Does HHV-6 Come From Pigs?
What about HHV-7 and HHV-8?
Is Porcine Lymphotropic Herpes Virus or Porcine CMV the HHV-6, 7 and 8 of Pigs? Or is HHV-6 actually the Human Adaptation of African Swine Fever Virus? Does HHV-6 act like African Swine Fever Virus? Are countries with ASFV epidemics at risk for ASFV infections in people? Should people be monitored for ASFV in countries with ASFV epidemics? Is there a higher incidence of HHV-8 in countries with African Swine Fever Virus? Is there a high incidence of HHV-8 triggered Kaposi's Sarcoma in countries with epidemics of African Swine Fever Virus?
Does SNL Know Something We don't?
Is Kaposi's Sarcoma in AIDS patients caused by a human form of ASFV? Are HHV-6, HHV-7 and HHV-8 related to African Swine Fever Virus? Should they be reclassified as Asfaviridae?
Are pigs the unrecognized reservoir of HHV-6 (or ASFV) that has been linked to AIDS, CFS and autism?
Breaking News:
African Swine Fever may cause human infections! Does this help support the notion that ASFV may be the real underlying cause of AIDS and Chronic Fatigue Syndrome and that HHV-6 is actually a novel form of ASFV?
Detection of Novel Sequences Related to African Swine Fever Virus in Human Serum and Sewage.
Loh J, Zhao G, Presti RM, Holtz LR, Finkbeiner SR, Droit L, Villasana Z, Todd C, Pipas JM, Calgua B, Girones R, Wang D, Virgin HW.
Departments of Pathology & Immunology and Molecular Microbiology, Department of Medicine and Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri; Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology, Faculty of Biology, University of Barcelona, Barcelona, Spain.
"The family Asfarviridae contains only a single virus species, African swine fever virus (ASFV). ASFV is a viral agent with significant economic impact due to its devastating effects on populations of domesticated pigs during outbreaks, but has not been reported to infect humans. We report here the discovery of novel viral sequences in human serum and sewage which are clearly related to the Asfarvirus family, but highly divergent from ASFV. Detection of these sequences suggests that greater genetic diversity may exist among Asfarviruses than previously thought, and raises the possibility that human infection by Asfarviruses may occur."
"HHV-6 and ASFV are very similar viruses; they are the same size, and they infect and kill the same kinds of cells. They both cause bleeding problems (they are "hemorrhagic" viruses), and both attack the immune system, allowing secondary (or "opportunistic") infections to become fatal. They both cause neurological disease. Both viruses mutate (change) easily. Furthermore, both HHV-6 and ASFV are characterized by having many strains, or types, of virus. ASFV, it has been known for dec ades, can cause a whole range of illness, from extremely acute (in which 100 percent of infected animals die within days) to chronic (in which animals develop immune system and neurological symptoms, but generally survive).HHV-6, also, has many strains that can cause different levels of disease, from an illness with mild fever and rash, to a fatal, hemorrhagic disease in which the patient rapidly bleeds to death. HHV-6 can cause a collapse of the immune system, and is associated with many kinds of neurological phenomena, including the development of brain lesions. Like ASFV, HHV-6 is found in people with acute illness -- that is, AIDS -- and chronic illness, CFS."
America's Biggest Cover-Up by Neenyah Ostrom
The controversial AIDS doctor, Matthias Rath has a patent on African Swine Fever Virus:
Polypeptides from African Swine Fever virus as vaccines for preventive and therapeutic use
United States Patent Application 20080131449
The present invention generally relates to the use of selected polypeptides from African Swine Fever virus for the prevention and therapy of African Swine Fever infections as well as other infections, including immune deficiencies in mammals and humans.
FIELD OF THE INVENTION
The present invention generally relates to the use of selected polypeptides from African Swine Fever virus for the prevention and therapy of African Swine Fever infections as well as other infections, including immune deficiencies in mammals and humans. BACKGROUND OF THE INVENTION
African Swine Fever is an endemic disease in sub-Saharan Africa and many other parts of the developing world. It is caused by the African Swine Fever virus that primarily replicates in macrophages and monocytes leading to the impairment of the structure and function of the immune system of the infected organisms. Until now the African Swine epidemic continues to spread despite all efforts to contain it. Thus, there is an objective need for effective, safe and affordable preventive and therapeutic approaches, in particular for effective vaccines, to control and eventually eradicate this disease.
