Baculovirus-expressed glycoprotein H of herpes simplex virus type 1 (HSV-1) induces neutralizing antibody and delayed type hypersensitivity responses, but does not protect immunized mice against lethal HSV-1 challenge

Ghiasi, Homayon; Kaiwar, Ravi; Nesburn, Anthony B.; Wechsler, Steven L.

doi:10.1099/0022-1317-73-3-719

Cited by 31 publications

(27 citation statements)

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“…In similar experiments, we have obtained significant protection against lethal HSV-1 challenge with baculovirus recombinants expressing HSV-1 gB, gC, gD, gE, gI, gG, or gK [8,9], but not with gH [11]. The lack of protection by baculovirus-gL could not be predicted based solely on its inability to induce neutralizing antibodies.…”

Section: Discussionmentioning

confidence: 70%

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Expression and characterization of baculovirus expressed herpes simplex virus type 1 glycoprotein L

Ghiasi

Kaiwar

Slanina

et al. 1994

Archives of Virology

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We have constructed a recombinant baculovirus expressing high levels of the herpes simplex virus type 1 (HSV-1) glycoprotein L (gL) in Sf9 cells. Sf9 cells infected with this recombinant virus synthesized three polypeptides of 26-27 kDa 28 kDa, and 31 kDa. The 28 and 31 kDa species were sensitive to tunicamycin and N-glycosidase F (PNGase F) treatment, suggesting that they were glycosylated. As shown by both indirect immunofluorescence and Western blot analysis, using polyclonal antibodies to synthetic gL peptides indicated that the baculovirus expressed gL was abundant on the surface of baculovirus gL infected Sf9 cells. A small fraction of the 31 kDa polypeptide was secreted into the extracellular medium as judged by Western blot analysis. The secreted form of gL was completely resistant to Endoglycosidase H (Endo-H), while the membrane associated form of gL was only partially resistant to Endo-H treatment, suggesting that the secreted gL represented a subpopulation of the membrane bound gL. Mice vaccinated with baculovirus expressed gL produced serum antibodies that reacted with authentic HSV-1 gL. However, these mice produced no HSV-1 neutralizing antibody (titer < 1:10) and they were not protected from lethal intraperitoneal or lethal ocular challenge with HSV-1. Thus, when used as a vaccine in the mouse model, gL, similar to our findings with HSV-1 gH, but unlike our results with the other 6 HSV-1 glycoproteins that we have expressed in this baculovirus system, did not provide any protection against HSV-1 challenge.

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Section: Discussionmentioning

confidence: 70%

“…Serum neutralization titers were measured by plaque reduction assays as we described previously [11]. The neutralizing antibody titers were expressed as the reciprocal of the serum dilution that produced a 50% plaque reduction.…”

Section: Serum Neutralization Assaymentioning

confidence: 99%

Expression and characterization of baculovirus expressed herpes simplex virus type 1 glycoprotein L

Ghiasi

Kaiwar

Slanina

et al. 1994

Archives of Virology

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“…HSV-1 gH expressed in insect cells using a recombinant baculovirus induced neutralizing antibody responses but did not protect mice against lethal challenge [8]. The gH gene homologue in equine herpesvirus 1 (EHV-1), the agent of abortion, respiratory and neurological disease in the horse, encodes a predicted polypeptide after signal cleavage of 90 kDa which shares an amino acid identity of approximately 85~ with EHV-4 gH and 20-35~o with gH homologues of other alphaherpesviruses [25,26].…”

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confidence: 99%

Expression and characterisation of equine herpesvirus 1 glycoprotein H using a recombinant baculovirus

McGowan

Hayden

Edwards

et al. 1994

Archives of Virology

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A recombinant baculovirus capable of expressing the glycoprotein H (gH) gene of equine herpesvirus 1 (EHV-1) was constructed. EHV-1 gH gene products in recombinant baculovirus infected insect cells were identified as 105 kDa and 110 kDa species compared with a 115 kDa product detected in EHV-1 infected mammalian cells. The extent of N-glycosylation of EHV-1 gH in both insect and mammalian cells was indicated by a shift in apparent molecular weights after PNGase F treatment to 90 kDa and 95 kDa forms, which compared with the predicted value of 90 kDa for the unglycosylated polypeptide. The recombinant EHV-1 gH was recognised by equine sera demonstrating that EHV-1 gH is a target for the immune system of the natural host. However, while the recombinant EHV-1 gH product from infected insect cells was immunogenic in mice, it did not induce a neutralizing antibody response against EHV-1.

