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Squalene References
As sufficiently known, squalene is and has been used as an adjuvant to boost the effec-
tiveness of vaccines. Yet, indications are that squalene in the form and concentrations used
in vaccines is very dangerous to human health and should be forbidden. The list that
follows has been established so that people in the field can check for themselves whether
the claims are justified. It has been taken from the groundbreaking book of Gary Matsu-
moto: “Vaccine A – The covert government experiment that’s killing our soldiers and Why
GI’s are only the first victims”, Basic Books, New York # 2004 (pp. 339-44).

Matsumoto’s book is about what caused the Gulf War Syndrome. When American troops
went to the Middle East to fight the Gulf War in 1991, they were immunized with the only
FDA-approved anthrax vaccine in existence – or at least they were supposed to be.
(Anthrax is an agent in biological warfare to which soldiers should be protected.) Many
received an experimental vaccine instead with, as is now well established, high doses of
squalene. Without their knowledge or consent, the militaries were used in a massive medi-
cal experiment, an experiment that went wrong disastrously. As a result many are now suf-
fering from what is commonly called the Gulf War Syndrome, which is associated with
diverse ailments.

The Gulf War Syndrome

The Gulf War Syndrome is a complex of multiple concurrent symptoms that affects
at least one fourth of the 700,000 U.S. veterans who served in that war. It is asso-
ciated with diverse biological alterations that most prominently affect the brain and
the nervous system. Research findings in veterans with the syndrome established
significant differences in brain structure and function, autonomic nervous system
function, neuro-endocrine and immune anomalies, and anomalies associated with
vulnerability to neurotoxic chemicals. This complex of disorders typically includes
persistent memory and concentration loss, chronic headaches, widespread pain,
gastrointestinal problems, and other chronic abnormalities not explained by well-
established diagnoses. No effective treatments have been identified for the Gulf War
illnesses, and studies indicate that few veterans have recovered over time.

Gulf War Syndrome fundamentally differs from trauma and stress-related syn-
dromes described after other wars. Studies consistently indicate that these are not
the result of combat or other stressors. Remarkably, Gulf War veterans have lower
rates of posttraumatic stress disorder than veterans of other wars. No similar wide-
spread, unexplained symptomatic illnesses have been identified in veterans who
have served in war zones since the Gulf War, including the Middle East deployments
in Afghanistan and Iraq in response to the Twin Towers debacle of 2001.

Source: Findings in Brief of the report: “Gulf War Illness and the Health of Gulf War Vete-
rans” by the Research Advisory Committee on Gulf War Veterans’ Illnesses, Washington,
D.C.: U.S. Government Printing Office # November 2008.
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• The following is excerpted from Matsumoto’s book (pp. 156-57) :

«« When Pam Asa first suggested that Gulf War Syndrome might be due to anti-squalene
antibodies, many scientists balked. They said it couldn’t happen: the immune system could
not form antibodies to a lipid or fat like squalene because these molecules were too small.
On the face of it, this was foolish because, for decades, anti-phospholipid antibodies had
been a key test for lupus. The scientists who didn’t know this were probably going by
something presumed to be true decades ago – that only relatively large molecules like pro-
teins could induce the production of an antibody.
The crux of the matter is that “although, under normal circumstances, the body will
not react to its own molecules - a phenomenon that immnologists called “tolerance” – tole-
rance can be broken. This is the essence of autoimmunity. Antibodies against self, or auto-
antibodies, are associated with debilitating and sometimes fatal autoimmune diseases.
There is at least one other reason why many smart, impressively credentialed scien-
tists presumed that the immuyne system would not produce antibodies to squalene. For the
better part of the twentieth century, scientists have believed that aduvants stimulate a non-
specific immune response. In other words, they supposedly activated a robust immune
response to everything but themselves. Scientists conceived this idea before they knew
where antibodies came from or how they worked, and then clung to that idea as scientific
gospel. (…) As molecules go, squalene was relatively light; it was small and it was an
adjuvant – qualities that theoretically added up to no immune resoponse or a non-specific
response. But if the human immune system made antibodies specific to squalene [as it
does] - if squalene activated the immune system’s whole cellular repertoire - then it would
be undeniably dangerous [and cause HAD or Human Adjuvant Disease]. »»

