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INTRODUTIONThe genetic aspect of autism is receivinga plethora of media attention in the US.Indeed, genetics play a role in determiningan individual’s susceptibility to autismspectrum disorders (ASD), celiac, canceror other complex diseases, but it is theenvironment that triggers the genes,which culminates in the disorder. Thepathophysiology of dysregulation in ASDcan be simplied into four understandablesteps. First, environmental triggers,such as viral infection, toxicants and/ordietary proteins and peptides, contributeto gut inammation. This results in steptwo, enhanced intestinal permeability,or “leaky gut.” This state of intestinaldysfunction brings on step three, immunedysregulation, which manifests itself inthe nervous system. In step four, thepatient has neuroinammation. Before anenvironmental factor triggers dysfunctionin the body, however, it must rstinltrate a network of immune defenses.

Basics of immunology

Working in concert, molecules, cells andtissues defend the body against non-selfantigens and pathogens. (Referencesfor this section, except where otherwisenoted, come from

Basic Concepts inImmunology: A Student’s Survival Guide

by John lancy, Jr., Ph.D., an easy tocomprehend guide to the immune systemas a whole.) Microbes attempting aninvasion must rst get through the skinor mucosal layer, which is composed ofscavenger cells and IgA antibodies. If themicrobe is successful in penetrating thebody’s protective shielding or epithelialsurfaces, it must still deal with two distinctbut complementary defense systems:innate immunity and adaptive immunity.

I (o-SpcIIc) d dpIv(cquIrd or SpcIIc) IuIy

Innate immunity is composed of a seriesof nonspecic defenses and substancesthat attack all invaders. It is so-calledbecause this kind of immunity is presentin the body from birth. It is also callednon-specic because it does not need tobe activated by a specic antigen, butwill send out specialized cells to engulfand destroy anything it doesn’t recognizeas being part of the “self.” omponentsof innate immunity include phagocytes,granulocytes, natural killer (NK) cells andthe complement system, a biochemicalcascade that helps clear pathogens froman organism. onsidered the rst lineof active defense, the innate immunesystem acts immediately upon attack.If the innate defense fails against themicrobe, the adaptive immune systemcomes into play. It is also called specicimmunity because, completely oppositeto the innate system, it must be triggeredby specic antigens, and then confrontsthe aliens with specic antibodies andcytotoxic T-cells tailored to effectivelydestroy the microbes. It is also unlikeinnate immunity in that there is a certaindelay in the adaptive system’s response.This is where the adaptive system gets the“acquired” part of its name. While it takesits time to assemble a specic adaptiveresponse to a particular antigen, adaptiveimmunity’s big gun is its immunologicalmemory. It “remembers” having previously

BIOMEDICAL

ARISTO VOJDANI, PhD, MSc, MT

Dr. Aristo Vojdani obtained his PhD inthe eld of microbiology and clinicalimmunology with postdoctoral studiesin tumor immunology. He is the EO andTechnical Director of ImmunosciencesLab., Inc. in Los Angeles, A; memberof the editorial board of three scientic journals; and has published more than100 articles in scientic journals. He isnoted for his papers on autoimmunityand neuroimmunological diseases. In2006, Dr. Vojdani was presented with theprestigious Herbert J. Rinkel Award bythe American Academy of EnvironmentalMedicine (AAEM) for excellence inteaching the techniques of environmentalmedicine.

JAMA LAMBERTImmunosciences Lab, Inc.

Jama Lambert earned her Bachelorof Arts and Sciences degree at theUniversity of Oregon. In 2005, she cameunder the tutelage of Aristo Vojdani,PhD, and she attributes her immuneexpertise to him. Jama is a technical andctional freelance writer, who makes herhome in Los Angeles, alifornia.

A GUT FEELING FOR

Immune DysregulatIon& neuroInflammatIon

IN AUTISM

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encountered an invading organism andreacts more rapidly and efciently thenext time, thus “acquiring” immunity tothat particular pathogen. This immunitycan be gained after recovering frommost pathogenic attacks and is generallylifelong, although it does not precludebeing subject to infection by previouslyunencountered pathogens.NK ELLS As part of innate immunity, NK cells havefascinated immunologists since theirdiscovery thirty years ago. These cellsmediate early non-adaptive responsesagainst viruses, intracellular bacteria,parasite-infected cells and malignancies.They mediate these effects through theproduction of cytokines and the directkilling of transformed, or infected, cellsby granule release as shown in Figure 1.Due to their involvement in regulatingimmune responses, NK cells can beassociated with autoimmune disorders. Asregulators of adaptive immune responses,NK cells inhibit autoreactive T-cells tocurb neuroinammation. To illustrateadaptive immunity, imagine that a glutenpeptide, or