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“…The finding that HSV-1 glycoproteins are the major inducers and targets of humoral and cell-mediated immune responses after infection (12)(13)(14)(15)(16) suggest that they may play a major role in the development of the immune responses that cause CS in infected individuals. We have investigated the role of the HSV-1 glycoproteins in protection from eye disease and have shown that immunization with glycoprotein B (gB), gC, gD, gE, or gI completely protected mice against lethal challenge with HSV-1 and eye disease, whereas no significant protection is seen on immunization with gG, gH, gL, or gJ (17)(18)(19)(20)(21). In marked contrast, immunization with gK leads to exacerbation of eye disease and facial dermatitis (14,20).…”

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Mutations within the Pathogenic Region of Herpes Simplex Virus 1 gK Signal Sequences Alter Cell Surface Expression and Neurovirulence

Matundan

Mott

Akhtar

et al. 2015

J Virol

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To investigate the role of the signal sequences of herpes simplex virus 1 (HSV-1) gK on virus replication and viral pathogenesis, we constructed recombinant viruses with or without mutations within the signal sequences of gK. These recombinant viruses expressed two additional copies of the mutated (MgK) or native (NgK) form of the gK gene in place of the latency-associated transcript with a myc epitope tag to facilitate detection at their 3= ends. The replication of MgK virus was similar to that of NgK both in vitro and in vivo, as well as in the trigeminal ganglia (TG) of latently infected mice. The levels of gB and gK transcripts in the corneas, TG, and brains of infected mice on days 3 and 5 postinfection were markedly virus and time dependent, as well as tissue specific. Mutation in the signal sequence of gK in MgK virus blocked cell surface expression of gK-myc in rabbit skin cells, increased 50% lethal dose, and decreased corneal scarring in ocularly infected mice compared to the NgK or revertant (RgK) virus. MgK and NgK viruses, and not the RgK virus, showed a reduced extent of explant reactivation at the lower dose of ocular infection but not at the higher dose. However, the time of reactivation was not affected by overexpression of the different forms of gK. Taken together, these results strongly suggest that the 8mer peptide (ITAYGLVL) within the signal sequence of gK promotes cell surface expression of gK in infected cells and ocular pathogenesis in infected mice. IMPORTANCEIn this study, we show for the first time that mutations within the signal sequence of gK blocked cell surface expression of inserted recombinant gK in vitro. Furthermore, this blockage in cell surface expression was correlated with higher 50% lethal dose and less corneal scarring in vivo. Thus, these studies point to a key role for the 8mer within the signal sequence of gK in HSV-1-induced pathogenicity. Herpes simplex virus 1 (HSV-1) infections are among the most frequent serious viral eye infections in the United States and are a major cause of virus-induced blindness (1-3). It is estimated that 70 to 90% of American adults have antibodies to HSV-1 and/or HSV-2 and ca. 25% of these individuals have clinical symptoms upon routine clinical inquiry. HSV-1 is responsible for Ͼ90% of ocular HSV infections (1, 2, 4-7). HSV-1-induced corneal scarring (CS), also broadly referred to as herpes stromal keratitis (HSK), can lead to blindness. Furthermore, HSV-1 is the leading cause of corneal blindness due to an infectious agent in developed countries (1, 2, 4-7). In addition to necrotizing HSK, ocular infection with HSV-1 can cause eye disease ranging in severity from blepharitis, conjunctivitis, and dendritic keratitis to disciform stromal edema (4, 7-11). In the United States, approximately 30,000 people per year suffer recurrent ocular HSV episodes, requiring doctor visits, medication, and in severe cases, corneal transplants.Although it is well established that HSV-1-induced CS is due to immune responses to the virus, the exact...

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Baculovirus-expressed glycoprotein H of herpes simplex virus type 1 (HSV-1) induces neutralizing antibody and delayed type hypersensitivity responses, but does not protect immunized mice against lethal HSV-1 challenge

Cited by 31 publications

References 10 publications

Expression and characterization of baculovirus expressed herpes simplex virus type 1 glycoprotein L

Expression and characterization of baculovirus expressed herpes simplex virus type 1 glycoprotein L

Expression and characterisation of equine herpesvirus 1 glycoprotein H using a recombinant baculovirus

Mutations within the Pathogenic Region of Herpes Simplex Virus 1 gK Signal Sequences Alter Cell Surface Expression and Neurovirulence

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