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From: Perl Molson on 19 Nov 2005 11:25
That is probably why,
"The HSV ICP47 inhibits CD8+ T cell recognition of HSV-infected cells
by blocking the function of the transporters associated with antigen
presentation (29) (30) (31). Our studies demonstrated that this protein
significantly increased HSV-1 neurovirulence in mice (32). However,
ICP47 is much less effective at blocking transporters associated with
antigen presentation function in mice than in humans (30) (33) (34).
Thus, mouse CD8+ T cells might be more responsive to HSV-1 antigens
than human CD8+ T cells, particularly in situations where both viral
protein and MHC class I antigen production are low. The increased
reactivity of mouse CD8+ T cells to HSV-1 antigens on sensory neurons
or surrounding cells might account for the inability of HSV to
spontaneously reactivate from latency in mouse sensory ganglia. "

http://www.jem.org/cgi/content/full/191/9/1459


furthermore, Prunella Vulgaris exibits such properties, overcoming the
gd protein similarity issue:

"Identification of Herpes Simplex Virus 1 Proteins Bound by the Novel
Anti-Herpes Prunella vulgaris Polysaccharide
G. DELANEY1, S. H. S. LEE1*, AND S. F. LEE1,2. 1Dept. of Microbiology &
Immunology, 2Dept. of Applied Oral Sciences, Dalhousie University,
Halifax, Nova Scotia.

Objective: We previously described a polysaccharide from the Chinese
herb Prunella vulgaris exhibiting activity against herpes simplex virus
1 and 2 (HSV-1 and 2). The polysaccharide apparently inhibited viral
binding as well as events post-viral binding and penetration. The
objective of this study is to identify the viral proteins bound by the
Prunella polysaccharide.

Methods: [35S]methionine-labeled proteins were obtained from
HSV-1-infected Vero cell lysates by freeze-thaw cycles and Noindet P-40
and sodium deoxycholate treatment. The proteins were applied to a
Prunella polysaccharide-Sepharose column. Bound proteins were eluted by
Prunella polysaccharide, immuno-precipitated by anti-gC and anti-gD
antibodies and analyzed by SDS-PAGE.

Results: Following chromatography on the Prunella polysaccharide
affinity column, two broad bands of 120-112 kDa and 69- 74 kDa were
obtained from the HSV-1 infected lysate; these protein bands were
absent from the mock infected lysate suggesting that the proteins were
of viral in origin. Similar proteins were obtained when
sucrose-gradient purified HSV-1 lysate was used. Two protein bands of
115 kDa and 74 kDa were recovered from the eluted proteins by
immuno-precipitation with the anti-gC antibody, while a 68 kDa protein
band was precipitated by the anti-gD antibody. The 115 kDa and 68 kDa
proteins corresponded to the reported size of gC and gD, respectively.
The nature of the 74 kDa is unclear but it could be a truncated version
of the gC or another HSV-1 protein which shared common antigenic
epitopes with gC. "
http://www.cacmid.ca/abstracts/a50.html


and then,
Recombinant Herpes Simplex Virus-2 gD
http://www.prospec.co.il/~prospec/cart/catalog/rHSV2_gD.html

furthermore:

Targeted delivery of DNA encoding herpes simplex virus type-1
glycoprotein D enhances the cellular response to primary viral
challenge.

Rogers JV, Bigley NJ, Chiou HC, Hull BE.

Biomedical Sciences Ph.D. Program, Wright State University, Dayton,
Ohio 45435, USA.

