'Acrylamide is a commonly used matrix for electrophoretic separation of nucleic acids and proteins'.
We will end this week's discussion seguing from DAN BROWN'S novel INFERNO-----with all that THE PROVOST and his MENDACIUM CORPORATION reeking havoc on 99% of global citizens by giving protection to all those DR NOs doing harmful and illegal MEDICAL RESEARCH.
We say often that this genome project which only a decade ago finished the mapping of a DNA molecule has several decades to a century of lab and mice research before the most basic of BASIC SCIENCE is understood as to how all those components of a DNA STRAND work individually, as neighboring codons, AND as support codons.
ONLY THEN WILL THE SCIENTIFIC METHOD allow for CONCLUSIONS with the ability PERHAPS to control for VARIABLES.
Below we see one process that has been used in diagnostic labs for decades. These kinds of molecule separation processes are fine to use separate from living tissue----they are proven MEDICAL PROCEDURES.
When we are working with GENOME PROJECT research scientists are doing just what we read below. They work to separate PROTEINS -----NUCLEIC ACIDS------and using DENATURING GELS to break down those components and INDEED-------these chemicals WILL BE CANCEROUS if medical procedures have these chemicals in daily PHARMA.
Acrylamide forms linear polymers and bis-Acrylamide introduces cross-links between polyacrylamide chains. By mixing the appropriate ratio of Acrylamide to bis-Acrylamide, the optimal pore size can be obtained for the protein being resolved. To avoid the toxicity of dry acrylamide, ready-to-use acrylamide solutions are available. Acrylamide Solution 40% (sc-3721) and bis-Acrylamide 2% (sc-3719) solutions offer convenience and eliminate the risks associated with the handling of acrylamide powder. Polymerization agents, TEMED (sc-29111) and APS (Ammonium persulfate, sc-202946) are also available. For denaturing gels, SDS (20% Solution) (sc-24950) is available'.
Today, we simply want to emphasize this: what are being called VIRUS VECTORS tied to bio-genetic procedures is not the same as those ordinary DISEASE VECTORS caused by environmental VIRUSES and bacteria. So, there will be no national VIRAL EPIDEMIC----no global PANDEMIC from these bio-genetic VIRUS VECTORS built to only function inside living body systems and cells.
The GORILLA-IN-ROOM problem in global banking 1% goals MOVING FORWARD is that these bio-genetic treatments will be MAINSTREAMED by OBAMA'S AFFORDABLE CARE ACT medical boards filled with global banking 5% freemason/Greek players creating policies around a 99% of citizens' HEALTH INSURANCE POLICIES--------and telling our local DOCTORS these bio-genetic treatments are REQUIRED.
THIS IS INDEED THE GOALS OF MOVING FORWARD AFFORDABLE CARE ACT------OF IGNORING AND ELIMINATING ALL US FEDERAL REQUIREMENTS FOR CLINICAL TRIALS AND SCIENTIFIC METHOD REAL DATA AND CONCLUSIONS.
Vaccines (Basel). 2014 Sep; 2(3): 624–641.
Published online 2014 Jul 29. doi: 10.3390/vaccines2030624
PMCID: PMC4494222
PMID: 26344749
Developments in Viral Vector-Based Vaccines
Takehiro Ura, Kenji Okuda, and Masaru Shimada*
Abstract
Viral vectors are promising tools for gene therapy and vaccines. Viral vector-based vaccines can enhance immunogenicity without an adjuvant and induce a robust cytotoxic T lymphocyte (CTL) response to eliminate virus-infected cells. During the last several decades, many types of viruses have been developed as vaccine vectors. Each has unique features and parental virus-related risks. In addition, genetically altered vectors have been developed to improve efficacy and safety, reduce administration dose, and enable large-scale manufacturing. To date, both successful and unsuccessful results have been reported in clinical trials. These trials provide important information on factors such as toxicity, administration dose tolerated, and optimized vaccination strategy. This review highlights major viral vectors that are the best candidates for clinical use.