Arlette Mercae explains that «« Squalene has been used extensively as an adjuvant in ani-
mal models to induce auto-immune diseases. (She also told that) recent research into Gulf
War Syndrome (…) showed that 95% of overtly ill troops who were deployed, and 100%
of those who were immunized but not deployed, had antibodies to squalene. Antibodies to
squalene were not detectable in the general public, or in Gulf War Veterans who were not
ill. Two control subjects who volunteered to participate in a vaccine trial at the United
States National Institutes of Health developed a multisystem disorder similar to that of
Gulf War Syndrome, which trial involved the use of a squalene-containing adjuvant. »»

Source: “From Immunology to Social Policy”, a thesis by Arlette Mercae, University of Tas-
mania # 2003 (pp. 83-84).
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Remark: Squalene related research has continued after the publication of the afore-
mentioned book and its list. The list is sufficiently broad to show by means of a study of
the research what is wrong with squalene as an adjuvant. And, at least, it gives a
springboard to look for further research.

• Squalene Induces Autoimmune Disease in Animals

1. Whitehouse MW, Orr KJ, Beck FW, Pearson CM [Division of Rheumatology, Department of
Medicine, University of California School of Medicine, Los Angeles, California], “Freund’s
Adjuvants: Relationship of Arhritogenicity and Adjuvanticity in Rats to Vehicle Composition,”
Immunology, (1974) Aug;27(2)311-30.
2. Beck FW, Whitehouse MW, Pearson CM [Division of Rheumatology, Department of
Medicine, University of California School of Medicine, Los Angeles, California],
“Improvements for consistently inducing experimental allergic encephalomyelitis (EAE) in
rats: I. without using mycobacterium. II. inoculating encephalitogen into the ear,” Proceedings
of the Society for Experimental Biology and Medicine, (1976) Mar;151(3):615-22.
3. Kohashi O, Pearson CM [Division of Rheumatology, Department of Medicine, University of
California School of Medicine, Los Angeles, California], “Arthritogenicity of Mycobacterium
smegmatis subfractions, related to different oil vehicle and different composition,”
International Archives of Allergy Applied Immunology, (1976); 51(4):462-70.
4. Beck FW, Whitehouse MW [Division of Rheumatology, Department of Medicine, University
of California School of Medicine, Los Angeles, California and Department of Experimental
Pathology, John Curtin School of Medical Research, The Australian National University,
Canberra A.C.T. 2600, Australia], “Modificatioons in the Establishment of Allergic
Encephalomyelitis (EAE) in Rats; an Improved Essay for Immunosuppressant Drugs,” Agents
Actions, (1976) July;6(4):460-7.
5. Zamma T [Department of Oral Surgery, School of Medicine, Nagoya University, Showa-Ku,
Nagoya, 466 Japan], “Adjuvant-Induced Arthritis in the Temporomandibular Joint of Rats,”
Infection and Immunity, (1983) Mar;39(3):1291-9.
6. Johnston BA, Eisen H, Fry D [Fred Hutchinson Cancer Research Center, Seattle, Washington],
“An Evaluation of Several Adjuvant Emulsion Regimens for the Production of Polyclonal
Antisera in Rabbits,” Laboratory Animal Science, (1991) Jan;41(1):15-21.
7. Lipman NS, Trudel LJ, Murphy JC, Sahali Y [Division of Comparative Medicine,
Massachusetts Institute of Technology, Cambridge, MA 02139], “Comparison of Immune
Response Potentiation and In Vivo Inflammatory Effects of Freund’s and Ribi Adjuvants in
Mice,” Laboratory Animal Science, (1992) Apr;42(2):193-7.
8. Leenaars PP, Hendriksen CF, Angulo AF, Koedam MA, Claasen E [National Institute of Public
Health and Environmental Protection (RIVM), P.O. Box 1, 3720 BA, Bilthoven, The
Netherlands], “Evaluation of several adjuvants as alternatives to the use of Freund’s adjuvant
in rabbits,” Veterinary Immunology and Immunopathology, (1994) Mar;40(3):225-41.
9. Leenaars M, Koedam MA, Hendriksen CF, Claassen E [National Institute of Public Health and
Environmental Protection (RIVM), P.O. Box 1, 3720 BA Bilthoven, The Netherlands],
“Immune responses and side effects of five different oil-based adjuvants in mice,” Veterinary
Immunology and Immunopathology, (1994) Mar;40(3):225-41.
10. Leenaars M, Koedam MA, Ester PW, Baumans V, Claassen E, Hendriksen CF [National
Institute of Public Health and Environmental Protection (RIVM), P.O. Box 1, 3720 BA
Bilthoven, The Netherlands], “Assessment of side effects induced by injection of different
adjuvant/antigen combinations in rabbits and mice,” Laboratory Animals (1998)
Oct;32(4):387-406.
11. Kleinau S, Erlandsson H, Klareskog L [Department of Clinical Immunology, University
Hospital, Uppsala, Sweden], “Percutaneous exposure of adjuvant oil causes arthritis in DA
rats,” Clinical Experimental Immunology, (1994) May;96(2):281-4 (* Refers to olive oil,
which contains squalene).
