Microarray analysis of amphotericin B-treated THP-1 monocytic cells identifies unique gene expression profiles among lipid and non-lipid drug formulations.

The effects of deoxycholate amphotericin B (DAmB), amphotericin B lipid complex (ABLC), and amphotericin B colloidal dispersion (ABCD) on mRNA profiles from 218 genes in treated THP-1 monocytes were compared. Sixty-one genes were up-regulated and 8 were down-regulated by one or more of the AmB formulations. Fifty-three genes were up-regulated by DAmB while 24 and 18 genes were up-regulated by ABCD and ABLC, respectively.

Distinct cytokine release profiles from human endothelial and THP-1 macrophage-like cells exposed to different amphotericin B formulations.

Amphotericin B(AmB) formulations, Fungizone, and Amphotec caused substantially greater proinflammatory cytokine release than AmBisome (L-AMB) and Abelcet in TPA differentiated THP-1 macrophages as determined by antibody based protein arrays. Lipopolysaccharide but not AmB induced significant pro-inflammatory cytokines in human endothelial cells.

Antibody array-generated profiles of cytokine release from THP-1 leukemic monocytes exposed to different amphotericin B formulations.

Cytokine antibody arrays were used to establish the profiles of cytokine release from THP-1 monocytes exposed to different amphotericin B (AMB) drug delivery systems. Fungizone (FZ) and Amphotec (ABCD) caused the release of significantly more inflammatory molecules and the release of inflammatory molecules at higher levels than either AmBisome (L-AMB) or Abelcet (ABLC) after 6 h of treatment. Specifically, tumor necrosis factor alpha (TNF-alpha), interleukin-8 (IL-8), GRO-(alphabetagamma), monocyte chemoattractant protein-1 (MCP-1), RANTES, IL-10, and IL-6 were detected and semiquantified with a chemiluminscence imaging system.

Selective permissiveness of TPA differentiated THP-1 myelomonocytic cells for human cytomegalovirus strains AD169 and Towne.

Two highly passaged laboratory strains of human cytomegalovirus (HCMV), AD169 and Towne, were tested for their ability to infect and replicate in THP-1 myelomonocytic cells differentiated with 12-O-tetradecanoylphorbol-13-acetate (TPA). TPA treatment of human THP-1 cells increased the number of cells that expressed HCMV immediate early (IE1) antigen from 0.06% prior to TPA treatment to 12% following cell differentiation.

Suppression of LPS-inducible cytotoxicity in cytomegalovirus-infected THP-1 monocytic cell cultures does not correlate with a decrease in TNF-alpha antigen.

We document suppression of tumor necrosis factor alpha (TNF-alpha)-associated cytotoxic activity in a human monocytic cell line (THP-1) infected with the mycoplasma free human cytomegalovirus (CMV) strain AD169. Addition of lipopolysaccharide (LPS) to cell cultures that had been infected with CMV for 24 h resulted in a significant reduction in released cytotoxic activity to mouse L929 cells at 3-22 h post-LPS compared with mock-infected cultures. However, using an ELISA to measure TNF-alpha antigen levels in these culture supernatants, we found infected cultures had significantly higher TNF-alpha antigen levels than in mock-infected cultures following LPS induction.

Increased monokines in cytomegalovirus infected myelomonocytic cell cultures.

In an investigation of the role of monokines in CMV associated immunosuppression we document modulation of both TNF and IL-1 beta in CMV infected THP-1 cells. CMV infected cultures released almost two-fold more IL-1 beta protein and contained significantly higher IL-1 beta mRNA levels than uninfected cultures for 72-96 h after induction. In both CMV infected and uninfected cultures, significant amounts of IL-1 beta protein were not detected until 24 h post induction, while maximum amounts of TNF were detected in culture supernatants by 3 h post induction, suggesting that TNF may play a role in IL-1 beta induction.

Interactions of human cytomegalovirus with leukocytes in vivo: analysis by in situ hybridization.

Reactivation of human cytomegalovirus (HCMV) from latency occurs in immunosuppressed individuals and infection is itself immunosuppressive. To better understand the basis for this virally induced impairment of immune function, we have analyzed virus-leukocyte interactions by in situ hybridization. We detected viral DNA in 12 viremic patients in the mononuclear cell population, predominantly in cells identified as monocytes by their morphology and by labelling the cells with a monocyte specific monoclonal antibody prior to in situ hybridization.

Estimation of the B lymphocyte precursor frequencies to herpes simplex type 1 glycoproteins by a limiting dilution assay.

