Is HPV Pathogenic?

In virology, in order to determine whether particles believed to be “viruses” are actually pathogenic, virologists must demonstrate pathogenicity in a suitable host in order to satisfy the last two of Koch’s four Postulates proving a microbe causes disease:

3. The cultured microorganism should cause disease when introduced into a healthy organism.

4. The microorganism must be reisolated from the inoculated, diseased experimental host and identified as being identical to the original specific causative agent.

These steps are performed through shockingly cruel animal models of experimentation which many times involves altering the animals in some way and using invasive injections of diseased animal tissues or toxic cultured materials into their skin, veins, stomach, nasal cavities, eyes, and/or brain to make the animal sick in some way. These grotesque forms of torture are utilized due to the “ethical” concerns regarding experimenting on human Guinea pigs (please disregard the human Guinea pig experiments currently going on with the “Covid” jabs). Granted, the ethical concerns are the go-to excuse to cover up for the fact that actual human-to-human pathogenicity studies using natural routes of “infection” failed repeatedly in the past. It turns out that it is much easier to get away with torturing and producing experimental disease in animals than using humans. “Ethics.”

Normally, virologists will go through trial and error until they find the perfect animal host which, upon injection with unpurified toxic cell culture soup said to contain the “virus,” sometimes comes down with something that kinda sorta somewhat resembles the human disease if looked at by tilting your head at the right angle. This is how they “prove” pathogenicity and then study how the “virus” works on these animal hosts.

With HPV, however, there are a few problems. For one, unlike other “viruses,” virologists simply can not grow it in the cell cultures they normally use, thus they can not shoot up the helpless animals with cultured HPV goo to determine pathogenicity. The other problem is that even if they could grow HPV in culture, it is supposedly a “virus” that only infects humans, thus they can not infect these animals with it either. Other issues include that it supposedly takes 15-20 years after infection for HPV (along with other co-factors) to cause cervical cancer as well as the fact that virologists can not find enough “virus” in humans to study it.  Since the discovery of the fragments of DNA (not “viral” particles) claimed to be HPV, virologists have had no way of testing “real HPV.”

Unfortunately for the animals (and eventually the vaccinated humans) involved, virologists came up with some, shall we say, “creative” ways to get around these problems. Below are highlights from a 2009 paper that details virology’s “solutions” but in order to fully grasp what these virologists did, a few definitions may help:

1. “VIRUS-LIKE” PARTICLES: molecules that closely resemble “viruses,” but are non-infectious because they contain no “viral” genetic material (i.e. not “real viruses”)
2. PSEUDOVIRUSES: synthetic chimeras that consist of a surrogate “viral” core, derived from a parent “virus,” and an envelope glycoprotein on its surface derived from a heterologous “virus” (i.e. not “real virus”)
3. TRANSGENIC MICE: a genetically modified mouse or genetically engineered mouse model; a mouse that has had its genome altered through the use of genetic engineering techniques (i.e. not a “real” mouse)

Without further ado, I present their “solutions:”

Animal models for human papillomavirus-associated cervical pathogenesis

“Human papillomavirus (HPV) infection is the second leading cause of cancer-associated morbidity and mortality among women worldwide due to its prerequisite role in cervical carcinogenesis. HPV is strictly species and tissue-restricted and cannot be propagated in vitro, which has retarded the development of in-vivo models for HPV infection. Much of our understanding on HPV, its life cycle, its oncogenicity and its synergy with co-factors was first established in vitro or by animal papillomaviruses in vivo. However, key differences between such systems and natural HPV infection have limited their use in addressing certain important mechanisms of HPV-associated carcinogenesis. With the development of sophisticated molecular techniques, the direct study of HPV in vivo has become less problematic. Although some uncertainty remains, the animal model system for HPV-associated cervical disease has been maintained as the most scientifically and economically powerful in-vivo model. So far, animal model systems, based on HPV virus-like particles, pseudoviruses, transgenic mice, HPV-transformed cervical cell lines, have been invaluable in the recognition of the natural history of HPV infection, the synergy between HPV and its co-factors, as well as evaluating the efficacy and safety of new prophylactic and therapeutic modalities for HPV-associated cervical precancers and invasive cancers.

