 |
|
| Science Forum Index » Chemistry Forum » Squalene in H1N1 vaccine... |
|
Page 1 of 1 |
|
| Author |
Message |
| Mike... |
 Posted: Thu Oct 29, 2009 7:54 am |
|
|
|
Guest
|
When the CDC/HHS determined that they would have
to start producing a H1N1 vaccine there was a
rather considerable cover-up as to the actual
number of influenza infection that were breaking
out, and both state and federal authorities
started actually classifying the actual numbers
of the confirmed cases in the name of political
expediency, with no priority for public health.
At around the same time the CDC and FDA started
talking about granting "authorization" for ""new
and untested drugs, based on a Presidential Declaration of
Emergency".
So here we are six months after the H1N1 outbreak
starts, and six months after the CDC/HHS and FDA
buttered up the medical community about a new and
untested vaccine that would be coming out right
around now so that immunizations could begin prior to Thanksgiving.
The CDC claims that the States now have million
of doses that they can deploy, but everything
seems to have gotten stuck on hold as it were,
until the President made the Emergency
Declaration on Friday based on a classified
briefing her was given on Thursday of last week.
It is good that we have a vaccine, it is just too
bad that we do not have enough to go around (hey, it sucks to be
you).
Those people who do take the vaccine are likely
to get a dose that has squalene, but the side
effects (pain and misery) from the adjuvant are
far less then actually getting deathly sick from the H1N1 virus..
The sad thing is that the government has been
lying to the public since the very first day of
this infection, and very little of what they have
told the public has been truthful.
The military has taken a serious thrashing over
the years for their use of squalene in
"classified vaccines" for some years, so you can
find out quite a bit about the use of squalene
and other vaccine adjuvants in the various
web-sites the military has set up to explain their position.
The use of adjuvants come from a profound lack of
preparedness by the government, and most
certainly from the public. They are used when you
really do not have enough of a vaccine to go
around, go you add a chemical to the vaccine to
trick the immune system of the person to whom you
are giving the vaccine to. Consider it to be a
type of virological "Hamburger Helper", so that
they can take when they have and squeeze more
doses out of a medication that is massively
diluted... possibly to the point where the vaccine is of little or no
value.
All things being considered this weak response is
the results of extremely poor planning on the
part of the government. They have had 8 months to
prepare, and they should have had the first few
million doses out and into the hands of the
medical specialists by late May 2009 at the latest.
It is one thing for them to not be prepared for
this situation, but it is completely unforgivable
that that have been lying the the public and
needlessly endangering the public for the past 6+
months. The only reason that H1N1 infections
numbers are exploding right now is simply because
school is back in session, and this illness is
rampantly spreading through the populations of
the children and young adults right now.
The CDC has been lying to the public by at least
a 25:1 ratio, so that when they say that there
has been only 40,000 cases, there really was an
actual 1,000,000+ cases. When they say that only
1000 chilren have died... there really was closer to 25,000 or more.
It will be important for anybody who takes the
H1N1 vaccine to get a copy of the sheet that is
supposed to be given out with each shot, and to
compare the writing on the vial to what is on the
piece of paper you are given. Then ask the person
who is giving it to you about it the vaccine has
any Squalene of not, if they do not know then
find someplace else to get the vaccine (most
medical people just give the medication, very few
actually have any idea what the medication actually is).
I have attached a snapshot of one of the military
pages where there is a whole bunch of tap dancing
by the military about the use of Squalene
adjuvants, along with a long list of references and sources.
Liquid chromatography, and a small group of
scientists who express vaccine from the injection
sites of recently immunized patients will have
some interesting announcements in a few weeks
after the vaccine is "available" and in wide spread use.
The interesting thing about the use of an
unapproved, and untested drugs and vaccines is
that if the President makes an Emergency
Declaration, then the companies who sold the
vaccine to the government, and the people,
hospitals, and agencies who give it can never be
sued or held responsible is even the slightest
way, even it it kills thousand of people. IT is
the ultimate "get out of jail free" card for the pharmaceutical
companies.
-jma
http://www.anthrax.osd.mil/resource/qna/qaAll.asp?cID=319
The Facts on Squalene
1) Executive Summary
2) What is squalene?
3) Does the anthrax vaccine use squalene as an adjuvant?
4) Does the anthrax vaccine contain squalene?
5) Should we be concerned about the presence
of trace quantities of squalene in tetanus, diphtheria, and anthrax
vaccines?
6) Can squalene cause harm?
7) If you wanted to use squalene as an adjuvant, what form would
it take?
 What do we know about the European
influenza vaccine that uses MF59 (an adjuvant containing squalene).
9) What testing has been done?
10) What did SRI find the first time?
11) References: (abstracts of many of these
articles can be viewed at www.ncbi.nlm.nih.gov ,
using the PubMed function of the National Library of Medicine)
12) What did the FDA find?
13) What did SRI find after it revised its test procedures?
14) Did DoD mislead or lie to anybody about
the squalene tests conducted by SRI?
15) Has anyone, anywhere found squalene
added as an adjuvant to any US-licensed vaccine?
16) Where did the squalene FDA found in its
anthrax vaccine tests come from?
17) What did the U.S. Senate say about squalene?
1 Did the British government test its anthrax vaccine for
squalene?
19) What are the claims about anti-squalene antibodies?
