scholarly journals HSV-1 Protein Expression Using Recombinant Baculoviruses

2014 ◽  
pp. 293-304 ◽  
Author(s):  
Lorry M. Grady ◽  
Ping Bai ◽  
Sandra K. Weller
Keyword(s):  
Protein Expression ◽  
Recombinant Baculoviruses ◽  
Hsv 1

Novel vectors for simultaneous high-level dual protein expression in vertebrate and insect cells by recombinant baculoviruses

2009 ◽  
Vol 160 (1-2) ◽  
pp. 132-137 ◽  
Author(s):  
Günther M. Keil ◽  
Constanze Klopfleisch ◽  
Katrin Giesow ◽  
Ulrike Blohm
Keyword(s):  
Protein Expression ◽  
Insect Cells ◽  
Recombinant Baculoviruses ◽  
High Level

Glial fibrillary acidic protein expression during HSV-1 infection in mouse cornea

Apmis ◽  
2013 ◽  
Vol 122 (2) ◽  
pp. 128-135 ◽  
Author(s):  
Ge Zhao ◽  
Hao Chen ◽  
Zicheng Song ◽  
Hongmei Yin ◽  
Yuanyuan Xu ◽  
...  
Keyword(s):  
Glial Fibrillary Acidic Protein ◽  
Protein Expression ◽  
Acidic Protein ◽  
Hsv 1

scholarly journals Newly Developed Oncolytic Herpes Simplex Viruses Expressing Multiple Immunomodulatory Transgenes Effectively Target AML Cells

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 1169-1169
Author(s):  
Yixin Zheng ◽  
Min Chen ◽  
Yanal Murad ◽  
Luke Bu ◽  
William Jia ◽  
...  
Keyword(s):  
Herpes Simplex ◽  
Protein Expression ◽  
Cell Lines ◽  
Cell Killing ◽  
Dependent Manner ◽  
Facs Analysis ◽  
Herpes Simplex Viruses ◽  
Infected Cells ◽  
Dose Dependent ◽  
Hsv 1

Abstract Acute myeloid leukemia (AML) is the most common human leukemia and is a major area of unmet medical need among hematologic malignancies. Progress has been made in identifying therapeutic targets and several approved therapies, but resistance to frontline chemotherapy remains a major cause of treatment failure, highlighting the need for new therapies. Oncolytic viruses (OV) are a promising new class of therapeutics that rely on tumor specific oncolysis and the generation of a potent adaptive anti-tumor immune response for efficacy. To investigate if our newly developed oncolytic herpes simplex viruses (oHSVs), designed to potentiate anti-leukemia immunity, effectively target primitive AML cells, we evaluated oHSV-VG161, which is engineered to express IL-12, IL-15 and the IL-15 receptor alpha subunit, along with a peptide fusion protein capable of disrupting PD-1/PD-L1 interaction. After screening several AML cell lines that expressed relatively high levels of a HSV entry receptor (HVEM), we demonstrated that VG161-infected OCIAML3 and MOLM13 cells significantly enhanced cell killing (IC 50: 0.4 & 1.8 multiplicity of infection (MOI) as compared to MV4-11 and U973 cells (IC 50: 3.0 & 9.5 MOI). These effects were 2-3 folds lower in control VG160-infected cells. We also observed that VG161-infected AML cells induced apoptosis in a dose-dependent manner (~50%) after 48 hours and cleaved PARP, Caspase-3 and Caspase-8 were increased in these cells, and to a lesser extent in control VG160-infected cells. Both VG160 and VG161 viruses replicated efficiently in OCIAML3 and MOLM13 cells in a timely, dose-dependent manner, evidenced by qPCR detection of HSV-1 ICP27 DNA copy numbers (>500-fold increase) over 48 hours of treatment. This result was supported by detection of protein expression of HSV-1 glycoprotein D in VG160 and VG161-infected cells (up to 40% of protein detected) by FACS analysis. Interestingly, IL-12 but not IL-15 protein expression was found in intracellular-stained VG161-infected OCIAML3 and MOLM13 cells in a dose-dependent manner (up to 13% of protein detected, P<0.01) but not in VG160-infected cells, as assessed by FACS analysis. Production of IL-12 was also detected in cultured media obtained from VG161-infected AML cells (up to 150 pg/mL) by ELISA. To investigate potential molecular mechanisms of VG161-mediated anti-leukemia response and specific signalling pathways, we have screened several potential candidates and found immune regulating genes, such as IRF3, IRF7, IRF9, NFkB and ISGs, as well as type I IFN to be highly increased in VG161-infected cells as compared to VG160-infected cells (2-4-folds, P<0.001) in a dose dependent manner over 48 hours of treatment, assayed by qRT-PCR. Western blot analysis demonstrated increased phosphorylation of p-STAT1 and its protein expression in VG161-infected cells compared to VG160 control cells (~2-fold). These results suggest that VG161 viruses expressing several engineered immunomodulatory transgenes, particularly IL-12, contribute to anti-leukemia responses by activating specific immune regulating pathways such as the JAK/STAT pathway. In addition, we detected an increase in both RNA and protein levels of PD-L1 in VG161-infected AML cells, suggesting the necessity of PD-L1 blocking peptide in the viral construct. To further investigate VG161's role in regulating innate and adaptive immune responses, we have examined the biological effects of VG160/VG161 in the presence of healthy peripheral blood mononuclear cells (PBMC) in both AML cell lines and primary AML patient cells in vitro. Most interestingly, VG160 or VG161-infected OCIAML3 and MOLM13 cells show enhanced cell killing when co-cultured with PBMC and this cell killing effect was greatly enhanced in VG161-infected cells as compared to VG160-infected cells, especially in the MOLM13 cell line (up to 90% killing). This observation was further supported when primitive AML patient cells were co-cultured with VG161 and PBMC as compared to VG160 control cells. Moreover, PD-L1 expression was highly increased in AML patient cells when cultured with VG161 as compared to VG160 (2.7-fold) and this was further enhanced when co-cultured with VG161 and PBMC. Thus, we have demonstrated that newly developed oHSVs engineered with several immunomodulatory transgenes effectively target primitive AML cells, suggesting a potential treatment strategy for AML. Disclosures No relevant conflicts of interest to declare.