Introduction

Cervical cancer is the second most common cancer (after breast cancer) among women worldwide, responsible for 510,000 cases and 288,000 deaths annually, almost 80% of them occurring in the developing countries [1]. Extensive epidemiological data have shown that human papillomavirus (HPV) DNA could be detected in 99.7% cases of cervical cancer, as well as a substantial proportion of other anogenital, head and neck cancers and skin cancers in immunosuppressed individuals [2]. Eighty-two percent of the female population would be infected with at least one type if not several of HPV in their sexual lifetime; about 95% of cervical infections are subclinical and self-limiting within 2 years due to the host immune response, and the remaining 5% that persist for more than 2 years are highly linked to cervical precancers and invasive cancers [3].

Human papillomavirus is strictly species and tissue-restricted, and even in experimental settings, it does not infect any other host than humans. The titer of infectious HPV virions isolated from naturally occurring human lesions is extraordinarily low. Meanwhile, HPV cannot be propagated in routine cell cultures. These characteristics have retarded the development of in-vivo models for HPV infection. Thus, much of our understanding on HPV, its life cycle, its oncogenicity and its synergy with
genetic and environmental co-factors was first established by animal papillomaviruses (reviewed in [4]), which possess considerable homology to HPV. Rhesus macaques are the only nonhuman model in which spontaneous papillomavirus-associated cervical dysplasia has been described [5]. They may serve as a highly relevant animal model for the study of HPV infection and cervical neoplasia. Many pathological mechanisms of cervical carcinogenesis may be evaluated in vitro or by animal papillomaviruses in vivo. However, key differences
between such systems and natural HPV infection have limited their use in addressing certain important events in HPV-associated carcinogenesis. The biological behavior of HPV is essentially somewhat different from that of animal papillomaviruses; meanwhile, in-vitro systems are usually unable to clarify important in-vivo phenomena, such as cell cycle control, signal cascade, and immune regulation. Thus, animal models for HPV-associated cervical pathogenesis have been employed to provide unique insights into analyzing HPV transmission, persistence, vaccination, and what is more important, the investigation of cervical carcinogenesis. In recent years, the number and variety of animal models continue to expand, which has led to considerable advances in our understanding of HPV.”

“Anogenital HPV infections are transmitted primarily through skin-to-skin or mucosa-to-mucosa routes. The reproductive life cycle of HPV is strongly dependent on
the differentiation status of its host keratinocyte. To initiate infection, the viral particle must, through tiny
tears in the genital skin or mucosa, bind to the basal epithelial cell surface. After entry into the host cell, the virion establishes its genome as the nuclear episome, which is coordinately replicated along with the host genome and maintained at low copy number. Notably, random integration of the HPV genome into the host is thought to be necessary for cervical carcinogenesis.”

Animal models using virus-like particles and pseudoviruses

To date, there are no studies indicating that HPV virions produced in vitro can infect keratinocytes and initiate a reproductive life cycle. The nonavailability of efficient techniques to produce high-titer HPV virions has hindered insight into the life cycle of HPV. However, highly efficient systems to produce HPV virus-like particles (VLPs)and pseudoviruses have been employed as a substitute for the study of its life cycle [13]. Thus, the mechanisms that regulate the binding and entry of HPV into cells are beginning to be elucidated.