20) Have any independent panels evaluated
the claims of researchers to find anti-squalene
antibodies in the blood of ill Gulf War veterans?
21) Are these panels really independent?
22) What did the GAO say about squalene
testing and what are DoD researchers doing?
23) What did the competitively funded
research project find regarding squalene antibodies?
24) Has DoD ever tested squalene-adjuvanted
vaccines in humans against any disease?
25) Could squalene concerns have anything to
do with various reported clusters of illnesses
among people given anthrax vaccine?
26) Bottom line, is there any reason for alarm here?
The Facts on Squalene
1) Executive Summary
A few people claim the Department of Defense
(DoD) added squalene to anthrax vaccine to
stretch the vaccine supply. Four civilian panels
have looked into these allegations since 1999 and
repeatedly found them groundless. Neither DoD nor
anybody else added squalene to anthrax vaccine
for our troops. DoD does not conduct illegal
experiments. Details and links to independent sources of data appear
below.
2) What is squalene?
Squalene is a naturally occurring substance found
in plants, animals, and humans. Squalene is
manufactured in the liver of every human body and
circulates in our bloodstreams. Squalene is
present in the oil left by human fingerprints
(Asano et al, 2002). Humans cannot live without
squalene, because we use squalene as an essential
building block to make hormones and other
substances in our bodies. Squalene is also found
in a variety of foods (for example: eggs, olive
oil (0.7%), cookies, yeast, meat), cosmetics (for
example: eye makeup, lipstick, baby powder),
over-the-counter medications, and health
supplements. Squalene in olive oil may contribute
to the low cholesterol levels of people who
consume Mediterranean-style diets (Smith, 2000).
People can purchase squalene at health food
stores. It is more commonly known as “shark liver oil.”
3) Does the anthrax vaccine use squalene as an adjuvant?
No, the adjuvant in the anthrax vaccine is
aluminum hydroxide. An adjuvant is a substance to
improve the body’s immune response to a vaccine
(Vogel et al, 1998; Burdin et al, 2004).
4) Does the anthrax vaccine contain squalene?
Maybe. Some lab tests come up positive for
squalene. Because of the difficulty of removing
squalene-containing fingerprint oils from
laboratory glassware, it is hard to know whether
the squalene is truly present in some lots of the
vaccine or is introduced by the testing process
itself. DoD, the Food & Drug Administration
(FDA), and several civilian advisory committees
agree that squalene at such low levels has no
adverse health consequences. In September 2000,
DoD became aware of FDA test results finding
trace amounts of squalene in three out of three
US vaccines tested: tetanus, diphtheria, and
anthrax. The level of squalene identified by the
FDA test is so minute that it is likely the
result of squalene in the oil of a fingerprint
not completely cleaned from lab glassware. It is
hard to completely remove fingerprint oils from
glassware. Before they go looking for squalene,
lab workers have to use a chemical solvent such
as hexane to completely remove their own
fingerprint oils from lab glassware. When lab
workers intentionally tested an extract of
fingerprint oil, the squalene reading went off
the chart. Before the FDA test results became
known, Stanford Research International (SRI),
under DoD contract, looked for squalene in
anthrax vaccine. At the limit of detection of its
test, 140 parts per billion, SRI found no
squalene in several lots of anthrax vaccine. The
FDA’s test, which was developed later, is more
sensitive. It is able to detect as little as 10
parts per billion. The FDA found squalene at 10
to 83 parts per billion in diphtheria toxoid,
tetanus toxoid, and anthrax vaccine. The trace
level of squalene found by the FDA in anthrax
vaccine is less than the concentration naturally
present in human blood (250 parts per billion)
(Miettinen, 1982; Nikkila et al, 1992). After the
FDA reported its results, DoD asked SRI to refine
its assay. Using an improved method that could
detect as little as 1 part per billion, SRI found
no squalene in 32 out of 33 lots of anthrax
vaccine tested (including lots in which FDA found
low levels of squalene). In one lot, they found
up to 9 parts per billion. The details appear below.
5) Should we be concerned about the presence of
trace quantities of squalene in tetanus, diphtheria, and anthrax
vaccines?
No. The trace level of squalene found by the FDA
and SRI in diphtheria, tetanus, and anthrax
vaccines is well below the concentration
naturally present in human blood (250 parts per
billion). Injecting trace amounts of squalene are
unlikely to have any biological effect, given
that it is already present in the body. In fact,
without squalene in the body to manufacture
hormones and other substances in our bodies, we
would die. In Congressional testimony on 3
October 2000, FDA official Mark Elengold said
that the trace quantities of squalene detected
were “both naturally occurring and safe."
6) Can squalene cause harm?
Some animal research to study arthritis used
injections of tuberculosis-like bacteria
(mycobacteria) dissolved in squalene (e.g.,
arthritis-prone rats, mice). Other studies
assessed 100% squalene injected into rat tails or
injected directly into joints. (Yoshino &
Yoshino, 1994; Lorentzen, 1999; Kuroda et al,
2004) The relevance of findings in susceptible
animal species to humans is unclear (IOM/Sox,
1999; Kuroda et al, 2004). Based on other
research, it is clear that whether squalene
causes harm or not is related to selected
conditions of concentration, dose, route of
application, and other factors (Benisek et al, 2004).
7) If you wanted to use squalene as an adjuvant, what form would it
take?