Since the characteristic feature of the African Swine Fever virus is to impair the immune system and to cause immune deficiencies in its hosts the development of vaccines and other therapeutic approaches against the African Swine Fever virus has implications for other immune deficiencies or diseases. Several other viruses are also known to cause immunodeficiency-like syndromes in humans, including cytomegalovirus, Epstein Barr Virus, HIV and others. Moreover, a series of cases of so-called “idiopathic” immunodeficiencies have been documented that display CD4+ T-lymphocytopenia with opportunistic infections, but show no evidence of HIV infection (1).
Since antibodies for the African Swine Fever virus have been detected in humans, the possibility of human infection with the African Swine Fever virus exists and may thus far have escaped any systematic screening. Thus, any preventive and therapeutic approach to African Swine Fever can have far-reaching implications to control immune deficiency conditions in humans. SUMMARY OF THE INVENTION
Synthetic oligopeptides prepared from African Swine Fever virus proteins are effective in prevention, treatment and diagnosis of African Swine fever as well as for immune deficiencies in humans.
Oligopeptides are identified and selected by means of suitable algorithms from the known amino acid sequence of pathogenicity-mediating African Swine Fever virus proteins. Subsequently, these oligopeptides are tested in vitro for their ability to decrease or completely block infection by the African Swine Fever virus (prevention and therapy) or for their ability to raise antibodies to detect the virus (diagnosis). Ultimately, the successfully tested African Swine Fever virus oligopeptides can be used in veterinary and clinical medicine. BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 shows the antigenicity scores derived from and according to the Hopp-Woods hydrophylicity algorithm for the protein p54, a 183 amino acid long structural protein of the African Swine Fever virus. Since p54 is involved in the pathogenesis of African Swine fever (2), interrupting this pathogenicity-mediating pathway will lead to a decrease or a complete block of infection by this virus.
The relative peaks of this algorithm, defined as amino acid sequence regions of either high hydrophilic characteristics or sequence regions of higher hydrophilic characteristics in relation to adjacent amino acid sequences or in comparison to hydrophobic regions of the protein, represent the likely candidate sequence regions (oligopeptides) serving as epitopes (antigens) for antibody formation. Moreover these defined oligopeptide sequences represent the likely region by which a protein interacts with other proteins and/or biological compounds in an organism, including those interactions that mediate infection or other forms of pathogenicity.
Producing synthetic oligopeptides, corresponding to these algorithm maxima allows the development of preventive and therapeutic agents to control African Swine Fever virus infections. The relative peaks of the Hopp-Woods algorithm for the p54 protein of the African Swine Fever virus selected for this patent application are marked by arrows. The synthetic oligopeptides for the other African Swine Fever Virus proteins specified in this disclosure and the claims are selected in an analogous way. DETAILED DESCRIPTION OF THE INVENTION
The African Swine Fever virus is a particular virus the pathogenicity of which is largely determined by targeting the immune system of the host and disabling it.
