Herpes Simplex Virus Type 2 Glycoprotein G-Negative Clinical Isolates Are Generated by Single Frameshift Mutations

ABSTRACT Herpes simplex virus (HSV) codes for several envelope glycoproteins, including glycoprotein G-2 (gG-2) of HSV type 2 (HSV-2), which are dispensable for replication in cell culture. However, clinical isolates which are deficient in such proteins occur rarely. We describe here five clinical HSV-2 isolates which were found to be unreactive to a panel of anti-gG-2 monoclonal antibodies and therefore considered phenotypically gG-2 negative. These isolates were further examined for expression of the secreted amino-terminal and cell-associated carboxy-terminal portions of gG-2 by immunoblotting and radioimmunoprecipitation. The gG-2 gene was completely inactivated in four isolates, with no expression of the two protein products. For one isolate a normally produced secreted portion and a truncated carboxy-terminal portion of gG-2 were detected in virus-infected cell medium. Sequencing of the complete gG-2 gene identified a single insertion or deletion of guanine or cytosine nucleotides in all five strains, resulting in a premature termination codon. The frameshift mutations were localized within runs of five or more guanine or cytosine nucleotides and were dispersed throughout the gene. For the isolate for which a partially inactivated gG-2 gene was detected, the frameshift mutation was localized upstream of but adjacent to the nucleotides coding for the transmembranous region. Thus, this study demonstrates the existence of clinical HSV-2 isolates which do not express an envelope glycoprotein and identifies the underlying molecular mechanism to be a single frameshift mutation.

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Monoclonal antibodies and human sera directed to the secreted glycoprotein G of herpes simplex virus type 2 recognize type-specific antigenic determinants

Journal of General Virology ◽

10.1099/0022-1317-83-1-157 ◽

2002 ◽

Vol 83 (1) ◽

pp. 157-165 ◽

Cited By ~ 20

Author(s):

Jan-Åke Liljeqvist ◽

Edward Trybala ◽

Johan Hoebeke ◽

Bo Svennerholm ◽

Tomas Bergström

Keyword(s):

Monoclonal Antibodies ◽

Herpes Simplex Virus ◽

Herpes Simplex ◽

Terminal Portion ◽

Glycoprotein G ◽

Infected Cells ◽

Simplex Virus ◽

Carboxy Terminal ◽

Hsv 1

Glycoprotein G-2 (gG-2) of herpes simplex virus type 2 (HSV-2) is cleaved to a secreted amino-terminal portion (sgG-2) and to a cell-associated carboxy-terminal portion which is further O-glycosylated to constitute the mature gG-2 (mgG-2). In contrast to mgG-2, which is known to elicit a type-specific antibody response in the human host, information on the immunogenic properties of sgG-2 is lacking. Here the sgG-2 protein was purified on a heparin column and used for production of monoclonal antibodies (mAbs). Four anti-sgG-2 mAbs were mapped using a Pepscan technique and identified linear epitopes which localized to the carboxy-terminal part of the protein. One additional anti-sgG-2 mAb, recognizing a non-linear epitope, was reactive to three discrete peptide stretches where the most carboxy-terminally located stretch was constituted by the amino acids 320RRAL323. Although sgG-2 is rapidly secreted into the cell-culture medium after infection, the anti-sgG-2 mAbs identified substantial amounts of sgG-2 in the cytoplasm of HSV-2-infected cells. All of the anti-sgG-2 mAbs were HSV-2 specific showing no cross-reactivity to HSV-1 antigen or to HSV-1-infected cells. Similarly, sera from 50 HSV-2 isolation positive patients were all reactive to sgG-2 in an enzyme immunoassay whilst no reactivity was seen in 25 sera from HSV-1 isolation positive patients or in 25 serum samples from HSV-negative patients suggesting that sgG-2 is a novel antigen potentially suitable for type-discriminating serodiagnosis.

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Faculty Opinions recommendation of Monoclonal antibodies and human sera directed to the secreted glycoprotein G of herpes simplex virus type 2 recognize type-specific antigenic determinants.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1003873.39455 ◽

2002 ◽

Author(s):

Rhoda Ashley Morrow

Keyword(s):

Monoclonal Antibodies ◽

Herpes Simplex Virus ◽

Herpes Simplex ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Antigenic Determinants ◽

Glycoprotein G ◽

Human Sera ◽

Simplex Virus

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Performance of the Focus and Kalon Enzyme-Linked Immunosorbent Assays for Antibodies to Herpes Simplex Virus Type 2 Glycoprotein G in Culture-Documented Cases of Genital Herpes

Journal of Clinical Microbiology ◽

10.1128/jcm.41.11.5212-5214.2003 ◽

2003 ◽

Vol 41 (11) ◽

pp. 5212-5214 ◽

Cited By ~ 36

Author(s):

R. A. Morrow ◽

D. Friedrich ◽

E. Krantz

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Genital Herpes ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Glycoprotein G ◽