Keywords: viral vector, vaccine, CTL, MVA, adenovirus
1. Introduction
Viral vectors are regarded as potential tools for gene therapy and vaccines. Their utility is based on the ability of viruses to infect cells. In general, the advantages of viral vectors are as follows: (a) high efficiency gene transduction; (b) highly specific delivery of genes to target cells; and (c) induction of robust immune responses, and increased cellular immunity.
The concept of viral vector vaccines is different from that of subunit vaccines, as the latter help prevent infectious diseases by eliciting a humoral response. Recombinant viral vectors have potential for therapeutic use because they enable intracellular antigen expression and induce a robust cytotoxic T lymphocyte (CTL) response, leading to the elimination of virus-infected cells. Despite their efficacy, viral vectors present unavoidable problems that need to be addressed. In the near future, viral vector-based vaccines may be increasingly used to fight major diseases, such as HIV-1 and malaria. In some vectors, stable expression of the interesting gene is achieved via viral integration mechanisms. Integration into the host genome can lead to cancer. Another obstacle to the clinical use of viral vectors is the presence of pre-existing immunity against the vector. This is caused by previous exposure to the virus and the production of neutralizing antibodies that reduce vaccine efficacy.
The development of viral vectors requires a high biological safety level in order to gain public acceptance. Therefore, non- (or low-) pathogenic viruses are often selected. In most cases, viruses are genetically engineered to reduce or eliminate pathogenicity. Additionally, most viral vectors are replication-defective. For example, in adenovirus-based vectors, the E1A and E1B encoding regions, which are needed for replication in infected cells, are deleted and replaced with the target gene.
When using a viral vector, it is important to assess the potential implications by understanding the epidemiological and virological characteristics. In this review, we describe several representative viral vectors with respect to risk and benefit, optimized vector-design attempts, and clinical results. Moreover, we discuss the combined use of viral vectors in prime-boost vaccination regimens.
2. Summary of Viral Vectors
The concept of the viral vector was introduced in 1972. Jackson et al. created recombinant DNA from the SV40 virus by genetic engineering [1]. Subsequently, Moss et al. reported the use of vaccinia virus as a transient gene expression vector in 1982 [2,3]. Several types of viral vectors have been developed, and they have been used in animal studies and clinical trials.
The specific properties of a vector are determined by the virus from which it derives. Each vector has distinct advantages and disadvantages (Table 1). Vaccinia virus and adenovirus are the most widely used vectors because they can induce a robust immune response, specifically involving CTL, against the expressed foreign antigens. Generally, viral vectors achieve high immunogenicity without an adjuvant. Viral components stimulate the innate immune response, leading to the production of interferons and inflammatory cytokines [4].
Table 1
Advantages and disadvantages of major viral vectors.
VirusAdvantagesDisadvantages
Major Clinical/Preclinical Studies
RetrovirusLong-term gene expression
Generation of replication-competent virus Potential for tumorigenesis
Infects dividing cells only[6]
Lentivirus
Long-term gene expression Infects non-dividing and dividing cells
Generation of replication-competent virus Potential for tumorigenesis[7,8]
Vaccinia virus
High immunogenicity Safety: used as a smallpox vaccine High titer productionPre-existing immunity[9]
Adenovirus
High immunogenicity Safety: used in many clinic trails High titer productionPre-existing immunity[10]
Adeno-associated virus
Long-term gene expression Non-pathogenic virusLow titer production[11]
Cytomegalovirus
Induces a unique CTL response Protects against SIV infection in an animal modelPre-existing immunity Risk of pathogenesis in specific individuals[12]
Sendai virus
High immunogenicity
Pre-existing immunity[13]Viral vector-based vaccines require assessment of efficacy and safety, including immunogenicity, genetic stability, ability to evade pre-existing immunity, replication deficiency or attenuation, and genotoxicity. Additionally, the cost-effectiveness must also be evaluated because infectious diseases are a problem in developing countries. Thus, it is necessary to consider the large-scale manufacturing of viral vectors. Typically, the manufacturing of viral vectors involves propagation in suitable cell lines.
