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Dec 6, 2013, 4:44 PM
(The ME/CFS patient community actually knew about this at the time it came out in 2011and discussed it on patient forums, fb, and with comments on media articles).
Robyn Erland shared a link.
I would like to point this out. More to follow:
Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Abstract
Purpose: To investigate the frequency of xenotropic murine leukemia virus (MLV) presence in human cell lines established from mouse xenografts and to search for the evidence of horizontal viral spread to other cell lines. Methodology: We examined xenograft tumor cell lines from 7 independent laboratories and 128 non-xenografted tumor cell lines. Cell line DNA was examined for mouse DNA contamination, and by three Taqman qPCR assays targeting the gag, env or pol regions of MLV. Sequencing was used for viral strain identification. Supernatant fluids were tested for reverse transcriptase (RT) activity. Results: Six of 23 (26%) mouse DNA free xenograft cultures were strongly positive for MLV and their sequences had greater than 99% homology to known MLV strains. Four of five available supernatant fluids from these viral positive cultures were strongly positive for RT activity. Three of these supernatant fluids were studied to confirm the infectivity of the released virions for other human culture cells. Of the 78 non-xenograft derived cell lines maintained in the xenograft culture-containing facilities, 13 (17%) were positive for MLV, including XMRV, a virus strain first identified in human tissues. By contrast, all 50 cultures maintained in a xenograft culture-free facility were negative for viral sequences. Conclusions: Human cultures derived after mouse xenografting frequently contain and release highly infectious xenotropic MLV viruses. Laboratories working with xenograft-derived human cultures should be aware of the risk of contamination with potentially biohazardous human-tropic mouse viruses and their horizontal spread to other cultures.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3218386/
Dec 6, 2013, 4:44 PM
(The ME/CFS patient community actually knew about this at the time it came out in 2011and discussed it on patient forums, fb, and with comments on media articles).
Robyn Erland shared a link.
I would like to point this out. More to follow:
Frequent detection of infectious xenotropic murine leukemia virus (XMLV) in human cultures established from mouse xenografts
Abstract
Purpose: To investigate the frequency of xenotropic murine leukemia virus (MLV) presence in human cell lines established from mouse xenografts and to search for the evidence of horizontal viral spread to other cell lines. Methodology: We examined xenograft tumor cell lines from 7 independent laboratories and 128 non-xenografted tumor cell lines. Cell line DNA was examined for mouse DNA contamination, and by three Taqman qPCR assays targeting the gag, env or pol regions of MLV. Sequencing was used for viral strain identification. Supernatant fluids were tested for reverse transcriptase (RT) activity. Results: Six of 23 (26%) mouse DNA free xenograft cultures were strongly positive for MLV and their sequences had greater than 99% homology to known MLV strains. Four of five available supernatant fluids from these viral positive cultures were strongly positive for RT activity. Three of these supernatant fluids were studied to confirm the infectivity of the released virions for other human culture cells. Of the 78 non-xenograft derived cell lines maintained in the xenograft culture-containing facilities, 13 (17%) were positive for MLV, including XMRV, a virus strain first identified in human tissues. By contrast, all 50 cultures maintained in a xenograft culture-free facility were negative for viral sequences. Conclusions: Human cultures derived after mouse xenografting frequently contain and release highly infectious xenotropic MLV viruses. Laboratories working with xenograft-derived human cultures should be aware of the risk of contamination with potentially biohazardous human-tropic mouse viruses and their horizontal spread to other cultures.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3218386/
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I found this paper back in 2011 and posted it on mecfsforums - a patient forum, where it was discussed at length!
Unintended spread of a biosafety level 2 recombinant retrovirus
Retrovirology 20096:86
Background
Contamination of vertebrate cell lines with animal retroviruses has been documented repeatedly before. Although such viral contaminants can beeasily identified with high sensitivity by PCR, it is impossible to screen for all potential contaminants. Therefore, we explored two novel methods to identify viral contaminations in cell lines without prior knowledge of the kind of contaminant.
Results
The first hint for the presence of contaminating retroviruses in one of our cell lines was obtained by electron microscopy of exosome-like vesicles released from the supernatants of transfected 293T cells. Random amplification of particle associated RNAs (PAN-PCR) from supernatant of contaminated 293T cells and sequencing of the amplicons revealed several nucleotide sequences showing highest similarity to either murine leukemia virus (MuLV) or squirrel monkey retrovirus (SMRV). Subsequent mass spectrometry analysis confirmed our findings, since we could identify several peptide sequences originating from monkey and murine retroviral proteins. Quantitative PCRs were established for both viruses to test currently cultured cell lines as well as liquid nitrogen frozen cell stocks. Gene fragments for both viruses could be detected in a broad range of permissive cell lines from multiple species. Furthermore, experimental infections of cells negative for these viruses showed that both viruses replicate rapidly to high loads. We decided to further analyze the genomic sequence of the MuLV-like contaminant virus. Surprisingly it was neither identical to MuLV nor to the novel xenotropic MuLV related retrovirus (XMRV) but showed 99% identity to a synthetic retrovirus which was engineered in the 1980s.
Conclusion
The high degree of nucleotide identity suggests unintended spread of a biosafety level 2 recombinant virus, which could also affect the risk assessment of gene-modified organisms released from contaminated cell cultures. The study further indicates that both mass spectrometry and PAN-PCR are powerful methods to identify viral contaminations in cell lines without prior knowledge of the kind of contaminant. Both methods might be useful tools for testing cell lines before using them for critical purposes.
https://retrovirology.biomedcentral.com/articles/10.1186/1742-4690-6-86?fbclid=IwAR0aU2Zk1vG7idj_NQZbqz6zzpyNmjuzqAOOvJmowb66WNCKNJAjv4clqCY
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Nov 16, 2013, 2:00 PM
Robyn Erland shared a link.
Check out the new paper just published listed below: This is what they found: "Altogether, our findings showing that in vitro infection by retrovirus vectors leads to replication perturbations raises new concerns regarding the safety of these vectors. This replication perturbation may increase genome instability and mutation rates that could lead to tumorigenicity. This risk is greater especially in patients with challenged immune systems, such as those with SCID. Hence, future studies should explore the molecular basis of this replication stress, and the ways in which it affects specific risks associated with gene therapy trials and treatments."
Infection with retroviral vectors leads to perturbed DNA replication increasing vector integrations into fragile sites Here's a link to the full paper:
http://www.nature.com/srep/2013/130715/srep02189/full/srep02189.html
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Robyn Erland shared a link.
Check this out here are the highlight of this abstract:
"We studied the ability of one of these approaches, based on degenerate oligonucleotide primer (DOP)-polymerase chain reaction (PCR), to detect viral sequences in cell lines known to express viral genes or particles."
It detected this in those cell lines: Detected viral sequences included nodavirus, bracovirus, and endogenous retroviruses in High Five cells, porcine circovirus type 1 and porcine endogenous retrovirus in PK15 cells, human T-cell leukemia virus 1 in MJ cells, human papillomavirus 18 in HeLa cells, human herpesvirus 8 in BCBL-1 cells, and Epstein–Barr Virus in Raji cells.
It is for "identifying contaminants in clinical and biological samples, including cell lines and reagents used to produce vaccines and therapeutic products."
THEY FOUND ALL THESE VIRUSES IN CELL LINES, AND THIS TEST CAN BE USED TO LOOK FOR RETROVIRUSES AND VIRUSES IN CELL LINES USED IN VACCINES AND THERAPEUTIC PRODUCTS!!!
