Human papillomavirus are miniscule double stranded DNA viruses which target cutaneous and mucosal epithelial cells, inducing lesions which may lead to hyper proliferation and tumorigenesis. There are over 100 identified strains of HPV which are classified into two types; Low-risk types which may lead to symptoms such as benign genital warts in contrast to high risk types which may lead to aggressive tumorigenesis and metastasis.
The genomes of all HPV strains contain nearly eight open reading frames (ORF). Each when transcribes produces a polycistronic mRNA( meaning that the mRNA can code for a multitude of different proteins). The regulatory sequences that are needed for the regulation of transcription of these polycistronic mRNAs
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Genther et al, 2003). E5 also appears to encode a membrane protein with a weak transforming activity (Leptak et al., 1991). E6 and E7 however are the main transformants involved which may lead to mutations that cause tumorigenesis. Thus the first step of this process is to obtain a high concentration of wild type HPV31 viral plasmid in order to be able to adequately re-engineer for the treatment.
A sample HPV31 will be bought an injected into several cultures of HeLa cells grown in DMEM+FBS, we will also have a control plate where no HPV31 will be injected. This will ensure that there is no contamination in our plates. Some of the live HPV31 will be set aside to extract their viral plasmid and sequence them either using new generation techniques or Sanger sequencing. The HeLa cultures infected with HPV and the control plate will be allowed to grow for two weeks to allow the proliferation of the HPV viral plasmid.
Once incubation is complete the viral DNA plasmid will be extracted from the cultures using a mini-prep protocol as follows. Cells will be washed twice with PBS and lysed in lysis solution (20 mM Tris-HCl (pH 8.0), 100 mM NaCl, 10 mM EDTA, and 0.2% SDS) for total genomic DNA extraction. The cell lysates will treated overnight with proteinase K (0.2 μg/ml), extracted with phenol-chloroform and precipitated with 2 volumes of 96% ethanol. DNA pellets will be dissolved in TE buffer containing 20 μg/ml RNase A and incubated for 1 hour at 37°C. The DNA will
II. (MP 2) HPV or human papilloma virus is an umbrella term for a group of over 150 related viruses. Of these 150 around 40 are genital HPV.
HPV is a name given to multiple types of one virus - Human Papilloma Virus. HPV has 150 different types, and each subtype is given a number. More commonly called Genital Warts, HPV is one of the most common infections worldwide, and 50% of the people who engage in sex acquire it at some point in their life.
The E6 and E7 proteins play and important rule in oncogenic property of human papillomavirus that can cause anogenital cancers as their high risk factor and warts as their low risk factor. E6 AND E7 proteins promote cell growth by inactivating the tumor suppressor proteins P53 and PRb (5).
HPV is typically spread through skin to skin contact with a person infected with the virus. It is classified as an STD because it can be transmitted sexually. It has been calculated that “at least 50 percent of sexually active men and women have been infected with genital HPV at some point in their lives” (Schoenstadt). HPV infects and replicates in the epithelial tissues which include the skin and mucous membranes (Panse). The life cycle of HPV is fairly easy to understand. It is important to understand that there are specific genes that are involved and expressed in the life cycle at different stages. The virus enters the body through micro-abrasions and it moves towards the basal epithelial cells. At this point the viral genome has entered and the genes E1 and E2 begin the genome replication as well as making mRNA for
The Human Papillomaviruses, more commonly known as HPV, are a group of more than 150 related viruses. Both men and women carry HPV and more than 40 of these viruses are easily transmitted sexually transmitted infections (STIs). Twenty HPV strains are known to cause cancers in the genital region.
