Tick-Borne Infections & Lyme

Disease Contributing to Autism

Spectrum Disorders

Robert C Bransfield, MD, DFAPA

Autism One Conference
Chicago, Ill
May 25, 2008
 Disclosure Statement
Robert Bransfield, MD, DFAPA, PC
• Most of my income is paid directly from patients
• No insurance, government or other payer contracts
that restrict patient care in return for referrals,
financial considerations or any other benefit
• No Lyme disease financial interests.
• Speakers Bureau: Abbott, Astra Zeneca, Forest,
GSK, Jazz, Lilly, Pfizer, Sanofi Aventis, Shire,
Takeda, UCB.
• President Elect International Lyme and Associated
Diseases Society
•
Outline
The association between LYD/TBD and ASD
• Background
• Clinical experience
• Gestational LYD/TBD
• Laboratory testing of ASD patients for TBD
• Biochemical similarities
• Brain imaging similarities
• Epidemiological findings
• Pathophysiology
• Theoretical issues: genes, infections and autism
• Klüver–Bucy Syndrome, infections and autism
• Neural networks, neurodevelopment, autism and borreliosis
• Borreliosis and Borrelia related complex
• Tick-borne/borreliosis infections and psychiatric illness
• Immune responses in ASD, Borreliosis and Mycoplasma infections
• Implications & Response
• Economic issues
• Assessment
• Treatment strategies
• Further evaluation of the hypothesis
• Summary
 Autism: A Syndrome of Many
Different Etiologies & Comorbidities
• Comorbidity with any DSM Dx
• Epilepsy
• Mental retardation (30% mild
to moderate, 40% serious to
profound)
• Hearing & visual impairments
• Fragile X syndrome & multiple
other genetic
• Tuberous sclerosis
• Cerebral palsey
• Phenylketonuria
• Neurofibromatosis
• Congenital rubella
• Rett’s syndrome
• Rasmussen’s encephalitis
• Lennox-Gastaut syndrome
• Post infectious
• Metabolic
• Pyruvate d hydrogenase
deficiency
• Impaired purine metabolism (uric
acid increased)
• Brain structural—cyst, etc.
• PKU phenylalanine
• Angelman’s syndrome
• Landau-Kleffner syndrome
• Prader-Willi
• Williams
• Multiple other genetic
impairments
• Associated with older paternal
age
 What Causes Autism?
• “Data suggest that autism results from
multiple etiologies with both genetic and
environmental contributions, which may
explain the spectrum of behaviors seen in
this disorder.”*

*Libbey, et al.
 The Neurobiology of Autism
• It is becoming clear that the normal trajectory of neurodevelopment
is altered in autism, with aberrations in brain growth, neuronal
patterning and cortical connectivity. Changes to the structure and
function of synapses and dendrites have also been strongly
implicated in the pathology of autism by morphological, genetic and
animal modeling studies.
• Environmental factors are likely to interact with the underlying
genetic profile, and foster the clinical heterogeneity seen in autism
spectrum disorders. In this review we attempt to link the molecular
pathways altered in autism to the neurodevelopmental and clinical
changes that characterize the disease.
• We focus on signaling molecules such as neurotrophin, Reelin,
PTEN and hepatocyte growth factor, neurotransmitters such as
serotonin and glutamate, and synaptic proteins such as neurexin,
SHANK and neuroligin.
• We also discuss evidence implicating oxidative stress, neuroglial
activation and neuroimmunity in autism.

Pardo CA, EberhartCG. Brain Pathology. Volume 17 Issue 4 Page 434-447, October 2007
 Personal Experience
• Primarily clinical practice
• 35 year work with autistic patients
• Interest in infectious causes of mental illness for
30+ years.
• About 2000+ patients in with infections and other
causes of inflammation, especially Lyme/tick-
borne disease. 1000 data points kept on each
patient. Many have progressive systemic
illnesses with encephalopathy.
• Moderator of Microbes and Mental Illness
Internet discussion group (9 years).
 • “Whereas Lyme disease and other tick-
borne diseases are a serious public health
threat;…Findings more common in
children include autism, Tourette’s
syndrome, attention deficit disorder,
dyslexia, lethargy, and a decline in grades,
tantrums;…”

Bransfield, Fallon, Raxlen, Shepler, Sherr“A Modest Proposal,” Psychiatric News

(Newspaper of the APA), Vol. XXXIII, Number 18, September 18, 1998, p. 16.
 A controlled study of cognitive deficits
in children with chronic Lyme disease
• Twenty children with a history of new-onset cognitive
complaints after Lyme disease were compared with 20
matched healthy control subjects. Each child was
assessed with measures of cognition and
psychopathology. Children with Lyme disease had
significantly more cognitive and psychiatric
disturbances. Cognitive deficits were still found after
controlling for anxiety, depression, and fatigue. Lyme
disease in children may be accompanied by long-term
neuropsychiatric disturbances, resulting in
psychosocial and academic impairments.
The association between tick-borne infections, Lyme
borreliosis and autism spectrum disorders
• Tick-borne infections, including Lyme disease contribute to
developing autism spectrum disorders (ASD) by direct effects,
promoting other infections and immune effects during fetal
development and infancy. Combined with other predisposing and
contributory factors these infections may provoke immune reactions
in susceptible individuals that result in inflammation, molecular
mimicry, kynurenine pathway changes, increased quinolinic acid
and decreased serotonin, oxidative stress, mitochondrial
dysfunction and excitotoxicity that impair the development of the
amygdala and other neural structures and neural networks resulting
in a partial Klüver–Bucy Syndrome and other deficits resulting in
autism spectrum disorders and/or exacerbating ASD from other
causes.
• Supporting data includes multiple cases of mothers with Lyme
disease and children with ASD; fetal neurological abnormalities
associated with tick-borne diseases; similarities between tick-borne
diseases and ASD regarding symptoms, pathophysiology, immune
reactivity, temporal lobe pathology, and brain imaging data; positive
reactivity in several studies with ASD patients for Lyme disease
(22%, 26% and 20–30%) and 58% for mycoplasma; similar
geographic distribution and improvement in autistic symptoms from
antibiotic treatment.
Bransfield RC, Wulfman JS, Harvey WT, Usman AI. Medical Hypotheses. 2007
 Chronic Bacterial and Viral Infections
in Neurodegenerative and
Neurobehavioral Diseases
• Often patients with neurodegenerative or neurobehavioral diseases
have chronic, neuropathic infections that could be important in disease
inception, progression or increasing the types/severities of
signs/symptoms. Although controversial, the majority of patients with
various neurodegenerative or neurobehavioral conditions, such as
amyotrophic lateral sclerosis, multiple sclerosis, Alzheimer’s disease,
Parkinson’s disease and autistic spectrum disorders, show evidence of
central nervous system and/or systemic bacterial and viral infections.
For example, using serology or polymerase chain reaction evidence of
Chlamydia pneumoniae, Borrelia burgdorferi, Mycoplasma species,
human herpesvirus-1 and -6 and other bacterial and viral infections
revealed high infection rates that were not found in control subjects.
Although chronic infections were not found in some studies and the
specific role of chronic infections in neurological disease pathogenesis
has not been determined or is inconclusive, the data suggest that
chronic bacterial and/or viral infections could be common features of
progressive neurodegenerative and neurobehavioral diseases.
Clinical Experience
 Clinical Observations
Autism & Lyme/Tick-Borne Disease
• A number of clinicians* and myself have noted an association
between autism & Lyme/tick-borne disease.
• This association is seen as:
– Infected mothers & children with autistic spectrum disorders (ASD)
– Infected infants & ASD or autistic symptoms
– Infected children & autistic symptoms
• Lyme/TBD infections in adults often have some symptoms
suggestive of ASD:
– Sound sensitivity & sensory hyperacusis
– Emotional detachment
– Mood instability
– Decline of speech & language
– Seizures
• More severe symptoms are associated with infections at younger
age, coinfections (Bartonella, Babesia & other TBD), genetic
vulnerability & lengthy misdiagnosis & under-treatment
• Multi-systemic symptoms is a diagnostic clue
• Antimicrobial (anti-bacterial & anti-viral) treatment can reverse these
symptoms
*Schaller, Corson, Levin, Berenbaum
 Opinion of Dr Burrascano
• “It is my contention that Autism is an
inflammatory encephalitis cause by a pathogen
such as Bartonella or Mycoplasma. I share the
view that Bartonella/BLO is a major infection that
may eclipse Bb as the ultimate cause of the
morbidity in chronic Lyme. Mycoplasma too is a
major concern of mine- in reviewing my 7000+
cases, those patients who were relentlessly
chronic all, at one point or another in their
illness, were PCR + for Mycoplasma.”
BI/TBI & ASD: Similar GI Symptoms
• Kugathasan S. Pediatric inflammatory bowel disease:
clinical and therapeutic aspects. Curr Opin
Gastroenterol. 2001;17(4):350-5.
• Zaidi AS, Singer C. Gastrointestinal and Hepatic
Manifestations of Tick-borne Diseases in the United
States. Clinical Infect Dis 2002;34:1206-12.
• Sherr VT. “Bell’s palsy of the gut” and other GI
manifestations of Lyme and associated diseases. Pract
Gastroent. 2006:74-91
• Fried MD, Abel M, Pietrucha D, Kuo YH, Bal A. The
Spectrum of Gastrointestinal Manifestations in Children
and Adolescents with Lyme Disease. JSTD 1999;6.
• Fried M. Gastrointestinal manifestations of Lyme
disease. Program and abstracts of the 14th Int Sc Conf
Lyme Disease and Other TBD; 2001
BI/TBI & ASD Respond to Similar Treatments