Recombinant L1 or L1 and L2 can be synthesized in a variety of expression systems to produce self-assembled VLPs, which possess morphological characteristics indistinguishable from authentic HPV virions and are still able to enter target cells. VLPs represent non-infectious HPV capsids without viral genome and are capable of inducing an immune response. On the basis of VLPs to elicit protective immunity against infectious viral challenge in animal models [14], two L1 VLP-based vaccines have been licensed for the prevention of HPV infection in humans since 2006 (reviewed in [15]). However, due to the HPV type-restriction of the protection from neutralizing antibodies, HPV 16 L1 VLP-based vaccine could not protect against any other type than HPV 16 [16]. Several animal experiments have shown chimeric VLPs containing cross-neutralizing L2-epitopes could be vaccine candidates providing broader protection than multivalent L1 VLP vaccines [17]. Chimeric VLPs containing a series of T-cell epitopes of E6 and/or E7 have also been used to induce neutralizing antibodies and a significant cytotoxic T-lymphocyte immune response against established HPV infections in mice [18].

On the contrary, VLPs with a molecule that emits fluorescence only after entry into a target cell have been used in research into the life cycle of HPV [19]. Experiments on tagged VLPs have demonstrated that cell-surface heparan sulfate may be the initial attachment receptor for the entry of most HPV types into the target cell, through its interaction with the carboxyl-terminal region of L1 [20]. a6 integrin may also act as a candidate receptor for HPV binding and entry [21]. Blocking the key receptor for HPV binding may offer a successful approach to inhibit the transmission of HPV. Since the tagged VLPs do not have any internal reporter gene, the
expression of DNA transported to the cell nucleus could not be detected, limiting the research into analyzing the uptake of HPV VLPs.

In contrast to VLPs, pseudovirus is composed of HPV structural proteins L1 and L2 and carries one or more encapsidated reporter genes (so-called pseudogenome) other than the HPV genome. The modified human embryonic kidney cell line, 293TTor 293FT cells, is co-transfected with plasmids expressing codon-modified L1, L2 and areporter plasmid, resulting in the self-assembly of the reporter plasmid into the L1/L2 capsids to generate infectious pseudoviruses. Unlike L1-VLPs, pseudovirus-mediated delivery and expression of the reporter gene in vivo are obviously enhanced by the presence of L2 [22]. Meanwhile, L2 has been shown to be necessary for DNA binding and encapsidation in some HPV types (reviewed in [23]). On the basis of pseudoviruses with L2 epitope-tagged (allowing tracking both the pseudogenome and L2), it is well established that uncoating is initiated in cytoplasmic vesicles, followed by L2 mediating the delivery of the viral genome into the nucleus to nuclear
domain 10 [24].

Human papillomavirus cannot be propagated in cell culture. Therefore, pseudoviruses have been used instead of authentic HPV virions to study the molecular biology of HPV and evaluate the protective efficacy of neutralizing antibodies derived from VLPs [25]. It has been demonstrated in vitro that lactoferricin [26] and sulfated polysaccharides [27,28], such as heparin, cellulose sulfate, dextran sulfate and carrageenan, can block the transmission of pseudoviruses by binding the viral capsid or heparan sulfate on the cell surface. However, such in-vitro systems fail to fully represent some aspects of HPV infection of keratinocytes in vivo. The genital transmission of HPV and the assembly and entry stages of its life cycle in vivo have been simplified by the recent development of an efficient method for in-vitro production of high-titer HPV pseudoviruses [13,29]. Although some uncertainty persists, pseudoviruses are thought to virtually mimic the HPV genital transmission phase, which are not species-restricted [27,29]. Thus, a pseudovirus-based cervico-vaginal challenge mouse model was recently constructed to imitate the establishment phase of HPV infection in vivo [30]. Systemic progesterone treatment 4 days before exposure to HPV pseudoviruses, chemical or mechanical disruption of the integrity of the genital epithelium immediately before inoculation and tissue analysis for reporter gene expression 3 days after pseudovirus challenge could all together contribute to the successful creation of this mouse model. Using the pseudovirus-based challenge model, it is revealed that genital transmission of HPV 16 is potentiated by nonoxynol-9
and inhibited by carrageenan. Similarly, heparinase treatment of the murine female genital tract significantly inhibited infection of HPV5, 16, 31 by greater than 90% and clearly inhibited HPV virion attachment to the basement membrane and the cell surface, clarifying that heparan sulfate is the primary attachment receptor for these HPV types in vivo [31]. The establishment of a pseudovirus-based mouse cervicovaginal challenge model will definitely lay a solid foundation for the investigation of anti-HPV intervention, the evaluation of HPV vaccines and other prophylactic reagents.”

