Immunology

of Autism
Aristo Vojdani, Ph.D., M.T.
Immunosciences Lab., Inc.
822 S. Robertson Blvd., Ste. 312
Los Angeles, California 90035
Phone (310) 657-1077
Fax (310) 657-1053
E-mail: drari@msn.com

Autism One
May 20-24, 2009
Chicago, Illinois
Understanding The Puzzle of Complex Diseases
♦ Understanding mechanisms
of action responsible for the
development of complex
diseases including
gastrointestinal,
cardiovascular and
autoimmune diseases.
♦ These diseases cannot
be ascribed to mutation
in a single gene; rather
they arise from the
combined action of
many genes,
environmental factors
and risk-conferring
behavior.
2
A model for the multifactorial nature of autoimmune disease.
Sex hormones represent an important modulatory factor in the immune
and autoimmune response. Sex hormones include the gonadal sex
steroids as well as other hormones that indicate differences between
men and women. Whitacre CC, Nature Immunol , 2:777-780, 2001
3
 

4
 Infections, Toxic Chemicals and Dietary Peptides
Binding to Lymphocyte Receptors and Tissue Enzymes
are Major Instigators of Autoimmunity in Autism
Aristo Vojdani, Jon B. Pangborn, Elroy Vojdani, Edwin L. Cooper;
International Journal of Immunopathology and Pharmacology, Vol. 16, 189-199 (2003)

“This study is the first to demonstrate that dietary peptides,

bacterial toxins and xenobiotics bind to lymphocyte
receptors and/or tissue enzymes, resulting in autoimmune
reaction in children with Autism.”
5
Fig. 2. Scattergram of serum titer of IgG, IgM and IgA antobodies against Gliadin peptides in
healthy control subjects  and autistic patients  and CD69 in healthy control subjects 
and autistic patients  expressed as optical density in ELISA test.
6
Fig. 3. Scattergram of serum titer of IgG, IgM and IgA antobodies against Streptokinase in
healthy control subjects  and autistic patients  and CD69 in healthy control subjects 
and autistic patients  expressed as optical density in ELISA test.
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Aristo Vojdani, Ph.D., M.T.
(310) 657-1077

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“We propose that superantigens (e.g. SK, HSP-60), dietary proteins
(eg. gliadin peptides) in individuals with predisposing HLA molecules
bind to aminopeptidases and induce autoantibodies against peptides
and tissue antigens.”

9
 120
Gliadin HSP 60 Gliadin HSP 60 Gliadin HSP 60
Percent Positive Gliadin or HSP 60 Peptide Antibodies

100

80

60

40

20

0
Elevated DPP IV Antibodies Elevated DPPI Antibodies Elevated CD 13 Antibodies

Percent Positive Sera From Patients with Autoimmune Disease for IgA , IgG ,
and IgM Antibodies against Gliadin and HSP 60 Peptides who are positive for
DPP IV, DPPI, or CD 13 Antibodies. 10
 100
Gliadin HSP 60 Gliadin HSP 60 Gliadin HSP 60
90
Percent Positive Gliadin or HSP 60 Peptide Antibodies

80

70

60

50

40

30

20

10

0
DPP IV Antibodies DPPI Antibodies CD 13 Antibodies

Percent Positive Sera From Patients with Autism for IgA , IgG , and IgM
Antibodies against Gliadin and HSP 60 Peptides who are positive for DPP IV, DPPI, or
CD 13 Antibodies. 11
12
13
 Stem Cell

Primitive cell produced in the

bone marrow and transformed
into T cells in the thymus and B
cells in the bone marrow. All
blood cells (the entire WBC family,
RBC’s, and platelets) are
produced from stem cell
transformation.

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 Macrophage
Housekeeper and frontline
defender, this cell engulfs and
digests debris that washes into
the bloodstream. Encountering a
foreign organism,
it summons
helper T cells to
the scene.

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16
17
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20
 T-Cell

Newly formed in the thymus

and ready for further
specific instruction and
development in the spleen
and lymph nodes.

21
 Mature T-Cell

Programmed with exquisite

specificity, the mature T cell
enters systemic circulation to
“read” a specific antigen and
orchestrate the body’s
defenses against it.

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 Memory Cell

Generated from an initial

infection, this defense cell may
circulate in the blood or lymph
for years, enabling the body to
respond more quickly to
subsequent infections.

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24
 Antibody
Engineered to target a specific
invader, this Y-shaped protein
molecule is rushed to the
infection site, where it either
neutralizes the enemy or tags
it far attack by other cells or
chemicals.