Intravenous injection of plasmid DNA encoding herpes simplex virus
type-1 glycoprotein D (gD-1) complexed with
asialoorosomucoid-poly-L-lysine (gD-ASOR) targets foreign DNA to the
liver, leading to hepatic expression of gD-1. BALB/c mice were given
two intravenous injections of gD-ASOR, pBK-ASOR (plasmid lacking the
gD-1 gene but complexed with ASOR), or PBS. The skin was inoculated
with 1 x 10(4) PFU of HSV-1 or sham-inoculated, and analyzed for
infectious virus and cellular infiltration 1, 3, and 5 days after
inoculation. Prior immunization with gD-ASOR led to significantly lower
(P < 0.05) viral titers in the skin 5 days after inoculation compared
with controls. Infiltration of the skin at the site of inoculation by
polymorphonuclear neutrophils (PMNs), T cells, B cells, dendritic
cells, and macrophages was monitored immunohistochemically.
Significantly higher numbers (P < 0.05) of CD4+ and CD8+ T cells,
dendritic cells, and macrophages responded to HSV-1 challenge in mice
immunized with gD-ASOR than in mice immunized with pBK-ASOR or PBS. The
response by PMNs and B cells was indistinguishable among the treatment
groups. These results suggest that BALB/c mice sensitized to gD-1
following gD-ASOR immunization develop an enhanced T-cell response to
primary HSV-1 infection.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11194892


free full article:
Identification of novel immunodominant CD4+ Th1-type T-cell peptide
epitopes from herpes simplex virus glycoprotein D that confer
protective immunity.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12915561&query_hl=2
The molecular characterization of the epitope repertoire on herpes
simplex virus (HSV) antigens would greatly expand our knowledge of HSV
immunity and improve immune interventions against herpesvirus
infections. HSV glycoprotein D (gD) is an immunodominant viral coat
protein and is considered an excellent vaccine candidate antigen. By
using the TEPITOPE prediction algorithm, we have identified and
characterized a total of 12 regions within the HSV type 1 (HSV-1) gD
bearing potential CD4(+) T-cell epitopes, each 27 to 34 amino acids in
length. Immunogenicity studies of the corresponding medium-sized
peptides confirmed all previously known gD epitopes and additionally
revealed four new immunodominant regions (gD(49-82), gD(146-179),
gD(228-257), and gD(332-358)), each containing naturally processed
epitopes. These epitopes elicited potent T-cell responses in mice of
diverse major histocompatibility complex backgrounds. Each of the four
new immunodominant peptide epitopes generated strong CD4(+) Th1 T cells
that were biologically active against HSV-1-infected bone
marrow-derived dendritic cells. Importantly, immunization of H-2(d)
mice with the four newly identified CD4(+) Th1 peptide epitopes but not
with four CD4(+) Th2 peptide epitopes induced a robust protective
immunity against lethal ocular HSV-1 challenge. These peptide epitopes
may prove to be important components of an effective immunoprophylactic
strategy against herpes.


A polysaccharide fraction from medicinal herb Prunella vulgaris
downregulates the expression of herpes simplex virus antigen in Vero
cells.

Chiu LC, Zhu W, Ooi VE.

Department of Biology, The Chinese University of Hong Kong, Shatin, New
Territories, Hong Kong, China. chimingchiu(a)graduate.hku.hk

Herpes simplex viruses (HSV) are pathogenic. With the emergence of
drug-resistant strains of HSV, new antiviral agents, especially those
with different modes of action, are urgently needed. Prunella vulgaris
L. (Labiatae), a perennial plant commonly found in China and Europe,
has long been used as a folk medicine to cure ailments. In this study,
a polysaccharide fraction was prepared from Prunella vulgaris (PPV),
and its effects on the expressions of HSV-1 and HSV-2 antigens in their
host Vero cells were investigated with flow cytometry. The HSV antigen
increased time-dependently in the infected cells, and PPV reduced its
expression. The effective concentrations of PPV with 50% reductions of
the HSV-1 and HSV-2 antigens were 20.6 and 20.1 microg/ml,
respectively. The novelty of PPV is that it also reduces the antigen
expression of acyclovir-resistant strain of HSV-1. After incubations
with 25-100 microg/ml of PPV the HSV antigen-positive cells were
reduced by 24.8-92.6%, respectively, showing that this polysaccharide
fraction has a different mode of anti-HSV action from acyclovir.
Results from this study show that PPV is effective against both the
HSV-1 and HSV-2 infections, and flow cytometry offers a quantitative
and highly reproducible anti-HSV drug-susceptibility assay.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15182906&query_hl=5