Transgenic mice

“Since the introduction of transgenic technologies in the early 1990s, an extremely valuable animal model system for HPV-induced carcinogenesis has been acquired through HPV transgenic mice, to explore the contribution of oncogenes regulating cervical neoplasia. Pilot studies of transgenic models have not targeted the keratinocyte for the transgene expression [32]. Targeted expression of HPV oncogenes to keratinocytes would address the role of these oncogenes in squamous epithelium, which is the site for clinical HPV disease in humans.”

It has been found that chronic estrogen exposure specifically induced a multistage neoplasia in the cervix and vagina in all of K14-HPV mice and developed invasive cancers in 60% of the treated transgenic mice, closely mimicking cervical cancer progression in humans [37]. Using estrogen titration, K14-HPV16 mice treated with 0.05 mg/60-day 17b-estradiol developed cervical neoplasia solely at the transformation zone without any other reproductive tract site, which closely imitates clinical cervical carcinogenesis in women [38]. Thus, chronic estrogen exposure is a leading co-factor of HPV oncogenes for cervical neoplasia in transgenic mice. In addition, K14-HPV16 mice developed tumors with increased efficiency when induced with chemical carcinogens, such as 7,12-dimethylbenz(a) anthracene and 12-O-tetradecanoylphorbol-13-acetate [33,39]. Genetic background is also significant in the determination of susceptibility to invasive cancers, which is in keeping with the effect of HLA genotype in the human disease.”

“To elucidate the individual roles of E6 and E7 in cervical carcinogenesis, further characterization of K14E6 and
K14E7 transgenic mice was performed. Although K14E6 and K14E7 mice developed tumors in the skin, no spontaneous reproductive malignancies arose without exogenous estrogen treatment [33,40]. Moderate cervical hyperplasia was noted in several adult K14E7 mice [40]. Within the 6-month treatment period with 17b-estradiol, the K14E7 transgenic mice developed high-grade cervical dysplasia and cervical cancer, whereas the K14E6 mice developed only low-grade cervical dysplasia without neoplastic progression [41]. The synergy between E7 and estrogen in inducing cervical cancer may in part reflect the ability of both factors to modulate TGF-b signal transduction, because mRNA and protein levels of TGF-b2 and TGF-b receptor were decreased in K14E7 mice [34]. DEK oncogene expression may also contribute to cervical neoplasia through increased proliferation and the retardation of differentiation [42].

Despite the compelling demonstration of E6 and E7, respectively, in the mouse cervix, these oncogenes are
acting cooperatively in humans. In the cervix of K14E6E7 double-transgenic mice, the coexpression of E6 and E7 contributes to the formation of larger and more invasive cancers, compared with K14E7 mice, revealing E6 modulated the malignant phenotype produced by E7. When the period of estrogen treatment was extended to 9 months, tumors arising in K14E7 and K14E6E7 mice were greatly increased in size [43]; K14E6 mice also developed cervical cancer, indicating E6 has a weaker oncogenic potential in the reproductive tract than E7 [44]. Analyzing the tumors arising in E6 transgenic mice, it is clear that E6-induced up-regulation of the E2F-responsive genes, Mcm7 and cyclin E, and down-regulation of p16 in the absence of E7 [44]. K14E7 and K14E6E7 mice treated 6 months with estrogen followed by 3 months without exogenous estrogen had significantly fewer tumors and the tumors were smaller and less aggressive than those arising in mice treated for full 9 months and treated for 6 months then immediately analyzed [43]. These results indicated the necessity of estrogen in the initiation, maintenance and progression of cervical cancer in combination with HPV oncogenes. Previously, estrogen was thought to up-regulate HPV oncogene trascription via the response elements in the viral URR. However, K14 HPV mice can maintain HPV oncogene expression in another unknown way, since the URR is replaced by K14, which cannot be up-regulated by estrogen [44].”