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E-mail I sent to another:
Sent: Thursday, July 24, 2014 9:09 PM
To: xxxxxx
Subject: cancer cell table and links
To: xxxxxx
Subject: cancer cell table and links
Hi, I stumbled across this paper (cell line table attached) again about the murine gammaretroviruses found in the cell lines. Here are some other links too for what's been going on in the cancer xenografting labs, and the warnings of potential problems that they've known about for years with the xenotransplantation. I wanted to send you some of the xenotransplantation and FDA stuff I've been finding. I've also glanced in the vaccine area of the FDA site. Some of it is very interesting:
http://www.frontiersin.org/Molecular_and_Cellular_Oncology/10.3389/fonc.2013.00156/abstract
Murine Endogenous Retroviruses in Human Cancer Cell Lines
The lesson of XMRV is a potentially important one. The fact that XMRV arose from xenotransplantation (i.e., xenografting) of CWR22Rv1 through mice – a very common practice when developing cancer cell lines – is particularly concerning. Although most researchers appear to be unaware of the potential contamination threat (Zhang et al., 2011), the infection of xenografted human cell lines with xenotropic retroviruses is well-established in the literature, dating back to the early 1970s (Takeuchi et al., 2008; Sfanos et al., 2011). In 1972, an endogenous feline retrovirus, RD114, was found to have infected human rhabdomyosarcoma cells that had been xenotransplanted through a fetal kitten brain (McAllister et al., 1972). In 1973, there was a similar finding in that a murine type-C retrovirus was isolated from rhabdomyosarcoma cells that had been xenotransplanted through NIH Swiss mice (Todaro et al., 1973). However, the implications of these and other findings have arguably not been fully realized. To date, and particularly after the XMRV controversy, several additional reports highlight the widespread issue of xenotropic MLV (XMLV) contamination of xenotransplanted cell lines.
The discovery and subsequent debunking of XMRV as a human pathogen brought back to light the issue of XMLV infection of xenotransplanted cell lines. This includes potentially artifactual experimental results due to virus-induced changes in cellular behavior, as well as the cross-contamination of uninfected cell lines grown in the same laboratory. With these dangers in mind, precautions, such as routine testing for XMLVs in human cell lines developed by xenotransplantation, or any cell lines cultured in laboratories concurrently growing xenotransplanted or known XMLV-infected cell lines, may warrant serious consideration.
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Here's information on John Coffin and Harold Varmus and animal product biotechnology and the IOM:
The IOM are the ones redefining ME/CFS and Coffin, Varmus and even Gallo have been members of the institute of Medicine/Academy of sciences since the 80's and 90's. Here's Coffins current membership:
IOM Directory: Committee Member - Dr. John M. Coffin
Dr. John M. Coffin
Tufts University School of Medicine
Massachusetts
•Other ParticipationThese activities reflect projects of the National Academy of Sciences, the National Academy of Engineering, and the National Research Council.
•Subcommittee on Defining Science-Based Concerns Associated with Products of Animal Biotechnology
•Committee on Transgenic Nomenclature
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Harold Varmus
A member of the National Academy of Sciences and the Institute of Medicine since 1993
President of Memorial Sloan Kettering Cancer Center
Director of the National Institutes of Health from 1993 to 1999. Director of NIH when worldwide ban on xenotransplantation was reinstated in 1999 for Porcine (and other) retroviruses
Director of the National Cancer Institute (NCI) in May 2010
recognized for his research on retroviruses
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This is still the guideline they use today and is still listed on the FDA xenotransplantation area:
1996 Xenotransplantation
Science, Ethics, and Public Policy
Committee on Xenograft Transplantation: Ethical Issues and Public Policy
Division of Health Sciences Policy
INSTITUTE OF MEDICINE
NATIONAL ACADEMY PRESS
NATIONAL ACADEMY PRESS
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Here's more:
Mouse cell therapy for brain tumors
Implantation of mouse cells releasing retrovirus vectors bearing a suicide gene has proven an effective treatment for brain tumors in experimental animals3,42,43. In clinical trials throughout the world, hundreds of patients have now been implanted with such mouse cells44. It is estimated that in the United States there are approximately 35,000 individuals with malignant gliomas. For these patients the prognosis is devastating. The time from diagnosis to death for glioblastoma multiforme usually ranges from a few months to a year, even with access to surgery, radiation and chemotherapy46. Thus an affected individual has much to gain from a few more months of life, and many of the risks to the individual undergoing this gene therapy protocol (hemorrhage and loss of cerebral function, for example) are already inherent in current treatments. However, the potential benefits of this procedure must be weighed against the potential risks of generating pathogens harmful to others.
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Genetic profiling reveals an alarming rate of cross-contamination among human cell lines used in China
US cancer institute to overhaul tumour cell lines
Cancer/ Genetics/ Life Science/ Medicine
From: Nature News - Full Text Published: February 24, 2016
After more than 25 years of heavy use by researchers around the world, the US National Cancer Institute (NCI) has decided to retire the NCI-60, its panel of 60 human cancer cell lines grown in culture, from its drug-screening programme. In late spring of this year, the institute will launch a rejuvenated repository of cancer models that are derived from fresh patient samples and tagged with details about their clinical past.
The NCI action responds to a widespread push for cancer models with a closer link to the patients they are intended to help. On 11 February, cancer researchers gathered in New Orleans, Louisiana, for a meeting hosted by the American Association for Cancer Research that focused on the creation of new cancer models from clinical samples.
Since 1990, industry and academic have screened more than 100,000 compounds using the NCI-60, in order to study the molecular details of cancers and find drugs to treat them.
When the NCI-60 was established, researchers had a very different conception of cancer, says James Doroshow, director of the Division of Cancer Treatment and Diagnosis at the NCI in Bethesda, Maryland. “Thirty years ago, the idea was that if you found a drug that worked on six breast cancer cell lines, then you could use it to treat breast cancer,” he says. “Well, it doesn’t work that way.”
https://scifeeds.com/social-media-item/us-cancer-institute-to-overhaul-tumour-cell-lines/
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This is what's currently on the FDA site:
FDA website for Xenotransplantation: Viral Safety Studies in Xenotransplantation and Gene Therapy Products Principal Investigator: Carolyn A. Wilson, Ph DOffice / Division / Lab: OCTGT / DCGT / GTIBGeneral Overview Gammaretroviruses are simple animal viruses that are related to HIV, the cause of AIDS.These viruses might pose risks in several regulated biologics products, both as contaminants and in their use for gene therapy, in which they act as carriers, or "vectors" to deliver genes that treat a disease by producing a therapeutic molecule.
Xenotransplantation is any procedure that involves the transplantation, implantation or infusion into a human recipient of either (a) live cells, tissues, or organs from a nonhuman animal source, or (b) human body fluids, cells, tissues or organs that have had ex vivo contact with live nonhuman animal cells, tissues or organs. The development of xenotransplantation is, in part, driven by the fact that the demand for human organs for clinical transplantation far exceeds the supply.
Although the potential benefits are considerable, the use of xenotransplantation raises concerns regarding the potential infection of recipients with both recognized and unrecognized infectious agents and the possible subsequent transmission to their close contacts and into the general human population. Of public health concern is the potential for cross-species infection by retroviruses, which may be latent and lead to disease years after infection. Moreover, new infectious agents may not be readily identifiable with current techniques.
Robyn Erland commented on her own FB link.
Dec 08, 2013 12:25pm
Well yes likely there were mistakes as well. They were using various animals to try and produce vaccines as far back as the late 1800's. Very doubtful they were careful or knew much about what could happen if injected into humans. And there's also the fact John coffin originally said that retroviruses were harmless. Then we know the biosafety level was changed to a higher risk level in 1995. And now Wallah! they are finding mlv's contaminated in cell lines and lines that are just housed in the same labs. Anyway it's looked at I agree though it wasn't good to open Pandora's box.
Robyn Erland shared a link. From facebook
Dec 06, 2013 4:44pm
Optimization of virus detection in cells using massively parallel sequencing
sciencedirect.com
Check this out here are the highlight of this abstract:
"We studied the ability of one of these approaches, based on degenerate oligonucleotide primer (DOP)-polymerase chain reaction (PCR), to detect viral sequences in cell lines known to express viral genes or particles."