The European Medicines Agency (EMA) recently provided guidelines on quality for non-clinical and clinical aspects [5]. These guidelines offer useful recommendations for the effective administration of live, recombinant viral vector-based infectious vaccines.
_____________________________________
'APHA is more than a professional association.
We're a community. Our members work in every discipline of public health, in every U.S. state and in countries across the globe'.
WE CAN'T STOP THIS shout those global banking 5% freemason/Greek players/pols MOVING FORWARD ONE WORLD ONE WORLD HEALTH ORGANIZATION inside US FOREIGN ECONOMIC ZONES!
OH, REALLY?????
Back in 1990s Clinton era we see below for whom these far-right wing global banking NEO-LIBERALS and NEO-CONS were working-----in 1990s 750 biotechnology corporations to include bio-genetics. 30 years ago many of these were regional businesses while today much of this industry has been consolidated into global banking 1% OLD WORLD KINGS medical corporations ----global MITRE CORPORATION THE HOSPITALLERS for example.
If we think today how many of these bio-tech/bio-genetic corporations exist and operate mostly OUTSIDE of US public health and safety ====that would be in the millions globally-----tens of thousands inside US.
'The Biotechnology Industry Organization (BIO) represents over 745 biotechnology companies, academic institutions, and state biotechnology centers in 46 states and 25 nations'.
Our local US city/county public health departments do not have to install these FEDERAL GUIDELINES-------our local PUBLIC HEALTH DEPARTMENTS CAN keep to our last century MEDICAL MORALS AND ETHICS.
Again, we must look to whom is appointed to VITAL public health positions. Our local public medical universities have THE PROVOST-----that DEAN who pushes OR NOT these health policies. We have that city public health department with a director who pushes OR NOT these health policies.
Below we see an association of what we KNOW are global banking 5% freemason/Greek players owning tech businesses or working for global medical corporations. These kinds of associations USED TO BE OUR COMMUNITY CITIZENS with businesses that HELPED AND KEPT SAFE our 99% of citizens.
IT MATTERS WHO IS APPOINTED----IT MATTERS FOR WHOM OUR CITY/COUNTY ASSOCIATIONS WORK. LET'S DO IT----WORK TO GET RID OF THESE GLOBAL BANKING 5% FREEMASON/GREEK PLAYERS WHO DO NOT CARE.
APHA is more than a professional association.
We're a community. Our members work in every discipline of public health, in every U.S. state and in countries across the globe.
And we're all united by a common goal: To create the healthiest nation in one generation.
Join APHA.
Together, we really can make a difference.
Why should you join?Exclusive benefits like a network of 25,000 peers who also are working to make a difference.
Special member savings on college tuition, professional certifications, insurance and more!
How much does it cost?That depends on your membership category. Learn more on our Member Types and Rates page.
How can I get the most out of my APHA membership?Watch our video series on such topics as advancing your career, navigating the Annual Meeting, earning CE credits and advocating for public health.
And while you're browsing APHA.org, notice the Member Perk image that highlights some of the great benefits of APHA membership, including access to our award-winning publications, opportunities for professional development and discounts on continuing education opportunities.
_________________________________________
GREEK TRAGEDIES --------MEDIEVAL MORALITY PLAYS-------AGE OF ENLIGHTENMENT SENSE AND SENSIBILITY PHILOSOPHY have told our 99% WE THE PEOPLE that selling our souls to the DEVIL which is what has MOVED FORWARD these few decades of CLINTON/BUSH/OBAMA staging to be PERMANENT in ONE WORLD ONE GOVERNANCE-----always ends badly for that PLAYER and often that PLAYER takes that family down as well.
youtube.com
Scratch's Song ( The Devil and Daniel Webster)
I Ringwald
Published on Apr 12, 2013
We will end this week's discussion seguing from DAN BROWN'S novel INFERNO-----with all that THE PROVOST and his MENDACIUM CORPORATION reeking havoc on 99% of global citizens by giving protection to all those DR NOs doing harmful and illegal MEDICAL RESEARCH.