streptokinase

(a bacterialantigen), penetrates the mucosal layer,passes through the intestinal barrierthrough open tight junctions and entersgeneral circulation. As shown in Figure2, a patrolling antigen-presenting cell(AP) such as a monocyte, macrophage ordendritic cell will either process or presentthe already processed antigen to receptorson T-cells. This begins an orchestrateddefense.NK ELLS IN AUTISMAlthough immune system abnormalitieshave been previously implicated in autismand reported in many articles, NK cellactivity has only been examined in onestudy that found reduced activity in 12of 31 patients. However, this study didnot provide evidence for the mechanismresponsible for reduced NK cell activity.Thus, we explored the measurement ofNK cell activity in 1027 blood samplesfrom autistic children obtained from tenclinics operated by Defeat Autism Now!practitioners and compared the results to113 healthy controls. The results of thisstudy were recently published in

Journalof Neuroimmunology

[Vojdani, 2008].We found that NK cell activity was low in41-81% of the patients from the differentclinics. This same low NK cell activity wasfound in only 8% of healthy subjects.Vojdani and colleagues concluded that45% of a subgroup of children withautism suffers from low NK cell activityand that low intracellular levels ofglutathione, interleukin (IL)-2 and IL-15may be responsible. From this study welearn that low NK cell activity is the mostprominent immunological abnormality inchildren with autism, which may be dueto suboptimal cytokine production andlow cellular level of glutathione. However,it remains to be proven whether or notsupplementation with glutathione andinducers of IL-2 and IL-15 will be able torestore low NK cell activity in childrenwith autism to normal levels. In an earlierstudy [Heuser, 1997], it was shown thatNK cell activity could be reversed byvitamin  supplementation in almost80% of adult patients who suffered fromchronic illnesses; this approach may alsobe considered for patients with autism.Although NK cell activity is a vital immunemarker, it remains to be investigatedwhether or not reversal of low NK cellactivity can contribute to improved clinicalmanifestations presented in patients withautism.THE T-ELL FAMILWorking in concert with NK cells in thekilling of targeted cells are the cytotoxicT-cells. T-cells are cells generated inthe thymus. The T-helper (Th) cells aremature D4 T-cells that provide helperfunctions maximizing the capabilities ofthe immune system and assist B-cells inhumoral immune responses; D8 cells arecytotoxic T-cells (TLs) that are involvedin recognizing foreign cells. The bindingof APs to D4 or D8 T-cells facilitatesthe activation of T-cells. In the primaryresponse, effector or activated T-cellsand memory T-cells are generated. Theseactivated T-cells will die by apoptosis,programmed cell death, within 2 to 3days after primary response. However,memory cells live months to years andrespond fervently to the same antigen insubsequent antigenic challenges knownas the secondary response. ActivatedT-cells will produce cytokines, chemicalmessengers which activate additional cells.NK and TL cells are alerted to attack theantigen. Upon a signal from the T-helpercell, the B-cell, which is generated in bonemarrow, makes immunoglobulin (Ig) totag the antigen. In other words, the B-cellplaces a bullseye on the antigen so thatkiller cells know what to target in theirattack. When the battle has been won, theT-suppressor cell calms the activities ofthe T-helper, thereby quelling the attack.The cell keeping a precarious balancebetween T-helper and T-suppressor isthe regulatory T-cell (T

reg

), or T-helper-3(Th3).The mysterious operations of T

regs

havenot been fully elucidated. Researchersknow T

regs

are the major arbiters ofimmune responses, mediating actionsthrough the suppression of inammatoryand destructive immune reactionsby balancing Th1, cell-mediated, and

Figure 1: NK Cell Targeting Tumor Cell

- on the left, the smalle K ell bins t a elage tm ell.  the ight, the K ell has elease its ganles, whih kill the tm ellb egaing the ell’s membane.