It detected this in those cell lines: Detected viral sequences included nodavirus, bracovirus, and endogenous retroviruses in High Five cells, porcine circovirus type 1 and porcine endogenous retrovirus in PK15 cells, human T-cell leukemia virus 1 in MJ cells, human papillomavirus 18 in HeLa cells, human herpesvirus 8 in BCBL-1 cells, and Epstein–Barr Virus in Raji cells.
It is for "identifying contaminants in clinical and biological samples, including cell lines and reagents used to produce vaccines and therapeutic products."
THEY FOUND ALL THESE VIRUSES IN CELL LINES, AND THIS TEST CAN BE USED TO LOOK FOR RETROVIRUSES AND VIRUSES IN CELL LINES USED IN VACCINES AND THERAPEUTIC PRODUCTS!!!
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Robyn Erland shared a link.
So these genius's want to insert the gammaretroviruses into the cancer genes insertion areas of humans to see how the genes react to to a cancer causing gammaretroviruses. How stupid do they think people are? Haven't they caused enough problems? JUST STOP MESSING WITH THESE RETROVIRUSES. A new paper just out and is this why many of us wind up with cancer? oh let's see gammaretroviruses love the cancer insertions gene areas of humans. And inserting them into the cancer causing areas in studies will help them learn how to come up with cancer treatments. Here's the paper:
Gamma-retroviruses preferentially integrate near cancer-associated genes
Profiling gamma-retrovirus integration sites may be a new tool to identify genes promoting specific cancers
Date: May 10, 2016
Source: PLOS
Summary: Identifying the sites where gamma-retroviruses commonly insert into the genome may help to identify genes associated with specific cancer types, according to a new study.
Identifying the sites where gamma-retroviruses commonly insert into the genome may help to identify genes associated with specific cancer types, according to a study published April 20, 2016 in the open-access journal PLOS ONE
Gamma-retroviruses, such as feline leukemia virus, tend to cause mutations when they insert into a host's genome, and have been used as a tool to discover genes associated with cancer. However, this discovery process can be time consuming, requiring the collection of multiple tumors from animals and comparative genomic analyses.
The authors of the present study sought to investigate the pattern of gamma-retrovirus insertion using deep sequencing to analyse common insertion sites for feline leukemia virus in cell culture. The study was also expanded to analyze published genome insertion profiles of other gamma-retroviruses.
The authors found that the gamma-retroviruses preferentially inserted into cancer-driving genes, regardless of transcription levels, in a cell type-specific manner.
https://www.sciencedaily.com/releases/2016/05/160510084207.htm
May 11, 2016, 3:05 PM
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Robyn Erland shared a link.
Incompetence even at the Bioterror Germ level of the CDC. Check out the USA Today article that came out today! Now do we wonder why we are all getting sick? When we have people like this running labs and facilities, who have also been creating and covering up data, and the fact these agencies have been contaminating labs all over the world with INFECTIOUS RETROVIRUSES too for decades!
CDC labs repeatedly faced secret sanctions for mishandling bioterror germs
"A laboratory operated by the Centers for Disease Control and Prevention is among the handful of facilities that have secretly had their permits suspended in recent years for serious safety violations while working with bioterror pathogens, according to documents obtained by USA TODAY after winning a Freedom of Information Act appeal."
And now we find out about SECRET SANCTIONS!
http://www.usatoday.com/story/news/2016/05/10/cdc-lab-secret-sanctions/84163590/
May 11, 2016, 3:05 PM
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Robyn Erland shared a link.
Incompetence even at the Bioterror Germ level of the CDC. Check out the USA Today article that came out today! Now do we wonder why we are all getting sick? When we have people like this running labs and facilities, who have also been creating and covering up data, and the fact these agencies have been contaminating labs all over the world with INFECTIOUS RETROVIRUSES too for decades!
CDC labs repeatedly faced secret sanctions for mishandling bioterror germs
"A laboratory operated by the Centers for Disease Control and Prevention is among the handful of facilities that have secretly had their permits suspended in recent years for serious safety violations while working with bioterror pathogens, according to documents obtained by USA TODAY after winning a Freedom of Information Act appeal."
And now we find out about SECRET SANCTIONS!
http://www.usatoday.com/story/news/2016/05/10/cdc-lab-secret-sanctions/84163590/
Jul 21, 2013, 8:14 PM
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Robyn Erland shared a link.
Let's remember that XMRV was a lab created retrovirus that came as a result of xenografting in cancer research. They didn't know until 2011 that XMRV and other infectious mlv retroviruses, that researchers were also lab creating in xenograft cancer research, were becoming airborne and getting into cell lines used in biologicals. For those who haven't heard about it yet this paper explains:
Identification of Replication Competent Murine Gammaretroviruses in Commonly Used Prostate Cancer Cell Lines
PLOS Published: June 17, 2011
DOI: 10.1371/journal.pone.0020874
"Our study demonstrates that prostate cancer cell lines appear to have a propensity for infection by murine gammaretroviruses. We propose that this tendency may be due to the fact that most of the established prostate cancer cell lines were created by passage through immunocompromised mice. In fact, Bxv-1 is present in SCID and nude mice and tumor cells passaged through these animals can become infected with xenotropic MLVs [22]. Although the infection of human cell lines with murine gammaretroviruses has been previously described in the literature, what makes our current study particularly important is that prior to this study it was not known that multiple commonly used prostate cancer cell lines are contaminated with MLVs. In fact, we discovered during the course of our study that other cell lines maintained in the laboratory (DU145, LNCaP) have apparently become infected with XMRV in the laboratory."
"There are several reported examples of retroviral infection modifying the biological properties of human cell lines. For example, the in vivo immunogenicity of cell lines has been shown to be strongly altered following retroviral infection after a single passage in nude mice [24]. Likewise, xenotropic MLVs acquired in cell lines used in HIV research after in vivo passage in immunocompromised mice were shown to alter the biological properties of HIV-1."
"It is not known what confounding effects the infection of commonly used prostate cancer cell lines with replication competent murine gammaretroviruses may have on experimental outcomes. Interestingly, it has been reported that XMRV proteins are more abundant in cell culture media of CWR22Rv1 cells than any human protein [4]. For reasons such as this, we suggest that cancer cell lines should undergo routine screening for contaminating MLVs, much like the current practice of routine testing of cultured cells for mycoplasma."
"The discovery of infectious XMRV in the prostate tumor cell line 22Rv1 prompted the examination of other prostate tumor cell lines for the presence of murine gammaretroviruses. Antisera against Moloney murine leukemia virus were used to screen 72 cell lines."
"How did these human prostate cancer cell lines become contaminated with murine gammaretroviruses? The authors believe this is a consequence of passage of the tumors through immunocompromised mice."
"Out of concern that prostate cancer cell lines producing replication competent viruses could cross-contaminate other cell lines in our laboratory which are negative for MLVs, we tested current cultures in our laboratory for the presence of MLVs. We were surprised to find two instances where MLV-negative cell lines maintained in the laboratory (DU145, LNCaP) have become contaminated with MLVs" http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0020874
Dec 21, 2015, 5:51 PM
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E-mail I sent to another on May 1, 2015, at 10:33 AM
Subject: RE: mlvs in cell lines
That's the link to the paper referenced below. They found human cell lines infected with mlvs at 3.3%. I've attached this full paper. http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0125622
In conclusion, we could demonstrate that a) 19 of 577 (3.3%) different human cell lines were contaminated with MLV; b) 17 of the MLV-positive cell lines produce active viruses; c) the infecting MLV constitute at least three evolutionary related groups, d) the retroviruses are transmissible from one infected cell culture to another. Other sources of infection could be heterotransplantation of the human cells and the use of murine feeder layer.