We say often that this genome project which only a decade ago finished the mapping of a DNA molecule has several decades to a century of lab and mice research before the most basic of BASIC SCIENCE is understood as to how all those components of a DNA STRAND work individually, as neighboring codons, AND as support codons.
ONLY THEN WILL THE SCIENTIFIC METHOD allow for CONCLUSIONS with the ability PERHAPS to control for VARIABLES.
Below we see one process that has been used in diagnostic labs for decades. These kinds of molecule separation processes are fine to use separate from living tissue----they are proven MEDICAL PROCEDURES.
When we are working with GENOME PROJECT research scientists are doing just what we read below. They work to separate PROTEINS -----NUCLEIC ACIDS------and using DENATURING GELS to break down those components and INDEED-------these chemicals WILL BE CANCEROUS if medical procedures have these chemicals in daily PHARMA.
Acrylamide forms linear polymers and bis-Acrylamide introduces cross-links between polyacrylamide chains. By mixing the appropriate ratio of Acrylamide to bis-Acrylamide, the optimal pore size can be obtained for the protein being resolved. To avoid the toxicity of dry acrylamide, ready-to-use acrylamide solutions are available. Acrylamide Solution 40% (sc-3721) and bis-Acrylamide 2% (sc-3719) solutions offer convenience and eliminate the risks associated with the handling of acrylamide powder. Polymerization agents, TEMED (sc-29111) and APS (Ammonium persulfate, sc-202946) are also available. For denaturing gels, SDS (20% Solution) (sc-24950) is available'.
Today, we simply want to emphasize this: what are being called VIRUS VECTORS tied to bio-genetic procedures is not the same as those ordinary DISEASE VECTORS caused by environmental VIRUSES and bacteria. So, there will be no national VIRAL EPIDEMIC----no global PANDEMIC from these bio-genetic VIRUS VECTORS built to only function inside living body systems and cells.
The GORILLA-IN-ROOM problem in global banking 1% goals MOVING FORWARD is that these bio-genetic treatments will be MAINSTREAMED by OBAMA'S AFFORDABLE CARE ACT medical boards filled with global banking 5% freemason/Greek players creating policies around a 99% of citizens' HEALTH INSURANCE POLICIES--------and telling our local DOCTORS these bio-genetic treatments are REQUIRED.
THIS IS INDEED THE GOALS OF MOVING FORWARD AFFORDABLE CARE ACT------OF IGNORING AND ELIMINATING ALL US FEDERAL REQUIREMENTS FOR CLINICAL TRIALS AND SCIENTIFIC METHOD REAL DATA AND CONCLUSIONS.
Vaccines (Basel). 2014 Sep; 2(3): 624–641.
Published online 2014 Jul 29. doi: 10.3390/vaccines2030624
PMCID: PMC4494222
PMID: 26344749
Developments in Viral Vector-Based Vaccines
Takehiro Ura, Kenji Okuda, and Masaru Shimada*
Abstract
Viral vectors are promising tools for gene therapy and vaccines. Viral vector-based vaccines can enhance immunogenicity without an adjuvant and induce a robust cytotoxic T lymphocyte (CTL) response to eliminate virus-infected cells. During the last several decades, many types of viruses have been developed as vaccine vectors. Each has unique features and parental virus-related risks. In addition, genetically altered vectors have been developed to improve efficacy and safety, reduce administration dose, and enable large-scale manufacturing. To date, both successful and unsuccessful results have been reported in clinical trials. These trials provide important information on factors such as toxicity, administration dose tolerated, and optimized vaccination strategy. This review highlights major viral vectors that are the best candidates for clinical use.