• Both BI/TBI and ASD respond to similar

treatments of psychiatric and cognitive
symptoms with psychotropics (including
modafinil, stimulants, memantine, mood
stabilizers, atypical agents and serotonin
reuptake inhibitors), diets, antimicrobials,
mitochondrial enhancers, immune
modulators, hyperbaric oxygen,
glutathione, chelation and allergen
elimination.
 BI/TBI & ASD Respond to Similar
Treatments: References
• Posey DJ, Kem DL, Swiezy NB, Sweeten TL, Wiegand
RE, McDougle C. Pilot study of D-cycloserine in subjects
with autistic disorder. Am J Psych. 2004;161(11):2115-7.
• Sandler RH, Finegold SM, Bolte ER, et al. Short-term
benefit from oral vancomycin treatment of regressive-
onset autism. J Child Neurol. 2000;15(7):429-35.
• Bransfield, RC. Update on the Neuropsychiatric
Management of Lyme and Associated Diseases. ILDS
Annual Meeting. 2004
• 27. Bransfield RC. Treatment of Autism with LYD/TBI.
LIA Conference; 2007
• 28. Erickson CA, Posey DJ, Stingler KA, McDougle CJ.
Pharmacotherapy of autism and related disorders.
Psychiatric Annals. 37:7
 Brain SPECT: Mother with Lyme
Disease & 3 ASD Children
• SPECT scans of a mother with TD/BI and
her three children with ASD which
demonstrate many of the points previously
described.
• Mother, 48: There is an extensive
hypoperfusion pattern, prominently
in the cerebral cortices and much
of the frontal lobes, with a lesser
degree in the temporal lobes and a
small degree hypoperfusion in the
cerebellum. The hypoperfusion is
moderately extensive and is likely
associated with toxic, inflammatory
and infectious processes. There is
hyperperfusion of the Basal
Ganglia, which is associated with
anxiety and mood dysregulation.
The diagnosis is chronic fatigue
syndrome, multiple sclerosis,
depression and possible
congenital Lyme disease. Lab
testing was positive for Borrelia
burgdorferi, Babesia WA-1,
Bartonella henselae, Mycoplasma
fermentans, HHV-6, Epstein Barr
virus, high anti-streptolysin o titre
and gamma Strep in stool.
• Son, 26: Some motion artifacts, however
significant hypoperfusion pattern is both
focal as well as generalized. The focal
pattern is throughout the cerebral cortex
bilaterally and the cerebellar hemispheres
(which demonstrate atrophy on MRI) There
is mild hyperperfusion of the basal ganglia
and a focally intense hypeperfusion area in
the deep white matter of the temporal lobe.
There is a hyperperfusion pattern involving
the temporal lobes and cerebellar
hemispheres. The focal decrease is more
suggestive of etiologies that would include
hypoxic, neuroimmune, traumatic factors,
infectious and inflammatory. There is a
hyperperfusion pattern of the basal ganglia
which may be associated with element of
anxiety, whereas the focal intense areas
can be associated with present interictal
seizure focus and is clinically significant as
the present dose of anticonvulsant is not
controlling this area. The patient is low
functioning with autism spectrum disorder
since two years, grand mal seizures,
movement disorder, ataxia, hypotonia,
megacolon, possible mitochondrial disorder,
mild hypergammaglobulenmia and
syncope.
• Lab testing was positive for Borrelia
burgdorferi, Babesia WA-1, Bartonella
henselae, Mycoplasma fermentans, HHV-6,
EBV and high strep titers; stool positive for
Citrobacter fundii, Klebsialla p. and gamma
Strep.
• Daughter, 23: There is an
extensive hyperperfusion pattern in
the cerebral cortices, temporal
lobes and cerebellum and
hypoperufsion of the frontal lobes
and is likely associated with toxic,
inflammatory and infectious
processes. The diagnosis is
Asperger’s, obsessive compulsive
disorder, generalized anxiety,
social anxiety disorder,
depression, posttraumatic stress
disorder from an auto accident,
possible narcolepsy, tremors,
cardiac disease, myocardial
infarction, osteopenia, arthritis and
psuedo rhematoid nodules since 5
years of age. Lab testing was
positive for Borrelia burgdorferi,
Anaplasma phagocytophilum,
Mycoplasma fermentans,
Homopholis, HHV-6, EBV,
elevated Strep titres; stool was
positive for Toxoplasmosis,
Cornybacteria and gamma Strep.
• Daughter, 20: there is extensive
hypoperfusion in the frontal lobes,
temporal lobes and to a lesser
degree to the occipital lobes and
slightly to the cerebellum. There
is hyperperufsion in the right
cerebellar hemisphere. The
hypoperfusion is likely associated
with neuroinflammatory,
neuroimmunological, infectious
and toxic substance exposure.
There is a seizure focus with
hyperperfusion in the right
cerebellar hemisphere. The
diagnosis is autism spectrum
disorder since 14 months, petit
mal seizure disorder, hypotonia,
perceptual impairments, and
anxiety. Lab testing was positive
for Borrelia burgdorferi, Bartonella
henselae, Mycoplasma
fermentans, HHV-6; stool positive
for Parvo/B-19, Klebsiella, p.,
Citrobacter f. and gamma Strep.
 Gestational Tick-Borne
Infections/Borreliosis Infections
 Maternal Fetal Issues
• Antibodies
– From mother
– From fetal immune response to intrauterine infection
• Pregnancy and the Maternal Immune System
– Down regulated
• Teratogenic Issues in the Developing Fetus
– Rubella infection model for intrauterine infections

Mac Donald
 Harvey, WT; Salvato, P: ‘Lyme Disease’: Ancient
Engine of an Unrecognized Borreliosis Pandemic?
Medical Hypotheses (2003) 60(5), 742-759; Elsevier Science Ltd.

Congenital

an ted

U Hu
ni m
Transfer

um ec
s

nf an
H inf

ec s
n

te
U

d
Humans
Infected With Sexual
Borrelia Transfer
burgdorferi

Nonarthropod- Arthropod-
Vectored Vectored
Zoonotic Zoonotic
Borreliosis Lyme
Disease

Figure 1. Revised model of human Borrelia burgdorferi infection

 Maternal Transmission of LYD
• Schlesinger PA, Duray PH, Burke BA, Steere AC, Stillman MT. Maternal-fetal transmission of
the LYD spirochete, B. burgdorferi. Ann Intern Med. 1985;103(1):67-8.
• Lampert F. Infantile multisystem inflammatory disease: another case of a new syndrome. Eur J
Pediatr. 1986;144(6):593-6.
• Lavoie PE, Lattner BP, Duray PH, et al. Culture positive, seronegative, transplacental Lyme
borreliosis infant mortality. Arthritis Rheum.1987; 3:S50.
• MacDonald AB. Gestational Lyme borreliosis. Implications for the fetus. Rheum Dis Clin North
Am. 1989;15(4):657-77.
• Markowitz LE, Steere AC, Benach JL, Slade JD, Broome CV. Lyme disease during pregnancy.
JAMA. 1986;255(24):3394-6.
• Nadal D, Hunziker UA, Bucher HU, Hitzig WH, Duc G. Infants born to mothers with antibodies
against Borrelia burgdorferi at delivery. Eur J Pediatr. 1989;148(5):426-7.
• Hercogova et al. Could borrelia found in the placenta influence the fetus? Study of 19 women
with EM during pregnancy. 6th Int Conf Lyme Borreliosis.1994
• Gardner T. Lyme disease In Infec Dis Fetus and Newborn Infant. Saunders, 1995
• Gardner T. Lyme disease. 66 Pregnancies complicates by Lyme Borreliosis. Infec Dis Fetus
and Newborn Infant. Saunders, 2000.
• Jones CR, Smith H, Gibb E, Johnson L. Gestational Lyme Disease Case studies of 102 Live
Births. Lyme Times. 2005;Summer:34-6.
• Harvey, WT; Salvato, P: ‘Lyme Disease’: Ancient Engine of an Unrecognized Borreliosis
Pandemic? Medical Hypotheses (2003) 60(5), 742-59.
• Bach G. Sexual Transmission of Lyme Disease. Microbes and Mental Illness Symp; Am Psych
Assn Inst Psych Services. 2000
• Jones, CR. Lyme Disease and Autism. LIA Conf;2007
• Schmidt BL, Aberer E, Stockenhuber C, Klade H, Breier F, Luger A. Detection of Borrelia
burgdorferi DNA by polymerase chain reaction in the urine and breast milk of patients with
Lyme borreliosis. Diagn Microbiol Infect Dis. 1995;21(3):121-8.
 Perinatal Transmission of the Agent
of Human Granulocytic Ehrlichiosis

• We describe a case of Human

Granulocytic Ehrlichiosis that
developed in a pregnant woman near
term and was transmitted perinatally
to her infant.
• (Hawaii has a high incidence of
Ehrlichiosis)

Horowitz HW et al. NEJM. Volume 339 Number 6. p

Laboratory testing of ASD
patients for tick-borne
diseases
 Testing ASD Patients for TBI
• Vojdani tested Autism samples from different clinics in Northern CA, NY, NJ and CT.
22% of (12/54) tested IgG and IgM positive for Bbsl by Immunosciences Lab (Note:
in this sample the Western Blot (WB) test used CDC surveillance criteria and did not
include the full complement of Bbsl specific bands.
• A LIAF study tested the blood of 19 children with an ASD diagnoses plus an
indication of immune dysfunction and five normal controls. Patients were not
screened for BI before study entry. WB and IFA IgG and IgM were performed by
IgeneX Laboratory. A result was considered Bbsl positive for exposure if there was
reactivity of one or more Bbsl specific bands. 26% of the ASD children were positive
compared to 0 controls.
• Levine tested nine consecutive ASD patients in Connecticut in 2003 and all nine
tested positive for Bbsl with WB by IGeneX Laboratory criteria.
• Nicolson tested 48 ASD patients with forensic PCR and Southern Blot confirmation.
20–30% (depending upon the lab) were positive for Bbsl. 58% were positive for
Mycoplasma species while 5% of 45 age matched controls were positive for
Mycoplasma (Odds ratio = 13.8) with 35% M. fermentans vs. 0% control, 33% M.
pneumoniae vs. 5% control, 10% M. homonis vs. 0% control, 2% M. penetrans vs.
0% control and 25% were M. fermentans and other species. Also 8% were positive
for C. pneumoniae vs. 2% of controls (Odds ratio = 5.6) and 29% were positive for
Human Herpes Virus-6 (HHV-6) vs. 8% of controls. 6.5% of healthy family members
were positive for Mycoplasma and 8% were positive for HHV-6 (P < 0.001) [18]. He
also reported WB positive BI patients had a 68% coinfection rate with Mycoplasma
(M. Fermentans was 70%), Bartonella, Ehrlichia, and Babesia.
 Gestational BI/TBI & Austism
• Jones et al. performed a comprehensive case history review on the
charts of 102 gestational BI/TBI cases.
• 9% had been diagnosed with autism and 56% with attention deficit
disorder. Psychiatric symptoms included irritability or mood swings
(54%), anger or rage (23%), anxiety (21%), depression (13%),
emotional (13%), OCD (11%) and suicidal thoughts (7%).
Neurological symptoms included headache (50%), vertigo (30%),
developmental delays (18%), tic disorders (14%), seizure disorders
(11%), involuntary athetoid movements (9%) and hypotonia (7%).
Sensory sensitivity symptoms included photophobia (43%),
hyperacuity (36%), motion sickness (9%) and other (tactile, taste or
smell) (23%). Cognitive symptoms included poor memory (39%),
cognitive impairments (27%), speech delays (21%), reading/writing
(19%), articulation (17%), auditory/visual processing (13%), word
selectivity (12%), and dyslexia (18%). GI symptoms were common
and included GERD (27%), abdominal pain (29%), diarrhea or
constipation (32%), and nausea (23%). As a control, 66 mothers
with Lyme disease who were treated with antibiotics prior to
conception and during the entire pregnancy; all gave birth to normal
healthy infants.
Jones CR, Smith H, Gibb E, Johnson L. Gestational Lyme Disease Case studies of 102 Live Births. Lyme Times 2005:34–6. Summer.
Biochemical Similarities
 Biochemical Similarities
• Testing patients with autism and BI/TBI also reveals
biochemical similarities. Disorders of an
oxidoreductive system in CSF and serum, increases of
superoxide dismutase, increased glutathione
peroxidase activity, increased concentration of serum
malondialdehyde and decreased glutathione have
been detected in neuroborreliosis and BI.
• In autism, several studies have suggested alterations
in the activities of antioxidant enzymes such as
superoxide dismutase and glutathione peroxidase,
altered glutathione levels and
homocysteine/methionine metabolism, increased
malondialdehyde levels and reduced glutathione.
• Impaired methylation & sulfation in both ASD & BI/TBI
 Biochemical Similarities: References
• Pancewicz SA, Skrzydlewska E, Hermanowska-Szpakowicz T,
Stankiewicz A, Kondrusik M. Evaluation of oxidoreductive
potential of patients with neuroborreliosis. Przegl Epidemiol.
2002;56(3):425-33.
• Pancewicz SA, Skrzydlewska E, Hermanowska-Szpakowicz T,
Zajkowska JM, Kondrusik M. Role of reactive oxygen species
(ROS) in patients with erythema migrans, an early
manifestation of Lyme borreliosis. Med Sci Monit.
2001;7(6):1230-5.
• Chauhan A, Chauhan V. Oxidative stress in autism.
Pathophysiol. 2006;13(3):171-81.
• Chauhan A, Chauhan V, Brown WT, Cohen I. Oxidative stress
in autism: increased lipid peroxidation and reduced serum
levels of ceruloplasmin and transferrin--the antioxidant proteins.
Life Sci. 2004;75(21):2539-49
• James SJ, Cutler P, Melnyk S, Jernigan S, Janak L, Gaylor
DW, Neubrander JA. Metabolic biomarkers of increased
oxidative stress and impaired methylation capacity in children
with autism. Am J Clin Nutr. 2004;80(6):1611-7.
Brain Imaging Tick-Borne
Diseases/Borreliosis
and Autism
 Brain Imaging Similarities
• Both BI/TBI and ASD patients demonstrate significant temporal lobe
dysfunction. In autism the cerebral cortex, hippocampus, and
amygdala showed trends toward being disproportionately smaller in
the developing autistic brain. In addition smaller amygdala volume
correlates with impairments in nonverbal social impairment in
autistic patients. Infectious encephalopathies associated with
autistic symptoms have demonstrated lesions of the temporal lobes.
PET scanning of neuroborreliosis patients demonstrates the most
striking finding was hypometabolism, which correlates with
decreased activity, in the temporal lobes in 74% patients.
• Both BI/TBI and ASD demonstrate predominately white matter
encephalopathy. Regional cerebral blood flow suggests that Lyme
encephalopathy may primarily affect cerebral white matter.
• Disruption of white matter tracts between regions implicated in
social functioning may contribute to impaired social cognition in
autism.
• Both ASD and BI/TBI patients demonstrate sensory hyperacusis
and this clinical observation is supported by brain imaging of
patients with BI that demonstrates increased thalamus activity and
increased activity in auditory and visual areas of cortex.
Maternal infection and white matter toxicity
• Studies examining maternal infection as a risk factor for
neurological disorders in the offspring have suggested that
altered maternal immune status during pregnancy can be
considered as an adverse event in prenatal development.
Infection occurring in the mother during the gestational period
has been implicated in multiple neurological effects. The
current manuscript will consider the issue of
immune/inflammatory conditions during prenatal development
where adverse outcomes have been linked to maternal
systemic infection. The discussions will focus primary on white
matter and oligodendrocytes as they have been identified as
target processes. This white matter damage occurs in very
early preterm infants and in various other human diseases
currently being examined for a linkage to maternal or early
developmental immune status. The intent is to draw attention
to the impact of altered immune status during pregnancy on
the offspring for the consideration of such contributing factors
to the general assessment of developmental neurotoxicology.