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Robyn Erland status from fb
The Vero cell line was derived from the kidney of a normal, adult, African green monkey (Cercopithecus) on March 27, 1962, by Y. Yasumura and Y. Kawakita at the Chiba University in Japan (Nippon Rinsho 21:1209, 1963). The cell line was brought to the
Laboratory of Tropical Vkology, NIAID, NIH, at passage Ilevel 93 from Chiba University by Dr. B. Simizu on June 15, 1964. There is documentation of the types of culture media and the concentration of bovine serum used in the media throughout its history. In addition to its use as a vaccine cell substrate, this cell line has been used extensively for virus replication studies and plaque assays. Vero cells are sensitive to infection with SV-40, SV-5, measles, arboviruses, reoviruses, rubella, simian adenoviruses, polioviruses, influenza viruses, parainfluenza viruses, respiratory syncytial viruses, vaccinia, and others. http://www.fda.gov/ohrms/dockets/ac/00/backgrd/3616b1a.pdf
August 29, 2016 12:10 PM--------------------------------------------------------------------
Robyn Erland shared a link.
Many cell lines have and still are contaminated with unidentified MLV infectious retroviruses.
Here's proof of the contaminated cell lines used in medicine and other biologicials. Database of Cross-contaminated or Misidentified Cell Lines -This database lists cell lines that are currently known to be cross-contaminated or otherwise misidentified. http://iclac.org/databases/cross-contaminations/
Here's another: Prevalence and Characterization of Murine Leukemia Virus Contamination in Human Cell Lines 2015, "Contaminations of cell cultures with microbiological organisms are well documented" However, the presence of viral contaminations in cell cultures is still a matter of debate and cannot be determined with general detection methods. In the present study we screened 577 human cell lines for the presence of murine leukemia viruses (MLV). Nineteen cell lines were found to be contaminated with MLV, including 22RV1 which is contaminated with the xenotropic murine leukemia virus-related virus variant of MLV. Of these, 17 cell lines were shown to produce active retroviruses determined by product enhanced reverse transcriptase PCR assay for reverse transcriptase activity. http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0125622,
And yet another. The ME/CFS patient community knew about all of these contaminated cell lines at the time they came out in 2011. We were discussing them at length on patients forums, facebook and as comments oon all major media aricles: Infection of Xenotransplanted Human Cell Lines by Murine Retroviruses: A Lesson Brought Back to Light by XMRV "Since many of the XMLV-infected cell lines in both the Sfanos et al. (2011) and Zhang et al. (2011) studies were found to be replication competent, the potential for cross-contamination of other cell lines grown in the same facilities is considerable. Indeed, Sfanos et al. (2011) reported that XMRV-negative cell lines that were being cultured in the laboratory at the same time as CWR22Rv1 had become contaminated. Similarly, Zhang et al. (2011) detected XMLV infection of 13 out of 78 non-xenografted cell lines from five different laboratories that were also culturing XMLV-infected xenografted cell lines. In contrast, all 50 cell lines gathered from non-xenograft culturing labs were XMLV-free.
The laboratories involved in both studies were using standard BSL2 aseptic technique, highlighting the infective potential of these XMLVs. Therefore, the dangers of XMLV contamination of human cell lines is not limited to xenografted lines, but potentially extends to all additional cell lines being cultured alongside infected lines. If contamination occurs and is not detected, there is significant potential for spreading of the retrovirus, even across laboratories sharing samples." https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3683812/
Apr 19, 2017, 1:47 PM
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Robyn Erland shared a link.
Viral Evaluation of Animal Cell Lines Used in Biotechnology
Abstract
Animal cell culture is being increasingly used for production of therapeutic reagents such as monoclonal antibodies, recombinant proteins, viral vaccines and replication incompetent viral vectors for gene therapy. Material derived from a variety of biological processes has been associated in the past with incidents involving the transmission of infectious agents, principally viruses (1). Thus, virological evaluation of animal cell substrates for use in the manufacture of biologicals is essential to ensure the safety of products for pharmaceutical use. http://link.springer.com/protocol/10.1385%2F0-89603-547-6%3A23
Mar 18, 2017, 12:09 AM
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Robyn Erland shared a link.
Does everyone know that the first Flu vaccine made from recombinant technology was put on the market in 2013? They actually have 4 types of flu vaccines this season and this Flublok is for those 18 and over.
This is what Recombinant technology is (it's also good to know that these cells are sometimes made from "mammalian cells"):
"A recombinant vaccine is a vaccine produced through recombinant DNA technology. This involves inserting the DNA encoding an antigen (such as a bacterial surface protein) that stimulates an immune response into bacterial or mammalian cells, expressing the antigen in these cells."
What is Recombinant DNA?":
Recombinant protein is a protein that whose code is carried through a recombinant DNA. The term recombinant DNA implies that two segments of DNA in a plasmid."
Recombinant DNA (rDNA) molecules are DNA molecules formed by laboratory methods of genetic recombination (such as molecular cloning) to bring together genetic material from multiple sources, creating sequences that would not otherwise be found in the genome.
My note: Now this new Flublok vaccine didn't work so well in the 2013 - 2014 season. So they came up with a new version for 2015 - 2017. Here is all the crap they put in it:
Vaccine - Antigenic Composition - Flublok Quadrivalent
45 mcg per antigen
H1N1:A/California/07/2009
H3N2: A/Texas/50/2012B/Massachusetts/2/2012
(B/Yamagata-lineage)
B/Brisbane/60/2008 (B/Victoria -lineage)
And they are in the process of making numerous Recombinant DNA new vaccines. They are calling them VLP's "virus like particles") They are basically taking a part of the Flu virus and mass producing it with nano- micro particles of the flu put in. They use something clalled hemagglutinin (HA) or haemagglutinin which is a glycoprotein found on the surface of influenza viruses.
Now The very first surface glycoprotein or shell DNA recombinant vaccines tried were the HIV vaccines. They did not work and it was said followup studies were needed to be done, to determine if the subjects were actually being given HIV. Knowing what we know now about what's happening in Asia with all the new HIV strains and recombinations, it's a safe bet more "lab created recombinations" of these particles and retroviruses are sure to come. People have no clue what is being injected in them. I asked a few who've been ill almost a month now if they knew what Recombinant DNA vaccine is? They looked at me and said No.
Here's the information where they are comparing it to another type of vaccination:
https://www.cdc.gov/vaccines/acip/meetings/downloads/slides-2016-02/flu-03-hachey.pdf
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Robyn Erland shared a link. (this was one of the FDA postings I originally found when I realized that full mouse brains had also been being used in JE vaccines in the US too)
WOW this is going to blow your minds. I found where FDA scientist Khan put out a presentation in 2009 and one of the cell lines listed was from mouse brains ( This info has since been removed from the FDA site):
U.S. Licensed Viral Vaccines:
Primary Cells or Tissues
Cell Substrate - Inactivated Vaccines
Mouse brain - Japanese Encephalitis
Now in 2011 they made a 2nd mouse brain vaccine for Japanese Encephalitis and they have been vaccinating people all over ASIA North America, Australia and various European countries!
A Vero cell-derived, inactivated and alum-adjuvanted JE vaccine based on the SA 14-14-2 strain was approved in 2009 in North America, Australia and various European countries. The primary two doses are administered 4 weeks apart. A booster dose is recommended 1–2 years after the primary immunization. This vaccine has been given concomitantly with hepatitis A vaccine without significant interference with the safety and immunogenicity of either vaccine. Data on concomitant administration with other vaccines frequently used in travellers are currently unavailable. The vaccine is licensed for use in individuals 17 years of age and older in the United States, and 18 years and above in other countries. Paediatric and post-marketing safety studies are under way.
Another Vero cell-derived inactivated JE vaccine was licensed by the Japanese authorities in February 2009 and a similar Japanese vaccine was licensed in 2011. These two vaccines use the same strain of JE virus (Beijing-1) as the mouse-brain-derived vaccine. Clinical trials have shown that the vaccines are safe and immunogenic, with seroconversion rates exceeding 95%.
In addition, a new live attenuated, JE–yellow fever chimeric vaccine has recently been licensed in Australia and Thailand. A single dose of this chimeric JE vaccine was found to be safe, highly immunogenic and capable of inducing long-lasting immunity in both preclinical and clinical trials.
http://www.who.int/ith/vaccines/japanese_encephalitis/en/
Jul 3, 2016, 9:13 PM
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Feb 27, 2017, 3:02 PM
Robyn Erland shared a link.