THE AUTISM FILE| www.autismle.com ISSUE 31 2009

BIOMEDICAL

Th2, humoral-mediated immunity.Inappropriate T

reg

cell functionalitypotentiates the pathogenesis of diseaseswith ranging magnitudes of severity.Lack of suppressive capability hindersrestraint on immune responses involved inautoimmunity, while suppressive capacityeffectively blocks processes necessaryfor tumor destruction (Vojdani, 2006A,2006B, 2006). In a subgroup of ASDpatients, the important balance of Th1/Th2 is abnormal, and thus contributes toimmune dysfunction associated with thespectrum disorder.ellular-mediated immunity involvesmacrophages, T-cells, natural killer cellsand cytokine production. ytokines areproduced by various immune cells, uponactivation, in order to communicatewith other cells. Th1 cytokines includeIL-2, interferon-gamma (IFN-

) andtumor necrosis factor-alpha (TNF-

).IL-2 and IFN-

are the two majorcytokines produced by Th1 cells. TheTh1 cells, NK cells and macrophagessecrete cytokines that are apparentlywell-coordinated to induce a number ofcytotoxic and inammatory functions.NK cells secrete IFN-

; Th1 cells secreteIFN-

, lymphotoxin and TNF-

; andmacrophages secrete TNF-

. Thesecytokines synergize to induce effectivelysis, destruction by disruption of themembrane, of target cells, particularlyvirus-infected cells. The same cytokinesactivate both macrophages andgranulocytes for increased killing function.There is a yin-yang dynamic betweenagonist and antagonist cytokines, whichmust be kept in balance for optimalimmune reactions.

T-HELPER-1 VERSUS T-HELPER-2 ountering Th1 is Th2 and its humoralimmune properties (see Figure 3).Humoral immunity refers to antibodyproduction and the accessory processesthat accompany it. omponents ofthis system include secretory IgA, acomplex of polymeric IgA and epithelialcell-derived secretory component,immunoglobulins, complement cascade,which is set off when circulatingcomplement encounters an antigen-antibody complex and each molecule ofthe complement performs a specialized job to destroy the target cell, and immunecomplexes, aggregates composed ofantigens and immunoglobulins, whichactivate complement cascade and blockcell-mediated immunity. Humoral-mediated immunity also uses cytokines tocommunicate. Th2 cytokines include IL-4,IL-5, IL-10 and IL-13. Th2 cytokines havethe ability to enhance allergic responses.IL-4 is a major switch factor that inducesheavy-chain class switching to IgEproduction by B-cells. As a result of Th2cell activation and the secretion of thecharacteristic Th2 cytokine pattern, mastcells and eosinophils become activated.Upon activation (see Figure 4), these cellsbind IgE molecules, respond to antigens,and release various mediators. IL-4 alsoenhances the differentiation of Th2cells and inhibits differentiation of Th1cells, thus encouraging the maintenanceof a Th2 response and continuedenhancement of allergy. In contrast toall these positive inuences of Th2 cellson the allergic process, Th1 cells secreteIFN-

, which inhibits the induction ofIgE switching by IL-4 in B-cells, theactivation of mast cells, the activation ofeosinophils, and the proliferation of Th2cells. Enhancement of the Th1 responseto balance Th1/Th2 is implied in allergytreatment.Evidence has been accumulating tosuggest that the pathogenesis of somehuman diseases may be classied interms of type-1 and type-2 T-helper cellcytokine proles. For example, multiplesclerosis, type-1 diabetes, and arthritis areTh1 phenotypes, while allergy, chemicalsensitivity, parasitic infection and lupuserythematosus are Th2 phenotypes (seeTable 1). Scientists theorize that at birththe immune system is immature and isskewed toward Th2 cytokine production.ertain stimuli such as infections andviruses usually contracted from oldersiblings, or peers in day care centers,can help immunological developmenttoward a healthy balance of Th1 andTh2 responses. hildren without suchexposures, living in a relative “sterile”environment, may perpetuate the Th2dominance resulting in increased riskof asthma, other atopic diseases and

Figure 2: Immune Response

- he antigen, a feign mlele, entes ilatin whee anantigen-esenting ell (pc) iks it  an takes it t, an atiates, the  ell. he  heleatiates the B ell t e a sei antib against the antigen, an whih tags theantigen f egnitin. he  hele als alets the ttxi  ell an the natal kille (K)ell, bth f whih attak the feign mateial. he  sess ell nties the  hele whenthe battle is e an ths shts wn the immne esnse. he eglat  ell (eg) keesa balane between the  sess an the  hele t maintain hmestasis.

CD8+CytoxicTCellB cellAntigenAPCNKT HelperT suppressorT reg