Harry GJ, Cindy L, Brunssen SH. Neurotoxicology. 2006 Sep;27(5):658-70.

 Brain Imaging Similarities
References
• Herbert MR, Ziegler DA, Deutsch CK, et al. Dissociations of cerebral cortex, subcort and cerebral white
matter volumes in autistic boys. Brain. 2003;126(5):1182-92.
• Nacewicz BM, Dalton KM, Johnstone T, et al. Amygdala volume and nonverbal social impairment in
adolescent and adult males with autism. Arch Gen Psych. 2006;63(12):1417-28
• DeLong GR, Bean SC, Brown FR 3rd. Acquired reversible autistic syndrome in acute encephalopathic illness
in children. Arch Neurol. 1981;38(3):191-4.
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chronic Lyme disease. J Neuropsychiatry Clin Neurosci. 2003;15(3):326-32.
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treatment Lyme disease syndrome. Neurol. 2001;57(11):1980-5.
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Neurol. 1997;49(6):1661-70.
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preliminary evidence from diffusion tensor imaging. Biol Psych. 2004;55(3):323-6.
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based morphometry. Neuroimage. 2004;23(1):242-51.
• Moeller, JR. Functional Neuroimaging findings in Chronic LYD; Research as a Tool to Solve Medical
Controversies. Lyme and TBD Research Cen Columbia Univ. 2007
Epidemiological Findings: Lyme
Disease/Tick-Borne
Disease and Autism
Number of Lyme disease cases
according to the CDC
 BI/TBI Epidemiology
• Acute BI/TBI is more endemic in humid temperate
locations with ecosystems that support populations of
ticks and other hosts that carry the infectious agents.
These locations are more commonly the Northeast;
coastal regions; areas near the Mississippi River, the
Mississippi tributaries, the Great Lakes region and the
lake region of the upper Mid West. These endemic
regions also coincide with the Atlantic, Mississippi and
Pacific migratory bird flyways and migratory birds have
been found to disseminate infected ticks. Veterinary data
is helpful in this analysis (www.dogsandticks.com).
 A Rapidly Changing Global
Environment Encourages the
Proliferation of Parasites, Including
Ticks and Tick-Borne Diseases
Is there as association between migratory bird flyways
(Atlantic, Pacific & Mississippi), TBD and ASD?
CDC: Lyme Disease --- United States, 2005
 Top 15 States for Autism & Lyme
Disease vs. Obesity (Control)
Autism • Lyme • Obesity
• 1, Minnesota • 1, Delaware • 1. Mississippi
• 2, Oregon • 2, Connecticut • 2. Alabama
• 3, Indiana • 3, New Jersey • 3. West Virginia
• 4, Maine • 4, Massachusetts • 4. Louisiana
• 5, Massachusetts • 5, Pennsylvania • 5. Kentucky
• 6, Michigan • 6, New York • 6. Tennessee
• 7, California • 7, Wisconsin • 7. Arkansas
• 8, Maryland • 8, Maryland • 8. Indiana (tie for 8th)
• 9, Connecticut • 9, New Hampshire • 8. South Carolina
• 10, Wisconsin • 10, Maine • 10. Texas
• 11, Rhode Island • 11, Minnesota • 11. Michigan
• 12, New Jersey • 12, Vermont • 12. Georgia
• • • 13. Oklahoma
13, Pennsylvania 13, Rhode Island
• • • 14. Missouri
14, Hawaii 14, Virginia
• 15. Alaska
• 15, Virginia • 15, West Virginia
©2006 Lyme disease info.(JRS)
 Informal Poll
• An informal poll by a show of hands was conducted at the LIAF
Conference. It has been calculated 6.7/1000 8 y/o children have
ASD (CDC). In addition, it has been calculated 11/100,000 5-9 y/o
children have LYD/year (CDC). According to the Infectious Disease
Society of America (IDSA) LYD is easily diagnosed & treated & not
chronic, therefore incidence equals prevalence. The statistical
probability of comorbid ASD + LYD is 6.7/1000 x 11/100,000 =
7/10,000,000, or approximately 1 in one million. If LYD + ASD are
comorbid more than 1/1,000,000 and CDC & IDSA are correct,
there is a causal association. As a control group, 18,000 have
hemophilia in the US: this number is similar to the approximately
20,000 yearly prevalence of LYD in the US. At the LIA conference
the author asked the 200 in attendance how many have seen ASD
associated with hemophilia. One attendee raised her hand. The
attendees were then asked how many have seen ASD associated
with LYD. Half raised their hands. In addition a significant number
have seen ASD associated with LYD in 10 or more cases, 6 have
seen ASD associated with LYD in 100 or more cases and one
physician has seen ASD associated with LYD in 500 or more cases.
Theoretical Issues: Genes,
Infections and
Autism
Physics, Math & Astrophysics:
Newton to Einstein
• Newton-Universal
gravitation and the
three laws of motion

• Einstein-Theory of
relativity, mass-
energy equivalence,
(E=mc²), nonuniform
motion & a new
theory of gravitation
Complex Human Diseases
Beyond Koch and Mendel
Mendel-Human traits are determined
by individual
genes which function independently of
other genes and of environmental
influences

Koch-Many human diseases are

caused by microbes which exert
their effect independently of other
microbes, environmental factors and
genes
Yolken
 Gene-Environmental Interactions
Beyond Koch’s Postulates
 Most common human diseases are caused by the
interaction of environmental insults and susceptibility
genes.
 Many of the susceptibility genes are diverse
determinants of human response to environmental
factors to infection.
 Informative laboratory methods for complex disorders
have to address both genetic and environmental factors.
 Prevention or treatment of the infections may result in
the effective treatment of complex disorders:
 Helicobacter-Peptic Ulcer
 HPV-Genital Cancer
 Chlamydia-Cardiac Disease?

Yolken
 Emerging Infectious Determinants of
Chronic Diseases
• Evidence now confirms that non-communicable
chronic diseases can stem from infectious
agents.
• Identifying the relationships can affect health
across populations, creating opportunities to
reduce the impact of chronic disease by
preventing or treating infection.
• Infectious agents likely determine more cancers,
immune-mediated syndromes,
neurodevelopmental disorders, and other
chronic conditions than currently appreciated.
• To capitalize on these opportunities, clinicians,
public health practitioners, and policymakers
must recognize that many chronic diseases may
indeed have infectious origins.
Siobh M. et al (CDC). Emerging Infectious Determinants of Chronic Diseases. Emerging Infectious Diseases. Vol. 12, No. 7
 Associations between
Chlamydophila infections,
schizophrenia and risk of HLA-A10
• Several microbes have been suspected as pathogenetic factors
in schizophrenia. We have previously observed increased
frequencies of chlamydial infections and of human lymphocyte
antigen (HLA)-A10 in independent studies of schizophrenia.
• We found chlamydial infection in 40.3% of the schizophrenic
patients compared to 6.7% in the controls. The association of
schizophrenia with Chlamydiaceae infections was highly
significant (P=1.39 10-10, odds ratio (OR)=9.43), especially with
Chlamydophila psittaci (P=2.81 10-7, OR=24.39).
• Schizophrenic carriers of the HLA-A10 genotype were clearly most
often infected with Chlamydophila, especially C. psittaci (P=8.03
10-5, OR=50.00). Chlamydophila infections represent the highest
risk factor yet found to be associated with schizophrenia. This risk is
even further enhanced in carriers of the HLA-A10 genotype.
Selected Infectious Agents and Risk
of Schizophrenia Among U.S.
Military Personnel

• The authors found significant associations

between increased levels of scaled T.
gondii IgG antibodies and schizophrenia
for antibodies measured both prior to and
after diagnosis.
 Maternal Exposure to Herpes
Simplex Virus and Risk of
Psychosis Among Adult Offspring
• Background: Viral exposure during gestation is thought to be a risk factor for
schizophrenia. Previous studies have indicated that prenatal exposure to
herpes simplex virus type 2 (HSV-2) may be a risk for the subsequent
development of schizophrenia in some populations. In this investigation, we
tested a large and diverse population to assess the risk of psychoses
among offspring of mothers with serological evidence of HSV-2 infection.
• Results: Offspring of mothers with serologic evidence of HSV-2 infection
were at significantly increased risk for the development of psychoses (odds
ratio [OR] = 1.6; 95% confidence interval [CI] = 1.1–2.3). This risk was
particularly elevated among women with high rates of sexual activity during
pregnancy (OR = 2.6; 95% CI = 1.4–4.6).
• Conclusions: Maternal exposure to herpes simplex virus type 2 is
associated with an increased risk for psychoses among adult offspring.
These results are consistent with a general model of risk resulting from
enhanced maternal immune activation during pregnancy.