Despite the DNA sequence of African Swine Fever virus having been determined (3), there is currently no effective vaccine available to control African Swine Fever as documented in the United Nations Food and Agricultural Organization's field handbook on this disease (4)
The present invention describes the identification and production of preventive and therapeutic agents, which—among others—can be used as vaccines against African Swine fever with the following specific steps being taken:
*
1. The identification of structural proteins and/or pathogenicity-mediating proteins and/or any other protein from the African Swine virus. 2. The analysis of the amino acid sequence of these proteins using specific algorithms allowing the determination of relative hydrophilic and/or polarity and/or charge and/or surface probability peaks and/or any other method allowing the determination of potential epitopes within these African Swine fever virus proteins. 3. The production of synthetic oligopeptides analogous to the epitope forming oligopeptides identified within the amino acid sequence of the African Swine fever virus proteins. 4. The modification of these synthetic oligopeptides to allow or improve antigencity and the formation of antibodies and/or to block pathogenicity of the African Swine fever virus in any other way by + a. adding one or several predetermined amino acids to the selected oligopeptide sequence; + b. subtracting one or several predetermined amino acids to the selected oligopeptide sequence; + c. replacing one or several predetermined amino acids within the selected oligopeptide sequence; + d. changing the linear topology of the selected oligopeptide to a cyclic topology; + e. forming a linear chain of covalently bound repeats of the selected oligopeptide sequence; + f. forming a cyclic chain of covalently bound repeats of the selected oligopeptide sequence; + g. coupling an originally selected and/or modified oligopeptide to one or more haptens; + h. to improve antigencity and enhance antibody formation in any other possible way; + i. producing natural and/or synthetic peptidomimetics mimicking the three dimensional srtructure of the natural or modified oligopeptide. 5. To conduct in vitro and in vivo tests with the selected oligopeptides and/or peptidomimetics in order to establish their efficacy and efficiency as a therapeutic or diagnostic agents. 6. To identify those originally selected and/or modified synthetic oligopeptides and/or peptidomimetics for therapeutic or diagnostic use that + a. display maximum and/or optimum ability to form antibodies against the African Swine Fever virus as potential therapeutic vaccines; + b. display maximum and/or optimum competitive inhibition of pathogenicity mediating pathways of the African Swine Fever virus as potential therapeutic agents used for—but not limited to—acute therapeutic treatment of African Swine fever; + c. display maximum and/or optimum antigenicity to raise antibodies for the development of tests to diagnose African Swine fever. 7. To use those originally selected and/or modified synthetic oligopeptides and/or peptidomimetics as therapeutic vaccines that display maximum and/or optimum ability to form antibodies against the African Swine virus. 8. To use those originally selected and/or modified synthetic oligopeptides and/or peptidomimetics as therapeutic vaccines that display maximum and/or optimum ability for competitive inhibition of pathogenicity mediating pathways of the African Swine Fever virus as potential therapeutic agents used for—but not limited to—the acute therapeutic treatment of African Swine fever. 9. To use those originally selected and/or modified synthetic oligopeptides and/or peptidomimetics that display maximum and/or optimum antigenicity to be used in the development of diagnostic tests or screening procedures for the African Swine virus.
The current invention also describes the application of the current invention for the diagnosis and treatment of immune deficiency conditions in mammals.
The following are the characteristics of the African Swine Virus:
- it targets the immune system of the host
- it has the following morphological features, in particular it structurally and functionally impairs the
- its hematological changes includes a significant decrease of CD4 and T-cell counts;
- it has clinical manifestations namely lymph node swelling, increased susceptibility to infections, and others;
- it has both an acute and chronic form of infectious states;
- it is known to display a high frequency of alteration of their genetic sequence in order to escape the host defense system;
- it is endemic in sub-Saharan Africa and few other regions.
While the African Swine Fever virus has been primarily detected in pigs and certain other animals, antibodies against the African Swine Fever virus have also been found in humans (5). The fact that there was no description of any finding of the African Swine Fever virus in humans may thus be attributable to oversight or a lack of understanding for the significance of African Swine fever virus for the pathogenicity of immune deficiencies in humans.
Thus, the inventions described in this patent application can have far reaching implications not only for the control of African Swine fever but also for the control of other immunodeficiency diseases.