PMID: 15182906 [PubMed - indexed for MEDLINE]



http://intimm.oxfordjournals.org/cgi/content/abstract/11/11/1763
Protective immune correlates can segregate by vaccine type in a murine
herpes model system
Jeong-Im Sin, Velpandi Ayyavoo, Jean Boyer, Jong Kim, Richard B.
Ciccarelli and David B. Weiner

Department of Pathology and Laboratory Medicine, 505 Stellar-Chance
Lab, University of Pennsylvania, 422 Curie Drive, Philadelphia, PA
19104, USA
1 WLV, Malvern, PA 19355, USA


Correspondence to: D. B. Weiner

A central tenet of vaccine development is to identify immune correlates
of protection. Both plasmid-encoded gD as well as recombinant protein
gD can protect mice from lethal herpes simplex virus (HSV) challenge.
It is known that different vaccine modalities should induce different
immune phenotypes. Yet, paradoxically, it is also thought that the
basis for protection should rely on exploitation of vulnerabilities of
the pathogen and therefore that the overlapping properties of these
different vaccines would reveal insight into common immune mechanisms
responsible for protection. We sought to investigate this question by
comparing two different vaccine modalities in the HSV-2 mouse model. We
observed that gD protein was a strong inducer of Th2-type immune
responses, and overall antibody titers of IgG, IgE and IgA were
significantly higher than those induced by plasmid gD vaccines. In
contrast, the plasmid gD vaccine induced a strong Th1 bias. Following
high-dose challenge the gD protein was most effective at providing
protection. However, at lower lethal dose challenge, while both
vaccines were protective with regards to survival, only the
plasmid-vaccinated animals were protected from HSV-2 infection-induced
morbidity. These studies suggest that these different vaccine
modalities induce protection through unique non-overlapping mechanisms,
supporting that vaccine correlates are associated with the types of
immunogen rather than solely the pathogen.



http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11449800&dopt=Citation
[Development and preparation of recombinant gD antigen of the herpes
simplex type 1 (HSV-1) virus]

[Article in Russian]

Susloparov IM, Pliasunov IV, Safronov PF, Bakhtina MM, Lokteva LA,
Grishaev MP, Susloparov MA.

The most potent antigen among HSV-1 proteins are glycoproteins gB(UL27)
and gD(US6). Multiple amino acid sequence alignment of these proteins
shows that gD protein is the most specific for HSV-1. Analysis of gD
protein epitopes detected the main antigenic determinants not
cross-reactive with antigens of other viruses. Virus was isolated and
genome DNA was prepared from morphological elements of a patient with
herpes simplex infection. US6 gene fragment was cloned in pUC19 vector.
Cloning in bacterial expression vectors helped obtain
beta-galactosidase-fused recombinant HSV-1 gD protein with 6-histidines
affine target for high-performance chromatography purification. ELISA
with a set of HSV-1-positive and negative donor sera and a commercial
panel of HSV-1 sera (Vektor-Best) showed that recombinant gD can be
used as an antigen to HSV-1-specific IgG.

free full text at:

http://jvi.asm.org/cgi/content/full/79/7/4540?view=long&pmid=15767456
The results reported here are consistent with and reconcile each of
these previous findings. Formation of a HVEM/gD/gH complex may require
the presence of a cellular receptor. Therefore, hetero-complexes would
not be found when purified virions are examined by electron microscopy.
Detected interactions of gD and the gH/gL heterodimer with each other
or with their respective cell receptors could be transient during entry
or could form after de novo protein synthesis in infected cells. The
interactions are consistent with what is known about viral glycoprotein
and receptor requirements for HSV infection and with models of
predicted events in its pH-independent entry, spread, or viral-induced
membrane fusion (1, 3, 10, 11, 21, 29). We detected associations of gD
and gH in infected human cells and also in infected HVEM-expressing
porcine cells where there is at least one human cellular receptor for
HSV. The interaction of gD and at least gH in a receptor complex is
consistent with predicted functions of HSV glycoproteins that are
required to mediate events of membrane fusion at virus spread or entry