12. Yoshino S, Yoshino J [Rheumatology Unit, Royal Adelaide Hospital, Adelaide, SA5000,
Australia], “Recruitment of pathologenic T cells to synovial tissues of rats injected
intraarticularly with nonspecific agents,” Cellular Immunology, (1994) Oct.15;158(2):305-13.
13. Smialek M, Gajkowska B, Ostrowski RP, Piotrowski P [Department of Neuropathology and
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Laboratory of the Ultrastructure of the Nervous System, Medical Research Centre, Polish
Academy of Sciences, Warszawa, Poland], “Experimental squalene encephaloneuropathy in
the rat,” Folia Neuropathologica, (1997);35(4):262-4.
14. Gajkowska B, Smialek M, Ostrowski RP, Piotrowski P, Frontczak-Baniewicz M [The
Laboratory of the Ultrastructure of the Nervous System, Medical Research Centre, Polish
Academy of Sciences, 5 Pawinskiego Street, 02-106 Warsaw, Poland], “The experimental
squalene encephaloneuropathy in the rat,” Experimental and Toxicologic Pathology, (1999)
Jan;5:75-80.
15. Lorentzen JC [Deparment of Medicine, Karolinska Hospital, Karolinska Institutet, Stockholm,
Sweden], “Identification of arhtritogenic adjuvants of self and foreign origin,” Scandinavian
Journal of Immunology, (1999) Jan;49(1):45-50.
16. Carlson BC, Jannson AM, Larsson A, Bucht A, Lorentzen JC [Department of Medicine,
Karolinska Institutet, Stockholm, Sweden], “The endogenous adjuvant squalene can induce a
chronic T-cell-mediated arthritis in rats,” American Journal of Pathology, (2000)
Jun;156(6):2057-65.
17. Holm BC, Zu HW, Jacobsson L, Larson A, Luthman H, Lorentzen JC [Center for Molecular
Medicine, Department of Medicine, Unit of Rheumatology, Karolinska Institutet, Stockholm,
Sweden], “Rats made congenic for Oia3 on chromosome 10 become susceptible to squalene-
induced arthritis,” Human Molecular Genetics, (2001) Mar;215(6):565-72.
18. Holmdahl R, L orentzen JC, Lu S, Olofsson P, Wester L., Holmberg J, Pettersson U, [Section
of Medidal Inflammation Research, Lund University, Sweden]. “Arthritis induced in rats with
nonimmunogenic adjuvants as models for rheumatoid arthritis” Immunological Reviews,
(2001) Dec;184:184-202.
19. Holm BC, Svelander L, Bucht A, Lorentzen JC [Department of Medicine, Unit of
Rheumatology, Karolinska Institutet, Stockholm and Department of Medical Countermeasures,
Division of NBC Defense, Defense Research Agency, Umea, Sweden], “The arthritogenic
adjuvant squalene does not accumulate in joints, but gives rise to pathogenic cells in both
draining and non-draining lymph nodes,” Clinical and Experimental Immunology, (2002)
Mar;127(3):430-5
20. Whitehouse MW, Beck FWJ, Matsumoto G [Department of Medicine, University of
Queensland, Princess Alexandra Hospital, Queensland, Australia; Wayne States University
Medical Center, Detroit, Michigan, U.S.A.], “Squalene is an Auto Toxicant Inducing
Polyarthritis in Rats and Immunopathies in Man, Abstract,” The Australian Health and
Medical Congress, 2002, no. 1143.
21. Gherardi RK [Groupe Nerf-Muscle, Departement de Pathologie, Hôpital Henri Mondor,
Créteil, Paris, France], “Lessons from macrophagic myofasciitis: towards definition of a
vaccine adjuvant-related sydrome,” Revue Neurologique, (2003) Feb;159(2):162-4.
22. Backdahl L, Ribbihammar U, Lorentzen JC [Center for Molecular Medicine, Karolinska
Institutet, Stockholm], “Mapping and functional characterization of rat chromosome 4 regions
that regulate arhritis models and phenotypes in congenic strains,” Arthritis and Rheumatism,
(2003) Feb;48(2):551-9.
23. Satoh M, Kuroda Y, Yoshida H, Behney KM, Mizutani A, Akaogi J, Nacionales DC,
Lorenson TD, Rosenbauer RJ, Reeves WH [Division of Rheumatology and Clinical
Immunology, Department of Medicine, University of Florida, Gainesville], “Induction
of lupus autoantibodies by adjuvants,” Journal of Autoimmunbity, (2003) Aug;21(1):1-9.
24. Kuroda Y, Akaogi J, Nacionales DC, Wasdo SC, Szabo NJ, Reeves WH, Satoh M [Division of
Freumatology and Clinical Immunology, Department of Medicine, University of Florida,
Gainesville], “Distinctive Patterns of Autoimmune Response Induced by Different Types of
Mineral Oil,” Toxicological Sciences, (2004) Apr;78(2):222-8.
25. Kuroda Y, Nacionales DC, Akaogi J, Reeves WH, Satoh M [Division of Rheumatology and
Clinical Immunology, Department of Medicine, University of Florida, Gainesville],
“Autoimmunity induced by adjuvant hydrocarbon oil components of vaccine,” Biomedicine &
Pharmflcotherapy, (2004) Jun;(58)5:325-37.
26. Holm BC, Lorentzen JC, Bucht A [Diabetes Research, Immunology Unit, Department of
Endocrinology, Lund University, Malmö University Hospital, Stockholm], “Adjuvant oil
induces waves of arthritogenic lymph node cells prior to arthritis on-set,” Clinical and
Experimental Immunology, (2004) Jul;137(1):59-64.
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• Adverse Reactions in Humans to Experimental Vaccines