Characterization of the DNA Polymerase Loci of Porcine Cytomegaloviruses from Diverse Geographic Origins
Journal Virus Genes Publisher Springer Netherlands ISSN 0920-8569 (Print) 1572-994X (Online) Issue Volume 21, Number 3 / October, 2000 http://www.springerlink.com/content/h681vq7567258755/
Michael Goltz1, Frederik Widen2, Malcolm Banks2, Sandor Belak3 and Bernhard Ehlers1 (1) Robert Koch-Institut, Nordufer 20, 13353 Berlin, Germany (2) Veterinary Laboratories Agency, New Haw, Addlestone, United Kingdom, KT15 3NB (3) The National Veterinary Institute, Department of Virology, Biomedical Center, Box 585, SE-751 23 Uppsala, Sweden
Abstract Porcine cytomegalovirus (PCMV) is an undesired pathogen in pigs intended for use as organ donors in xenotransplantation. In the present work, we characterized the first set of genes of PCMV. From a German isolate, the DNA polymerase (DPOL) locus was amplified and two complete open reading frames (ORF) as well as two partial ORFs including the complete DPOL gene and the 3prime-end of the glycoprotein gB gene were sequenced. The deduced amino acid sequences showed the highest identities with the respective proteins of the betaherpesviruses, in particular those (ORFs 36–39) of the human herpesviruses 6 and 7 (HHV-6 and -7). In phylogenetic analysis, PCMV clustered also with HHV-6 and HHV-7. On this basis, PCMV could be firmly classified to the Betaherpesvirinae and tentatively assigned to the genus Roseolovirus. In addition to the German isolate, the DPOL gene was analysed from a British and a Japanese strain as well as a Spanish isolate. Differences of 0.4 to 1% were found on the nucleotide and the amino acid level. On the basis of the conserved regions, primer pairs were selected for PCR which detected PCMV in blood and tissue samples from four European countries. Therefore, these are the first nucleic acid-based test systems which were shown to universally detect PCMV. The application of these assays to organs of domestic pigs from Germany revealed a PCMV prevalence of >50%.
Characterization of the DNA polymerase loci of the novel porcine lymphotropic herpesviruses 1 and 2 in domestic and feral pigs
Sven Ulrich1, Michael Goltz1 and Bernhard Ehlers1
Robert Koch-Institut, Nordufer 20, 13353 Berlin, Germany1
Author for correspondence: Bernhard Ehlers. Fax +49 30 4547 2598. e-mail ehlersb@rki.de
Two novel porcine gammaherpesviruses, porcine lymphotropic herpesviruses 1 and 2 (PLHV-1 and -2), have been detected by amplification of short DNA polymerase (DPOL) sequences from blood and spleen of domestic pigs while searching for unknown herpesviruses in pigs as possible risk factors in xenotransplantation. In the present study, the DPOL genes of the two viruses and the open reading frames (ORFs) that follow in the downstream direction were amplified by PCR-based genome walking from adaptor-ligated restriction fragment libraries of porcine spleen samples. The sequences determined for the two PLHVs exhibited a very low G+C content (37 mol%) and a marked suppression of the CpG dinucleotide frequency. The DPOL proteins encoded were 95% identical and showed a close relationship (60% identity) to the DPOL protein of a ruminant gammaherpesvirus, alcelaphine herpesvirus 1 (AlHV-1). This was confirmed by phylogenetic analyses of the conserved regions of the two PLHV DPOL proteins. The PLHV ORFs downstream of DPOL exhibited 83% identity to each other and >>50% similarity to ORF A5, the position equivalent of AlHV-1. From these data, the PLHVs can be firmly classified to the subfamily Gammaherpesvirinae. To find a natural reservoir for the PLHVs, organs of feral pigs were screened with five different PCR assays, targetting either the DPOL gene or 3'-flanking sequences. In all samples, PLHV sequences were detected that originated predominantly from PLHV-2, suggesting the possibility of virus transfer between feral and domestic pig populations. http://vir.sgmjournals.org/cgi/content/abstract/80/12/3199
Early weaning of piglets fails to exclude porcine lymphotropic herpesvirus.
Mueller NJ, Kuwaki K, Knosalla C, Dor FJ, Gollackner B, Wilkinson RA, Arn S, Sachs DH, Cooper DK, Fishman JA.
Infectious Diseases Division, Massachusetts General Hospital, Harvard Medical School, Massachusetts General Hospital, Boston, MA 02114, USA.
