7 research outputs found
A 3D Model of the Membrane Protein Complex Formed by the White Spot Syndrome Virus Structural Proteins
Outbreaks of white spot disease have had a large negative economic impact on cultured shrimp worldwide. However, the pathogenesis of the causative virus, WSSV (whit spot syndrome virus), is not yet well understood. WSSV is a large enveloped virus. The WSSV virion has three structural layers surrounding its core DNA: an outer envelope, a tegument and a nucleocapsid. In this study, we investigated the protein-protein interactions of the major WSSV structural proteins, including several envelope and tegument proteins that are known to be involved in the infection process.In the present report, we used coimmunoprecipitation and yeast two-hybrid assays to elucidate and/or confirm all the interactions that occur among the WSSV structural (envelope and tegument) proteins VP51A, VP19, VP24, VP26 and VP28. We found that VP51A interacted directly not only with VP26 but also with VP19 and VP24. VP51A, VP19 and VP24 were also shown to have an affinity for self-interaction. Chemical cross-linking assays showed that these three self-interacting proteins could occur as dimers.From our present results in conjunction with other previously established interactions we construct a 3D model in which VP24 acts as a core protein that directly associates with VP26, VP28, VP38A, VP51A and WSV010 to form a membrane-associated protein complex. VP19 and VP37 are attached to this complex via association with VP51A and VP28, respectively. Through the VP26-VP51C interaction this envelope complex is anchored to the nucleocapsid, which is made of layers of rings formed by VP664. A 3D model of the nucleocapsid and the surrounding outer membrane is presented
Efficacy of Recombinant Herpes Simplex Virus 1 Glycoprotein D Candidate Vaccines in Mice
To compare the
immunogenity of the herpes simplex virus 1 (HSV-1/HHV-1) recombinant
glycoprotein D (gD1), as a potential protective vaccine, Balb/c mice were
immunized with either gD1/313 (the ectodomain of the gD1 fusion protein
consisting of 313 amino acid residues), or the plasmid pcDNA3.1-gD (coding for
a full length gD1 protein, FLgD1). A live attenuated HSV-l (deleted in the gE
gene), and a HSV-1 (strain HSZP)-infected cell extract served as positive
controls, and three non-structural recombinant HSV-1 fusion proteins (ICP27,
UL9/OBP and thymidine kinase - TK) were used as presumed non-protective
(negative) controls. Protection tests showed that the LD
50
value of
the challenging infectious virus increased 90-fold in mice immunized with
ICP27, but remained unchanged in other control mice immunized with TK and OBP
polypeptides. A significant protection (the LD
50
value of
challenging virus increased 800-fold) was noted following immunization with
gD1/313, while immunization with the gE-del virus and/or the gD1 DNA vaccine
resulted in a more than 4,000-fold increase of the challenging virus dose
killing 50% of the animals. Using ELISA, elevated antibody titers were detected
following immunizations with gD1/313, gE-del virus, and/or HSV-1-infected-cell
extract. In addition, all of the three non-structural proteins elicited a good
humoral response (with titres ranging from 1:16,000 to 1:128,000). The lowest
IgG response (1:8,000) was noted after immunization with the gD1 DNA vaccine.
Peripheral blood leukocytes (PBLs) as well as splenocytes from mice immunized
with gD1/313, gE-del virus, and gD1-plasmid responded in lymphocyte
transformation test (LTT) to the presence of purified gD1/313 antigen. For
PBLs, the most significant stimulation of thymidine incorporation was
registered at a gD1/313 concentration of 5 µg/100 µl, while the splenocytes
from DNA vaccine-immunized mice responded already at a concentration of 1
µg/100 µl