Stephen L. Buka, Tyrone D. Cannonc, E. Fuller Torrey, Robert H. Yolken.

Biological Psychiatry. Volume 63, Issue 8, 15 April 2008, Pages 809-815
 Disease Contributors Change with
Time
Predisposing Factors
Precipitating Factors
Perpetuating Factors

Symptom
Threshold
“Intensity”

1 2 3 4
Preclinical Onset Short-Term Chronic
Course

Adapted from Spielman AJ et al. Psychiatr Clin North Am. Course of Insomnia; 1987;10:541-553.
Schema of Etiologic and Pathogenetic Factors That Have Been Implicated in Cell Death in
Parkinson Disease and Possible Neuroprotective Approaches

Schapira, A. H. V. et al.
JAMA 2004;291:358-364.

Copyright restrictions may apply.
 The Biotoxin Pathway High cytokine levels in the capillaries attract white blood
cells , leading to restricted blood flow , and lower oxygen
Capillaries levels. Reduced VEGF leads to fatigue, muscle cramps,
In genetically susceptible people , biotoxin binds to fat -cell receptors, and shortness of breath (may be over-ridden by
causing continuing, unregulated production of cytokines . replacement with erythropoietin).

Bo
d Immune system symptoms
b ya
t ox iot o c qu Surface
org in-p xins ires Biotoxin (“Toll”)
Increased Cytokines
foo anis r odu or (HLA susceptible) Fat Cell Patients with certain HLA genotypes (immunity-
c receptor
or d, wa ms f ing related genes) may develop inappropriate
ins te rom immunity. Most common are antibodies to:
ec , a r
t b ir, -- Myelin basic protein (often from fungal
ite
s biotoxins; affects nervous-system functions)
-- Gliadin (affects digestion)

Inc
Fat cells then -- Cardiolipins (affects blood clotting )

rea
I nc
(H Bi produce more The “complement” alternative immune pathway

s
LA oto

rea

ed
su x in leptin, leading to may be triggered (detectable as an increase in

se

Le
s ce obesity (which levels of the proteins C3a C4a).

dC
pt i

pti
ble doesn’t respond to

n
yto
)
exercise and diet ).

k in
Cytokine-related symptoms

es
Nerve cell Excessive
cytokine levels
can damage leptin Leptin High levels of cytokines produce flu -like
receptors in the receptor Damaged leptin symptoms: Headaches, muscle aches , fatigue,
Biotoxins have direct effects, including hypothalamus. receptors lead to unstable temperature, difficulty concentrating .
impairment of nerve cell function. One reduced production by High levels of cytokines also result in increased
Hypothalamus levels of several other immune -response related
result is poor performance on contrast the hypothalamus of
Bio t HL le)

sensitivity test . MSH, a hormone with substances, including TNF, MMP-9, IL-1B, and
su
(no ept i

VIP AVP PAI-1. MMP-9 delivers inflammatory elements

t ox A

many functions.
sc

in

from blood to brain, nerve, muscle, lungs, and

joints. It combines with PAI-1 in increasing clot
b

formation and arterial blockage.

Removal from
the body
In most people, biotoxins
are either removed from
the blood by the liver or
attacked by the immune
system, broken down,
and excreted harmlessly.
In people who don’t have
the right immune-system
genes, however,
biotoxins can remain in
the body indefinitely.
© G. Alexander 2004
Sleep
disturbance
Production of
melatonin is reduced,
leading to light , non-
restorative sleep.
Chronic pain
Endorphin production is
suppressed. This can lead
to chronic, sometimes
unusual, pain.
Gastrointestinal
problems
Lack of MSH can cause
malabsorption in the
gut, resulting in diarrhea.
This is sometimes called
“leaky gut” and
resembles (but is not)
celiac disease . Patients
must avoid gluten, whey,
and amylose.
Prolonged illness
White blood cells lose
regulation of cytokine
response, so that
recovery from other
illnesses , including
infectious diseases , may
be slowed.
Reduced
MSH
Resistant Staph bacteria
Colonies of Staph bacteria with
resistance to multiple antibiotics
may develop in mucous
membranes. The bacteria
produce substances that
aggravate both the high cytokine
levels and low MSH levels .
Reduced ADH
Stage 1:
Reduced MSH can cause the Biotoxin effects
pituitary to produce lower levels
of anti-diuretic hormone (ADH),
Stage 2:
leading to thirst , frequent Cytokine effects
urination, and susceptibility to
shocks from static electricity .
Stage 3:
Reduced VEGF
Reduced sex hormones
Reduced MSH can cause the Stage 4:
pituitary to lower its Immune effects
production of sex hormones.
Stage 5:
Changes in cortisol and Low MSH
ACTH levels
Stage 6:
The pituitary may produce elevated
Resistant Staph bacteria
levels of cortisol and ACTH in early
stages of illness , then drop to
excessively low levels later . Stage 7:
(Patients should avoid steroids such Pituitary hormone effects
as prednisone, which can lower
levels of ACTH.)
Rev. 10, 12-12-05
Heavy Metal Toxicity
 Summarizing the Theories
• Most commonly human diseases are caused by the
interaction of environmental insults and susceptibility
genes. Many of the susceptibility genes respond to
environmental factors and infection. Environmental
insults contributing to ASD may include a complex
interaction with infections, heavy metals, biotoxins,
allergens, nutritional excesses/deficits and possibly
vaccines resulting in a pathogenic interaction that
includes inflammation, oxidative stress, mitochondrial
dysfunction and excitotoxicity resulting in neuronal
dysfunction.
Klüver–Bucy Syndrome,
Infections and
Autism
http://www.athealth.com
 Limbic System
• The limbic system is the mammalian brain
• The creation & evolution of the limbic
system parallels the creation & evolution
of the family
 The Amygdala Theory of Autism
• There is a network of neural regions that
comprise the "social brain", which includes
the amygdala
• Since the childhood psychiatric condition
of autism involves deficits in "social
intelligence", it is plausible that autism
may be caused by an amygdala
abnormality
• This includes reference to the Klüver-Bucy
syndrome
Baron-Cohen S, et. Al. Neurosci Biobehav Rev. 2000 May;24(3):355-64.
 Klüver-Bucy Syndrome
• The syndrome is named for Heinrich Klüver and Paul Bucy, who
removed the temporal lobe bilaterally in rhesus monkeys in an attempt
to determine its function. This caused the monkeys to develop visual
agnosia, emotional changes, altered sexual behavior,
hypermetamorphosis and oral tendencies.
• Though the monkeys could see, they were unable to recognize even
previously familiar objects, or their use. They would examine their
world with their mouths instead of their eyes ("oral tendencies") and
developed a desire to explore everything ("hypermetamorphosis").
• Their overt sexual behavior increased dramatically
("hypersexualism"), and the monkeys indulged in indiscriminate
sexual behavior including masturbation, heterosexual acts and
homosexual acts.
• Emotionally, the monkeys became dulled, and their facial expressions
and vocalizations became far less expressive. They were also less
fearful of things that would have instinctively panicked them in their
natural state, such as humans or snakes. Even after being attacked
by a snake, they would willingly approach it again. This aspect of
change was termed "placidity".
• People with bilateral lesions in their temporal lobes show similar
behaviors. They may display oral or tactile exploratory behavior
(socially inappropriate licking or touching); hypersexuality; bulimia;
memory disorders; flattened emotions (placidity); and an inability to
 Infantile Autism and the Temporal
Lobes of the Brain
• Studies are reviewed that support the hypothesis that
infantile autism results from a neuropathology of the
temporal lobes of the brain. First, there are parallels
between symptoms noted in autism and those found in
the Kluver-Bucy and amnesic syndromes. Second, there
is a similarity between developmental dysphasia and
autism. Third, the formation of cross-modal associations
may be deficient in autistic children, a symptom
resembling aspects of Geschwind's disconnection
syndromes. Finally, a large number of organic factors
have been associated with the development of autism,
some of these having specific implications for temporal
lobe involvement. It is concluded that the main autistic
symptoms are most consistent with a neurological model
involving bilateral dysfunction of the temporal lobes.
Individual differences in the extent of bilateral
involvement and/or other coexistent neuropathologies
could contribute to the heterogeneity of the autistic
population. Hetzler BE, Griffin JL. J Autism Dev Disord. 1981 Sep;11(3):317-30.
 Autism, the superior temporal
sulcus and social perception
– Based on recent brain-imaging results, our hypothesis is that
abnormalities in the superior temporal sulcus (STS) are highly
implicated in ASD. STS abnormalities are characterized by
decreased gray matter concentration, rest hypoperfusion and
abnormal activation during social tasks. STS anatomical and
functional anomalies occurring during early brain development
could constitute the first step in the cascade of neural
dysfunction underlying ASD. We will focus this review on the
STS, which has been highly implicated in social cognition. We
will review recent data on the contribution of the STS to normal
social cognition and review brain-imaging data implicating this
area in ASD.