Here is a slideshow from the FDA rep Khan. from 2009. This slide 23 he explains what it needed to reduce the risk associated CBER products. These are the products used in vaccines allergy shots, blood products and other biologicals. Notice where it says "REPLACE ANIMAL-DERIVED MATERIALS". That is something they have still NOT done. And they DID not have adequate technology to detect
Efforts to minimize the risk associated with Biological Raw
Materials
• Replace animal-derived materials – Serum-free cell cultures
• Use of new technologies for virus inactivation and removal
• Development of more sensitive and broad detection assays for adventitious agent detection and characterization to identify source
http://c.ymcdn.com/sites/www.casss.org/resource/resmgr/imported/WCBPCMC09KhanSlides.pdf
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Jul 3, 2016, 12:46 PM
Robyn Erland shared a link.
From the FDA site and this notice came out on October 12 2009. from the attached announcement: "We are also evaluating the risk of retrovirus infections in humans. (Retroviruses are RNA viruses that use an enzyme called reverse transcriptase (RT) to replicate; RNA is the de-coded form of DNA)". Here's the announcement:
Investigating Viruses in Cells Used to Make Vaccines; and Evaluating the Potential Threat Posed by Transmission of Viruses to Humans
Virus-based vaccines are made in living cells (cell substrates). Some manufacturers are investigating the use of new cell lines to make vaccines. The continual growth of cell lines ensures that there is a consistent supply of the same cells that can yield high quantities of the vaccine.
In some cases the cell lines that are used might be tumorigenic, that is, they form tumors when injected into rodents. Some of these tumor-forming cell lines may contain cancer-causing viruses that are not actively reproducing. Such viruses are hard to detect using standard methods. These latent, or "quiet," viruses pose a potential threat, since they might become active under vaccine manufacturing conditions. Therefore, to ensure the safety of vaccines, our laboratory is investigating ways to activate latent viruses in cell lines and to detect the activated viruses, as well as other unknown viruses, using new technologies. We will then adapt our findings to detect viruses in the same types of cell substrates that are used to produce vaccines. We are also trying to identify specific biological processes that reflect virus activity.
These methods will enable FDA scientists to help manufacturers to determine whether their specific cell substrate is safe to use for vaccine production. The methods our laboratory are developing and testing will help to ensure the production of safe and effective vaccines in two ways: 1) FDA will be able to develop testing guidelines for manufacturers who use new cell substrates for producing vaccines; and 2) FDA will publish the new methods it develops in peer-reviewed scientific journals, thus making them readily accessible to all manufacturers. http://web.archive.org/web/20091012050025/http://www.fda.gov/BiologicsBloodVaccines/ScienceResearch/BiologicsResearchAreas/ucm127327.htm
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Robyn Erland shared a link.
Let's remember that XMRV was a lab created retrovirus that came as a result of xenografting in cancer research. They didn't know until 2011 that XMRV and other infectious mlv retroviruses, that researchers were also lab creating in xenograft cancer research, were becoming airborne and getting into cell lines used in biologicals. For those who haven't heard about it yet this paper explains:
Identification of Replication Competent Murine Gammaretroviruses in Commonly Used Prostate Cancer Cell Lines
PLOS Published: June 17, 2011
DOI: 10.1371/journal.pone.0020874
"Our study demonstrates that prostate cancer cell lines appear to have a propensity for infection by murine gammaretroviruses. We propose that this tendency may be due to the fact that most of the established prostate cancer cell lines were created by passage through immunocompromised mice. In fact, Bxv-1 is present in SCID and nude mice and tumor cells passaged through these animals can become infected with xenotropic MLVs [22]. Although the infection of human cell lines with murine gammaretroviruses has been previously described in the literature, what makes our current study particularly important is that prior to this study it was not known that multiple commonly used prostate cancer cell lines are contaminated with MLVs. In fact, we discovered during the course of our study that other cell lines maintained in the laboratory (DU145, LNCaP) have apparently become infected with XMRV in the laboratory."
"There are several reported examples of retroviral infection modifying the biological properties of human cell lines. For example, the in vivo immunogenicity of cell lines has been shown to be strongly altered following retroviral infection after a single passage in nude mice [24]. Likewise, xenotropic MLVs acquired in cell lines used in HIV research after in vivo passage in immunocompromised mice were shown to alter the biological properties of HIV-1."
"It is not known what confounding effects the infection of commonly used prostate cancer cell lines with replication competent murine gammaretroviruses may have on experimental outcomes. Interestingly, it has been reported that XMRV proteins are more abundant in cell culture media of CWR22Rv1 cells than any human protein [4]. For reasons such as this, we suggest that cancer cell lines should undergo routine screening for contaminating MLVs, much like the current practice of routine testing of cultured cells for mycoplasma."
"The discovery of infectious XMRV in the prostate tumor cell line 22Rv1 prompted the examination of other prostate tumor cell lines for the presence of murine gammaretroviruses. Antisera against Moloney murine leukemia virus were used to screen 72 cell lines."
"How did these human prostate cancer cell lines become contaminated with murine gammaretroviruses? The authors believe this is a consequence of passage of the tumors through immunocompromised mice."
"Out of concern that prostate cancer cell lines producing replication competent viruses could cross-contaminate other cell lines in our laboratory which are negative for MLVs, we tested current cultures in our laboratory for the presence of MLVs. We were surprised to find two instances where MLV-negative cell lines maintained in the laboratory (DU145, LNCaP) have become contaminated with MLVs" http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0020874
Dec 21, 2015, 5:51 PM
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E-mail I sent to another on May 1, 2015, at 10:33 AM
Subject: RE: mlvs in cell lines
That's the link to the paper referenced below. They found human cell lines infected with mlvs at 3.3%. I've attached this full paper. http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0125622
In conclusion, we could demonstrate that a) 19 of 577 (3.3%) different human cell lines were contaminated with MLV; b) 17 of the MLV-positive cell lines produce active viruses; c) the infecting MLV constitute at least three evolutionary related groups, d) the retroviruses are transmissible from one infected cell culture to another. Other sources of infection could be heterotransplantation of the human cells and the use of murine feeder layer.
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Robyn Erland status from fb
The Vero cell line was derived from the kidney of a normal, adult, African green monkey (Cercopithecus) on March 27, 1962, by Y. Yasumura and Y. Kawakita at the Chiba University in Japan (Nippon Rinsho 21:1209, 1963). The cell line was brought to the
Laboratory of Tropical Vkology, NIAID, NIH, at passage Ilevel 93 from Chiba University by Dr. B. Simizu on June 15, 1964. There is documentation of the types of culture media and the concentration of bovine serum used in the media throughout its history. In addition to its use as a vaccine cell substrate, this cell line has been used extensively for virus replication studies and plaque assays. Vero cells are sensitive to infection with SV-40, SV-5, measles, arboviruses, reoviruses, rubella, simian adenoviruses, polioviruses, influenza viruses, parainfluenza viruses, respiratory syncytial viruses, vaccinia, and others. http://www.fda.gov/ohrms/dockets/ac/00/backgrd/3616b1a.pdf
August 29, 2016 12:10 PM--------------------------------------------------------------------
Robyn Erland shared a link.
Many cell lines have and still are contaminated with unidentified MLV infectious retroviruses.