“Meanwhile, tumor-bearing animals can also be employed as unique, preclinical, experimental systems to elucidate the mechanisms of other anticancer interventions, such as chemotherapy, radiotherapy, surgery, immunotherapy and combination therapy. Subcutaneous transplantation of the C57Bl/6 TC-1 tumor cell line expressing HPV 16 E6 and E7 is the most widely employed animal model. Another alternative C57Bl/6 tumor cell line is the C3 line, which expresses the entire HPV 16 genome. Although these two cell lines are not true cervical cancer cell lines, they serve as a suitable model for the development of therapeutic strategies since they depend on E6 and E7 for their oncogenicity. Furthermore, human cervical cancer cell lines, such as HPV 16 (CaSki and SiHa), HPV 18 (HeLa) and HPV 68 (ME-180)-transformed cell lines, can also subcutaneously be transplanted in the immunodeficient nude mice, to investigate the biological behavior of cervical cancer and anticancer treatments [60–62]. However, the conclusions drawn from these animal models are unable to reflect the interactions between tumor cells and the human immune system. Thus, the humanized-severe combined immunodeficient (hu-SCID) mouse model, produced by engrafting human peripheral blood lymphocytes, lymphoid tissues or bone marrow cells from healthy adult female volunteers into SCID mice, has been designed to mimic the human immune system and induce specific antitumor immunity [63].

These experimental models involving implanting human cervical cancer cells subcutaneously in nude mice, although relatively easy to perform, fail to yield metastases, partially due to the poor blood supply and the lack of surrounding visceral organs. The rapid spread of cervical cancer cells to the regional lymph nodes and the surrounding organs is a common phenomenon, and it is the leading cause of death among patients with cervical cancer. Therefore, elucidating the mechanism of metastasis in this disease is essential for improving prognosis and designing therapeutic strategies.”

Conclusion

“Human papillomavirus is strictly species and tissue-restricted and cannot be propagated in vitro, which has hampered our recognition of HPV as a pathogen. Nevertheless, the animal model system for HPV associated pathogenesis in the cervix has been important in the recognition of the natural history of HPV infection, the synergy between HPV and its co-factors (genetic and environmental), as well as evaluating the efficacy and safety of new prophylactic and therapeutic modalities for cervical cancers. Although some uncertainty remains, animal model systems will still lead the way and contribute significantly to unsolved issues on HPV-associated cervical pathogenesis in the future.”

http://dx.doi.org/10.1097/MRM.0b013e328331ad65

In Summary:

• HPV is strictly species and tissue-restricted and cannot be propagated in vitro, which has retarded the development of in-vivo models for HPV infection
• Much of our understanding on HPV, its life cycle, its oncogenicity and its synergy with co-factors was first established in vitro or by animal papillomaviruses in vivo
• However, key differences between such systems and “natural” HPV infection have limited their use in addressing certain important mechanisms of HPV-associated carcinogenesis
• Although some uncertainty remains, the animal model system for HPV-associated cervical disease has been maintained as the most scientifically and economically powerful in-vivo model
• Animal model systems have been based on:
  1. HPV “virus-like” particles
  2. Pseudoviruses
  3. Transgenic mice
  4. HPV-transformed cervical cell lines
• Eighty-two percent of the female population would be infected with at least one type if not several of HPV in their sexual lifetime; about 95% of cervical infections are subclinical and self-limiting within 2 years due to the host immune response, and the remaining 5% that persist for more than 2 years are highly linked (i.e. not proven) to cervical precancers and invasive cancers
• Even in experimental settings, HPV does not “infect” any other host than humans
• The titer of infectious HPV “virions” isolated from naturally occurring human lesions is extraordinarily low
• HPV cannot be propagated in routine cell cultures
• Many pathological mechanisms of cervical carcinogenesis may be evaluated in vitro (i.e. within the lab) or by animal “papillomaviruses” in vivo (i.e. within an animal)
• However, key differences between such systems and “natural HPV” infection have limited their use in addressing certain important events in HPV-associated carcinogenesis
• The biological behavior of HPV is essentially somewhat different from that of animal “papillomaviruses”
• In-vitro systems are
  usually unable to clarify important in-vivo phenomena, such as cell cycle control, signal cascade, and immune regulation
• Random integration of the HPV genome into the host is thought to be necessary for cervical carcinogenesis (in other words, they do not know and they are guessing)
• To date, there are no studies indicating that HPV “virions” produced in vitro can infect keratinocytes and initiate a reproductive life cycle
• The nonavailability of efficient techniques to produce high-titer HPV “virions” has hindered insight into the life cycle of HPV
• However, highly efficient systems to produce HPV “virus-like” particles (VLPs) and “pseudoviruses” have been employed as a substitute for the study of its life cycle (i.e. they are using admittedly fake “viruses” to study their actual fake “viruses” that they can not “grow”)
• Recombinant L1 or L1 and L2 can be synthesized  in a variety of expression systems to produce self-assembled VLPs, which possess morphological characteristics indistinguishable from authentic HPV “virions” and are still able to enter target cells (i.e. they looked the same as the particles assumed to be HPV which have never been proven to be a “virus” through purification/isolation)
• VLPs represent “non-infectious” HPV capsids without “viral” genome and are capable of inducing an immune response
• On the basis of VLPs to elicit protective immunity against infectious “viral” challenge in animal models, two L1 VLP-based vaccines have been licensed for the prevention of HPV infection in humans since 2006
• Since the tagged VLPs do not have any internal reporter gene, the expression of DNA transported to the cell nucleus could not be detected, limiting the research into analyzing the uptake of HPV VLPs
• As human “papillomavirus” cannot be propagated in cell culture, “pseudoviruses” have been used instead of authentic HPV “virions” to study the molecular biology of HPV and evaluate the protective efficacy of neutralizing antibodies derived from VLPs
• In other words, they use the fake “infectious virus” to test the fake antibodies derived from the fake noninfectious “virus-like” particles
• It has been demonstrated in vitro that lactoferricin and sulfated polysaccharides, such as heparin, cellulose sulfate, dextran sulfate and carrageenan, can block the transmission of “pseudoviruses” (i.e. not HPV) by binding the “viral” capsid or heparan sulfate on the cell surface
• However, such in-vitro systems fail to fully represent some aspects of HPV infection of keratinocytes in vivo
• Although some uncertainty persists, “pseudoviruses” are thought to virtually mimic the HPV genital transmission phase, which are not species-restricted
• Thus, a “pseudovirus-based” cervico-vaginal challenge mouse model was recently constructed to imitate the establishment phase of HPV infection in vivo
• The establishment of a “pseudovirus-based” mouse cervicovaginal challenge model will definitely lay a solid foundation for the investigation of anti-HPV intervention, the evaluation of HPV vaccines and other prophylactic reagents
• Since the introduction of transgenic technologies (relating to or denoting an organism that contains genetic material into which DNA from an unrelated organism has been artificially introduced) in the early 1990s, an extremely valuable animal model system for HPV-induced carcinogenesis has been acquired through HPV transgenic mice, to explore the contribution of oncogenes regulating cervical neoplasia
• Pilot studies of transgenic models have not targeted the keratinocyte for the transgene expression
• It has been found that chronic estrogen exposure specifically induced a multistage neoplasia in the cervix and vagina in all of K14-HPV mice and developed invasive cancers in 60% of the treated transgenic mice, closely mimicking cervical cancer progression in humans
• Using estrogen titration, K14-HPV16 mice treated with 0.05 mg/60-day 17b-estradiol developed cervical neoplasia solely at the transformation zone without any other reproductive tract site, which closely imitates clinical cervical carcinogenesis in women
• Thus, chronic estrogen exposure is a leading co-factor of HPV oncogenes for cervical neoplasia in transgenic mice
• Genetic background is also significant in the determination of susceptibility to invasive cancers
• In other words, blasting genetically altered mice wtth high levels of estrogen can cause cervical cancer, thus showing unnatural estrogen levels and mutated genetics influence cancer development
• Although K14E6 and K14E7 mice developed tumors in the skin, no spontaneous reproductive malignancies arose without exogenous estrogen treatment
• When the period of estrogen treatment was extended to 9 months, tumors arising in K14E7 and K14E6E7 mice were greatly increased in size; K14E6 mice also developed cervical cancer
• K14E7 andK14E6E7 mice treated 6 months with estrogen followed by 3 months without exogenous estrogen had significantly fewer tumors and the tumors were smaller and less aggressive than those arising in mice treated for full 9 months and treated for 6 months then immediately analyzed
• These results indicated the necessity of estrogen in the initiation, maintenance and progression of cervical cancer in combination with HPV oncogenes
• Human cervical cancer cell lines, such as HPV 16 (CaSki and SiHa), HPV 18 (HeLa) and HPV 68 (ME-180)-transformed cell lines, can also subcutaneously be transplanted in the immunodeficient nude mice, to investigate the biological behavior of cervical cancer and anticancer treatments
• In other words, they use human cervical cancer cells (not HPV) to create cervical cancer in IMMUNODEFICIENT mice to study it
• However, the conclusions drawn from these animal models are unable to reflect the interactions between tumor cells and the human immune system
• Thus, the humanized-severe combined immunodeficient (hu-SCID) mouse model, produced by engrafting human peripheral blood lymphocytes, lymphoid tissues or bone marrow cells from healthy adult female volunteers into SCID mice, has been designed to mimic the human immune system and induce specific antitumor immunity
• These experimental models involving implanting human cervical cancer cells subcutaneously in nude mice, although relatively easy to perform, fail to yield metastases, partially due to the poor blood supply and the lack of surrounding visceral organs
• Human “papillomavirus” is strictly species and tissue-restricted and cannot be propagated in vitro, which has hampered our recognition of HPV as a pathogen
• Nevertheless, the animal model system for HPV associated pathogenesis in the cervix has been important in the recognition of the natural history of HPV infection and the synergy between HPV and its co-factors (genetic and environmental)
• Although some uncertainty remains, animal model systems will still lead the way and contribute significantly to unsolved issues on HPV-associated cervical pathogenesis in the future