http://www.skinchoice.com/herpasil.htm Herpasil (5% Prunella Topical
Balm)


http://www.uq.edu.au/vdu/HSV.htm


Over 100 different herpesviruses have been described in various animal
species. Herpesviruses are readily identified by the distinctive
architecture of the virus virion. Their are four major structural
components of the virus:
An electron dense core harboring the dsDNA viral genome
A protein capsid surrounding the virus core, the capsid is comprised
of 162 capsomeres.
An amorphous layer surrounding the capsid termed the tegument.
An envelope (lipid bilayer) containing spikes that probably represent
viral glycoproteins.
The herpesviruses are distinguished by their biological properties: a)
they encode many enzymes involved in nucleic acid metabolism, b) their
replication and assembly occur in the nucleus, c) the cell is killed
(lysed) as an outcome of virus infection, d) they have the capacity to
enter a latent state in which only a small subset of the viral gene
complement is expressed.


The herpesviruses have been classified into three subfamilies.


Alphaherpesvirinae


Are characterized by a broad host range and are highly lytic in
culture.


Betaherpesvirinae


Have a restricted host range, grow more slowly in culture and cells
infected with this subclass often show enlargement.


Gammaherpesvirinae


Infect lymphoblastoid cells.


Human Herpesviruses


At least six human herpesviruses have been described. These include:
Herpes simplex virus type 1 (HSV-1), Herpes simplex virus type two
(HSV-2), Varicella zoster virus (VZV), Cytomegalovirus (CMV),
Epstein-Barr virus (EBV), and Human herpesvirus six (HHV - 6). This
handout will focus on HSV-1 and HSV-2. These two viruses share
extensive nucleic acid sequence homology (50%) and for the purposes of
this discussion can be considered together.


Herpes simplex virus diseases


Primary infection


Infection of individuals who have not been previously exposed to HSV.


Recurrent infection


Reactivation of latent virus that results in a second infection


Initial infection


Infection of an individual with HSV-1 who was previously exposed to
HSV-2 (or the reverse)


Exogenous reinfection


Infection of an individual with the a second strain of the same HSV
type.


Herpes labialis


(orolabial herpes). Usually occurs in children under five years of
age, the primary infection usually presents as a gingivostomatitis.
Pharyngitis may also be present. Predominantly associated with HSV-1.


Genital herpes


Sexually transmitted disease associated mainly with HSV-2.


Neonatal infection


Life threatening infection by HSV usually acquired by child during
birth. Systemic infection.


Keratoconjunctivitis


HSV infection of conjunctiva and eye proper which can lead to
blindness.


Skin infections


Usually acquired by health care workers or researchers by direct
inoculation of virus from contaminated materials to a cut on the hand
(whitlow).


CNS infections


Herpes encephalitis is a rare but devastating infection of the brain
with high mortality and morbidity.


The virion


HSV virions contain over 30 proteins (virion polypeptides, VPs)
including eight glycoproteins (gB, gC, gD, gE, gG, gH and gI) some of
which are components of the envelope spikes. The tegument contains at
least two proteins of known function: àTIF (alpha trans-inducing
factor, also known as VP16 and vmw65) and VHS (virion host shut off).


Viral DNA


The viral genome is 150 kbp in size and contains single stranded nicks
and gaps. It consists of two components, a long and short region
flanked by inverted repeats. The structure can be written like this
ab-UL-b'a'c'-Us-ca. The "a" sequence is highly conserved and consists
of variable numbers of repeat elements. The long and short components
can invert relative to each other yielding four linear isomers of the
viral genome. The importance of inversion of viral genomes is
uncertain in that mutants which do not invert grow normally in
culture.