Keywords: viral vector, vaccine, CTL, MVA, adenovirus
1. Introduction
Viral vectors are regarded as potential tools for gene therapy and vaccines. Their utility is based on the ability of viruses to infect cells. In general, the advantages of viral vectors are as follows: (a) high efficiency gene transduction; (b) highly specific delivery of genes to target cells; and (c) induction of robust immune responses, and increased cellular immunity.
The concept of viral vector vaccines is different from that of subunit vaccines, as the latter help prevent infectious diseases by eliciting a humoral response. Recombinant viral vectors have potential for therapeutic use because they enable intracellular antigen expression and induce a robust cytotoxic T lymphocyte (CTL) response, leading to the elimination of virus-infected cells. Despite their efficacy, viral vectors present unavoidable problems that need to be addressed. In the near future, viral vector-based vaccines may be increasingly used to fight major diseases, such as HIV-1 and malaria. In some vectors, stable expression of the interesting gene is achieved via viral integration mechanisms. Integration into the host genome can lead to cancer. Another obstacle to the clinical use of viral vectors is the presence of pre-existing immunity against the vector. This is caused by previous exposure to the virus and the production of neutralizing antibodies that reduce vaccine efficacy.
The development of viral vectors requires a high biological safety level in order to gain public acceptance. Therefore, non- (or low-) pathogenic viruses are often selected. In most cases, viruses are genetically engineered to reduce or eliminate pathogenicity. Additionally, most viral vectors are replication-defective. For example, in adenovirus-based vectors, the E1A and E1B encoding regions, which are needed for replication in infected cells, are deleted and replaced with the target gene.
When using a viral vector, it is important to assess the potential implications by understanding the epidemiological and virological characteristics. In this review, we describe several representative viral vectors with respect to risk and benefit, optimized vector-design attempts, and clinical results. Moreover, we discuss the combined use of viral vectors in prime-boost vaccination regimens.
2. Summary of Viral Vectors
The concept of the viral vector was introduced in 1972. Jackson et al. created recombinant DNA from the SV40 virus by genetic engineering [1]. Subsequently, Moss et al. reported the use of vaccinia virus as a transient gene expression vector in 1982 [2,3]. Several types of viral vectors have been developed, and they have been used in animal studies and clinical trials.
The specific properties of a vector are determined by the virus from which it derives. Each vector has distinct advantages and disadvantages (Table 1). Vaccinia virus and adenovirus are the most widely used vectors because they can induce a robust immune response, specifically involving CTL, against the expressed foreign antigens. Generally, viral vectors achieve high immunogenicity without an adjuvant. Viral components stimulate the innate immune response, leading to the production of interferons and inflammatory cytokines [4].
Table 1
Advantages and disadvantages of major viral vectors.
VirusAdvantagesDisadvantages
Major Clinical/Preclinical Studies
RetrovirusLong-term gene expression
Generation of replication-competent virus Potential for tumorigenesis
Infects dividing cells only[6]
Lentivirus
Long-term gene expression Infects non-dividing and dividing cells
Generation of replication-competent virus Potential for tumorigenesis[7,8]
Vaccinia virus
High immunogenicity Safety: used as a smallpox vaccine High titer productionPre-existing immunity[9]
Adenovirus
High immunogenicity Safety: used in many clinic trails High titer productionPre-existing immunity[10]
Adeno-associated virus
Long-term gene expression Non-pathogenic virusLow titer production[11]
Cytomegalovirus
Induces a unique CTL response Protects against SIV infection in an animal modelPre-existing immunity Risk of pathogenesis in specific individuals[12]
Sendai virus
High immunogenicity
Pre-existing immunity[13]Viral vector-based vaccines require assessment of efficacy and safety, including immunogenicity, genetic stability, ability to evade pre-existing immunity, replication deficiency or attenuation, and genotoxicity. Additionally, the cost-effectiveness must also be evaluated because infectious diseases are a problem in developing countries. Thus, it is necessary to consider the large-scale manufacturing of viral vectors. Typically, the manufacturing of viral vectors involves propagation in suitable cell lines.