Containing Squalene
27. Keitel W, Couch R, Bond N., Adair S, Van Nest G, Dekker C [Baylor College of Medicine,
Department of Microbiology and Immunology, One Baylor Plaza, Houston, Texas 77030],
“Pilot evaluation of influenza virus vaccine (IVV) combined with adjuvant,” Vaccine,
(1993);11(9):909-13; *[See also Nos. 27 & 31].

• Squalene Stimulates the Immune System

28. Ott G, Barchfield GL, Chernoff D, Radhakrishnan R, van Hoogevest P, Van Nest G
[Chiron Corporation, Emeryville, California 94608], “MF59 - Design and evaluation of a safe
and potent adjuvant for human vaccines,” Pharmaceutical Biotechnology, (1995);6:277-96.
29. Ott G, Barchfield GL, Chernoff D, Radhakrishnan R, van Hoogevest P, Van Nest G [Chiron
Corporation, Emeryville, CA 94608], “MF59 - Design and Evaluation of a Safe and Potent
Adjuvant for Human Vaccines,” Vaccine Design: The Subunit and Adjuvant Approach
(Monograph), (1995) Ch.10:277-311.
30. Ott G, Barchfield GL, Van Nest G [Chiron Corporation, EmeryvilIe, CA 94608],
“Enhancement of humoral response against human influenza vaccine with the simple
submicron oil/water emulsion adjuvant MF59,” Vaccine, (1995) Nov;13(16):1557-62.
31. O’Hagan DT, Ott GS, Van Nest G [Chiron Corporation, Emeryville, CA 94704], “Recent
advances in vaccine adjuvants: the development of MF59 emulsion and polymeric
microparticles,” Molecular Medicine Today, (1997) Feb;3(2):69-75.
32. Allison AC [Suromed Corporation, 1060 East Meadow Circle, Palo Alto, California 94303],
“Squalene and squalene emulsions as adjuvants,” Methods, (1999) Sept;19(1):87-93.

• How the Immune System Processes Squalene

33. Depuis M, Murphy TJ, Higgins D, Ugozzoli M, Van Nest G, Ott G, McDonald DM
[Cardiovascular Research Institute, University of California, San Francisco, CA 94143],
“Dendritic cells internalize vaccine adjuvant after intramuscular injection,” Cellular
Immunology, (1998) May 25;186(1):18-27.
34. Depuis M, McDonald OM, Ott G [Cardiovascular Research Institute, University of California,
San Francisco, CA 94143], “Distribution of adjuvant MF59 and antigen gD2 after
intramuscular injection in mice,” Vaccine, (1999) Oct 14;18(5-6):434-9.
35. Depuis M, Denis-Mize K, LaBarbara A, Peters W, Charo IF, McDonald DM, Ott G
[Cardiovascular Research Institute and Department of Anatomy, University of California, San
Francisco, CA 94143], “Immunization with the adjuvant MF59 induces macrophage trafficking
and apoptosis,” European Journal of Immunology, (2001) Oct 31;(10):2910-18.