Virus Growth
Attachment and penetration


Five of eight viral glycoproteins are dispensable for virus growth in
culture (gC, gE, gG, gI, gJ). Three glycoproteins (gB, gD, and gH) are
essential and represent the minimal set of surface proteins necessary
to sustain and carry out the dominant flow of events. Heparin sulfate
proteoglycans appear to be the receptor molecules which are recognized
by either gB or gC and which permit initial attachment of the virus.
gB and gD are essential for virus penetration. Penetration occurs by
direct fusion of the viral envelope with the cell membrane. Virions
which attach to the plasma membrane which cannot fuse are internalized
and degraded in endocytotic vesicles. Capsids are transported by the
cellular cytoskeleton to nuclear pores and viral DNA is released into
the nucleus where it accumulates. Virion components mediate the shut
off of host macromolecular synthesis and àTIF acts to induce initial
gene expression. HSV has the potential to encode at least 70
polypeptides but detectable HSV polypeptides do not exceed 50.


Gene expression


HSV transcription and protein synthesis is highly ordered. Although
the absolute levels of viral protein synthesis may vary, different
genes can be grouped on the basis of their requirements for synthesis.
Hence, HSV genes have been subdivided into 3 broad groups based on
their time and requirements for expression (alpha, beta and gamma).


Alpha genes


There are five alpha genes which have been identified and described as
ICPs (infected cell proteins), these include ICP0, ICP4, ICP22, ICP27
and ICP47. The à genes are by definition expressed in the absence of
viral protein synthesis and contain the sequence
GyATGnTAATGArATTCyTTGnGGG upstream of their coding regions. Their peak
synthesis occurs 2-4 hours post infection, but they continue to
accumulate until late in infection. All alpha genes appear to function
as regulatory proteins with the possible exception of ICP47.


Beta genes


These genes are not expressed in the absence of alpha proteins and
their expression is enhanced in the presence of drugs which block DNA
synthesis. They reach peak rates of synthesis 5-7 hr post infection.
The genes have been subdivided into the beta 1 and beta 2 subclasses.
beta 1 genes appear early after infection, but require the presence of
à 4 protein for their synthesis. Examples of beta 1 genes include the
large component of ribonucleotide reductase and the major DNA binding
protein (ICP8). beta 2 genes include viral thymidine kinase (TK) and
the viral DNA polymerase. beta gene synthesis immediately precedes the
onset of viral DNA synthesis and most viral genes involved in viral
nucleic acid metabolism appear to be beta genes.


Gamma genes


This class of genes is for convenience also separated into two groups.
gamma 1 genes are expressed early in infection and are only minimally
affected by inhibitors of DNA synthesis (example, major capsid
protein). In contrast, gamma 2 genes are expressed late in infection
and are not expressed in the presence of inhibitors of viral DNA
synthesis.


The location of the gene classes within the HSV genome is of interest.
alpha genes map at the termini of the long and short components and
tend to cluster together. In particular, alpha genes surround the HSV
origin of replication in the short region (oris). Each alpha gene has
its own promoter-regulatory region and transcription initiation and
termination sites. beta and gamma genes are scattered in both the long
and short components. Interestingly, the beta genes specifying the DNA
polymerase and the DNA binding protein flank the origin of replication
in the long region (oriL). There is little gene overlap and few
instances of gene splicing for any of the HSV gene classes.


Essential and nonessential genes


Large numbers of viral mutants have been generated and have led to
identification of genes that are essential or nonessential for HSV
growth in tissue culture. essential: gB, gD, major DNA binding protein
(ICP8), alpha 27 and alpha 4. nonessential: all genes in the unique
short region (except for gD), dUTPase, gC, alkaline DNAse, thymidine
kinase, ribonucleotide reductase, uracil DNA glycosylase.


Synthesis of viral DNA


HSV specifies a large number of enzymes involved in viral DNA
synthesis. Viral DNA synthesis begins 3 hrs postinfection and
continues for 9-12 hrs. DNA replication occurs in the nucleus and
evidence suggests that late in infection HSV DNA replicates by the
rolling circle mechanism.


