Comparative Inhibitory Effects of Nucleoside Analogues on Different Clinical Isolates of Human Cytomegalovirus In Vitro

ABSTRACT A recombinant human cytomegalovirus (AD169-GFP) expressing green fluorescent protein was generated by homologous recombination. Infection of human fibroblast cultures with AD169-GFP virus produced stable and readily detectable amounts of GFP signals which were quantitated by automated fluorometry. Hereby, high levels of sensitivity and reproducibility could be achieved, compared to those with the conventional plaque reduction assay. Antiviral activities were determined for four reference compounds as well as a set of putative novel cytomegalovirus inhibitors. The results obtained were exactly in line with the known characteristics of reference compounds and furthermore revealed distinct antiviral activities of novel in vitro inhibitors. The fluorometric data could be confirmed by GFP-based flow cytometry and fluorescence microscopy. In addition, laboratory virus variants derived from the recombinant AD169-GFP virus provided further possibilities for study of the characteristics of drug resistance. The GFP-based antiviral assay appeared to be very reliable for measuring virus-inhibitory effects in concentration- and time-dependent fashions and might also be adaptable for high-throughput screenings of cytomegalovirus-specific antiviral agents.

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In vitro Inhibitory Effects of Combinations of anti-Influenza Agents

Antiviral Chemistry and Chemotherapy ◽

10.1177/095632029500600205 ◽

1995 ◽

Vol 6 (2) ◽

pp. 109-113 ◽

Cited By ~ 36

Author(s):

L. K. Madren ◽

C. Shipman ◽

F. G. Hayden

Keyword(s):

Viral Replication ◽

Influenza A ◽

H1n1 Virus ◽

Mdck Cells ◽

Clinical Isolates ◽

H1n1 Subtype ◽

Additive Effects ◽

Inhibitory Effects ◽

Influenza A H1n1

To assess the possible interactions among candidate anti-influenza agents, dual combinations of rimantadine, ribavirin, 2′-deoxy-2′-fluoroguanosine (2-FDG) and 4-guanidino-Neu5Ac2en (GG167) were tested against clinical isolates of influenza A H3N2 and H1N1 subtype viruses in MDCK cells by ELISA. Each of the dual combinations showed additive effects, except for the combination of 2-FDG and ribavirin which was synergistic against the influenza A (H1N1) virus. However, this combination also showed enhanced cytotoxicity. In this assay system, influenza agents with differing mechanisms of antiviral interaction interacted in an additive fashion with respect to inhibition of viral replication.

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In VitroCharacterization of Human Cytomegalovirus-Targeting Therapeutic Monoclonal Antibodies LJP538 and LJP539

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00382-16 ◽

2016 ◽

Vol 60 (8) ◽

pp. 4961-4971 ◽

Cited By ~ 11

Author(s):

Hetalkumar D. Patel ◽

Pavel Nikitin ◽

Thomas Gesner ◽

James J. Lin ◽

David T. Barkan ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Human Cytomegalovirus ◽

Target Genes ◽

Hcmv Infection ◽

Clinical Isolates ◽

Hcmv Disease ◽

Life Threatening ◽

Approved Drugs ◽

Loss Of Susceptibility

ABSTRACTHuman cytomegalovirus (HCMV) infection is usually benign in healthy individuals but can cause life-threatening disease in those with compromised immune systems. Approved drugs available to treat HCMV disease, including ganciclovir, cidofovir, and foscarnet, have significant toxicities that limit their use in certain patient populations. LJP538 and LJP539 are human monoclonal antibodies that are being evaluated as immunoglobulin therapeutics. The antibodies target glycoproteins gB and the gH/gL/UL128/UL130/UL131a pentameric complex, respectively. Here we present anin vitrocharacterization of these antibodies. We show that LJP538 and LJP539 are more potent than a marketed immunoglobulin at inhibiting HCMV infection of various cell lines relevant to pathogenesis. We find that LJP538 and LJP539 are active against a panel of clinical isolatesin vitroand demonstrate minor-to-moderate synergy in combination. Passage of HCMV in the presence of LJP538 or LJP539 alone resulted in resistance-associated mutations that mapped to the target genes. However, no loss of susceptibility to the combination of antibodies was observed for >400 days in culture. Finally, the binding regions of LJP538 and LJP539 are conserved among clinical isolates. Taken together, these data support the use of LJP538 and LJP539 in combination for clinical trials in HCMV patients.

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Effects of Certain Nucleoside Analogues on Human Cytomegalovirus Replication in vitro

Journal of General Virology ◽

10.1099/0022-1317-65-1-47 ◽

1984 ◽

Vol 65 (1) ◽

pp. 47-53 ◽

Cited By ~ 23

Author(s):

E.-C. Mar ◽

P. C. Patel ◽

Y.-C. Cheng ◽

J. J. Fox ◽

K. A. Watanabe ◽

...

Keyword(s):

Human Cytomegalovirus ◽

Nucleoside Analogues

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Experimental study on the action of allitridin against human cytomegalovirus in vitro: Inhibitory effects on immediate-early genes

Antiviral Research ◽

10.1016/j.antiviral.2006.03.017 ◽

2006 ◽

Vol 72 (1) ◽

pp. 68-74 ◽

Cited By ~ 22

Author(s):

Hong Zhen ◽

Feng Fang ◽

Du-yun Ye ◽

Sai-nan Shu ◽

Yu-feng Zhou ◽

...