The European Medicines Agency (EMA) recently provided guidelines on quality for non-clinical and clinical aspects [5]. These guidelines offer useful recommendations for the effective administration of live, recombinant viral vector-based infectious vaccines.
_____________________________________
'APHA is more than a professional association.
We're a community. Our members work in every discipline of public health, in every U.S. state and in countries across the globe'.
WE CAN'T STOP THIS shout those global banking 5% freemason/Greek players/pols MOVING FORWARD ONE WORLD ONE WORLD HEALTH ORGANIZATION inside US FOREIGN ECONOMIC ZONES!
OH, REALLY?????
Back in 1990s Clinton era we see below for whom these far-right wing global banking NEO-LIBERALS and NEO-CONS were working-----in 1990s 750 biotechnology corporations to include bio-genetics. 30 years ago many of these were regional businesses while today much of this industry has been consolidated into global banking 1% OLD WORLD KINGS medical corporations ----global MITRE CORPORATION THE HOSPITALLERS for example.
If we think today how many of these bio-tech/bio-genetic corporations exist and operate mostly OUTSIDE of US public health and safety ====that would be in the millions globally-----tens of thousands inside US.
'The Biotechnology Industry Organization (BIO) represents over 745 biotechnology companies, academic institutions, and state biotechnology centers in 46 states and 25 nations'.
Our local US city/county public health departments do not have to install these FEDERAL GUIDELINES-------our local PUBLIC HEALTH DEPARTMENTS CAN keep to our last century MEDICAL MORALS AND ETHICS.
Again, we must look to whom is appointed to VITAL public health positions. Our local public medical universities have THE PROVOST-----that DEAN who pushes OR NOT these health policies. We have that city public health department with a director who pushes OR NOT these health policies.
Below we see an association of what we KNOW are global banking 5% freemason/Greek players owning tech businesses or working for global medical corporations. These kinds of associations USED TO BE OUR COMMUNITY CITIZENS with businesses that HELPED AND KEPT SAFE our 99% of citizens.
IT MATTERS WHO IS APPOINTED----IT MATTERS FOR WHOM OUR CITY/COUNTY ASSOCIATIONS WORK. LET'S DO IT----WORK TO GET RID OF THESE GLOBAL BANKING 5% FREEMASON/GREEK PLAYERS WHO DO NOT CARE.
APHA is more than a professional association.
We're a community. Our members work in every discipline of public health, in every U.S. state and in countries across the globe.
And we're all united by a common goal: To create the healthiest nation in one generation.
Join APHA.
Together, we really can make a difference.
Why should you join?Exclusive benefits like a network of 25,000 peers who also are working to make a difference.
Special member savings on college tuition, professional certifications, insurance and more!
How much does it cost?That depends on your membership category. Learn more on our Member Types and Rates page.
How can I get the most out of my APHA membership?Watch our video series on such topics as advancing your career, navigating the Annual Meeting, earning CE credits and advocating for public health.
And while you're browsing APHA.org, notice the Member Perk image that highlights some of the great benefits of APHA membership, including access to our award-winning publications, opportunities for professional development and discounts on continuing education opportunities.
_________________________________________
GREEK TRAGEDIES --------MEDIEVAL MORALITY PLAYS-------AGE OF ENLIGHTENMENT SENSE AND SENSIBILITY PHILOSOPHY have told our 99% WE THE PEOPLE that selling our souls to the DEVIL which is what has MOVED FORWARD these few decades of CLINTON/BUSH/OBAMA staging to be PERMANENT in ONE WORLD ONE GOVERNANCE-----always ends badly for that PLAYER and often that PLAYER takes that family down as well.
youtube.com
Scratch's Song ( The Devil and Daniel Webster)
I Ringwald
Published on Apr 12, 2013
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