Origins of replication


Three origins of replication have been identified within the HSV
genome. One origin is present in each "c" component of the short
region and one origin is present in the unique long region between the
major DNA binding protein (ICP8) and the DNA polymerase. oriL has A/T
rich sequences with a near perfect palindrome. oriL and one copy of
oris can be deleted without affecting the ability of the virus to
multiply. Both origins are situated between transcriptional initiation
sites.


Functional requirements for viral replication.


Proteins essential for viral origin dependent amplification
Enzymes involved in nucleic acid metabolism (thymidine kinase,
ribonucleotide reductase, dUTPase, uracil DNA glycosylase, alkaline
exonuclease.
Using the Challberg assay seven genes have been identified which are
necessary for origin dependent replication: viral DNA polymerase, ICP8
(single stranded DNA binding protein), origin binding protein, dsDNA
binding protein ,and three other proteins which may be involved in
primase and helicase activities.


The importance of the virally encoded enzymes in HSV replication is
detailed below.


Alkaline DNase: essential for viral growth and DNA replication
Thymidine kinase (TK): broad substrate specificity
Ribonucleotide reductase: reduces ribonucleotides to
deoxyribonucleotides creating a pool of substrates for DNA synthesis.
It is comprised of two subunits 140kd and 38kd . Both subunits are
required for activity.
Uracil DNA glycosylase: involved in DNA repair and proof reading,
corrects insertion of dUTP into DNA.
dUTPase: converts dUTP to dUMP preventing dUTP incorporation into DNA
and providing a pool of dUMP for conversion to dTMP.
Capsid assembly occurs in the nucleus. Cleavage and packaging of HSV
DNA are probably linked processes. The packaged DNA can be defined by
the distance between two directly repeated "a" sequences. Evidence
suggests that a head full mechanism may also operate once a minimal
amount of DNA has been packaged. Virus matures at nuclear membranes at
"patches" where cellular proteins have been displaced by viral
glycoproteins. How glycoproteins are targeted to and enter the nuclear
membranes is unclear.


Regulation of viral gene expression


RNA polymerase II is responsible for viral mRNA synthesis. In general
viral mRNAs are capped and polyadenylated just like cellular mRNAs.
Only a small portion are spliced. DNA sequences upstream of HSV genes
determine their capacity to be expressed as alpha or beta genes. alpha
gene expression is directed by alpha-TIF (VP16, vmw65) which complexes
with cellular proteins to bind a TAATGArATT motif. Despite controversy
over the ability of ICP4 to bind DNA their appears to be support for
specific ICP4 binding sites upstream of the TK gene (beta gene) and of
ICP4 itself. In general expression of HSV genes appear to be
controlled by three means:(a) cis-acting sites for both viral
transacting factors and cellular factors. (b) trans-acting signal
proteins specified by the virus and (c) viral and cellular factors
involved in viral DNA synthesis and post-translational modification of
viral proteins.


Thus far there are three "top guns" for viral gene regulation.


Alpha-TIF: alpha trans-inducing factor (also known as VP16 and vmw65)
is a structural component of the HSV virion (located in the tegument).
In conjunction with cellular proteins alpha-TIF binds the TAATGArATT
motif upstream of alpha genes and induces their transcription.
alpha-TIF does not bind DNA directly and there are about 500-1000
copies of alpha-TIF per virus particle.


ICP0: also known as IE110, it is an alpha gene which can promiscuously
transactivate transfected genes. Its function in infected cells is not
known. ICP0 deletions mutants are still viable in cell culture.


ICP4: (also known as IE175) This is the big guy. It is the major
transactivator of HSV genes and is an essential gene product. ICP4 is
autoregulatory and probably turns off its own synthesis and ICP0
synthesis as well. ICP4 functions as both a transactivator and a
repressor and may be regulated by post-translational modifications,
position of the DNA binding site or strength of binding to DNA.



My conclusion?

It seems that, indeed, Prunella Vulgaris is a cure-like treatment for
Herpes Simplex Virus

Folks, drink lots of tea (infussion) with Prunella Vulgaris (Self
Heal) and you will forget you have herpes.
Perl von Molson

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