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Interactions of cellular and humoral immunity as defense against invaders 26
27
 60

50
% ABNORMAL NK, T AND B CELL FUNCTION

40 38

34
31
30

20
15

10 8
6
3 *
2
0
NK T-CELL FUNCTION B-CELL FUNCTION CYTOKINES

% ABNORMAL LYMPHOCYTE FUNCTION AND CYTOKINE LEVELS IN CONTROLS AND CHILDREN

WITH AUTISM
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NK Cell Cytotoxic Activity

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 journal of
Neuroimmunolog
y Journal of Neuroimmunology 205 (2008) 148-154

Low natural killer cell cytotoxic activity in autism: the role of 
glutathione, IL-2 and IL-15
A. Vojdani, et al.

Although many articles have reported immune abnormalities in autism, NK cell activity has only been examined in 
one study of 31 patients, of whom 12 were found to have reduced NK activity. The mechanism behind this low NK 
cell  activity  was  not  explored.  For  this  reason,  we  explored  the  measurement  of  NK  cell  activity  in  1027  blood 
samples  from  autistic  children  obtained  from  ten  clinics  and  compared  the  results  to  113  healthy  controls.  This 
counting of NK cells and the measurement of their lytic activity enabled us to express the NK cell activity/100 cells. 
At  the  cutoff  of  15-50  LU  we  found  that  NK  cell  activity  was  low  in  41-81%  of  the  patients  from  the  different 
clinics.  This  NK  cell  activity  below  15  LU  was  found  in  only  8%  of  healthy  subjects  (p  <  0.001).  Low  NK  cell 
activity in both groups did not correlate with percentage and absolute number of CD16+/CD56+ cells. When the NK 
“The induction of NK cell activity by IL-2, IL-15
cytotoxic activity was expressed based on activity/100 CD16 + and
/CD56 + glutathione was more
 cells, several patients who had displayed NK 
pronounced in 15 
cell  activity  below  a subgroup
LU  exhibited with very
normal  NK low
cell  NK cellOverall, 
activity.  activity.  We
after  this conclude that 45%
correction  factor,  ofthe 
45%  of 
children  with  autism  still  exhibited  low  NK  cell  activity,  correlating  with  the  intracellular  level  of  glutathione. 
a subgroup of children with autism suffers from low NK cell activity, and that low
Finally, we cultured lymphocytes of patients with low or high NK cell activity/cell with or without glutathione, IL-2 
intracellular levels of glutathione, IL-2 and IL-15 may be responsible.”
and IL-15. The induction of NK cell activity by IL-2, IL-15 and glutathione was more pronounced in a subgroup 
with very low NK cell activity. We conclude that 45% of a subgroup of children with autism suffers from low NK 
cell activity, and that low intracellular levels of glutathione, IL-2 and IL-15 may be responsible. 
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 120 n = 289
n = 113
n = 189
n = 129
100
NK Cell Activity in Lytic Units

n = 226
n = 49

n = 21
80 n = 37
n = 25
n = 36

60

40 n = 26
32.5

23.3 24.4
22.2 22.4
20 18.8
16.8
20
16.8
13.8
9.6

0
0 C 0.5
1 2 13 4 5
1.5 6 27 8 9
2.5 10
Figure
Figure1 1 -–Distribution
Distributionof NKof NK
cell cellmeasured
activity activity inmeasured
1027 patientsin 1027
with patients
autism with
from 10 different
clinics (1-10) in comparison to Controls (C). n = number of subjects in each group; box point
autism
showsfrom
mean 10 different clinics (1-10) in comparison to Controls (C).
value.
n = number of subjects in each group; box point shows mean value. 
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  100

90
81%
80
70% 71%
70 64%
56% 56%
60
50% 53%
50
46%
41%
40

30

20
8%
10

0
Controls 1 2 3 4 5 6 7 8 9 10
Figure 2 - % NK cell activity below 15 lytic units in controls and patients with autism
obtained from 10 different clinics
Figure 2 – Percentage of NK cell activity below 15 lytic units in controls
and patients with autism obtained from 10 different clinics.

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 70% 6-12

60%
% regulatory T cells

50%

40% 0-5
30%
13-25
20%

10%

0%

Classification of controls  and children with autism

based on % regulatory T cells (CD4+CD25+) in blood.
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% with different levels of TGF-β 60%

50%

40%

30%

20%

10%

0%
10-30 ng/mL 31-60 ng/mL 61-100 ng/mL

Classification of controls  and children with autism

based on TGF-β production by PMBC.
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Levels of cytokine production in controls 
and in children with autism. 36
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 80 50% 47.50%
45%

Individuals w/ IL-17 greater than RR of 57 µg/mL

70

40%
60
Interleukin-17 Level (Pg/mL)