Here's proof of the contaminated cell lines used in medicine and other biologicials. Database of Cross-contaminated or Misidentified Cell Lines -This database lists cell lines that are currently known to be cross-contaminated or otherwise misidentified. http://iclac.org/databases/cross-contaminations/
Here's another: Prevalence and Characterization of Murine Leukemia Virus Contamination in Human Cell Lines 2015, "Contaminations of cell cultures with microbiological organisms are well documented" However, the presence of viral contaminations in cell cultures is still a matter of debate and cannot be determined with general detection methods. In the present study we screened 577 human cell lines for the presence of murine leukemia viruses (MLV). Nineteen cell lines were found to be contaminated with MLV, including 22RV1 which is contaminated with the xenotropic murine leukemia virus-related virus variant of MLV. Of these, 17 cell lines were shown to produce active retroviruses determined by product enhanced reverse transcriptase PCR assay for reverse transcriptase activity. http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0125622,
And yet another. The ME/CFS patient community knew about all of these contaminated cell lines at the time they came out in 2011. We were discussing them at length on patients forums, facebook and as comments oon all major media aricles: Infection of Xenotransplanted Human Cell Lines by Murine Retroviruses: A Lesson Brought Back to Light by XMRV "Since many of the XMLV-infected cell lines in both the Sfanos et al. (2011) and Zhang et al. (2011) studies were found to be replication competent, the potential for cross-contamination of other cell lines grown in the same facilities is considerable. Indeed, Sfanos et al. (2011) reported that XMRV-negative cell lines that were being cultured in the laboratory at the same time as CWR22Rv1 had become contaminated. Similarly, Zhang et al. (2011) detected XMLV infection of 13 out of 78 non-xenografted cell lines from five different laboratories that were also culturing XMLV-infected xenografted cell lines. In contrast, all 50 cell lines gathered from non-xenograft culturing labs were XMLV-free.
The laboratories involved in both studies were using standard BSL2 aseptic technique, highlighting the infective potential of these XMLVs. Therefore, the dangers of XMLV contamination of human cell lines is not limited to xenografted lines, but potentially extends to all additional cell lines being cultured alongside infected lines. If contamination occurs and is not detected, there is significant potential for spreading of the retrovirus, even across laboratories sharing samples." https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3683812/
Apr 19, 2017, 1:47 PM
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Robyn Erland shared a link.
Viral Evaluation of Animal Cell Lines Used in Biotechnology
Abstract
Animal cell culture is being increasingly used for production of therapeutic reagents such as monoclonal antibodies, recombinant proteins, viral vaccines and replication incompetent viral vectors for gene therapy. Material derived from a variety of biological processes has been associated in the past with incidents involving the transmission of infectious agents, principally viruses (1). Thus, virological evaluation of animal cell substrates for use in the manufacture of biologicals is essential to ensure the safety of products for pharmaceutical use. http://link.springer.com/protocol/10.1385%2F0-89603-547-6%3A23
Mar 18, 2017, 12:09 AM
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Robyn Erland shared a link.
Does everyone know that the first Flu vaccine made from recombinant technology was put on the market in 2013? They actually have 4 types of flu vaccines this season and this Flublok is for those 18 and over.
This is what Recombinant technology is (it's also good to know that these cells are sometimes made from "mammalian cells"):
"A recombinant vaccine is a vaccine produced through recombinant DNA technology. This involves inserting the DNA encoding an antigen (such as a bacterial surface protein) that stimulates an immune response into bacterial or mammalian cells, expressing the antigen in these cells."
What is Recombinant DNA?":
Recombinant protein is a protein that whose code is carried through a recombinant DNA. The term recombinant DNA implies that two segments of DNA in a plasmid."
Recombinant DNA (rDNA) molecules are DNA molecules formed by laboratory methods of genetic recombination (such as molecular cloning) to bring together genetic material from multiple sources, creating sequences that would not otherwise be found in the genome.
My note: Now this new Flublok vaccine didn't work so well in the 2013 - 2014 season. So they came up with a new version for 2015 - 2017. Here is all the crap they put in it:
Vaccine - Antigenic Composition - Flublok Quadrivalent
45 mcg per antigen
H1N1:A/California/07/2009
H3N2: A/Texas/50/2012B/Massachusetts/2/2012
(B/Yamagata-lineage)
B/Brisbane/60/2008 (B/Victoria -lineage)
And they are in the process of making numerous Recombinant DNA new vaccines. They are calling them VLP's "virus like particles") They are basically taking a part of the Flu virus and mass producing it with nano- micro particles of the flu put in. They use something clalled hemagglutinin (HA) or haemagglutinin which is a glycoprotein found on the surface of influenza viruses.
Now The very first surface glycoprotein or shell DNA recombinant vaccines tried were the HIV vaccines. They did not work and it was said followup studies were needed to be done, to determine if the subjects were actually being given HIV. Knowing what we know now about what's happening in Asia with all the new HIV strains and recombinations, it's a safe bet more "lab created recombinations" of these particles and retroviruses are sure to come. People have no clue what is being injected in them. I asked a few who've been ill almost a month now if they knew what Recombinant DNA vaccine is? They looked at me and said No.
Here's the information where they are comparing it to another type of vaccination:
https://www.cdc.gov/vaccines/acip/meetings/downloads/slides-2016-02/flu-03-hachey.pdf
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Robyn Erland shared a link. (this was one of the FDA postings I originally found when I realized that full mouse brains had also been being used in JE vaccines in the US too)
WOW this is going to blow your minds. I found where FDA scientist Khan put out a presentation in 2009 and one of the cell lines listed was from mouse brains ( This info has since been removed from the FDA site):
U.S. Licensed Viral Vaccines:
Primary Cells or Tissues
Cell Substrate - Inactivated Vaccines
Mouse brain - Japanese Encephalitis
Now in 2011 they made a 2nd mouse brain vaccine for Japanese Encephalitis and they have been vaccinating people all over ASIA North America, Australia and various European countries!
A Vero cell-derived, inactivated and alum-adjuvanted JE vaccine based on the SA 14-14-2 strain was approved in 2009 in North America, Australia and various European countries. The primary two doses are administered 4 weeks apart. A booster dose is recommended 1–2 years after the primary immunization. This vaccine has been given concomitantly with hepatitis A vaccine without significant interference with the safety and immunogenicity of either vaccine. Data on concomitant administration with other vaccines frequently used in travellers are currently unavailable. The vaccine is licensed for use in individuals 17 years of age and older in the United States, and 18 years and above in other countries. Paediatric and post-marketing safety studies are under way.
Another Vero cell-derived inactivated JE vaccine was licensed by the Japanese authorities in February 2009 and a similar Japanese vaccine was licensed in 2011. These two vaccines use the same strain of JE virus (Beijing-1) as the mouse-brain-derived vaccine. Clinical trials have shown that the vaccines are safe and immunogenic, with seroconversion rates exceeding 95%.
In addition, a new live attenuated, JE–yellow fever chimeric vaccine has recently been licensed in Australia and Thailand. A single dose of this chimeric JE vaccine was found to be safe, highly immunogenic and capable of inducing long-lasting immunity in both preclinical and clinical trials.
http://www.who.int/ith/vaccines/japanese_encephalitis/en/
Jul 3, 2016, 9:13 PM
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Feb 27, 2017, 3:02 PM
Robyn Erland shared a link.
Here is a slideshow from the FDA rep Khan. from 2009. This slide 23 he explains what it needed to reduce the risk associated CBER products. These are the products used in vaccines allergy shots, blood products and other biologicals. Notice where it says "REPLACE ANIMAL-DERIVED MATERIALS". That is something they have still NOT done. And they DID not have adequate technology to detect
Efforts to minimize the risk associated with Biological Raw
Materials
• Replace animal-derived materials – Serum-free cell cultures
• Use of new technologies for virus inactivation and removal
• Development of more sensitive and broad detection assays for adventitious agent detection and characterization to identify source
http://c.ymcdn.com/sites/www.casss.org/resource/resmgr/imported/WCBPCMC09KhanSlides.pdf
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Jul 3, 2016, 12:46 PM
Robyn Erland shared a link.
From the FDA site and this notice came out on October 12 2009. from the attached announcement: "We are also evaluating the risk of retrovirus infections in humans. (Retroviruses are RNA viruses that use an enzyme called reverse transcriptase (RT) to replicate; RNA is the de-coded form of DNA)". Here's the announcement:
Investigating Viruses in Cells Used to Make Vaccines; and Evaluating the Potential Threat Posed by Transmission of Viruses to Humans
Virus-based vaccines are made in living cells (cell substrates). Some manufacturers are investigating the use of new cell lines to make vaccines. The continual growth of cell lines ensures that there is a consistent supply of the same cells that can yield high quantities of the vaccine.