Zibovicius M et al. Trends Neurosci. 2006 Jul;29(7):359-66. Epub 2006 Jun 27.
 Amygdala volume and nonverbal social impairment in
adolescent and adult males with autism

• In study 1, individuals with autism who had small amygdalae were

slowest to distinguish emotional from neutral expressions (P=.02)
and showed least fixation of eye regions (P=.04). These same
individuals were most socially impaired in early childhood, as
reported on the Autism Diagnostic Interview-Revised (P<.04).
• Study 2 showed smaller amygdalae in individuals with autism than in
control subjects (P=.03) and group differences in the relation
between amygdala volume and age. Study 2 also replicated findings
of more gaze avoidance and childhood impairment in participants
with autism with the smallest amygdalae. Across the combined
sample, severity of social deficits interacted with age to predict
different patterns of amygdala development in autism (P=.047).
• CONCLUSIONS: These findings best support a model of amygdala
hyperactivity that could explain most volumetric findings in autism.
Further psychophysiological and histopathological studies are
indicated to confirm these findings

Nacewicz, et. al. Arch Gen Psychiatry. 2006 Dec;63(12):1417-28

Medline Cited Infectious Causes of Autism
• Rubella, • Herpes virus family
• Herpes simplex • Malaria
• Mycoplasma • Blastocystis
pneumoniae • Cytomegalovirus
• Shigella • Syphilis
• Neurocysticercosis • Varicella
• Unknown • Toxoplasmosis
• Various viral infection • Yet unrecognized
• Borna infectious
• HHV-6 • Tick-Borne/Lyme
• Chlamydia
Microbes that Can Cause Mental Symptoms
• Syphilis •- Anaplasmas (related to the genera • Pandemic Influenza of 1918
• Malaria Rickettsia and Ehrlichia) • Influenza virus
• Toxoplasmosis - Hepatitis-C • Measles
• Bartonellosis: •
Candidiasis Papovirus
• - Bartonella henselae (cat scratch •
Other spirochetes Poliovirus
• fever), Bartonella rochalimae •
1. Borrelia burgdorferi sensu Rabies
stricto(USA,UK,Europe) - Bartonella quintana (trench fever) • Toga virus
2. Borrelia garinii (UK, Europe) - Viral Meningitis •
3. Borrelia afzelii (UK, Europe) Toxoplasmosis
Candida dubliniensis •
"CHRONIC LYME DISEASE" or "NEW Cryptococcus
Asfarviridae, Reoviridae, •
LYME DISEASE" Coccidiomycosis
is a combination of LYME DISEASE Rhabdoviridae, Orthomyxoviridae,
• Histoplasmosis
and Bunyaviridae, Flaviviridae &
one or more of the following Co- • Cysticercosis
Arenaviridae family viruses (38+
infections: • Rubella,
species
Relapsing Fever caused by the • Herpes simplex
spirochetes: Streptococcus
• Mycoplasma pneumoniae
- Borrelia hermsii Japanese B encephalitis
- Borrelia turicatae • Shigella
HHV-1
Mycoplasmas: • Neurocysticercosis
HHV-2
- Mycoplasma fermentans • Unknown
- Mycoplasma pneumoniae Borna virus
• Various viral infection
Babesiosis: Epstein-Barr virus
- Babesia microti • Borna
Pandemic Influenza of 1918
- Babesia WA and other Babesia • Herpes virus family
species Hong Kong flu
• Malaria
• Coxackie virus
- Chlamydia pneumoniae • Blastocystis
Rickettsial Diseases: Pneumococcus
• Cytomegalovirus
- Rocky Mountain Spotted Fever Haemophilus
- Coxiella burnetti (Q-Fever and "Post- • Syphilis
Meningococcus
Q Fever Fatigue Syndrome") • Varicella
- Colorado Tick Fever Leptospira
• Toxoplasmosis
- Eastern tick-borne Rickettsiosis Mycobacterium tuberculosis
• Yet unrecognized infectious
- Rickettsialpox Cytomegalovirus
- Tularemia (rabbit fever) Enterovirus
- Ehrlichiosis (caused by Ehrlichia, HIV
and rickettsia-like bacteria)
 Infections, Klüver-Bucy & Autism I
• Baron-Cohen S, Ring HA, Bullmore ET, Wheelwright S, Ashwin C, Williams
SC. The amygdala theory of autism. Neurosci Biobehav Rev.
2000;24(3):355-64.
• Hetzler BE, Griffin JL. Infantile autism and the temporal lobe of the brain. J
Autism Dev Disord. 1981;11(3):317-30.
• Libbey JE, Sweeten TL, McMahon WM, Fujinami RS. Autistic disorder and
viral infections. J Neurovirol. 2005;11(1):1-10
• Stubbs EG, Crawford ML. Depressed lymphocyte responsiveness in autistic
children.
• J Autism Child Schizophr. 1977;7(1):49-55.
• De Tiege X, De Laet C, Mazoin N, et. al. Postinfectious immune-mediated
encephalitis after pediatric herpes simplex encephalitis. Brain Dev.
2005;27(4):304-7.
• DeLong GR, Bean SC, Brown FR 3rd. Acquired reversible autistic syndrome
in acute encephalopathic illness in children. Arch Neurol. 1981;38(3):191-4.
• Libbey JE, Sweeten TL, McMahon WM, Fujinami RS. Autistic disorder and
viral infections. J Neurovirol. 2005;11(1):1-10
• Lancaster K, Dietz DM, Moran TH, Pletnikov MV.Abnormal social behaviors
in young and adult rats neonatally infected with Borna disease virus. Behav
Brain Res. 2007;176(1):141-8.
• Stubbs EG, Crawford ML. Depressed lymphocyte responsiveness in autistic
children. J Autism Child Schizophr. 1977;7(1):49-55.
 Infections, Klüver-Bucy & Autism II
• Markowitz PI. Autism in a Child with Congenital Cytomegalovirus Infection. J
Autism Dev Disord. 1983;13(3)
• Stubbs EG, Ash E, Williams CPS. Autism and Congenital Cytomegalovirus.
J Autism Dev Disord. 1984;14(2).
• Auvichayapat N, Auvichayapat P, Watanatorn J, Thamaroj J, Jitpimolmard
S. Kluver-Bucy syndrome after mycoplasmal bronchitis. Epilepsy Behav.
2005;8(1):320-2.
• Guedalia JS, Zlotogorski Z, Goren A, Steinberg A. A reversible case of
Klüver-Bucy syndrome in association with shigellosis. J Child Neurol.
1993;8(4):313-5.
• Stubbs EG, Crawford ML. Depressed lymphocyte responsiveness in autistic
children. J Autism Child Schizophr. 1977;7(1):49-55.
• Patel R, Jha S, Yadav RK. Pleomorphism of the clinical manifestations of
neurocysticercosis. Trans R Soc Trop Med Hyg. 2006;100(2):134-41.
• Mankoski RE, Collins M, Ndosi NK, Mgalla EH, Sarwatt VV, Folstein SE.
Etiologies of autism in a case-series from Tanzania. J Autism Dev Disord.
2006;36(8):1039-51
• Thong YH. Reptilian behavioural patterns in childhood autism. Med
Hypotheses. 1984;13(4):399-405.
• Boorom KF. Is this recently characterized gastrointestinal pathogen
responsible for rising rates of inflammatory bowel disease (IBD) and IBD
associated autism in Europe and the United States in the 1990s? Med
Hypotheses. 2007;69(3):652-9.
 Infections, Klüver-Bucy & Autism III
• Singh VK and Jensen RL. Elevated levels of measles antibodies in children with
autism. Pediatric Neurol. 2003;28:292-294.
• Singh VK. Autism, vaccines, and immune reactions. Accessed 8-3-07. http://
vacinfo.org/vijendra_singh.htm
• Halsey NA, Hyman SL. Measles-mumps-rubella vaccine and ASD: report from the
New Challenges in Childhood Immunizations Conf. Pediatrics. 2001;107(5):E84
• Bransfield RC, Wulfman JS, Harvey WT, Usman AI. The association between tick-
borne infections, Lyme borreliosis and autism spectrum disorders Medical
Hypotheses. 2007
• Nicolson GL, Gan R, Nicolson NL, et al. Evidence for Mycoplasma, Chlamydia
pneunomiae and HHV-6 Co-infections in the blood of patients with Autism Spectrum
Disorders. J Neuroscience Res. 2007;85:1143-48.
• Takahashi H, Arai S, Tanaka-Taya K, et al. Autism and infection/immunization
episodes in Japan. Jpn J Infect Dis. 2001;54:78-79.
• Yamashita Y, Fujimoto C, Nakajima E, et al. Possible association between
congenital cytomegalovirus infection and autistic disorder. J Autism Dev Disord.
2003;33:355-59.
• Rosen NJ, Yoshida CK, Croen LA. Infection in the first 2 years of life and autism
spectrum disorders. Pediatrics. 2007;119:61-9.
• Nicolson GL, Berns P, Gan R, et al. Chronic mycoplasmal infections in Gulf War
veterans’ children and autism patients. Med Veritas. 2005;2:383-87.
• Nicolson GL. Chronic Bacterial and Viral Infections in Neurodegenerative and
Neurobehavioral Diseases. Laboratory Medicine 2008. In Press
 Neural Networks,
Neurodevelopment,
Autism and Borreliosis
 The neurodevelopmental impact of
prenatal infections at different times of
pregnancy: the earlier the worse?
• Infection associated immunological events
in early fetal life can have adverse effects
on cell proliferation and differentiation;
predispose the developing nervous
system to undergo additional failures in
subsequent cell migration, target selection,
and synapse maturation, eventually
leading to multiple brain and behavioral
abnormalities apparent later in life.
 Critical periods of vulnerability for the
developing nervous system: evidence
from humans and animal models
• Brain developmental processes (i.e. cell
proliferation, migration, differentiation,
synaptogenesis, myelination, and
apoptosis) occur at vulnerable periods
during the development of the nervous
system and are sensitive to environmental
insults that can contribute to autism.

Rice D, Barone S. Environ Health Perspect 2000;108(3):511–33.

 Immune Reactions to LYD Disrupt
Nerve Fibers
• Younger has demonstrated on biopsies
that small nerve fiber disruption can occur
in Lyme vaccine recipients and BI/TBI
patients who subsequently may heal in
response to anti-infective treatment

Younger D. Small nerve fiber disruption in OspA vaccine

recipients. Am Acad Neurol. Meeting 2007.
 Oxidative Stress
• “Chronic intracellular infections (Mycoplasma,
Borrelia, Chlamydia, etc.) cause increased
oxidative stress by their release of ROS and
stimulation of ROS in mitochondria. This causes
increased oxidation of mitochondrial lipid
membranes and proteins and loss of mitochondrial
function. We see the same thing in CFS patients
who also have these infections, although in fully
developed brains we do not, of course, see the
autism and other developmental disorders.”
Prof. Garth Nicolson
 Evidence of toxicity, oxidative stress,
and neuronal insult in autism
• This article discusses the evidence for the case that
some children with autism may become autistic from
neuronal cell death or brain damage sometime after birth
as result of insult; and addresses the hypotheses that
toxicity and oxidative stress may be a cause of neuronal
insult in autism. The article first describes the Purkinje
cell loss found in autism, Purkinje cell physiology and
vulnerability, and the evidence for postnatal cell loss.
Second, the article describes the increased brain volume
in autism and how it may be related to the Purkinje cell
loss. Third, the evidence for toxicity and oxidative stress
is covered and the possible involvement of glutathione is
discussed. Finally, the article discusses what may be
happening over the course of development and the
multiple factors that may interplay and make these
children more vulnerable to toxicity, oxidative stress, and
neuronal insult.
Kern & Jones. J Toxicol Environ Health B Crit Rev. 2006 Nov-Dec;9(6):485-99
 Oxidative stress, nitric oxide, and the
mechanisms of cell death in Lurcher
Purkinje cells
– Oxidative stress is postulated to play a role in cell death in many neurodegenerative
diseases. As a model of neonatal neuronal cell death, we have examined the role of
oxidative stress in Purkinje cell death in the heterozygous Lurcher mutant (+/Lc).
Lurcher is a gain of function mutation in the delta2 glutamate receptor (GluRdelta2)
that turns the receptor into a leaky membrane channel, resulting in chronic
depolarization of +/Lc Purkinje cells starting around the first week of postnatal
development. Virtually, all +/Lc Purkinje cells die by the end of the first postnatal
month. To investigate the role of oxidative stress in +/Lc Purkinje cell death, we have
examined nitric oxide synthase (NOS) activity and the expression of two markers for
oxidative stress, nitrotyrosine and manganese super oxide dismutase (MnSOD), in
wild type and +/Lc Purkinje cells at P10, P15, and P25. The results show that NOS
activity and immunolabeling for nitrotyrosine and MnSOD are increased in +/Lc
Purkinje cells. To determine whether peroxynitrite formation is a prerequisite for +/Lc
Purkinje cell death, +/Lc mutants were crossed with an alpha-nNOS knockout mutant
(nNOSalpha(-/-)) to reduce the production of NO. Analysis of the double mutants
showed that blocking alpha-nNOS expression does not rescue +/Lc Purkinje cells.
However, we present evidence for sustained NOS activity and nitrotyrosine formation
in the GluRdelta2(+/Lc):nNOS(-/-) double mutant Purkinje cells, which suggests that
the failure to rescue GluRdelta2(+/Lc):nNOS(-/-) Purkinje cells may be explained by
the induction of alternative nNOS isoforms.
 Major Histocompatibility Complex