If you wanted to use squalene as an adjuvant (to
boost immune responses) you would have to
multiply the amount of squalene found by the FDA
about 1 million times, as well as change it from
a simple liquid (its natural state) to an
emulsion. An emulsion is a stable suspension of
tiny droplets, like an oil-and-vinegar mixture
that doesn’t separate. This double difference is
like the difference between a teaspoon of oil and
2,000 pounds of mayonnaise. [If you emulsify oil
with eggs, you get mayonnaise.] Squalene in the
form of an emulsion (emulsified squalene, such as
an adjuvant called MF59) has been added as an
adjuvant to some investigational vaccines in the
U.S. (Burdin et al., 2004) There is no squalene
adjuvant in any US-licensed vaccine. Whatever the
arguments for or against squalene as a vaccine
adjuvant, the fact is that none of the anthrax
vaccine administered to U.S. troops contained
squalene as an adjuvant. Based on manufacturing
records, FDA can verify that no squalene was
added to any vaccine formulation used during the
Gulf War. This includes the anthrax vaccine. To
date, the FDA has licensed, and US manufacturers
have used, only aluminum salts (for example,
aluminum hydroxide, aluminum phosphate, aluminum
potassium sulfate) as adjuvants.
What do we know about the European influenza
vaccine that uses MF59 (an adjuvant containing squalene).
In 1997, European health agencies approved
emulsified squalene (with influenza virus in the
center of each droplet) for use as an adjuvant in
an influenza vaccine (Fluad, Chiron Corporation,
Marburg, Germany, and Siena, Italy,
http://www.forumimpfen.de/impfnavigator/packungsbeilage/5205fluad.pdf;
Sesardic & Dobbelaer, 2004). Some clinicians
consider influenza vaccine with MF59 adjuvant to
be better able to induce immunity in elderly
people (Banzhoff et al, 2003). To make this
influenza vaccine work, researchers needed a
squalene concentration of 1.95% (about 2 parts
per hundred or 20 million parts per billion) to
boost the immune response. This squalene had to
be in the form of an emulsion (a mixture of tiny
droplets) to be recognized by the immune system.
Squalene in its oily state is naturally present
inside the human body. Tens of millions of doses
of this European influenza vaccine have been administered safely since
1997.
9) What testing has been done?
Three sets of US tests have been performed:
Initial tests by SRI, tests by FDA, and improved
tests by SRI. Each is described below.
10) What did SRI find the first time?
To determine whether squalene was present in
anthrax vaccine, the DoD contracted with an
independent civilian laboratory, Stanford
Research Institute (SRI) International of Menlo
Park, California www.sri.com, to test for the
presence of squalene in anthrax vaccine. SRI
developed a laboratory method to detect squalene
as dilute as 140 parts per billion (ppb). At this
level of detection, extraordinary measures must
be taken to avoid contaminating samples,
glassware, and equipment with squalene from the
skin, because squalene is a natural component of
the oils in our skin. The SRI test used a
technique called high-pressure liquid
chromatography (HPLC) with ultraviolet detection
at a wavelength of 203 nanometers. SRI tested 17
lots of anthrax vaccine: FAV008, FAV017, FAV019,
FAV020, FAV024, FAV030, FAV031, FAV033, FAV034,
FAV036, FAV037, FAV038, FAV041, FAV043, FAV044,
FAV047, and FAV048B. SRI reported "based on
triplicate analysis, no squalene was detected in
the sample. The limit of detection is 70
nanograms per 0.5 milliliter dose (140 ppb)." (Spanggord et al.,
2002)
11) References: (abstracts of many of these
articles can be viewed at www.ncbi.nlm.nih.gov ,
using the PubMed function of the National Library of Medicine)
Asa PB, Cao Y, Garry RF. Antibodies to squalene
in Gulf War syndrome. Experimental & Molecular Pathology 2000;68(Feb):
55-64.
Asa PB, Wilson RB, Garry RF. Antibodies to
squalene in recipients of anthrax vaccine.
Experimental & Molecular Pathology 2002;73(Aug):19-27.
Alving CR, Grabenstein JD. Letter to the editor.
Experimental & Molecular Pathology 2000;68 (Jun):196-7 (letter).
Armed Forces Epidemiological Board.
Recommendations regarding review of the paper
“Antibodies to Squalene in Gulf War Syndrome by
P. B. Asa, Y. Cao and R. F. Garry. 11 Jul 2000.
http://www.ha.osd.mil/afeb/reports/squalene.pdf
Asano KG, Bayne CK, Horsman KM, Buchanan MV.
Chemical composition of fingerprints for gender
determination. Journal of Forensic Science 2002;47(Jul):805-807.
Banzhoff A, Nacci P, Podda A. A new
MF59-adjuvanted influenza vaccine enhances the
immune response in the elderly with chronic
diseases: Results from an immunogenicity
meta-analysis. Gerontology. 2003;49(May-Jun):177-84.
Benisek Z, Suli J, Elias D, Lenhardt L,
Ondrejkova A, Ondrejka R, Svrcek S, Bajova V.
Experimental squalene adjuvant. II. Harmlessness
and local reactogenity. Vaccine. 2004;22(Sep 3):3470-4.
Burdin N, Guy B, Moingeon P. Immunological
foundations to the quest for new vaccine adjuvants. BioDrugs 2004;18
(2):79-93.
