (dmPGE2) has previously been shown to inhibit the development of disease in this model and in parallel, inhibit induction of PCA, a macrophage effector molecule which has previously been shown to correlate with resistance/susceptibility to MHV-3 infection. These studies were undertaken to determine if inhibition of PCA was a specific property of dmPGE2 or if this property was shared by other eicosanoids including prostacyclin (PGI2), PGF2a and leukotriene B4 (LTB4). Furthermore, using the recently developed anti-PCA monoclonal antibody 3D4.3 (IgG2ak) which reacted with and inhibited functional PCA, studies were then undertaken to determine the mechanism by which PCA was inhibited by dmPGE2 (transcriptional, post-transcriptional or post-translational). Treatment with dmPGE2 resulted in inhibition of PCA induction compared with vehicle control over a range of 10(-12) to 10(-6) M. Utilizing the monoclonal antibody 3D4.3, it was demonstrated by Western immunoblot and immunofluorescence studies that although PCA was functionally inhibited by dmPGE2, it was still antigenically expressed as proteins of molecular weights 74 and 70 kd. Treatment of macrophages with PGI2, PGF2a or LTB4 failed to inhibit or augment PCA induction to MHV-3 stimulation at all concentrations tested (10(-12) to 10(-6) M). These results suggest that inhibition of PCA by dmPGE2 is a specific property of this eicosanoid and that its actions occur at a post-translational level.

Topics: 16,16-Dimethylprostaglandin E2; Animals; Blood Coagulation Factors; Dinoprost; Eicosanoids; Epoprostenol; Fluorescent Antibody Technique; Hepatitis, Viral, Animal; Immunoblotting; Kinetics; Leukotriene B4; Macrophages; Male; Mice; Mice, Inbred BALB C; Murine hepatitis virus; Peritoneal Cavity

Topics: 16,16-Dimethylprostaglandin E2; Animals; Dinoprostone; Disease Susceptibility; Factor VIII; Hepatitis, Viral, Animal; Immunity, Innate; In Vitro Techniques; Kinetics; Leukotriene B4; Macrophages; Mice; Mice, Inbred A; Mice, Inbred BALB C; Models, Biological; Murine hepatitis virus; Species Specificity; Tumor Necrosis Factor-alpha

Topics: 16,16-Dimethylprostaglandin E2; Animals; Blood Coagulation Factors; Cytopathogenic Effect, Viral; Hepatitis, Viral, Animal; In Vitro Techniques; Macrophages; Mice; Mice, Inbred BALB C; Murine hepatitis virus; Prostaglandins E, Synthetic

16, 16 Dimethyl prostaglandin E2 (dmPGE2), a known cytoprotective agent, was examined for its ability to alter the course of fulminant hepatitis in an experimental model of fulminant viral hepatitis, murine hepatitis murine hepatitis type 3 (MHV-3). Fully susceptible BALB/cJ mice, infected with 100 50% lethal doses (LD50) of MHV-3 developed histologic and biochemical evidence of fulminant hepatitis, as evidenced by massive hepatic necrosis with hypoglycemia, metabolic acidosis, and a markedly elevated serum alanine aminotransferase (ALT) (mean, 1,402 +/- 619 IU/liter). In contrast, animals treated with dmPGE2 either before or after infection (up to 48 h) demonstrated a marked reduction in both histologic and biochemical evidence of liver damage as characterized by normal blood glucose, total CO2, and ALT determinations (mean ALT, 63 +/- 40 IU/liter). Treatment of infected mice with PGF2 alpha demonstrated no cytoprotective effects. High titers of infectious virus were recovered from the livers of both dmPGE2-treated and -untreated animals throughout the course of infection. In a parallel in vitro study, dmPGE2 (10(-4)-10(-8) M) demonstrated a similar cytoprotective effect on monolayers of isolated cultured hepatocytes from fully susceptible BALB/cJ mice infected at a multiplicity of infection of 0.1, 1.0, and 10.0. In addition, splenic macrophages recovered from infected and untreated BALB/cJ mice demonstrated a marked augmentation in procoagulant activity (PCA) from a basal 10 +/- 5 mU/10(6) splenic macrophages to a maximum of 615 +/- 102 mU/10(6) splenic macrophages, whereas no increase in macrophage PCA was detected in infected animals treated with dmPGE2. These results suggest that dmPGE2, without detectably altering viral replication or infectivity in vivo, confers a marked cytoprotective effect on hepatocytes both in vivo and in vitro, and prevents the induction of macrophage PCA in vivo in fully susceptible BALB/cJ mice after murine hepatitis virus type 3 infection.

Topics: 16,16-Dimethylprostaglandin E2; Animals; Blood Coagulation Factors; Cells, Cultured; Dinoprost; Hepatitis Viruses; Hepatitis, Viral, Animal; Liver; Mice; Mice, Inbred BALB C; Prostaglandins E, Synthetic; Prostaglandins F, Synthetic