• An infection during pregnancy or early

development in a genetically predisposed
individual may evoke an immune response that
that disrupts fetal brain development by altering
major histocompatibility complex molecules that
impact glia and microgila cells, glutamate
functioning and synaptic development and
plasticity and contribute to the pathophysiology
of autism.
 Cerebellar atrophy in temporal lobe
epilepsy
– The goal of this work was to determine the presence
and degree of cerebellar atrophy in chronic temporal
lobe epilepsy
– CONCLUSIONS: The presence of cerebellar atrophy
is a reflection of the extra-temporal abnormalities that
can be observed in localization-related temporal lobe
epilepsy, which may be due, at least in part, to
factors associated with epilepsy chronicity.
Borreliosis and Borrelia
Related Complex
 Tick-Borne Pathogens

• Humans have 400 different species of

bacteria in their mouth.
• What pathogens, known and unknown,
can be transmitted from a tick that lives
in filth and sucks on the blood of
rodents?
 POLYMICROBIAL INFECTIONS IN
ANIMALS AND HUMANS
• Polymicrobial diseases represent the clinical and pathological manifestations
induced by the presence of multiple microorganisms. These are serious
diseases whose etiologic agents are sometimes difficult to diagnose and
difficult to treat. They are often called complex infections, complicated
infections, dual infections, mixed infections, secondary infections, co-
infections, synergistic infections, concurrent infections, or polymicrobial
infections. These diseases in animals and humans are induced by multiple
viral infections, multiple bacterial infections, viral and bacterial infections,
multiple mycotic and parasitic infections, and opportunistic infections
secondary to microbe-induced immunosuppression. There are five common
underlying mechanisms of disease pathogenesis. First, physical, physiologic,
or metabolic abnormalities and stress predispose the host to polymicrobial
disease. Second, one microorganism induces changes in the mucosa that
favors the colonization of other microorganisms. Third, microorganisms or
their products can trigger proinflammatory cytokines that increase the severity
of disease, reactivate latent infections, or favor the colonization of other
microorganisms. Fourth, microorganisms share determinants among each
other allowing them the collective ability to damage tissue. Finally, one
microorganism alters the immune system, which allows the colonization of
the host by other microorganisms. Many areas of study in polymicrobial
diseases are at their infancy and it is our hope that this Conference will
stimulate interest and work in this evolving area.
•
What is Chronic LYD/TBD?
LYME DISEASE caused by three types of Spirochete Borrelia bacteria (300 strains):
1. Borrelia burgdorferi sensu stricto(USA,UK,Europe)
2. Borrelia garinii (UK, Europe)
3. Borrelia afzelii (UK, Europe)
"CHRONIC LYME DISEASE" or "NEW LYME DISEASE"
is a combination of LYME DISEASE and
one or more of the following Co-infections:
Relapsing Fever caused by the spirochetes:
- Borrelia hermsii
- Borrelia turicatae
Mycoplasmas:
- Mycoplasma fermentans
- Mycoplasma pneumoniae
Babesiosis:
- Babesia microti
- Babesia WA and other Babesia species
• - Chlamydia pneumoniae
Rickettsial Diseases:
- Rocky Mountain Spotted Fever
- Coxiella burnetti (Q-Fever and "Post-Q Fever Fatigue Syndrome")
- Colorado Tick Fever
- Eastern tick-borne Rickettsiosis
- Rickettsialpox
- Tularemia (rabbit fever)
- Ehrlichiosis (caused by Ehrlichia, a rickettsia-like bacteria)
- Anaplasmas (related to the genera Rickettsia and Ehrlichia)
- Hepatitis-C
Bartonellosis:
- Bartonella henselae (cat scratch fever)
- Bartonella quintana (trench fever)
- Viral Meningitis
• Candida dubliniensis
• Asfarviridae, Reoviridae, Rhabdoviridae, Orthomyxoviridae, Bunyaviridae, Flaviviridae &
Arenaviridae family viruses (38+ species) Sean and Leslee Dudley & Labuda and Nuttall & Nunn
 Borrelia Pathophysiology
• Invade, penetrate, injure or kill host cells
• Indirect injury at a distance (coagulation cascade
of proteins, activation of coagulation system,
blebs, microthrombi, septic emboli)
• Biological amplification-cascade of injury
• Reservoir inside of host
• Leeching-”saps nutrients”
• Toxins
• Incorporate gene sequences into host genome
• Immune effects—inflammation,
immunosupression, molecular mimicry
• Herxheimer pathophysiology
Adapted from Mac Donald
Fighting Back: How B burgdorferi Persists

• That spirochetes tend to persist in the human body has been demonstrated in both syphilis, caused by
Treponema pallidum, and Lyme disease, caused by Borrelia burgdorferi. What accounts for this ability to evade or
suppress an effective immune response? According to Charles Pavia, PhD,[1] of the New York College of
Osteopathic Medicine, New York Institute of Technology, Old Westbury, New York, there are at least 6 potential
explanations:
• antigenic variation (this is seen with the Borrelia species that cause tick-borne relapsing fever) or differential
expression of antigens (especially the outer surface proteins; with B burgdorferi, only OspC is expressed during
mammalian infection)
• production of an outer protective coat (eg, capsule, as seen with T pallidum)
• atypical forms (eg, cyst-like variants)
• incomplete immune response (eg, insufficient antibody , T-cell , or phagocytic response)
• deranged host immune response (eg, host-, tick-, or spirochete-derived immunosuppressive factors)
• other evasive factors (eg, motility)
• Immune Suppression
• Is there evidence that any of these mechanisms allow B burgdorferi to persist in the human body? As of now, not
much. However, there have been a few suggestive studies in animals that support immune suppression as a
possible explanation. For instance, a study by Chiao and colleagues[2] showed that B burgdorferi is capable of
suppressing the immune response. When sonicated Borrelia were added to lymphocytes, the ability of the
lymphocytes to proliferate -- a measure of the immune system's ability to respond to an infectious challenge --
was inhibited. A similar study by Giambartolomei and coworkers[3] showed that Borrelia can stimulate interleukin-
10 (IL-10) production, a downregulator of the immune system. In this series of experiments, heat-killed B
burgdorferi caused peripheral blood mononuclear cells of humans and rhesus monkeys to produce this cytokine.
Another study, by Keane-Myers and Nickell,[4] found that B burgdorferi could suppress T-cell responses in mice,
specifically T-helper cells.
• Even the tick itself may play a role in immunosuppression. Urioste and colleagues[5] showed that the saliva of
Ixodes dammini ticks contains an uncharacterized substance that can suppress the immune response,
specifically suppressing lymphocyte proliferation and other markers of immune system activity.
• Looking at the issue of immune suppression from the other side -- that is, by boosting the immune response with
the use of cytokines -- Zeidner and colleagues[6] showed that tumor necrosis factor alpha (TNF-alpha), IL-2, and
interferon-gamma could suppress B burgdorferi infection in mice.
• By contrast, it appears that infection with B burgdorferi can also overstimulate the immune system, and this may
explain many of the symptoms of both acute and chronic Lyme disease. For instance, Lim and colleagues[7]
showed that CD4+ T cells play a role in the arthritis seen in the hamster model of Lyme disease.
TBD: Borrelia Burgdorferi, Babesia
Cause Immunosupression
• Borrelia burgdorferi-induced tolerance
as a model of persistence via
immunosuppression. (Diterich et. al.
Infect Immun. 2003 Jul;71(7):3979-87)
• Immunodepression in Babesia microti
infections. (Purvis AC. Parasitology. 1977
Oct;75(2):197-205)
• Other tick-borne pathogens may also be
immunosupressant
Tick Saliva Causes Immunospression
• Kyckova & Kopecky. Effect of tick saliva on mechanisms
of innate immune response against Borrelia afzelii.
J Med Entomol. 2006 Nov;43(6):1208-14.
• Holden K, Hodzic E, Feng S, Freet KJ, Lefebvre RB,
Barthold SW. Coinfection with Anaplasma
phagocytophilum alters Borrelia burgdorferi population
distribution in C3H/HeN mice.
Infect Immun. 2005 Jun;73(6):3440-4.
• Hannier S, Liversidge J, Sternberg JM, Bowman AS.
Characterization of the B-cell inhibitory protein factor in
Ixodes ricinus tick saliva: a potential role in enhanced
Borrelia burgdoferi transmission.Immunology. 2004
Nov;113(3):401-8.
• Wikel SK. Tick modulation of host immunity: an
important factor in pathogen transmission. Int J Parasitol.
1999 Jun;29(6):851-9.
 Propensity to excessive proinflammatory
response in chronic Lyme borreliosis

• All the clinical manifestations, acute or chronic,

are characterized by strong inflammation.
Borrelia burgdorferi can induce the production of
several proinflammatory and anti-inflammatory
cytokines
• We conclude that chronic forms of Lyme
borreliosis can evolve due to an aberrant innate
proinflammatory response.

Kisand et. al. APMIS, Volume 115 Issue 2 Page 134-141 - February 2007
 Invasion of human neuronal and glial cells by an
infectious strain of Borrelia burgdorferi
• Human infection by Borrelia burgdorferi, the etiological agent for Lyme
disease, can result in serious acute and late-term disorders including
neuroborreliosis, a degenerative condition of the peripheral and
central nervous systems. To examine the mechanisms involved in the
cellular pathogenesis of neuroborreliosis, we investigated the ability of
B. burgdorferi to attach to and/or invade a panel of human neuroglial
and cortical neuronal cells. In all neural cells tested, we observed B.
burgdorferi in association with the cell by confocal microscopy.
Further analysis by differential immunofluorescent staining of external
and internal organisms, and a gentamicin protection assay
demonstrated an intracellular localization of B. burgdorferi. A non-
infectious strain of B. burgdorferi was attenuated in its ability to
associate with these neural cells, suggesting that a specific borrelial
factor related to cellular infectivity was responsible for the association.
Cytopathic effects were not observed following infection of these cell
lines with B. burgdorferi, and internalized spirochetes were found to
be viable. Invasion of neural cells by B. burgdorferi provides a
putative mechanism for the organism to avoid the host's immune
response while potentially causing functional damage to neural
cells during infection of the CNS.
Livengood & Gilmore. Microbes Infect. 2006 Nov-Dec;8(14-15):2832-40. Epub 2006 Sep 22.
 Lyme Disease: The Quest for
Magic Bullets
• Borrelia burgdorferi is one of the most
complex bacteria known to man.
• Two major clinical hurdles are the absence
of a therapeutic endpoint in treating Lyme
disease and the presence of tick-borne
coinfections that may complicate the
course of the illness.