Epstein JE, Charoenvit Y, Kester KE, Wang R,
Newcomer R, Fitzpatrick S, Richie TL, Tornieporth
N, Heppner DG, Ockenhouse C, Majam V, Holland C,
Abot E, Ganeshan H, Berzins M, Jones T, Freydberg
CN, Ng J, Norman J, Carucci DJ, Cohen J, Hoffman
SL. Safety, tolerability, and antibody responses
in humans after sequential immunization with a
PfCSP DNA vaccine followed by the recombinant
protein vaccine RTS,S/AS02A. Vaccine 2004;22(Apr 16):1592-603.
General Accounting Office. Questions about the
presence of squalene antibodies in veterans can
be resolved. GAO/NSIAD099-5, March 1999.
http://www.gao.gov/archive/1999/ns99005.pdf
Hoffman SL, Edelman R, Bryan JP, Schneider I,
Davis J, Sedegah M, Gordon D, Church P, Gross M,
Silverman C. Safety, immunogenicity, and efficacy
of a malaria sporozoite vaccine administered with
monophosphoryl lipid A, cell wall skeleton of
mycobacteria, and squalane as adjuvant. American
Journal of Tropical Medicine & Hygiene 1994;51(Nov):603-12.
Institute of Medicine Committee on Health Effects
Associated with Exposures During the Gulf War.
Gulf War & Health: Volume I: Depleted Uranium,
Sarin, Pyridostigmine Bromide, Vaccines. [Harold
C. Sox, chair] Fulco CE, Liverman CT, Sox HC,
editors. Washington, DC: National Academy of
Sciences, September 2000. See
http://stills.nap.edu/books/030907178X/html,
pages 307-313 (especially 311-312).
Institute of Medicine Committee to Assess the
Safety and Efficacy of the Anthrax Vaccine. The
Anthrax Vaccine: Is it Safe? Does it Work? [Brian
L. Strom, chair] Joellenbeck LM, Zwanziger L,
Durch JS, Strom BL, editors. Washington, DC:
National Academy of Sciences, March 2002. See
http://www.nap.edu/catalog/10310.html, especially pages 96-97 and
147.
Kuroda Y, Nacionales DC, Akaogi J, Reeves WH,
Satoh M. Autoimmunity induced by adjuvant
hydrocarbon oil components of vaccine. Biomed Pharmacother 2004;58
(Jun):325-37.
Lorentzen JC. Identification of arthritogenic
adjuvants of self and foreign origin.
Scandinavian Journal of Immunology 1999;49:45-50.
Matyas GR, Wassef NM, Rao M, Alving CR. Induction
and detection of antibodies to squalene. Journal
of Immunologic Methods 2000;245(Nov 1):1-14.
http://www.vaccines.mil/documents/library/Squalene1.pdf
Matyas GR, Rao M, Alving CR. Detection of
antibodies to squalene. II. Optimization of the
assay for murine antibodies. Journal of
Immunologic Methods 2001;267(Sep 15):119-129.
http://www.vaccines.mil/documents/library/Squalene2.pdf
Matyas GR, Rao M, Pittman PR, Burge R, Robbins
IE, Wassef NM, Thivierge B, Alving CR. Detection
of antibodies to squalene. III. Naturally
occurring antibodies to squalene in humans and
mice. Journal of Immunologic Methods
2004;286(Mar):47-67. htthttp://www.vaccines.mil/documents/library/Squalene3..pdf
Miettinen TA. Diurnal variation of cholesterol
precursors squalene and methyl sterols in human
plasma lipoproteins. Journal of Lipid Research
1982;23:466-73. http://www.jlr.org/cgi/reprint/23/3/466
Nikkila K, Hockerstedt K, Miettinen TA. Serum and
hepatic cholestanol, squalene and noncholesterol
sterols in man: A study on liver transplantation. Hepatology
1992;15:863-70.
Nitayaphan S, Khamboonruang C, Sirisophana N,
Morgan P, Chiu J, Duliege AM, Chuenchitra C,
Supapongse T, Rungruengthanakit K, deSouza M,
Mascola JR, Boggio K, Ratto-Kim S, Markowitz LE,
Birx D, Suriyanon V, McNeil JG, Brown AE, Michael
RA. A phase I/II trial of HIV SF2 gp120/MF59
vaccine in seronegative Thais: Armed Forces
Research Institute of Medical Sciences--Research
Institute for Health Sciences Vaccine Evaluation
Group. Vaccine 2000;18(Feb 14):1448-55.
Pitisuttithum P, Nitayaphan S, Thongcharoen P,
Khamboonruang C, Kim J, de Souza M, Chuenchitra
T, Garner RP, Thapinta D, Polonis V, Ratto-Kim S,
Chanbancherd P, Chiu J, Birx DL, Duliege AM,
McNeil JG, Brown AE; Thai AIDS Vaccine Evaluation
Group. Safety and immunogenicity of combinations
of recombinant subtype E and B human
immunodeficiency virus type 1 envelope
glycoprotein 120 vaccines in healthy Thai adults.
Journal of Infectious Diseases 2003;188(Jul 15):219-27.
Sesardic D, Dobbelaer R. European Union
regulatory developments for new vaccine adjuvants
and delivery systems. Vaccine 2004;22(Jun 23):2452-6.
Sever JL, Brenner AI, Gale AD, Lyle JM, Moulton
LH, West DJ. Safety of anthrax vaccine: A review
by the Anthrax Vaccine Expert Committee (AVEC) of
adverse events reported to the Vaccine Adverse
Event Reporting System (VAERS).