The precursor frequency of B lymphocytes from Balb/c mice producing HSV-1 glycoprotein B (gB), glycoprotein C (gC), and glycoprotein D (gD) antibody was determined by limiting dilution analysis under conditions to detect antibody from the clonal progeny of a single B cell precursor. In spleens of naive mice the average gC frequency was 1/48,917 +/- 5,550, while gD was 1/73,330 +/- 15,898, and gB frequency was in excess of 1/100,000. Immunization with live HSV-1 (KOS) increased the B cell frequencies of all three glycoproteins to approximately 1:3,000; however, the serum gB antibody ELISA titer was fivefold higher than gC or gD.

Augmentation of immunity to herpes simplex virus by in vivo administration of interleukin 2.

The immune mechanisms responsible for recovery from herpesvirus infections are multiple and include a principle role for aspects of T cell immunity. Our investigations add further support for this notion. We show that the ability of immune lymphocytes from animals infected i.

A new field strain of equine abortion virus (equine herpesvirus-1) among Kentucky horses.

From restriction endonuclease characterization of the DNA of 317 isolates of equine abortion virus (equine herpesvirus-1; EHV-1) from 176 epizootically unrelated outbreaks of equine virus abortion occurring over 24 years in Kentucky, an epizootic pattern and variation of the virus have emerged. Two electropherotypes of EHV-1 (1P and 1B) accounted for greater than 90% of the nonvaccine-related abortion isolates examined. From 1960 to 1981, EHV-1 1P was the predominant isolate circulating in the central Kentucky area and the cause of greater than 80% of EHV-1-related abortions.

Identification of the envelope surface glycoproteins of equine herpesvirus type 1.

The structural polypeptides of purified enveloped virions of the Army 183 strain of equine herpesvirus type 1 (EHV-1) were examined by different analytical techniques to identify the envelope glycoproteins. Glycoproteins were identified by electrophoretic analysis in polyacrylamide slab gels of virus labelled in vivo with [3H]glucosamine or labelled enzymically in vitro with either UDP-[14C]galactose or sodium [3H]borohydride. Fluorograms revealed eleven glycoproteins (mol.

Assessment of the base sequence homology between the two subtypes of equine herpesvirus 1.

The magnitude of the genetic relatedness of the two antigenic subtypes of equine herpesvirus 1 (EHV-1) was determined by DNA-DNA reassociation kinetics. Denatured, labeled viral DNA from one EHV-1 subtype was allowed to reassociate in the presence or absence of the unlabeled heterologous viral DNA. The initial rate of reassociation of either labeled viral DNA was increased by the presence of the heterologous viral DNA to an extent indicating 10 to 20% homology between the two EHV-1 genomes.

Serologic and molecular comparisons of several equine herpesvirus type 1 strains.

The molecular and serologic relatedness of 2 recent respiratory tract isolates of equine herpesvirus type 1, designated T1 and T2, were compared with the Army 183, Kentucky-A hamster-adapted (KyA-ha), and L-M cell-adapted (KyA-LM) strains. Electrophoresis in polyacrylamide gels revealed differences in virion structural proteins among 4 purified strains. Seven envelope glycoproteins (molecular weight of 93,000, 65,000, 62,000, 60,000, 36,000, 20,000, and 18,000) corresponding to virion proteins 13, 16, 17, 18, 23, 25, and 26a, respectively, found in both the Army 183 and KyA-ha strains had slightly different molecular weight counterparts in both the T1 and T2 isolates, which had identical structural protein profiles.

Vasodilation, fibrinolysis, and thrombolysis with intraarterial infusion of urokinase in the canine superior mesenteric artery.

Urokinase, the plasminogen activator from human urine, produces a dose-dependent increase in blood flow in the canine superior mesenteric artery when injected intraarterially at doses from 10(-1) to 10(3) units kg-1. This vasodilation persists despite blockade of beta-adrenergic and histamine H1 and H2 receptors as well as inhibition of plasminogen activation, suggesting that these mechanisms are not involved. Infusion of urokinase at 10(2) CTA (Committee on Thrombolytic Agents) units kg-1 min-1 does not produce a sustained vasodilation, but is effective in achieving complete lysis of thrombi within 100 min in the superior mesenteric arterial circulation.

The role of acid and ischemia in production of stress ulcers during canine hemorrhagic shock.

Hemorrhage to a mean arterial pressure of 41 mm. Hg in ten dogs decreased Heidenhain pouch blood flow to 6 ml. per minute and aminopyrine clearance to 0.

Stress ulceration and the gastric mucosal barrier.

The role of the gastric mucosal barrier in the pathogenesis of post-traumatic stress ulcerations is far from clear. Clinical studies on critically ill patients have shown disrupted gastric mucosal barriers with hydrogen ion back diffusion, but no correlation has been made between these findings and gastric erosions. In addition, numerous assumptions concerning gastric secretions, pyloric loss and esophageal contributions to the assayed gastric juice have to be made in these patients.