Stricker RB, Lautin A, Burrascano JJ. Chemotherapy. 2006 Feb 22;52(2):53-59

Tick-Borne/Borreliosis Infections
and
Psychiatric Illness
 Higher prevalence of antibodies to
Borrelia burgdorferi in psychiatric
patients than in healthy subjects
• 166 (33%) of the psychiatric patients and 94
(19%) of the healthy comparison subjects were
seropositive in at least one of the four assays
for Borrelia burgdorferi.
• These findings support the hypothesis that
there is an association between Borrelia
burgdorferi infection and psychiatric morbidity.
In countries where this infection is endemic, a
proportion of psychiatric inpatients may be
suffering from neuropathogenic effects of
Borrelia burgdorferi.
Hajek T et al. Am J Psychiatry 2002 Feb;159(2):297-301
BI/TBI & Neuropsychiatric Disorders
• Fallon BA, Schwartzberg M, Bransfield R, Zimmerman B, Scotti A, Weber CA,
Liebowitz MR. Late-Stage Neuropsychiatric Lyme Borreliosis: Differential
Diagnosis and Treatment. Psychosomatics 1995;36:295-300.
• Adams WV, Rose CD, Eppes SC, Klein JD. Long-term cognitive effects of
Lyme disease in children. Appl Neuropsychol 1999;6(1):39-45.
• Fallon BA, Nields JA. Lyme Disease: A Neuropsychiatric Illness. Am J Psych
1994;151(11):1571-83.
• Fallon BA, Bird H, Hoven C, Cameron D, Liebowitz MR, Shaffer S. Psychiatric
aspects of Lyme disease in children and adolescents: A community
epidemiologic study in Westchester, New York. JSTD 1994;1:98-100.
• Waniek C, Prohovnik I, Kaufman MA, Dwork AJ. Rapidly progressive frontal-
type dementia associated with Lyme disease J Neuropsych Clin Neurosci
1995;7(3):345-7.
• Fallon BA, Kochevar JM, Gaito A, Nields JA. The Underdiagnosis of
Neuropsychiatric LYD in Children and Adults. Psych Clinics No Am, 1998;
21:693-703.
• Bransfield RC. Case Report: LYD and Complex Seizures. JSTD 1999;6:123-
5.
• Massei F, Gori L, Macchia P, Maggiore G. The expanded spectrum of
bartonellosis in children. Infect Dis Clin North Am. 2005;19(3):691-711
• Murakami K, Tsukahara M, Tsuneoka H, et al. Cat scratch disease: analysis
 Neuroactive Kynurenines in Lyme
Borreliosis
• We conclude that CSF quinolinic acid is
significantly elevated in B burgdorferi
infection-- dramatically in patients with CNS
inflammation, less in encephalopathy.
• The presence of this known agonist of
NMDA synaptic function--a receptor
involved in learning, memory, and synaptic
plasticity--may contribute to the neurologic
and cognitive deficits seen in many Lyme
disease patients.
Halperin JJ, Heyes MP. Neuroactive kynurenines in Lyme borreliosis.
Neurology 1992 Jan;42(1):43-50
 Proinflammatory Cytokines Increase
Indoleamine 2,3-dioxygenase (IDO)
• The IDO enzyme converts tryptophan into kynurenine,
because IDO activation leads to reduced levels of
tryptophan, the precursor of serotonin (5-HT), and thus
to reduced central 5-HT synthesis.
• Kynurenine metabolites such as 3-hydroxy-kynurenine
(3-OH-KYN) and quinolinic acid (QUIN) have toxic
effects on brain function. 3-OH-KYN is able to produce
oxidative stress by increasing the production of reactive
oxygen species (ROS), and QUIN may produce
overstimulation of hippocampal N-methyl-D-aspartate
(NMDA) receptors, which leads to apoptosis and
hippocampal atrophy. Both ROS overproduction and
hippocampal atrophy caused by NMDA overstimulation
have been associated with depression.
Wichers MC, Maes M. J Psychiatry Neurosci. 2004 Jan;29(1):11-7.
 Tryptophan metabolites and brain disorders

• Tryptophan is metabolised primarily along the

kynurenine pathway, of which two components are now
known to have marked effects on neurons in the central
nervous system. Quinolinic acid is an agonist at the
population of glutamate receptors which are sensitive to
N-methyl-D-aspartate (NMDA), and kynurenic acid is an
antagonist at several glutamate receptors. Consequently
quinolinic acid can act as a neurotoxin while kynurenic
acid is neuroprotectant. A third kynurenine, 3-
hydroxykynurenine, can generate free radicals and
contribute to, or exacerbate, neuronal damage. Changes
in the absolute or relative concentrations of these
kynurenines have been implicated in a variety of central
nervous system disorders such as the AIDS-dementia
complex and Huntington's disease
Stone TW, Mackay GM, Forrest CM, Clark CJ, Darlington LG.
Clin Chem Lab Med. 2003 Jul;41(7):852-9
Immune Responses in ASD,
Borreliosis and
Mycoplasma Infections
 Natural Killer Cell CD-57
• Autistic children are known to have many
metabolic dysfunctions which are shared
by victims of LD, in particular, chronically
low counts of CD57 natural-killer (NK)
cells.
 Gene expression changes in children
with autism
– The objective of this study was to identify gene expression differences in blood
differences in children with autism (AU) and autism spectrum disorder (ASD)
compared to general population controls. Transcriptional profiles were compared with
age- and gender-matched, typically developing children from the general population
(GP). The AU group was subdivided based on a history of developmental regression
(A-R) or a history of early onset (A-E without regression). Total RNA from blood was
processed on human Affymetrix microarrays. Thirty-five children with AU (17 with
early onset autism and 18 with autism with regression) and 14 ASD children (who did
not meet criteria for AU) were compared to 12 GP children. Unpaired t tests (corrected
for multiple comparisons with a false discovery rate of 0.05) detected a number of
genes that were regulated more than 1.5-fold for AU versus GP (n=55 genes), for A-E
versus GP (n=140 genes), for A-R versus GP (n=20 genes), and for A-R versus A-E
(n=494 genes). No genes were significantly regulated for ASD versus GP. There were
11 genes shared between the comparisons of all autism subgroups to GP (AU, A-E,
and A-R versus GP) and these genes were all expressed in natural killer cells and
many belonged to the KEGG natural killer cytotoxicity pathway (p=0.02). A subset of
these genes (n=7) was tested with qRT-PCR and all genes were found to be
differentially expressed (p<0.05). We conclude that the gene expression data support
emerging evidence for abnormalities in peripheral blood leukocytes in autism that
could represent a genetic and/or environmental predisposition to the disorder.

•Gregg JP, Sharp FR et al. Genomics. 2008 Jan;91(1):22-9. Epub 2007 Nov 14.
 Gene Expression Profile Distinctions In Autistic Children
Identified: Genomic analysis could add biological
certainty to behavioral diagnosis
• A group of genes with known links to natural-killer cells -- the first to attack viruses,
bacteria and malignancies -- are expressed at high levels in the blood of children with
autism when compared to children without the disorder.
• researchers also found gene expression distinctions in children with early onset and
regressive forms of the disorder. "What we found were 11 specific genes with
expression levels that were significantly higher in the blood of children with autism when
compared to the blood of typically developing children," Those 11 genes are all known to
be expressed by natural-killer cells, which are cells in the immune system necessary for
mounting a defense against infected cells.
• There is a pattern of 140 genes differentially expressed in children with the early onset
form of the disorder and a pattern of 20 genes differentially expressed in children with
the regressive form of the disorder. These separate experiences offers biological
evidence that there are at least two types of autism -- early onset and regressive.
• In addition to being expressed by natural-killer cells, some of the 11 genes found to be
expressed at higher levels in children with autism are also expressed by CD8+ T
lymphocytes -- cells that target infected cells and, once bound to them, destroy them.
• "What we are seeing can reflect something in the environment that is triggering the
activation of these genes…” "Such an immune response could be caused by exposure to
a virus, another infectious agent or even a toxin. Another possibility is that these
changes represent a genetic susceptibility factor that predisposes children to autism
when they are exposed to some environmental factor.“
• "If the natural-killer cells are dysfunctional, this might mean that they cannot rid a
pregnant mother, fetus or newborn of an infection, which could contribute to autism."
 Invasion and cytopathic killing of
human lymphocytes by spirochetes
• Lyme disease is a persistent low-density spirochetosis caused by
Borrelia burgdorferi sensu lato. Although spirochetes causing Lyme
disease are highly immunogenic in experimental models, the onset
of specific antibody responses to infection is often delayed or
undetectable in some patients. The properties and mechanisms
mediating such immune avoidance remain obscure. To examine the
nature and consequences of interactions between Lyme disease
spirochetes and immune effector cells, we coincubated B.
burgdorferi with primary and cultured human leukocytes. We found
that B. burgdorferi actively attaches to, invades, and kills human B
and T lymphocytes. Significant killing began within 1 hour of mixing.
Cytopathic effects varied with respect to host cell lineage and the
species, viability, and degree of attenuation of the spirochetes. Both
spirochetal virulence and lymphocytic susceptibility could be
phenotypically selected, thus indicating that both bacterial and host
cell factors contribute to such interactions. These results suggest
that invasion and lysis of lymphocytes may constitute previously
unrecognized factors in Lyme disease and bacterial pathogenesis.
 Balanced Inflammation
• Inflammation could have a protective role and promote
regeneration of damaged neurons. We do not yet know
how to achieve a "balanced" inflammation. Because
some novel anti-inflammatory treatment might have
detrimental consequences, carefully monitoring disease
progress in patients treated with this category of drugs is
indispensable
• A variety of neurological diseases the initial triggers differ
significantly, while the subsequent pathways involving
inflammatory processes and causing brain damage
share certain pathological mechanisms

Aktas, O. et al. Arch Neurol 2007;64:185-189.

 ASD & BI/TBI both have:
• Been associated with a combination of
inflammatory and autoimmune
pathophysiology.
• Elevated TNF and IL-6 & reduced NKC
• Antibodies against neural tissue
• Microglial activation
• Oxidative stress
• Greater susceptibility to herpes and other
viral infections
• HLA-DR4 genotypes frequently
 ASD & BI/TBI & Inflammation
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• Patterson P. International neuroscience conference. 2007. Melbourne. Accessed 8-3-07
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 Inflammation and Cognitive Deficits IV
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 Inflammation and Cognitive Deficits V
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 Maternal Immune Activation Alters Fetal
Brain Development through Interleukin-6
• Schizophrenia and autism are thought to result from the interaction between
a susceptibility genotype and environmental risk factors. The offspring of
women who experience infection while pregnant have an increased risk for
these disorders. Maternal immune activation (MIA) in pregnant rodents
produces offspring with abnormalities in behavior, histology, and gene
expression that are reminiscent of schizophrenia and autism, making MIA a
useful model of the disorders. However, the mechanism by which MIA
causes long-term behavioral deficits in the offspring is unknown. Here we
show that the cytokine interleukin-6 (IL-6) is critical for mediating the
behavioral and transcriptional changes in the offspring. A single maternal
injection of IL-6 on day 12.5 of mouse pregnancy causes prepulse inhibition
(PPI) and latent inhibition (LI) deficits in the adult offspring. Moreover,
coadministration of an anti-IL-6 antibody in the poly(I:C) model of MIA
prevents the PPI, LI, and exploratory and social deficits caused by poly(I:C)
and normalizes the associated changes in gene expression in the brains of
adult offspring. Finally, MIA in IL-6 knock-out mice does not result in several
of the behavioral changes seen in the offspring of wild-type mice after MIA.
The identification of IL-6 as a key intermediary should aid in the molecular
dissection of the pathways whereby MIA alters fetal brain development,
which can shed new light on the pathophysiological mechanisms that
predispose to schizophrenia and autism.