Pharmacoepidemiology & Drug Safety 2002;11
(Apr-May):189-202. http://www.vaccines.mil/documents/library/AVEC_ms.pdf
Sever JL, Brenner AI, Gale AD, Lyle JM, Moulton
LH, Ward BJ, West DJ. Safety of anthrax vaccine:
An expanded review and evaluation of adverse
events reported to the Vaccine Adverse Event
Reporting System (VAERS). Pharmacoepidemiology &
Drug Safety 2004;13:in press. Epub version on
Internet. http://www.vaccines.mil/documents/library/SeverArticle.pdf
Smith TJ. Squalene: Potential chemopreventive
agent. Expert Opinion Investigational Drugs 2000;9(Aug):1841-8.
Spanggord RJ, Wu B, Sun M, Lim P, Ellis W.
Enhancement of an analytical method for the
determination of squalene in anthrax vaccine
adsorbed formulations. Journal of Pharmaceutical
and Biomedical Analysis 2002;29(Jun):183-193.
http://www.vaccines.mil/documents/library/JPB42(2006)494–499.pdf
United States Senate, Committee on Veterans’
Affairs, 105th Congress. Report of the Special
Investigation Unit on Gulf War Illnesses. Chapter
3: Evaluation of Wartime Exposures, Gulf War
Veteran Health Concerns and Related Research, and
Unanswered Questions. Washington, DC: 1998, pages
123 and 303.
http://veterans.senate.gov/Reports/siu.htm and
http://veterans.senate.gov/Reports/chapt3.pdf
Vogel FR, Powell MF, Alving CR. A Compendium of
Vaccine Adjuvants and Excipients, 2nd ed.
Bethesda, MD: National Institute of Allergy &
Infectious Diseases, 1998.
http://www.niaid.nih.gov/daids/vaccine/pdf/compendium.pdf
Wang R, Epstein J, Charoenvit Y, Baraceros FM,
Rahardjo N, Gay T, Banania JG, Chattopadhyay R,
de la Vega P, Richie TL, Tornieporth N, Doolan
DL, Kester KE, Heppner DG, Norman J, Carucci DJ,
Cohen JD, Hoffman SL. Induction in humans of CD8+
and CD4+ T cell and antibody responses by
sequential immunization with malaria DNA and
recombinant protein. Journal of Immunology 2004;172(May 1):5561-9.
Yoshino S, Yoshino J. Recruitment of pathogenic T
cells to synovial tissues of rats injected
intraarticularly with nonspecific agents. Cellular Immunology
1994;158:305-313.
Related to Influenza and Other Vaccines with MF59
Adjuvant (which contains squalene as one component of the adjuvant):
DeDonato S, Granoff D, Minutello M, Lecchi G,
Faccini M, Agnello M, Senatore F, Verweij P,
Fritzell B, Podda A. Safety and immunogenicity of
MF59-adjuvanted influenza vaccine in the elderly.
Vaccine 1999;17(Aug 6):3094-101.
Frey S, Poland G, Percell S, Podda A. Comparison
of the safety, tolerability, and immunogenicity
of a MF59-adjuvanted influenza vaccine and a
non-adjuvanted influenza vaccine in non-elderly
adults. Vaccine 2003;21(Oct 1):4234-7.
Giudice GD, Fragapane E, Bugarini R, Hora M,
Henriksson T, Palla E, O’Hagan D, D.y J,
Rappuoli R, Podda A. Vaccines with the MF59
Adjuvant Do Not Stimulate Antibody Responses
against Squalene. 2006. Clinical and Vaccine
Immunology, Vol. 13, No. 9. http://www.vaccines.mil/documents/library/MF59.pdf
Heineman TC, Clements-Mann ML, Poland GA,
Jacobson RM, Izu AE, Sakamoto D, Eiden J, VanNest
GA, Hsu HH. A randomized controlled study in
adults of the immunogenicity of a novel hepatitis
B vaccine containing MF59 adjuvant. Vaccine 1999;17:2769-2778.
Martin JT. Development of an adjuvant to enhance
the immune response to influenza vaccine in the
elderly. Biologicals 1997;25:209-13.
Minutello M, Senatore F, Cecchinelli G, Bianchi
M, Andreani T, Podda A, Crovari P. Safety and
immunogenicity of an inactivated subunit
influenza virus vaccine combined with MF59
adjuvant emulsion in elderly subjects, immunized
for three consecutive influenza seasons. Vaccine 1999;17(Jan):99-104.
Podda A, Del Giudice G. MF59-adjuvanted vaccines:
increased immunogenicity with an optimal safety
profile. Expert Review of Vaccines 2003;2(Apr):197-203.
Podda A. The adjuvanted influenza vaccines with
novel adjuvants: experience with the
MF59-adjuvanted vaccine. Vaccine 2001;19(Mar 21):2673-80.
Squarcione S, Sgricia S, Biasio LR, Perinetti E.
Comparison of the reactogenicity and
immunogenicity of a split and a
subunit-adjuvanted influenza vaccine in elderly
subjects. Vaccine 2003;21(Mar 7):1268-74.
12) What did the FDA find?