35%
51+29
50
30%

40 36+21 25%

20%
30
15%
15%
20
10%

10
5%

0 0%
Healthy Controls Children w/ Autism Healthy Controls Children w/ Autism

PBMCs from healthy controls  and children with autism  were

stimulated with 10 µg/mL PHA and incubated for 48 hours at 37C.
Aliquots of cell culture supernatant were removed and assayed for
level of IL-17. The percentage of IL-17 elevation above the reference
range is calculated.
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40
 journal of
Neuroimmunolog
y

♦ We measured autoantibodies against nine different neuron-specific antigens and three cross-
reactive peptides in the sera of autistic subjects and healthy controls by means of enzyme-
linked immunosorbent assay (ELISA) testing.
♦ The antigens were myelin basic protein (MBP), ganglioside (GM1), sulfatide (SULF), chondroitin
sulfate (CONSO4), myelin oligodendrocyte glycoprotein (MOG), α-B-crystallin (α-B-CRYS),
neurofilament proteins (NAFP), tubulin and three cross-reactive peptides, Chlamydia
pneumoniae (CPP), Streptococcal M protein (STM6P) and milk butyrophilin (BTN).
♦ Autistic children showed the highest levels of IgG, IgM and IgA antibodies against all
neurologic antigens as well as the three cross-reactive peptides. These antibodies are
specific because immune absorption demonstrated that only neuron-specific antigens or their
cross-reactive epitopes could significantly reduce antibody levels.
“These results suggest a mechanism by which bacterial
♦ These antibodies may have been synthesized as a result of an alteration in the blood-brain
barrier. This barrier promotes access of preexisting T-cells and central nervous system
infection and milk antigens may modulate autoimmune
antigens to immunocompetent cells, which may start a vicious cycle.
responses in autism.”
♦ These results suggest a mechanism by which bacterial infection and milk antigens may
modulate autoimmune responses in autism.
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Chapter 19, Neuropsychiatric Disorders and Infection,
pp 171-186, S.H. Fatemi (ed.) Taylor & Francis Ltd., London, 2005

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 Immune Response to Dietary Proteins, Gliadin and
Cerebellar Peptides in Children with Autism
Vojdani et al. Nutritional Neuroscience, 2004; vol. 7: 151-161

♦ A significant percentage of autism patients showed

elevations in antibodies agaisnt gliadin and cerebellar
peptides simultaneously.
♦ The reaction of anti-cerebellar peptide to gliadin peptide
was 22%.
♦ Binding of anti-myelin basic protein, anti-milk, anti-egg and
anti-soy to gliadin was less than 10%.
♦ We conclude that a subgroup of patients with
autism produce antibodies against Purkinje cells
and gliadin peptides, which may be responsible
for some of the neurological symptoms in autism.

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 3.0
O.D. FOR ANTIBODY BINDING TO GLIADI

2.5
CEREBELLAR PEPTIDE

2.0

1.5

1.0

0.5

0.0
Rabbit Anti-Gliadin 8 A.A. Peptide Rabbit Anti-Cerebellar 8 A.A. Peptide

(EQVPLVQQ) (EDVPLLED)
Reaction of rabbit anti-gliadin or anti-cerebellar 8 amino acid (A.A.) peptides
with non-specific protein (HSA) non-specific peptide and specific peptides
gliadin 18 A. A. and cerebellar 22 A.A. peptides measured by O.D. in
ELISA. 47
 Science, 2002, 298:1424-1427

 Activation-induced cytidine deaminase (AID) plays an essential role in

class switch recombination (CSR) and somatic hypermutation (SHM) of
immunoglobulin genes.
 Deficiency in AID results in the development of hyperplasia of isolated
lymphoid follicles (ILF) associated with a 100-fold expansion of
anaerobic flora in the small intestine.
 Reduction of bacterial flora by antibiotic treatment of AID-/- mice
abolished ILF hyperplasia as well as the germinal center enlargement
seen in secondary lymphoid tissues.
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Abnormal somatic hypermutation and class switch recombination is
responsible for production of pathogenic multi-reactive antibodies.
% elevation of antibodied at 2 SD above the

70

58
60 56
54
50 51
48
50 45
42
40 36
mean

32
30
20
20 16 15
12
10 10 10
8 7
10
3
0
Gliadin DPPIV HSP-60 SK VIP MBP NFP Tubulin STM6P Milk BTN
Peptide

Detection of Antibodies in Patients with Autism

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MUCOSAL IMMUNE ABNORMALITIES

IMBALANCED GUT FLORA

INTESTINAL BARRIER DYSFUNCTION

SYSTEMIC INFLAMMATION

NEUROINFLAMMATION

NEUROINVASION

NEURODEGENERATION

From mucosal immune abnormalities to neuroinflammation and neurodegeneration. 53

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