Stephen EP et al. The Journal of Neuroscience, October 3, 2007, 27(40):10695-10702

 Stereotypies and hyperactivity in
rhesus monkeys exposed to IgG from
mothers of children with autism
• Four control rhesus monkeys were exposed to
human IgG collected from mothers of multiple
typically developing children. Five additional
monkeys were untreated controls.
• Rhesus monkeys gestationally exposed to IgG
class antibodies from mothers of children with
ASD consistently demonstrated increased
whole-body stereotypies across multiple testing
paradigms. These monkeys were also
hyperactive compared to controls.
 Maternal Immune Activation Alters Fetal
Brain Development through Interleukin-6
• Schizophrenia and autism are thought to result from the interaction between
a susceptibility genotype and environmental risk factors. The offspring of
women who experience infection while pregnant have an increased risk for
these disorders. Maternal immune activation (MIA) in pregnant rodents
produces offspring with abnormalities in behavior, histology, and gene
expression that are reminiscent of schizophrenia and autism, making MIA a
useful model of the disorders. However, the mechanism by which MIA
causes long-term behavioral deficits in the offspring is unknown. Here we
show that the cytokine interleukin-6 (IL-6) is critical for mediating the
behavioral and transcriptional changes in the offspring. A single maternal
injection of IL-6 on day 12.5 of mouse pregnancy causes prepulse inhibition
(PPI) and latent inhibition (LI) deficits in the adult offspring. Moreover,
coadministration of an anti-IL-6 antibody in the poly(I:C) model of MIA
prevents the PPI, LI, and exploratory and social deficits caused by poly(I:C)
and normalizes the associated changes in gene expression in the brains of
adult offspring. Finally, MIA in IL-6 knock-out mice does not result in several
of the behavioral changes seen in the offspring of wild-type mice after MIA.
The identification of IL-6 as a key intermediary should aid in the molecular
dissection of the pathways whereby MIA alters fetal brain development,
which can shed new light on the pathophysiological mechanisms that
predispose to schizophrenia and autism.

Stephen EP et al. The Journal of Neuroscience, October 3, 2007, 27(40):10695-10702

 Stereotypies and hyperactivity in
rhesus monkeys exposed to IgG from
mothers of children with autism

• Rhesus monkeys gestationally exposed to

IgG class antibodies from mothers of
children with ASD consistently
demonstrated increased whole-body
stereotypies across multiple testing
paradigms. These monkeys were also
hyperactive compared to controls.
 Autism: Maternally derived antibodies specific for
fetal brain proteins
– Autism is a profound disorder of neurodevelopment with poorly understood biological
origins. A potential role for maternal autoantibodies in the etiology of some cases of
autism has been proposed in previous studies. To investigate this hypothesis,
maternal plasma antibodies against human fetal and adult brain proteins were
analyzed by western blot in 61 mothers of children with autistic disorder and 102
controls matched for maternal age and birth year (62 mothers of typically developing
children (TD) and 40 mothers of children with non-ASD developmental delays (DD)).
We observed reactivity to two protein bands at approximately 73 and 37kDa in
plasma from 7 of 61 (11.5%) mothers of children with autism (AU) against fetal but
not adult brain, which was not noted in either control group (TD; 0/62 p=0.0061 and
DD; 0/40 p=0.0401). Further, the presence of reactivity to these two bands was
associated with parent report of behavioral regression in AU children when compared
to the TD (p=0.0019) and DD (0.0089) groups. Individual reactivity to the 37kDa band
was observed significantly more often in the AU population compared with TD
(p=0.0086) and DD (p=0.002) mothers, yielding a 5.69-fold odds ratio (95%
confidence interval 2.09-15.51) associated with this band. The presence of these
antibodies in the plasma of some mothers of children with autism, as well as the
differential findings between mothers of children with early onset and regressive
autism may suggest an association between the transfer of IgG autoantibodies during
early neurodevelopment and the risk of developing of autism in some children.

Braunschweig D et al. Neurotoxicology. 2007 Nov 6 [Epub ahead of print]

 Regressive Autism Reactivity:
28% to 37-kDa & 25% to 73-kDa
Table 2
Summary and significant associations of maternal
autoantibody reactivity patterns for human fetal brain
proteins in autism
Prevalence 37 & 73 (kD) 37 (kD) 73 (kD)
Total (n = 61) 7 (12%)* 15 (25%)* 10 (17%)
Regressive (36) 6 (17%)* 10 (28%)* 9 (25%)*
Early onset (25) 1 (4%) 5 (21%) 1 (4%)

Braunschweig D et al. Neurotoxicology. 2007 Nov 6 [Epub ahead of print]

What is associated with 37-kDa &
73-kDa antibodies?
 Medline Search for 37 & 73-kDa Antibodies
• 37-kDa
– Mycoplasma agalactiae [1]
– Mycoplasma gallisepticum [2]
– Mycoplasma arthritidis [3]
• 73-kDa
– Chlamydia [4]
– Streptococcus pneumoniae [5]
– Mycoplasma conjunctivae [6]
• 37-kDa & 73-kDa
– Bartonella henselae and Bartonella quintana [7]
– Borrelia burgdorferi
[1] Fleury B et al. Infect Immun. 2002 Oct;70(10):5612-21.
[2] Gorton TS et al. FEMS Microbiol Lett. 1997 Oct 1;155(1):31-8.
[3] Hasebe A et al. Infect Immun. 2007 Apr;75(4):1820-6.
[4] Kanamoto Y et al. Microbiol Immunol. 1993;37(6):495-8.
[5] Choi IH et al. Microbiol Immunol. 1999;43(8):807-12.
[6] Degiorgis MP et al. J Wildl Dis. 2000 Apr;36(2):265-71.
[7] Haimerl M et al. J Med Microbiol. 1999 Sep;48(9):849-56.
 73-kDa & 37-kDa Amtibodies & LYD
• 73-kDa proteins of Borrelia burgdorferi are
dominant immunogens and expressed in
all strains of B. burgdorferi. The humoral
response to this antigen occurs relatively
early during the course of infection.[1]
• A 37-kDa protein from Borrelia burgdorferi
(the agent of Lyme disease) was identified
as a target for immune-mediated
resolution of Lyme arthritis. [2]
• 37-kDa is a marker for neuroborreliosis. [3]
[1] Luft BJ et al. J Immunol. 1991 Apr 15;146(8):2776-82.
[2] Feng S et al. Immun. 2000 Jul;68(7):4169-73.
[3] Cinco M et al. Immunol Med Microbiol. 1996 Jun;14(2-3):159-66.
 Antibodies against fetal brain in sera of
mothers with autistic children
• Serum antibodies in 100 mothers of children with autistic
disorder (MCAD) were compared to 100 age-matched
mothers with unaffected children (MUC) using as antigenic
substrates human and rodent fetal and adult brain tissues,
GFAP, and MBP. MCAD had significantly more individuals
with Western immunoblot bands at 36 kDa in human fetal
and rodent embryonic brain tissue. The density of bands
was greater in fetal brain at 61 kDa.
• MCAD plus developmental regression had greater
reactivity against human fetal brain at 36 and 39 kDa.
Data support a possible complex association between
genetic/metabolic/environmental factors and the placental
transfer of maternal antibodies in autism.

Singer HS et al. Neuroimmunol. 2008 Feb;194(1-2):165-72. Epub 2008 Feb 21.

 Residual serologic reactivity in children
with resolved Lyme arthritis

• “The 41, 39, and 60 kDa were the most

commonly observed reactive bands.”

Rose CD et al. J Rheumatol. 1996 Feb;23(2):367-9.

 The antibody response in Lyme
disease

• “These polypeptides had molecular

weights of 62, 60, 47, 37, 22, 18, and 15
kDa, and were not recognized by control
sera.”

Craft JE et al. Yale J Biol Med. 1984 Jul-Aug;57(4):561-5

 What is the significance of these
bands?
• 39 kDa is highly specific for Lyme disease
• 36/37 kDa is a Lyme disease band
• 60/62 kDa is a Lyme disease band
 The MMR Debate & TBD
• In 1998, Dr. Wakefield "postulated in the
Lancet that the vaccine might cause
autism."
• Patients with TBD often report symptom
flares following vaccinations.
• Is there an association between vaccines,
patients infected with TBD’s & ASD?
Vaccines a risk for pregnant women?

• “Vaccinating a pregnant woman may be

risky if her immune response interferes
with neuronal growth in her unborn baby’s
brain.”
 Economic Issues
• It may cost $3.2 million to care for one autistic
person in their lifetime and the preliminary data
suggests Borreliosis may be a contributor in 20–
30% of ASD, and pathogenic Mycoplasma may
be a contributor in 58%. If 20% or 58% of the
560,000 recognized cases of ASD in the US can
be prevented or more effectively treated, this
could result in a savings of $358 billion to $1
trillion in addition to incalculable human impact
of this disease.

Bransfield RC, Wulfman JS, Harvey WT, Usman AI. Medical Hypotheses. 2007
 Assessment
• When a patient has been diagnosed with
childhood bipolar illness, ADHD, autism
and other comorbidity, consider the
presence of Tick-borne disease/Lyme
disease and perform an adequate
evaluation.
• Evaluating the possibility of TBI/BI should
be considered in the evaluation of autism
 Treatment Strategies
• Since ASD is caused by an interaction of
genes and environment
• Should treatment be focused upon
changing:
– Genes?
– Environment contributors?
• When TBI/BI is a possibility, consider a
course of antibiotic treatment
 Antibiotic Treatment
• “In our work with children with LD, we
have encountered a few children with
autistic-like disorders,” says Dr Fallon.
“When they received intensive antibiotic
therapy, the autistic syndromes
dramatically improved and, in some cases,
resolved.”
 Further evaluation of the
hypothesis
• It is important to research this association further
and to address the other environmental
contributors that increase the impact of these
infections diseases.
• Narrow and restrictive opinions on the diagnosis
and treatment of Lyme disease may have
contributed to the increased incidence of ASD.
• It is imperative to research all possible causes,
prevent every preventable case and treat every
treatable case of ASD.
 Summary
• A broad base of research & clinical
observations supports the conclusion that
Lyme disease, other tick-borne diseases
and other infectious diseases are
significantly associated with a growing
epidemic of autism spectrum disorder.
• Now greater attention needs to be focused
upon the pathophysiology, prevention,
early diagnosis and treatment.
Thanks for Your Attention