BACKGROUND: Xenotransplantation using pigs as source species carries a risk for the activation of latent herpesviruses from the porcine donor and potential transmission to the recipient. In pig-to-baboon xenotransplantation, activation of porcine cytomegalovirus (PCMV) has been associated with xenograft injury and an increased incidence of consumptive coagulopathy and graft loss. Activation of porcine lymphotropic herpesvirus (PLHV)-1 was not observed in pig-to-baboon solid organ xenotransplantation, but was associated with a syndrome of post-transplantation lymphoproliferative disorder (PTLD) after allogeneic stem cell transplantation in pigs. MATERIAL and METHODS: Early weaning of piglets was used to try to reduce the viral burden of xenograft donors. This consisted of separating the piglets of a litter from the sow within the first 2 weeks after birth and raising them in isolation from the remaining herd. RESULTS: We have previously demonstrated that PCMV could be excluded from source animals by early weaning of piglets. However, early weaning failed to exclude PLHV-1 from source pigs. CONCLUSIONS: This disparity between PCMV and PLHV-1 reflects differing pathogenesis of infection of these herpesviruses. New approaches will be needed to exclude PLHV-1 from pig colonies. http://www.ncbi.nlm.nih.gov/pubmed/15598274?dopt=Abstract
Porcine lymphotropic herpesviruses 1 and 2
Herpesviruses are widely distributed and have been found in insects, reptiles, amphibians and every species of bird and mammal. One important characteristic of herpesvirus infection is that the virus persists in the infected host for life and is frequently reactivated and shed. In pigs, five herpesviruses have been identified: pseudorabies virus, porcine cytomegalovirus (PCMV) and three recently identified lymphotrophic herpesviruses, PLHV-1, PLHV-2 and PLHV-3.
PLHV-1 and -2 are highly homologous to each other but not to PLHV-3. The two viruses are widespread in domestic pigs and are closely related to several ruminant gammaherpesviruses, most of which are etiologically implicated in the occurrence of malignant catarrhal fever (MCF), a lymphoproliferative inflammatory disease with an often fatal outcome. PLHV-1 and -2 are also related to Epstein-Barr Virus, Human Herpesvirus-8 and other gammaherpesviruses.
A recent study (McMahon et al., 2006) in domestic pigs has shown that PLHV1 infections are most common, being found in 74% of animals tested, followed by PLHV3 at 45% and PLHV2 at 21%. Infections with multiple PLHV species were frequently detected.
Like porcine CMV, antibodies to PLHV have been found in a high percentage of swine herds worldwide. Because of the high prevalence of positive serology, serological identification of infected pigs is not possible. Many of these latent carriers remain unidentified, posing serious problems for research using the pig as a model. In xenotransplantation between pig and human, reactivation of these latent viruses can cause postransplantation failure. Molecular detection of these viruses is an important tool to provide rapid, sensitive and specific detection of the viral nucleic acid in suspected animals. http://www.zoologix.com/zoo/Datasheets/PorcineLymphotropicHerpesvirus.htm
These links provide background on ASFV and its possible relationship with HHV-6, AIDS and Chronic Fatigue Syndrome. Needless to say, not all of it is accurate.
http://www.blackherbals.com/a_virus_that_causes_an_aids_in_pigs.htm
Excerpt from a very important article on AIDS, HHV-6 and ASFV
BELDEKAS MEETS GALLO AT NCI AND PRESENTS HIS FINDINGS.
In August, 1986, John Beldekas was invited to go to the NCI and present his findings on the link between ASFV and AIDS, which he did. Beldekas gave samples of all his lab work to Gallo. Later, the government asked Beldekas to turn over all his reagents and lab work to the government, which he did. Beldekas had found ASFV presence in nine of 21 AIDS patients using two standard procedures. At the meeting, Gallo was reported saying: "we know it is not ASFV." How could Gallo know this as he hadn’t done any of his own tests to look for ASFV?
Two months later, Gallo published an article in Science (Oct 31, 1986) that he discovered a new possible co-factor in AIDS, a virus he called Human B Cell Lymphotropic Virus which he named HBLV. Like ASFV, HBLV infected B cells and also lived in macrophages. Did Gallo steal Beldekas’s ASF virus he found in AIDS patients and rename it HBLV? Later on, when Gallo found that HBLV could also infect other immune cells, he changed the name of HBLV to HHV-6. Eventually, Gallo identified his HBLV as the variant A strain of HHV-6 and called it a human herpesvirus.
http://www.keephopealive.org/report10.html
http://www.nytimes.com/1986/03/07/us/link-to-aids-is-seen-in-virus-affecting-pigs.html
http://www.fms-help.com/aids.htm
http://www.nytimes.com/1986/05/28/us/us-to-test-florida-herd-for-swine-fever.html?&pagewanted=all
http://www.nytimes.com/1986/09/23/science/no-swine-fever-link-to-aids-seen.html?&pagewanted=all
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