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- Member of: Embryo Project Encyclopedia Articles
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Methods: We have designed a multiplexed magnetics programmable bead ELISA (MagProBE) to profile the immune responses of the proteins from 11 high-risk HPV types and 2 low-risk types—106 genes in total. HPV genes were optimized for human expression and either built with PCR or commercially purchased, and cloned into the Gateway-compatible pANT7_cGST vector for in vitro transcription/translation (IVTT) in a MagProBE array. Anti-GST antibody (Ab) labeling was then used to measure gene expression.
Results: 53/106 (50%) HPV genes have been cloned and tested for expression of protein. 91% of HPV proteins expressed at levels above the background control (MFI = 2288), and the mean expression was MFI = 4318. Codon-optimized genes have also shown a 20% higher expression over non-codon optimized genes.
Conclusion: Although this research is ongoing, it suggests that gene optimization may improve IVTT expression of HPV proteins in human HeLa lysate. Once the remaining HPV proteins have been expression confirmed, the cDNA for each gene will be printed onto slides and tested in serologic assays to identify potential Ab biomarkers to CIN3.
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Henrietta Lacks, born Loretta Pleasant, had terminal cervical cancer in 1951, and was diagnosed at The Johns Hopkins University in Baltimore, Maryland, where researchers collected and stored her cancer cells. Those cells went on to become the first immortal human cell line, which the researchers named HeLa. An immortal cell line is an atypical cluster of cells that continuously multiply on their own outside of the organism from which they came, often due to a mutation. Lacks’s cancer cells enabled scientists to study human cells outside of the human body, though that was controversial since she did not voluntarily donate her cells for such research. Science writer Rebecca Skloot chronicled Lacks’s life in her book, The Immortal Life of Henrietta Lacks, which became a movie in 2017. Lacks’s HeLa cell line has contributed to numerous biomedical research advancements and discoveries and her story has prompted legal and ethical debates over the rights that an individual has to their genetic material and tissue.
The Human Papillomavirus (HPV) strains 16 and 18 are the two most common HPV strains that lead to cases of genital cancer. HPV is the most commonly sexually transmitted disease, resulting in more than fourteen million cases per year in the United States alone. When left untreated, HPV leads to high risks of cervical, vaginal, vulvar, anal, and penile cancers. In 1983 and 1984 in Germany, physician Harald zur Hausen found that two HPV strains, HPV-16 and HPV-18, caused cervical cancer in women. In the early twenty first century, pharmaceutical companies Merck & Co. and GlaxoSmithKline created HPV vaccines protecting against HPV-16 and HPV-18, which have reduced the number of HPV infections by fifty-six percent in the US. Discovering HPV strains 16 and 18 allowed physicians to test for those cancer-causing cell populations using Pap smears, a diagnostic tool that collects cells from the woman's cervix to identify cancerous cases of HPV infection. By identifying the cancerous strains of HPV-16 and HPV-18 and utilizing preventative measures such as the Pap smear and HPV vaccines, the rates of cervical cancer and other HPV-related cancers have reduced.
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From 1977 to 1987, Harald zur Hausen led a team of researchers across several institutions in Germany to investigate whether the human papillomavirus (HPV) caused cervical cancer. Zur Hausen's first experiment tested the hypothesis that HPV caused cervical cancer rather than herpes simplex virus type 2 (HSV-2), the then accepted cause. His second and third experiments detailed methods to identify two previously unidentified HPV strains, HPV 16 and HPV 18, in cervical cancer tumor samples. The experiments showed that HPV 16 and 18 DNA were present in cervical tumor samples. Zur Hausen concluded that HPV, not HSV-2, caused cervical cancer, which enabled researchers to develop preventions, such as the HPV vaccine.
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Richard Doll was an epidemiologist and public figure in the twentieth and early twenty-first centuries. Working primarily at the University of Oxford, in Oxford, England, Doll established a definitive correlation between cigarette smoking and lung cancer. Furthermore, Doll’s work helped legitimize epidemiology as a scientific discipline. Doll’s research also helped establish modern guidelines for oncological studies, as well as for contemporary and future research on the effect of smoking on pregnancy and fetal development. In addition to studying the health effects of smoking, Doll also studied cervical cancer and contraceptives.
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During the twentieth century, Austin Bradford Hill researched diseases and their causes in England and developed the Bradford Hill criteria, which comprise the minimal requirements that must be met for a causal relationship to be established between a factor and a disease. Hill also suggested that researchers should randomize clinical trials to evaluate the effects of a drug or treatment by monitoring large groups of people. In addition, Hill advocated for case-control studies, in which researchers compare a group of people with a medical condition to a group without that condition to investigate the condition's possible causes. Hill's own work with clinical trials and case-control studies helped him prove that smoking caused lung cancer. The Bradford Hill criteria have also been used to establish causal links between factors and cancer, including reproductive cancers such as human papillomavirus that causes cervical cancer.
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George Nicholas Papanicolaou developed the Pap test in the United States during the twentieth century. The Pap test is a diagnostic procedure used to test for cervical cancer in women. Papanicolaou’s work helped improve the reproductive health of women by providing an effective means of identifying precancerous cells and improving the likelihood of early treatment and survival of cervical cancer.