Keyword(s):

Experimental Study ◽

Human Cytomegalovirus ◽

Immediate Early Genes ◽

Immediate Early ◽

Inhibitory Effects ◽

Early Genes

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The attenuated Towne strain of human cytomegalovirus may revert to both endothelial cell tropism and leuko- (neutrophil- and monocyte-) tropism in vitro

Journal of General Virology ◽

10.1099/0022-1317-83-8-1993 ◽

2002 ◽

Vol 83 (8) ◽

pp. 1993-2000 ◽

Cited By ~ 25

Author(s):

Giuseppe Gerna ◽

Elena Percivalle ◽

Antonella Sarasini ◽

Fausto Baldanti ◽

M. Grazia Revello

Keyword(s):

Endothelial Cell ◽

Cell Cultures ◽

Human Cytomegalovirus ◽

Biological Properties ◽

Clinical Isolates ◽

Viral Gene ◽

Cell Tropism ◽

Human Embryonic Lung ◽

Healthy Human

The Towne strain of human cytomegalovirus (HCMV), originally recovered from the urine of a congenitally infected newborn, was attenuated through 125 passages in human embryonic lung fibroblast cell cultures. Although reliable markers of attenuation were not identified, the virus was shown to be attenuated by inoculation of both healthy human volunteers and immunocompromised patients. More recently, Towne (like other laboratory-adapted strains) was shown not to have two biological properties typical of recent clinical isolates: endothelial cell tropism and polymorphonuclear leukocyte tropism. These markers of attenuation are lost by all clinical isolates on extensive propagation in cell cultures and are apparently associated with one another. Here, we show that Towne may reacquire both endothelial cell tropism and leuko- (polymorphonuclear- and monocyte-) tropism on adaptation to growth in endothelial cell cultures. However, reversion to endothelial cell tropism is dissociated from reversion to leukotropism, since the latter was reacquired 10–20 passages later. Thus, these two biological properties, which were considered to be encoded by the same viral gene(s), appear to be distinct. Both restriction fragment length polymorphism and Southern blot analysis demonstrated the identity of the attenuated and endothelial cell tropic variants of Towne, thus suggesting that only minor variations (mutations) of the viral genome may be responsible for loss or reacquisition of the two biological properties. Viral genes involved in endothelial cell tropism and leukotropism remain to be identified. However, reversion of attenuated strains to pathogenicity in vivo cannot be excluded a priori.

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Inhibitory effects of piceatannol on human cytomegalovirus (hCMV) in vitro

The Journal of Microbiology ◽

10.1007/s12275-020-9528-2 ◽

2020 ◽

Vol 58 (8) ◽

pp. 716-723

Author(s):

San-Ying Wang ◽

Jing Zhang ◽

Xiao-Gang Xu ◽

Hui-Li Su ◽

Wen-Min Xing ◽

...

Keyword(s):

Human Cytomegalovirus ◽

Inhibitory Effects

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Inhibitory effects of bioaccessible anthocyanins and procyanidins from apple, red grape, cinnamon on α-amylase, α-glucosidase and lipase

International Journal for Vitamin and Nutrition Research ◽

10.1024/0300-9831/a000652 ◽

2020 ◽

pp. 1-9

Author(s):

Pınar Ercan ◽

Sedef Nehir El

Keyword(s):

Inhibitory Activity ◽

Digestive Enzymes ◽

Positive Control ◽

Anthocyanin Content ◽

Inhibitory Effects ◽

Total Anthocyanin ◽

Total Anthocyanin Content ◽

Before And After ◽

Red Grape

Abstract. The goals of this study were to determine and evaluate the bioaccessibility of total anthocyanin and procyanidin in apple (Amasya, Malus communis), red grape (Papazkarası, Vitis vinifera) and cinnamon (Cassia, Cinnamomum) using an in vitro static digestion system based on human gastrointestinal physiologically relevant conditions. Also, in vitro inhibitory effects of these foods on lipid (lipase) and carbohydrate digestive enzymes (α-amylase and α-glucosidase) were performed with before and after digested samples using acarbose and methylumbelliferyl oleate (4MUO) as the positive control. While the highest total anthocyanin content was found in red grape (164 ± 2.51 mg/100 g), the highest procyanidin content was found in cinnamon (6432 ± 177.31 mg/100 g) (p < 0.05). The anthocyanin bioaccessibilities were found as 10.2 ± 1%, 8.23 ± 0.64%, and 8.73 ± 0.70% in apple, red grape, and cinnamon, respectively. The procyanidin bioaccessibilities of apple, red grape, and cinnamon were found as 17.57 ± 0.71%, 14.08 ± 0.74% and 18.75 ± 1.49%, respectively. The analyzed apple, red grape and cinnamon showed the inhibitory activity against α-glucosidase (IC50 544 ± 21.94, 445 ± 15.67, 1592 ± 17.58 μg/mL, respectively), α-amylase (IC50 38.4 ± 7.26, 56.1 ± 3.60, 3.54 ± 0.86 μg/mL, respectively), and lipase (IC50 52.7 ± 2.05, 581 ± 54.14, 49.6 ± 2.72 μg/mL), respectively. According to our results apple, red grape and cinnamon have potential to inhibit of lipase, α-amylase and α-glucosidase digestive enzymes.

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