Using a more sensitive test, developed after the
initial SRI test, the Food & Drug Administration
(FDA) found trace amounts of squalene in three
out of three US vaccines tested in Jun 1999:
diphtheria toxoid, tetanus toxoid, and anthrax
vaccine
(http://www.vaccines.mil/documents/library/Squalene1.pdf).
The FDA test used a technique called gas
chromatography with flame-ionization detection.
The FDA method could detect squalene as dilute as
10 parts per billion (ppb). Testing five lots of
anthrax vaccine and two lots each of diphtheria
and tetanus vaccines, FDA concluded, "there were
only trace amounts of squalene in the lots
tested." Based on manufacturing records, FDA
verified that no squalene was added to any
vaccine formulation used during the Gulf War. The
amounts of squalene identified in the specific lots were:
Anthrax lot FAV020 11.3 ppb
Anthrax lot FAV030 10.1 ppb
Anthrax lot FAV038 27.1 ppb
Anthrax lot FAV043 40.0 ppb
Anthrax lot FAV047 82.9 ppb
Diphtheria lot 3710 22.5 ppb
Tetanus lot 7271 28.7 ppb
Squalene is constantly present in the human blood
stream at 250 ppb (250 nanograms per milliliter),
a concentration 3 to 25 times higher than the
level detected in the FDA test. The amount of
squalene added as an adjuvant to a
European-approved influenza vaccine is 4 grams
per 100 ml (4 parts per hundred), which is about
1,000,000 times more than the concentration of
squalene detected in the FDA test. This European
influenza vaccine has been administered safely to
hundreds of thousands of people.
13) What did SRI find after it revised its test procedures?
After the FDA released its findings in September
2000, SRI revised its squalene test, lowering its
limit of detection of 1 ppb or 0.5 nanograms per
0.5 ml. With this more sensitive test, SRI found
no squalene in 32 out of 33 lots tested. SRI
found squalene in each of three vials of lot
FAV008, at 1, 7, and 9 ppb. SRI found no squalene
in lots 12, 13, 18, FAV001, FAV002, FAV003,
FAV004, FAV005, FAV006, FAV007, FAV009, FAV012,
FAV016, FAV017, FAV018, FAV019, FAV020, FAV022,
FAV024, FAV030, FAV031, FAV032, FAV033, FAV034,
FAV036, FAV037, FAV038, FAV041, FAV043, FAV044,
FAV047, and FAV048B. SRI also tested some
non-vaccine injectable pharmaceuticals. SRI found
no squalene in human insulin regular U-100, human
insulin isophane (NPH) U-100, lidocaine 2%
solution, sodium chloride 0.9% solution, or
potassium chloride 2 mEq/ml solution.
14) Did DoD mislead or lie to anybody about the
squalene tests conducted by SRI?
No. DoD truthfully and fully reported its
findings at each step since May 1999, when SRI
first developed its squalene test. DoD did not
know of FDA’s findings until they were publicly
released. At the initial limit of detection of
its test, 140 parts per billion, SRI found no
squalene in anthrax vaccine (Spanggord et al.,
2002). It was scientifically proper to say ‘no
squalene was found to the limit of detection of
the assay,’ which DoD officials sometimes
oversimplified to say ‘there is no squalene present.’
15) Has anyone, anywhere found squalene added as
an adjuvant to any US-licensed vaccine?
No
16) Where did the squalene FDA found in its anthrax vaccine tests come
from?
The most likely source of the trace squalene in
the FDA tests is the result of squalene in the
oil of a fingerprint not cleaned from lab
glassware. Squalene is not added to anthrax
vaccine or any US-licensed vaccine. It is hard to
completely remove fingerprint oils from
glassware. Lab workers have to use a chemical
solvent such as hexane to completely remove
fingerprint oils from lab glassware.
17) What did the U.S. Senate say about squalene?
In its investigations of illnesses among Gulf War
veterans, the Senate Special Investigations Unit
(SIU) found no credible information indicating
that vaccines used during the Gulf War contained
squalene (1998, page 123)
http://veterans.senate.gov/Reports/chapt3.pdf
(chapter 3, page 23 of 55) In its report, the SIU
stated that according to the Food and Drug
Administration (FDA), squalene can be contained
in a vaccine due to two different processes: 1)
as an adjuvant, which is an agent to enhance the
immune response; or 2) in minute quantities in
certain vaccines manufactured using eggs, since
eggs are rich in squalene and cholesterol. The
FDA verified that none of the vaccines used
during the Gulf War contained squalene as an adjuvant.
1 Did the British government test its anthrax vaccine for squalene?
Yes, The United Kingdom’s Ministry of Defence
arranged for an independent laboratory to test 11
lots of the British anthrax vaccine manufactured
at Porton Down, as well as other vaccines. No
squalene was detected in those lots of vaccine,
with a limit of detection of 0.1 microgram/ml (100 parts per
billion).
19) What are the claims about anti-squalene antibodies?