In some cases the cell lines that are used might be tumorigenic, that is, they form tumors when injected into rodents. Some of these tumor-forming cell lines may contain cancer-causing viruses that are not actively reproducing. Such viruses are hard to detect using standard methods. These latent, or "quiet," viruses pose a potential threat, since they might become active under vaccine manufacturing conditions. Therefore, to ensure the safety of vaccines, our laboratory is investigating ways to activate latent viruses in cell lines and to detect the activated viruses, as well as other unknown viruses, using new technologies. We will then adapt our findings to detect viruses in the same types of cell substrates that are used to produce vaccines. We are also trying to identify specific biological processes that reflect virus activity.
These methods will enable FDA scientists to help manufacturers to determine whether their specific cell substrate is safe to use for vaccine production. The methods our laboratory are developing and testing will help to ensure the production of safe and effective vaccines in two ways: 1) FDA will be able to develop testing guidelines for manufacturers who use new cell substrates for producing vaccines; and 2) FDA will publish the new methods it develops in peer-reviewed scientific journals, thus making them readily accessible to all manufacturers. http://web.archive.org/web/20091012050025/http://www.fda.gov/BiologicsBloodVaccines/ScienceResearch/BiologicsResearchAreas/ucm127327.htm
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Jul 3, 2016, 1:02 PM
Robyn Erland shared a link.
"The first-generation of JEV vaccines have been available since the 1950s and were in routine use for decades: (1) Mouse brain–derived inactivated vaccines (MBJEVs). MBJEVs were made from either Nakayama or Beijing-1 virus strains propagated in mouse brain tissue. Due to the strong protective efficacy, MBJEVs (e.g. JE-VAX) were extensively employed worldwide especially in Asia, Europe and the USA."
Hum Vaccin Immunother. 2014 Dec; 10
Published online 2015 Feb 10
the overall of JE vaccine evaluation is disorganized, the large variation in study designs, vaccine types, schedules, doses, population and few hand-to-hand trails, make direct comparisons difficult.
My note: In other words they have NO CLUE what they were doing and it went on from the 1950's until 2009 when they discontinued the MOUSE BRAIN version of the vaccines in the US! (which are known to contain pathogenic cancer and neuro diseases causing retroviruses) in vaccines!!!
This is really unbelievable everyone: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4514081/
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Robyn Erland shared a link.
"The first-generation of JEV vaccines have been available since the 1950s and were in routine use for decades: (1) Mouse brain–derived inactivated vaccines (MBJEVs). MBJEVs were made from either Nakayama or Beijing-1 virus strains propagated in mouse brain tissue. Due to the strong protective efficacy, MBJEVs (e.g. JE-VAX) were extensively employed worldwide especially in Asia, Europe and the USA."
Hum Vaccin Immunother. 2014 Dec; 10
Published online 2015 Feb 10
the overall of JE vaccine evaluation is disorganized, the large variation in study designs, vaccine types, schedules, doses, population and few hand-to-hand trails, make direct comparisons difficult.
My note: In other words they have NO CLUE what they were doing and it went on from the 1950's until 2009 when they discontinued the MOUSE BRAIN version of the vaccines in the US! (which are known to contain pathogenic cancer and neuro diseases causing retroviruses) in vaccines!!!
This is really unbelievable everyone: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4514081/
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Jul 5, 2016, 10:10 AM
Robyn Erland updated her status.
1954 Mouse brain vaccine with the approval of the NIH and CDC still in use today! Gulf War vets took this vaccine.
This document sets out the recommendations for manufacture and quality assessment in Part A. Guidance specific to the nonclinical and clinical evaluation of inactivated JE vaccines is provided in Parts B and C, respectively. Part D provides recommendations for national regulatory agencies. This document should be read in conjunction with all relevant WHO guidelines including those on nonclinical (3) and clinical evaluation (4) of vaccines.
A mouse brain derived inactivated JE vaccine was first licensed in Japan in 1954. This type of vaccine is manufactured in a similar fashion using the Nakayama-NIH strain in Japan (for export only) and also in India, Republic of Korea, Taiwan Province of China, Thailand, and Viet Nam. Since 1989, the JE vaccine that is actually used in Japan has contained the Beijing-1 strain. The efficacy of mouse brain-derived, inactivated JE vaccines was evaluated in two field trials in endemic areas. A study in Taiwan Province of China, in 1966, showed that the efficacy of the Nakayama-NIH strain vaccine was 80% after two doses. A later study in Thailand demonstrated that the efficacy of both a monovalent vaccine containing the Nakayama-NIH strain and a bivalent vaccine containing the Nakayama-NIH strain and the Beijing-1 strain was 91% over two transmission seasons. The Centers for Disease Control and Prevention (CDC), USA later pointed out that the level of protective efficacy observed in this study might in part reflect past exposure to JE virus and/or other flaviviruses. It was considered that the regimen of two doses given 7 days apart could not be assumed to give similar results in non-immune travellers. Therefore, the US licence for the Nakayama-NIH strain vaccine (approved in the USA in 1992) recommends a three-dose schedule based on immunogenicity data from non-immune US soldiers who received two or three doses. The duration of protection in non-immune people after a three-dose primary regimen remains unknown. Booster doses of the US-licensed Nakayama-NIH strain vaccine are recommended after 2 years although some studies done in Japan indicate that protective antibody levels persist for at least 4 year There is considerable information available on the adverse events associated with use of mouse brain-derived inactivated JE vaccines. Local reactions at the injection site and fever each occur in approximately 10% of vaccinated children in Japan. Severe allergic reactions characterized by generalized urticaria, respiratory symptoms and cardiovascular symptoms have been reported. Severe neurological disorders including acute disseminated encephalomyelitis (ADEM) have been reported following vaccination with mouse brain-derived JE vaccine. Eighteen cases of ADEM were reported in Japan after vaccination with mouse brain-derived inactivated JE vaccines from 1996 to 2005, which corresponds to approximately two cases per year following about 3 million inoculations. It is, however, assumed that there are 60–120 cases of ADEM per year in children in Japan whatever the cause (2). Inactivated JE vaccine prepared from the Beijing-3 strain in primary hamster kidney cells has been produced exclusively in China since 1968. Approximately 75 million doses were distributed annually in China up to 1988. http://www.who.int/biologicals/vaccines/Annex_1_WHO_TRS_963.pdf?ua=1
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Jul 3, 2016, 10:34 PM
Robyn Erland shared a link.
Do you really know what are in the vaccines? Would anyone like some VLP's "Virus Like Particles" that can insert the virus into their cells? Oh and they've been using them in Hep B and HPV vaccines. And FYI some VLP technology comes from the NCI/NIH and is the technology the NIH licensed to Merck and Glaxo for the HPV vaccine that is giving young girls ME/CFS, and a newly named pain syndrome which is really Fibromyalgia. And isn't it interesting that the FDA manufacturing guidelines allow for "virus like particles" in their finished products!
Here's another thought, with the recent postings about the over 400 mostly cancer cell lines that are contaminated or mislabeled. And the fact some cancer xenograft cell lines were tested in 2011, and were found to contain several mlv retroviruses some of which were "unidentified".
See the problem here is the Virus Like particles are also being made in labs that are not bio-hazard safety level 3. So any animal retroviruses say floating around in cell lines and these labs can recombine with these supposed non-infectious virus like particles. Oh and they have been making several types of these VLP's to include "mammal" ones. That would include mice! Quite the mess they are making in these labs. No wonder they had to shut down the mlv retroviral research which threatened a trillion dollar vaccine business, gene therapy, and xeno cancer research. Not to mention paying all those they've sickened restitution for the suffering and deaths!