Immunosorbent Assays ◽

Simplex Virus

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Herpes Simplex Virus Type 2 Mucin-Like Glycoprotein mgG Promotes Virus Release from the Surface of Infected Cells

Viruses ◽

10.3390/v13050887 ◽

2021 ◽

Vol 13 (5) ◽

pp. 887

Author(s):

Edward Trybala ◽

Nadia Peerboom ◽

Beata Adamiak ◽

Malgorzata Krzyzowska ◽

Jan-Åke Liljeqvist ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Culture Medium ◽

Glycoprotein G ◽

Infected Cells ◽

Virus Interaction ◽

Simplex Virus

The contribution of virus components to liberation of herpes simplex virus type 2 (HSV-2) progeny virions from the surface of infected cells is poorly understood. We report that the HSV-2 mutant deficient in the expression of a mucin-like membrane-associated glycoprotein G (mgG) exhibited defect in the release of progeny virions from infected cells manifested by ~2 orders of magnitude decreased amount of infectious virus in a culture medium as compared to native HSV-2. Electron microscopy revealed that the mgG deficient virions were produced in infected cells and present at the cell surface. These virions could be forcibly liberated to a nearly native HSV-2 level by the treatment of cells with glycosaminoglycan (GAG)-mimicking oligosaccharides. Comparative assessment of the interaction of mutant and native virions with surface-immobilized chondroitin sulfate GAG chains revealed that while the mutant virions associated with GAGs ~fourfold more extensively, the lateral mobility of bound virions was much poorer than that of native virions. These data indicate that the mgG of HSV-2 balances the virus interaction with GAG chains, a feature critical to prevent trapping of the progeny virions at the surface of infected cells.

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Variability of the Glycoprotein G Gene in Clinical Isolates of Herpes Simplex Virus Type 1

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.6.6.826-831.1999 ◽

1999 ◽

Vol 6 (6) ◽

pp. 826-831 ◽

Cited By ~ 12

Author(s):

Elham Rekabdar ◽

Petra Tunbäck ◽

Jan-Åke Liljeqvist ◽

Tomas Bergström

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Herpes Simplex Virus Type ◽

Clinical Isolates ◽

Missense Mutations ◽

Sequence Variability ◽

Glycoprotein G ◽

Simplex Virus ◽

Hsv 1

ABSTRACT Glycoprotein G (gG) of herpes simplex virus type 1 (HSV-1) has been used as a prototype antigen for HSV-1 type-specific serodiagnosis, but data on the sequence variability of the gene coding for this protein in wild-type strains are lacking. In this study, direct DNA sequencing of the gG-1 genes from PCR products was performed with clinical HSV-1 isolates from 11 subjects as well as with strains Syn 17+, F, and KOS 321. The reference strains Syn 17+ and F showed a high degree of conservation, while KOS 321 carried 13 missense mutations and, in addition, 12 silent mutations. Three clinical isolates showed mutations leading to amino acid alterations: one had a mutation of K122 to N, which is a gG-1–to–gG-2 alteration; another contained all mutations which were observed in KOS 321 except two silent mutations; and the third isolate carried five missense mutations. Two clinical isolates as well as strain KOS 321 showed a mutation (F111→V) within the epitope of a gG-1-reactive monoclonal antibody (MAb). When all viruses were tested for reactivity with the anti-gG-1 MAb, the three strains with the F111→V mutation were found to be unreactive. Furthermore, gG-1 antibodies purified from sera from the two patients carrying strains mutated in this epitope were less reactive when they were tested by an HSV-1-infected-cell assay. Therefore, our finding that the sequence variability of the gG-1 gene also affects B-cell epitope regions of this protein in clinical isolates may have consequences for the use of this protein as a type-specific antigen for serodiagnosis.

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Typing of Clinical Herpes Simplex Virus Type 1 and Type 2 Isolates with Monoclonal Antibodies

Journal of Clinical Microbiology ◽

10.1128/jcm.37.8.2717-2718.1999 ◽

1999 ◽

Vol 37 (8) ◽

pp. 2717-2718 ◽

Cited By ~ 13

Author(s):

Jan-Åke Liljeqvist ◽

Bo Svennerholm ◽

Tomas Bergström

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Herpes Simplex Virus ◽

Herpes Simplex ◽

Glycoprotein C ◽

Glycoprotein G ◽

Simplex Virus ◽

Hsv 1

The purpose of this study was to evaluate the performance of a herpes simplex virus (HSV) type 1-specific anti-glycoprotein C-1 monoclonal antibody (MAb) and a type 2-specific anti-glycoprotein G-2 MAb for typing of 2,400 clinical HSV-1 isolates and 2,400 clinical HSV-2 isolates, respectively, using an enzyme immunoassay. The anti-HSV-1 MAb showed sensitivity and specificity of 100%, and the anti-HSV-2 MAb showed a sensitivity of 99.46% and 100% specificity, indicating that these MAbs are suitable for typing of clinical HSV isolates.

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