As can be seen from this paper, everything we know about HUMAN “papillomaviruses” actually comes from ANIMAL “papillomaviruses.” Virologists have just equated what they “know” about the animal version to humans. However, knowing that this was not enough due to key differences between animal and human systems, the biological behavior of the “virus,” and the modes of HPV infection, they decided to synthetically create “viruses” to inject into genetically modified mice to mimic what they think HPV infection looks like. They also utilized “pseudoviruses” for their mimickery purposes. Then the completely “ethical” virologists cranked up the estrogen levels into these genetically-altered mice and upon producing tumors, state that they were successful with their synthetic “virus” while relegating the unnaturally high estrogen levels and the genetic modifications as “co-factors.” Is the creation of “pseudoviruses” and/or “virus-like” particles mimicking so-called HPV, genetically altering mice, and then overloading the altered mice with estrogen proof of a pathogenic human “papillomavirus?” Would this mad-science experiment in any way satisfy any of Koch’s Postulates? Keep in mind that these synthetic VLP’s are used in the vaccines as well as to judge vaccine efficacy based on challenge trials using “pseudoviruses” and not HPV. There is nothing that reflects reality or nature in the HPV pathogenicity studies.

Creating synthetic “virus-like” particles to inject into genetically engineered mice while using high levels of estrogen in order to “prove” the existence/pathogenicity/mechanisms of hybridized cloned DNA fragments based off of grounded up “virus-like” particles taken from warts.

This is viroLIEgy.