As mentioned earlier, HPV is a non-enveloped, icosahedral capsid, double-stranded circular DNA virus (Morshed, 2014). Infection of HPV occurs in the cervix, glans of the penis, penile shaft, scrotom and anal verge by interactions with putative “host cell surface receptors such as heparin sulfate proteoglycans and alpha-6 integrins” Schafer, 2015; Letian, 2010). Most studies suggest that HPV16 enters the cell via clathrin-mediated endocytosis (Day, 2003, Bousarghin, 2003, Smith, 2007, Hindmarsh, 2007), however, notably, a few contradictory studies suggest that HPV16 is clathrin and caveolaen independent (Spoden, 2008). Once the virus has entered the cell, it then un-coats and delivers its genome (8 genes) to the host cell nucleus to be expressed. These eight genes are named for when they are expressed during the virus’s occupation of its host: early (E) and late (L). The proteins E1, E2, E4 and E5 play an important role in genome replication (Beutner, 1997; Gnanamony, 2007) while E6 and E7 are oncogenes and are capable of causing cancerous growth of host cell (Gnanamony, 2007; Sikorski, 1998). E6 is thought to complex with p53 which is a cellular tumor repressor (Werness, 1990); when E6 complexes with P53, it is thought to promote the degradation of p53 via the ubiquitin pathway (Schffner et al., 1990). This degradation of p53 contributes to the oncogenic potential of high-risk HPVs (Stewart, 2005, Thomas, 1999). Crook showed that E6 can bind with p53
The human papillomavirus (HPV) is the most common sexually transmitted infection (STI) in the United States (Centers for Disease Control & Prevention [CDC], 2015). Scientists predict that nearly all sexually active men and women will be infected with HPV at one point during their lifetime (Centers for Disease Control & Prevention [CDC], 2010, Centers for Disease Control & Prevention [CDC], 2015; International Agency for Research on Cancer, 2007, Riguad, 2015). Although HPV is common, it usually resolves on its own within a year or two of infection. It does not pose a risk until it has remained in the body for many years, in which it can lead to pre-cancerous cells and genital warts (Centers for Disease Control & Prevention [CDC], 2013; Human Papillomavirus (HPV), 2015; National Cancer Institute [NCI], 2012). HPV can be categorized into two groups, high risk HPV and low risk HPV. High risk HPV is responsible for the development of cancers, such as cervical, oropharyngeal, anal, vaginal, vulvar, and penile, whereas low risk HPV is associated with the development of genital warts (International Agency for Research on Cancer, 2007). Currently, there are nearly 26,000 new HPV-related cancers in the United States and roughly 18,000 occur in women, and 8,000 occur in men (President’s Report).
Human Papillomavirus (HPV) is a sexually transmitted disease (STD). “Approximately 20 million Americans are infected with HPV as of 2009, and another 6.2 million people become newly infected each year” (Langwith, 2013, p.16). There are many causes for HPV including vaginal sex, oral sex, anal sex, abrasions of skin, and passing of HPV between mother and child during a vaginal birth. The effects of HPV after being contracted include cancer, genital warts, planter warts, and palmer warts.
Human papillomavirus (HPV) is a group of around 150 viruses. Some HPV types can lead to wart formation and other types can lead to cancer; primarily cervical, vaginal, vulvar, penile, anal, and Oropharyngeal cancer (1,2)
The human papillomavirus (HPV) is one of the most common causes of sexually transmitted disease in both men and women around the world, especially in developing countries, where the occurrence of asymptomatic infection varies from 2 to 44%, depending on the population and studied region. Most HPV infection is transient and some studies show that the majority of sexually active individuals are exposed to and acquire infection from this virus at some phase in their lives. HPV infection is more prevalent in young adults, at the beginning of their sexual activity, which is more important to detect the virus at the early edge to prevent from the most damages in the later of life that leads to less impact in the economy in their country and family too. with a subsequent decline in the prevalence rate with increasing age, likely as a result of development of an immune response against the virus and reduction of sexual activity.
The human papillomavirus is a viral disease that can cause anything from warts that we see on the skin of our hands and feet and genitalia to cancer of the cervix, anus or oral orifices. Although anybody can be infected with the human papillomavirus, it is more common in adolescent and young women. We will see how the human papillomavirus is unique in the way that it infects cell as well as its symptoms and its current treatment.
Human papillomaviruses (HPVs) are a group of more than 200 related viruses. More than 40 HPV types can be easily spread through direct sexual contact via vaginal, anal, and oral sex (NIH-NCI, 2015). While high-risk HPV types cause approximately 5% of all cancers worldwide, in the United States, high-risk HPV types cause approximately 3% and 2% of all cancer cases among women and men respectively. Cervical cancer, vaginal cancer, vulvar cancer, anal cancer, penile cancer, and oropharyngeal cancers can be caused by high-risk HPV (de Martel et al., 2012; Jemal et al., 2013).
Human papillomavirus (HPV) is a non-enveloped virus with double-stranded circular genome. The genome encodes for 6 early proteins, which are necessary for virus replication; and 2 late proteins, which are required to build viral structural proteins. Mucosal tissues are the target for HPV to infect, especially through anogenital region. Indeed, Infection with HPV is the most common sexually transmitted infection. Up to 80% of sexually active adults are infected once in their lifetimes with HPV (McNeil vaccine). There are numerous types of HPV, with 40 types are known to infect human through genital tract. HPV can be designated as high-risk (oncogenic virus), which means they are highly potential to develop cancer. Specifically, HPV 16 and
Clearly, the development of cell culture systems for the newer HPyVs is important, in order to advance research into their biology, for example, the possibility of isolating and growing the infectious polyomavirus particles in the laboratory or experimental system.
Specific Aim 2: Determine the risk of different blood products in the outcome of viral infection Blood products contain red blood cells, white blood cells, sera, and plasma. HPV DNA has been detected in different blood products including PBMCs, sera, and plasma of HPV-positive patients and healthy donors. Further studies demonstrate that CD20+ and CD56+,