In an effort to explain the health problems of
some Gulf War veterans, a few people have
theorized that a vaccine adjuvant may have caused
an autoimmune disease in veterans. A Vanity Fair
article by Gary Matsumoto, "The Pentagon’s Toxic
Secret" (May 1999), alleges that the DoD possibly
used "an illicit and secret anthrax vaccine" on
its own soldiers. The writer’s interpretation and
presentation of the facts regarding the
Department’s use of anthrax vaccine are
speculative, inflammatory, and wrong. His
allegations and the reported "clinical evidence"
are not new. Since 1997, reports in the
Washington Times, its magazine Insight on the
News, and the (Wilmington) Delaware News Journal,
have made similar allegations regarding “secret
medical experiments” and the like. Investigators
cited in these articles (Pamela Asa, Ph.D.,
Memphis, TN, and Robert Garry, Ph.D., Tulane
University School of Medicine, New Orleans, LA)
report they developed in 1997 and patented a test
for anti-squalene antibodies (ASA). Autoimmune
Technologies, LLC, of New Orleans, has an
exclusive license on the use of this test. The
investigators report that they detected
anti-squalene antibodies in the blood of ill Gulf
War veterans. Their methods were published in the
February 2000 and August 2002 issues of the
journal Experimental and Molecular Pathology. In
the February 2000 article, the authors themselves
conclude: "It is important to note that our
laboratory-based investigations do not establish
that squalene was added as adjuvant to any
vaccine used in military or other personnel who
served in the Persian Gulf War era." Asa and
colleagues published a second article in the
August 2002 issue of Experimental and Molecular
Pathology, but it also provides no validation of
the original assay. As a result, the findings of
the second article are also in question. The
authors' comment that the Matyas article of Nov
2000 supports their findings is mistaken.
20) Have any independent panels evaluated the
claims of researchers to find anti-squalene
antibodies in the blood of ill Gulf War veterans?
Yes, four independent civilian panels considered
the February 2000 article by Asa and colleagues
and other allegations related to squalene and
anti-squalene antibodies. When the Institute of
Medicine (part of the National Academy of
Sciences) Committee on Gulf War and Health (the
“Sox committee”) evaluated the 2000 Asa claims of
anti-squalene antibodies in the blood of ill Gulf
War veterans, it concluded that the paper
contains shortcomings, some serious, that combine
to invalidate the authors’ conclusions. The
report says: "The committee does not regard this
study as providing evidence that the
investigators have successfully measured
antibodies to squalene." See
http://www.nap.edu/books/030907178X/html, pages
311-312. The civilian experts on the Armed Forces
Epidemiological Board (AFEB) said in July 2000,
"the research reported in this paper does not
support this claim; … it remains unclear if the
assay actually measures antibodies to squalene,
as the authors assert…"
http://www.ha.osd.mil/afeb/reports/squalene.pdf
Regarding assertations that Service Members who
received anthrax vaccination from the five lots
cited in the FDA squalene tests experienced more
or more severe adverse events after vaccination,
the civilian physicians on the Anthrax Vaccine
Expert Committee (AVEC) evaluated adverse events
by lot and geographic location. They found no
meaningful differences based on lot or on
geographic location. (Sever et al. 2002
http://www.vaccines.mil/documents/library/AVEC_ms.pdf,
especially pages 198-200, and Sever et al, 2004
http://www.vaccines.mil/documents/library/SeverArticle.pdf,
especially pages 13-15) Of note, the five lots
cited in the FDA squalene tests were shipped to
multiple DoD installations. In addition, Dover
AFB received lots other than the five lots
mentioned above. After the comprehensive review
of anthrax vaccine safety by the National Academy
of Sciences (the “Strom committee,” March 2002,
www.nap.edu/catalog/10310.html), which included
hearing from personnel from Dover AFB and
elsewhere concerned that they suffered adverse
events after anthrax vaccination, the civilian
physicians and scientists concluded that “The
[SRI] study report, dated August 14, 2001, found
that 1 lot of over 30 lots tested contained
measurable levels of squalene. Three samples from
that lot [FAV008] contained squalene at 7, 9, and
approximately 1 parts per billion, respectively.
Use of vaccine from that lot has not been
associated with elevated rates of adverse events.
… Because the available data ... demonstrate that
the presence of trace amounts of squalene is not
associated with an increase in the rates of
adverse events following vaccination with AVA,
the committee concludes that further
investigation of possible AVA contamination is not warranted at this
time.”
21) Are these panels really independent?
The IOM committee members were selected by the
National Academy of Sciences to be fully
independent of both the Department of Defense and
the Department of Veterans Affairs. The AVEC
committee members were selected by the Department
of Health & Human Services to be fully
independent of the Department of Defense. The DHB
is appointed by the Secretary of the Army to
advise the Surgeons General of the military
Services. These civilians constitute a highly
accomplished and widely respected scientific
advisory board. These civilians are free to
render whatever opinions they wish, and their
candidness is important to ensuring that DoD is
using the best possible medical information.
22) What did the GAO say about squalene testing
and what are DoD researchers doing?
In March 1999, the U.S. General Accounting Office
(GAO, now the Government Accountability Office)
released a report "Gulf War Illnesses: Questions
about the Presence of Squalene Antibodies in
Veterans Can be Resolved" (GAO/NSIAD-99-5). The
Department of Defense disagreed with the GAO’s
opinion that "the first step is to determine the
extent to which they [antibodies to squalene] are
present in a larger group of sick Gulf War-era
veterans." The proper first step is to show that
the test for squalene antibodies measures what it claims to measure.
Further, the medical significance and the origin
of antibodies to squalene, even if their
existence is corroborated, remain unknown.