• Specificity of Antibody Response to Squalene

36. Asa PB, Cao Y, Garry RF [Department of Microbiology and Immunology, Tulane Medical
School, 1430 Tulane Avenue, New Orleans, Louisiana 70112], “Antibodies to Squalene in
Gulf War Syndrome,” Experimental and Molecular Pathology, (2000) Feb;68(1):55-64.
37. Matyas GR, Wasseff NM, Rao M, Alving CR [Department of Membrane Biochemistry, Walter
Reed Army Institute of Research, 20910-7500, Silver Spring, MD], “Induction and detection of
antibodies to squalene,” Journal of Immunological Methods (2000) Nov 1;245(1-2):1-14.
38. Alving CR, Grabenstein JD [Walter Reed Army Institute of Research and Anthrax Vaccine
Immunization Program Office], “RE: Antibodies to squalene in Gulf War Syndrome,”
Experimental and Molecular Pathology, (2000) Jun;68(3):196-8.
39. Asa PB, Cao Y, Garry RF [Department of Microbiology and Immunology, Tulane Medical
School, 1430 Tulane Avenue, New Orleans, Louisiana 70112], “Reply,” Experimental and
Molecular Pathology, (2000) Jun;68(3):197-8.
40. Asa PB, Wilson RB, Garry RF [Department of Microbiology and Immunology, Tulane
Medical School, 1430 Tulane Avenue, New Orleans, Louisiana 70112], “Anti-bodies to
Squalene in recipients of anthrax vaccine,” Experimental and Molecular Pathology, (2002)
Aug;73(1):19-27.
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41. Matyas G, Rao M, Alving C [Department of Membrane Biochemistry, Walter Reed Army
Institute of Research, 503 Robert Grant Avenue, 20910-7500, Silver Spring,
MD, USA], “Induction and detection of antibodies to squalene. II. Optimization of the assay
for murine antibodies,” Journal of Immunological Methods, (2002) Sep 15; 267(2):119.
42. Matyas GR, Rao M, Pittman PR, Burge R, Robbins IE, Wassef NM, Thivierge B, Alving CR
[Department of Membrane Biochemistry, Walter Reed Army Institute of Research], “Detection
of antibodies to squalene: III. Naturally occurring antibodies to squalene in humans and mice,”
Journal of Immunological Methods, (2004) Mar;286(102):47-67.

• Ingested Squalene Is Processed Differently from Injected Squalene

43. Tilvis RS, Miettinen TA [Department of Medicine, University of Helsinki, Helsinki, Finland],
“Absorption and metabolic fate of dietary 3H-squalene in the rat,” Lipids, (1983)
Mar;18(3):233-38.
44. Gylling H, Miettinen TA [Second Department of Medicine, University of Helsinki, Helsinki,
Finland], “Postabsorptive metabolism of dietary squalene,” Artherosclerosis, (1994)
Apr;106(2):169-78.
45. Relas H, Gylling H, Miettinen TA [Department of Medicine, University of Helsinki, Helsinki,
Finland], “Effect of stanol ester on postabsorptive squalene and retinyl palmitate,” Metabolism,
(2000) April;49(4):473-78.

• Detecting Squalene in Anthrax Vaccine Adsorbed [BioThrax]

and U.K. Anthrax Vaccine
46. Spangood RJ, Wu B, Sun M, Lim P, Ellis WY [SRI International, 333 Ravenswood Avenue,
Menlo Park, CA 94025], “Development and application of an analytical method for the
determination of squalene in formulations of anthrax vaccine adsorbed,” Journal of
Pharmaceutical and Biomedical Analysis, (2002) June 20;29(1-2):183-93.
47. May JC, Del Grosso A, Swartz L, Progar JJ, “Chemical Test Results for Michigan Department
of Public Health, Anthrax Vaccine Adsorbed, Lots FAV020 and FAV030,” Department of
Health and Human Services, Food and Drug Administration, CBER, Lab Report, Personal
Communication to Neil Goldman, Ph.D., June 25;1999:1-6.
48. Wood D [Scientific Analysis Laboratories Ltd., Medlock House, New Elm Road, Manchester
M3 4JH, United Kingdom], “The Determination of Squalene in a Vaccine Sample, Scientific
Analysis Laboratories, SAL Report 34120E,” Scientific Analysis Laboratories Ltd., Lab
Report, Personal Communication to Asif Hasan, Granada Television, Febr 26;2003:1-6.