scholarly journals Effect of promoter strength on protein expression and immunogenicity of an HSV-1 amplicon vector encoding HIV-1 Gag

Vaccine ◽  
2007 ◽  
Vol 25 (9) ◽  
pp. 1634-1646 ◽  
Author(s):  
Kathlyn Santos ◽  
Cindy M.P. Duke ◽  
Sol M. Rodriguez-Colon ◽  
Anthony Dakwar ◽  
Shongshan Fan ◽  
...  
Keyword(s):  
Protein Expression ◽  
Promoter Strength ◽  
Hiv 1 ◽  
Hsv 1

Assembly of VP26 in herpes simplex virus-1 capsid implicated by 3-D structure of recombinant capsid

1995 ◽  
Vol 53 ◽  
pp. 840-841
Author(s):  
Z. Hong Zhou ◽  
Jing He ◽  
Joanita Jakana ◽  
J. D. Tatman ◽  
Frazer J. Rixon ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
3D Structure ◽  
Structure Study ◽  
Herpes Simplex Virus 1 ◽  
Shell Proteins ◽  
Life Threatening ◽  
Infected Cells ◽  
Simplex Virus ◽  
Hsv 1

Herpes simplex virus-1 (HSV-1) is a ubiquitous virus which is implicated in diseases ranging from self-curing cold sores to life-threatening infections. The 2500 Å diameter herpes virion is composed of a glycoprotein spike containing, lipid envelope, enclosing a protein layer (the tegument) in which is embedded the capsid (which contains the dsDNA genome). The B-, and A- and C-capsids, representing different morphogenetic stages in HSV-1 infected cells, are composed of 7, and 5 structural proteins respectively. The three capsid types are organized in similar T=16 icosahedral shells with 12 pentons, 150 hexons, and 320 connecting triplexes. Our previous 3D structure study at 26 Å revealed domain features of all these structural components and suggested probable locations for the outer shell proteins, VP5, VP26, VP19c and VP23. VP5 makes up most of both pentons and hexons. VP26 appeared to bind to the VP5 subunit in hexon but not to that in penton.

CFTR protein expression in healthy men spermatozoa is not correlated with fertilization rate of ova

2010 ◽  
Vol 34 (8) ◽  
pp. S12-S12
Author(s):  
Hong‑Ge Li ◽  
Chen Min Xu ◽  
Kun Li ◽  
Ya Ni ◽  
Wen‑Ying Chen ◽  
...  
Keyword(s):  
Protein Expression ◽  
Fertilization Rate ◽  
Healthy Men ◽  
Cftr Protein

Bone sialoprotein mRNA and protein expression in human multiple myeloma cell lines and patients

2000 ◽  
Vol 111 (4) ◽  
pp. 1118-1121 ◽  
Author(s):  
A. Bellahcene ◽  
I. Van Riet ◽  
C. de Greef ◽  
N. Antoine ◽  
M. F. Young ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Protein Expression ◽  
Cell Lines ◽  
Myeloma Cell ◽  
Bone Sialoprotein ◽  
Multiple Myeloma Cell ◽  
Human Multiple Myeloma ◽  
Mrna And Protein Expression
Sign in / Sign up

Export Citation Format

Share Document