APVO210: A Bispecific Anti-CD86-IL-10 Fusion Protein (ADAPTIR™) to Induce Antigen-Specific T Regulatory Type 1 Cells

ABSTRACT Herpes simplex virus type 1 glycoprotein K (gK) and the UL20 protein (UL20p) are coordinately transported to the trans-Golgi network (TGN) and cell surfaces and are required for cytoplasmic virion envelopment at the TGN. In addition, cell surface expression of gK and UL20p is required for virus-induced cell fusion. Previously, confocal microscopy colocalization and intracellular transport experiments strongly suggested direct protein-protein interactions between gK and UL20p. Direct protein-protein interactions between gK and UL20p were demonstrated through reciprocal coimmunoprecipitation experiments, as well as with glutathione S-transferase (GST) pull-down experiments. A fusion protein consisting of the amino-terminal 66 amino acids of UL20p fused in-frame with GST was expressed in Escherichia coli and purified via glutathione column chromatography. Precipitation of GST-UL20p from mixtures of GST-UL20p fusion protein with cellular extracts containing gK specifically coprecipitated gK but not other viral glycoproteins. The purified UL20p-GST fusion protein reacted with all gK-associated protein species. It was concluded that the amino terminus of UL20p, most likely, interacted with gK domain III, which is predicted to lie intracellularly. UL20p-gK domain-specific interactions must serve important functions in the coordinate transport of UL20p and gK to the TGN, because retention of UL20p in the endoplasmic reticulum (ER) via the addition of an ER retention signal at the carboxyl terminus of UL20p forced the ER retention of gK and drastically inhibited intracellular virion envelopment and virus-induced cell fusion.

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Inhibition of Cyclic AMP-Dependent Kinase by Expression of a Protein Kinase Inhibitor/Enhanced Green Fluorescent Fusion Protein Attenuates Angiotensin II-Induced Type 1 AT1Receptor mRNA Down-Regulation in Vascular Smooth Muscle Cells

Molecular Pharmacology ◽

10.1124/mol.54.3.514 ◽

1998 ◽

Vol 54 (3) ◽

pp. 514-524 ◽

Cited By ~ 10

Author(s):

Xiaofei Wang ◽

T. J. Murphy

Keyword(s):

Smooth Muscle ◽

Angiotensin Ii ◽

Protein Kinase ◽

Cyclic Amp ◽

Fusion Protein ◽

Smooth Muscle Cells ◽

Vascular Smooth Muscle Cells ◽

Kinase Inhibitor ◽

Green Fluorescent

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Construction of Recombinant Lactococcus lactis Strain Expressing VP1 Fusion Protein of Duck Hepatitis A Virus Type 1 and Evaluation of Its Immune Effect

Vaccines ◽

10.3390/vaccines9121479 ◽

2021 ◽

Vol 9 (12) ◽

pp. 1479

Author(s):

Xiaoting Zhang ◽

Ruihua Zhang ◽

Jingyu Wang ◽

Nana Sui ◽

Guige Xu ◽

...

Keyword(s):

Immune System ◽

Fusion Protein ◽

Lactococcus Lactis ◽

Virus Type ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Interleukin 2 ◽

Interleukin 4 ◽

Duck Hepatitis

With the continuous development of duck farming and the increasing breeding density, the incidence of duck hepatitis A virus type 1 (DHAV-1) has been on the rise, seriously endangering the development of duck farming. To reduce the use of antibiotics in duck breeding, susceptibility risks and mortality, and avoid virulence recovery and immune failure risk, this study aims to develop a new type of mucosal immune probiotics and make full use of molecular biology techniques, on the level of genetic engineering, to modify Lactococcus lactis (L. lactis). In this study, a secretory recombinant L. lactis named MG1363-VP1 with an enhanced Green Fluorescent Protein (eGFP) and translation enhancer T7g10L was constructed, which could express the VP1-eGFP fusion protein of DHAV-1. The animal experiment in ducklings was performed to detect the immune response and protection effect of oral microecologics by recombinant L. lactis. The results showed that oral L. lactis MG1363-VP1 significantly induced the body’s humoral immune system and mucosal immune system to produce specific anti-VP1 IgG antibodies and mucosal secretory immunoglobulin A (sIgA) for DHAV-1 in ducklings, and cytokines including interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10), and interferon gamma (IFN-γ). The mortality rate was monitored simultaneously by the natural infestation in the process of production and breeding; notably, the ducklings vaccinated with L. lactis MG1363-VP1 were effectively protected against the nature infection of DHAV-1. The recombinant L. lactis MG1363-VP1 constructed in this study provides a new means of preventing and controlling DHAV-1 infection in the future.

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