Without such information, Gulf War veterans get
only speculation about the meaning of the test
result and its implication for their health. Gulf
War veterans deserve objective evidence and
recommendations based on sound science. To
investigate the anti-squalene antibody theory, a
scientifically proven test for squalene
antibodies is needed to assess whether Gulf War
veterans have antibodies to squalene. In response
to a DoD solicitation for research on illnesses
among Gulf War veterans, a DoD investigator and
nationally recognized expert on antibodies to
cholesterol and other lipids submitted a research
proposal to determine the feasibility of
developing a test for antibodies to squalene. The
competitively funded research project to
determine whether antibodies to squalene exist has five main
objectives:
1) Development and validation of an enzyme-linked
immunosorbant assay (ELISA) for antibodies against squalene.
2) Evaluation and potential development of other
assays for antibodies to squalene.
3) Development of a positive control antibody to squalene.
4) Production of the positive control antibody to
squalene for use in the assays.
5) Testing of normal human serum for antibodies
to squalene by ELISA and other methods.
23) What did the competitively funded research
project find regarding squalene antibodies?
In April 2000, the research project published its
first peer-reviewed report, describing an
enzyme-linked immunosorbent assay (ELISA) that
could detect antibodies to squalene induced in
mice. Use of squalene alone did not produce a
significant amount of anti-squalene antibodies. A
special chemical was needed to induce the
antibodies against squalene in mice. After
injecting mice with liposomes (fat globules)
containing 71% squalene (710 million parts per
billion), plus a second chemical called lipid A,
antibodies to squalene were readily induced in
mice. The validity of the method was established
using positive and negative controls to preclude
false positive and false-negative test results.
The investigators concluded that squalene is a
weak antigen (a weak inducer of antibodies). (Matyas et al., 2000).
By September 2001, researchers reported improving
the assay and ensuring these tests were
reproducible and sensitive enough to detect 80
ng/ml of anti-squalene antibody. The test was
also reproducible from experiment to experiment
(Matyas et al., 2001). The third study from this
research effort, published in 2004, adapts the
test described above so that it could detect
anti-squalene antibodies if present in human
serum. Serum from three groups of people were
tested: retired employees of the U.S. Army
Medical Research Institute of Infectious Diseases
(average 68 years of age, 88% of whom received
anthrax vaccine, mean = 26 doses per person) ,
civilian volunteers of similar age from
Frederick, Maryland (none of whom received
anthrax vaccine), and random blood donors from
Fort Knox, Kentucky (vaccination status unknown),
This next study indicates that anti-squalene
antibodies are found in 7.5% of the vaccinated
USAMRIID alumni, 15% of the unvaccinated
Frederick civilians, and in 0% of the Fort Knox
blood donors. The antibodies described in the
previous sentence were a type of antibody called
IgG. Researchers found another type of
anti-squalene antibody called IgM in all three
groups (37%, 32%, and 19%). The researchers found
that anti-squalene antibodies are more common
with increasing age (a characteristic also found
in mice). The presence of anti-squalene
antibodies was unrelated to anthrax vaccination
status. They concluded that anti-squalene
antibodies occur naturally in humans (Matyas et al., 2004).
24) Has DoD ever tested squalene-adjuvanted
vaccines in humans against any disease?
Yes. The DoD conducted several human clinical
trials exploring the value of investigational
vaccines containing squalene-based adjuvants to
prevent malaria and HIV infection. The
squalene-containing adjuvants principally
involved products known as MF59 (licensed from
Chiron Corporation) and AS02A (licensed from
GlaxoSmithKline). Each of these studies involved
an FDAapproved scientific plan in human
volunteers told the contents of the vaccine.
Malaria: Hoffman et al, 1994; Epstein et al,
2004; Wang et al, 2004. HIV: Nitayaphan et al,
2000; Pitisuttithum et al, 2003. The Department
of Defense (DoD) has never exposed any military
member or civilian to any squalene-adjuvanted
investigational product without the person’s
informed consent, abiding by FDA regulations.
Civilian researchers, including some funded by
the National Institutes of Health, have conducted
clinical trials of these and other
squalene-adjuvanted vaccines on human volunteers,
ranging from infants to the elderly.
25) Could squalene concerns have anything to do
with various reported clusters of illnesses among
people given anthrax vaccine?
A panel of civilian physicians selected by the
Department of Health & Human Services reviewed
all reports of adverse events after anthrax
vaccination from 1998 to 2001 (Sever et al, 2002;
Sever et al, 2004). This panel was known as the
Anthrax Vaccine Expert Committee (AVEC).
To evaluate assertations that Service Members who
received anthrax vaccination from the five lots
cited in the FDA squalene tests experienced more
or more severe adverse events after vaccination,
these civilian physicians evaluated adverse
events by lot and geographic location. They found
no meaningful differences based on lot or on
geographic location. Of note, the five lots cited
in the FDA squalene tests were shipped to
multiple DoD installations. In addition, Dover
AFB received lots seven lots other than the five
test-positive lots mentioned above.
26) Bottom line, is there any reason for alarm here?
No. Squalene is not added to any US-licensed
vaccine, including anthrax vaccine. The
background level of squalene found by the FDA is
less than the concentration normally present in
human blood. The FDA confirms that these trace
levels are "naturally occurring and safe."
Improved tests found no squalene in the lots
where FDA found it. Nonetheless, DoD continues to
compile additional knowledge about squalene and anti-squalene
antibodies. |
|
|
| Back to top |
|
|
|
|
|
All times are GMT - 5 Hours
The time now is Fri Dec 11, 2009 12:00 am
|
|