From the attached: "CONCLUSIONS
Virus-like particles are multimeric nanostructures
assembled from viral structural proteins responsible for
cell penetration by the virus, ensuring efficient cell
entry and thus tissue-specific targeting."
http://www.actabp.pl/pdf/3_2014/531.pdf
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Jul 26, 2016, 9:13 AM
Robyn Erland updated her status.
"400 cell lines either lack evidence of origin or have become cross-contaminated with human or other animal cells at some point in their laboratory lineage. Cell lines are often chosen for their ability to reproduce and be bred for long periods of time, so they’re hardy buggers that can move around a lab if they end up on a researcher’s gloves, for example. “It’s astonishingly easy for cell lines to become contaminated."
My note: This is not good Since retroviruses can recombine with others in the lab to produce infectious versions! And they also found they horizontally transmit to other cell lines housed in the same lab.
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Jul 23, 2016, 12:10 AM
Robyn Erland shared a link.
More Cell line contamination information From 2007
Cross-Contamination and Misidentification
Increasing data demonstrate that cellular cross-contamination, misidentified cell lines, and the use of cultures at high-passage levels contrib-ute to the generation of erroneous and misleading results as well as wasted research funds. Contamination of cell lines by other lines has been recognized and documented back to the 1950s. Based on submissions to major cell repositories in the last decade, it is estimated that between 18% and 36% of cell lines may be contaminated or misidentified.
The problem of intraspecies and inter-species cross-contamination among cell lines has been recognized for half a century, and although reviews have been published, evidence of continued use of misidentification and cellular cross-contamination of cell cultures has not declined (23–36). Masters et al. found as much as 20%
of all ostensible cell lines in use today are not as they are purported (37), and other results support these findings (23). An additional
report estimates that more than one-third of cell cultures are cross-contaminated either with cells from other species (inter-species contamination) or with unrelated cells from the same species (intraspecies contamination) (32). MacLeod et al. http://www.biotechniques.com/multimedia/archive/00003/BTN_A_000112598_O_3282a.pdf
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Jul 22, 2016, 10:54 PM
Robyn Erland updated her status.
Now about those mlv retroviruses found in cell lines! They have no clue what they've been doing in labs for decades. And they make vaccines using cell lines. One story is reporting that they are finding human cell lines contaminated with animal cells. Imagine that!! They never knew if the contamination was from a pig, rabbit, or mice in their research. It's being reported that many scientific papers are wrong because the cell lines some used are full of the wrong cells. This is incredible: http://iclac.org/databases/cross-contaminations/
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Jul 22, 2016, 8:17 AM
Information on VLP's Virus Like Particles
Michał Zdanowicz
Jadwiga Chroboczek
DOI: https://doi.org/10.18388/abp.2016_1275
AbstractVirus-like particles (VLPs) assemble spontaneously during the viral cycle or in heterologous systems during expression of viral structural protein. Depending on the complexity of the VLPs, they can be obtained by expression in prokaryotic or eukaryotic expression system from the suitable recombinant vectors, or formed in cell-free conditions. Moreover, they can be built from proteins of a single virus, or can present the proteins or peptides derived from a virus or cell on a platform derived from any other single virus, thus forming chimeric VLPs. VLPs are best known for their immunogenic properties, but the versatility of VLPs allows a wide variety of applications. They are lately in the centre of investigations in vaccinology, drug delivery and gene therapy. This review focuses on utilization of VLPs for drug delivery.
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Delivery systems for gene therapy
Abstract
The structure of DNA was unraveled by Watson and Crick in 1953, and two decades later Arber, Nathans and Smith discovered DNA restriction enzymes, which led to the rapid growth in the field of recombinant DNA technology. From expressing cloned genes in bacteria to expressing foreign DNA in transgenic animals, DNA is now slated to be used as a therapeutic agent to replace defective genes in patients suffering from genetic disorders or to kill tumor cells in cancer patients. Gene therapy provides modern medicine with new perspectives that were unthinkable two decades ago. Progress in molecular biology and especially, molecular medicine is now changing the basics of clinical medicine. A variety of viral and non-viral possibilities are available for basic and clinical research. This review summarizes the delivery routes and methods for gene transfer used in gene therapy.
Retroviruses
Retroviruses are RNA viruses that carry a gene for a reverse transcriptase that transcribes the viral genetic material into a double stranded DNA intermediate. This DNA intermediate is then incorporated into the host DNA allowing the host cell machinery to produce all the necessary viral components. Additionally, because the viral genome is stably integrated into the host DNA, any modification that has been made will be passed to all daughter cells that are derived from the transfected cell currently; the most common retrovirus used is derived from the murine leukemia virus.
In general, retroviruses have been used for ex vivo gene therapy applications as they are unable to efficiently infect non-dividing cells.
Go to:
Limitations of Retroviral Vectors
(a) low vector titer, (b) low transfection efficiency, demonstrated in in vitro experiments, (c) particle instability and difficulty to concentrate, and (d) inability to transduce non-dividing post-mitotic cells, particles infecting only proliferating cells.
The limitation of retroviral infection has been overcome, in part, by the use of lentiviral vectors, which have been shown to activate the immune system in pre-clinical animal models of oral cancer.
Using different viral envelope proteins that recognize different receptors can vary the range of cells that can be transduced, but still does not provide much specificity. The difficulty is that, because retroviral vectors cannot be generated at a high titer, it is not possible to get a large number of vector particles to the desired cell type in vivo. The viral particles would bind to many cells they encounter and therefore, would be diluted out before reaching their target.
The envelope protein has two functions: Binding to its receptor (by the surface [SU] moiety) and enabling the entry of the viral nucleoprotein core (carried out primarily by the trans membrane [TM] moiety). The SU protein binds to its receptor on the target cell surface and as a result, the SU/TM complex undergoes a conformational change that allows fusion of the viral and cellular membranes, followed by entry of the viral core (which carries the virus's genetic information) into the target cell's cytoplasm.
Two broad approaches to providing target cell specificity have been followed. First, the natural receptor-binding domain of the SU protein has been replaced with a ligand or single-chain antibody that recognizes a specific cell surface receptor.
Although, several creative systems have been designed, the most successful approach at present appears to be insertion of a ligand that recognizes an extracellular matrix (ECM) component into a part of the SU protein that does not disturb the natural receptor-binding domain. This tethering concentrates the vector in the ECM in the vicinity of the target cells. Receptor binding and core entry can then occur through the natural envelope-receptor mechanism. Two ligands that appear particularly useful for tethering are those specific for fibronectin for collagen. Fibronectin is present in normal ECM and exposed collagen is present in areas of damage, for example after wound injury as in the cardiovascular system after angioplasty.[2,3,4,7,8]
Lentiviruses (e.g., human immunodeficiency virus type 1: HIV-1) are a special group of retroviruses with the ability to infect both proliferating and quiescent cells. To expand the spectrum of target cells, it is possible to replace the genes for surface glycoproteins by genes from another viral genome in the packaging cell lines packaging cell lines (PCL) of the vector.
Lentiviral transfer systems ensure long-term expression and efficient transfer without inflammatory responses.[4,5,6,9]
The adenovirus does not integrate its genome into the host genome. Instead, the adenoviral genome remains in the nucleus as an episomalelement after the infection of the host cell. The advantages common to all adenoviral vectors include, the ease of purification and concentration and the high efficiency rate of host cell infection of various cell types, dividing or non-dividing. These advantages make adenoviral vectors a good candidate for direct in vivo gene transfer. The usefulness of these vectors is limited by 2 factors. In most tissues, the duration of transgene expression is limited to a few days to a week and viral genes are also transduced and expressed, eliciting an immune response to the transduced cells that ultimately results in their clearance.
Adenoviruses are DNA viruses that infect a cell, lose their protein coat, and transfer DNA into the nucleus, where it is transcribed. This DNA does not integrate into the host genome, and thus, its effects are transient (range: 7-42 days).
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3722627/
More to come.....
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