Hypersensitivity

Reactions

Dr. Devinder Toor

Assistant Professor
Amity Institute of Virology &
Immunology
Amity University
• Effector molecules act  remove Ag by various mechanisms.
• Generally, effector molecules induce localized inflammatory rxn that
eliminates Ag without extensively damaging host’s tissue.
• Under certain circumstances, however, this inflammatory response can have
deleterious effects, resulting in significant tissue damage or even death.
• This inappropriate immune response is termed hypersensitivity or allergy.
• Although the word hypersensitivity implies an increased response, the
response is not always heightened but may, instead, be an inappropriate
immune response to an antigen.
• Hypersensitive reactions may develop in the course of either humoral or cell-
mediated responses.
 History
• The ability of immune system to respond inappropriately to antigenic challenge was
recognized when Prince of Monaco became aware of bathers in the Mediterranean reacting
violently to the stings of Portuguese Man of War jellyfish.
• Prince engaged help of Two French scientists, Paul Portier and Charles Richet.
• Portier & Richet concluded that the localized reaction of the bathers was the result of toxins.
• To counteract this reaction, the scientists experimented with the use of isolated jellyfish
toxins as vaccines.
• Their first attempts met with disastrous results. Portier & Richet injected dogs with the
purified toxins, followed later by a booster of toxins.
• Instead of reacting to the booster by producing Abs against the toxins, the dogs immediately
reacted with vomiting, diarrhea, asphyxia and in some instances, death.
• Clearly this was an instance where the animals “overreacted” to the antigen.
• Portier and Richet coined the term anaphylaxis, loosely
translated from Greek to mean the opposite of prophylaxis, to
describe this overreaction.
• Richet was subsequently awarded the Nobel Prize in
Physiology or Medicine in 1913 for his work on anaphylaxis.
• We currently refer to anaphylactic reactions within the humoral branch
initiated by antibody or antigen-antibody complexes as immediate
hypersensitivity, because the symptoms are manifest within minutes or
hours after a sensitized recipient encounters antigen.
• Delayed-type hypersensitivity (DTH) is so named in recognition of the
delay of symptoms until days after exposure.

• We will examine the mechanisms and consequences of the four primary

types of hypersensitive reactions.
 Gell and Coombs Classification
• As it became clear that several different immune mechanisms
give rise to hypersensitive reactions, P. G. H. Gell and R. R. A.
Coombs proposed a classification scheme in which
hypersensitive reactions are divided into four types.
• Three types of hypersensitivity occur within the humoral
branch and are mediated by antibody or antigen-antibody
complexes: IgE-mediated (type I), antibody-mediated (type II),
and immune complex–mediated (type III).
• A fourth type of hypersensitivity depends on reactions within
the cell-mediated branch, and is termed delayed-type
hypersensitivity, or DTH (type IV).
• Each type involves distinct mechanisms, cells, and mediator
molecules (Figure)
 IgE-Mediated (Type I) Hypersensitivity

• Type I rxn  induced by certain types of Ags referred as

allergens, and has all the hallmarks of a normal humoral
response.
• Difference b/w type I hypersensitive response from normal
humoral response  plasma cells secrete IgE.
• IgE - binds with high affinity to Fc receptors - on surface of
tissue mast cells & blood basophils.
 • Mast cells & basophils coated by IgE - said to be sensitized.
• Later exposure to same allergen cross-links membrane-bound
IgE on sensitized mast cells & basophils, causing
degranulation.
• Pharmacologically active mediators - released from granules
 act on the surrounding tissues.
• Principal effects - vasodilation & smooth-muscle contraction
- may be either systemic or localized, depending on the extent
of mediator release.

Vasodilation refers to widening of blood vessels. It results from relaxation of smooth muscle cells within the vessel walls, particularly in the
large veins, large arteries, and smaller arterioles. When blood vessels dilate, the flow of blood is increased due to a decrease in
vascular resistance.
 Type I Reactions –
Systemic or Localized

Clinical manifestations of type I rxns 

range from life-threatening conditions
(systemic anaphylaxis and asthma), to hay
fever and eczema, which are merely
annoying.
 Systemic anaphylaxis
• Systemic anaphylaxis - shock-like and often fatal state whose
onset occurs within minutes of type I hypersensitive reaction.
• Was observed by Portier & Richet in dogs after antigenic
challenge.
• Systemic anaphylaxis - can be induced in a variety of
experimental animals.
• Animal model of choice  guinea pig.
• Why? Anaphylaxis can be induced - with ease & its symptoms
closely parallel those observed in humans.
• Active sensitization in guinea pigs - induced by single injection
of foreign protein (eg. egg albumin).
• After incubation period of ~2 weeks, animal is challenged with
intravenous injection of the same protein.
• Within 1 min, animal becomes restless, its respiration
becomes labored and blood pressure drops.
• As smooth muscles of the GI tract & bladder contract, guinea
pig defecates and urinates.
• Finally - bronchiole constriction results in death by
asphyxiation within 2–4 min of the injection.
• These events - stem from systemic vasodilation & smooth-
muscle contraction - by mediators.
• Postmortem examination reveals - massive edema, shock &
bronchiole constriction - major causes of death.
• Systemic anaphylaxis in humans - characterized by similar
sequence of events.
• Wide range of Ags - shown to trigger this reaction in
susceptible humans.
• Egs. - venom from bee, wasp, hornet, ant stings
- drugs - penicillin, insulin, and antitoxins; - Food
- seafood and nuts.
• If not treated quickly, these reactions can be fatal.
• Epinephrine - drug of choice for systemic anaphylactic Rxns.
• Epinephrine counteracts the effects of mediators like
histamine & leukotrienes by relaxing smooth muscles and
reducing vascular permeability.
• Epinephrine - also improves cardiac output which is necessary
to prevent vascular collapse during an anaphylactic reaction.
• In addition, epinephrine increases cAMP levels in mast cell,
thereby blocking further degranulation.
 on s
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 REAGINIC ANTIBODY (IGE)

• Existence of human serum factor that reacts with allergens

was first demonstrated by K. Prausnitz & H. Kustner in 1921.

• Local wheal & flare response that occurs when an allergen is

injected into a sensitized individual is called the P-K reaction.
• Because the serum components responsible for the P-K
reaction displayed specificity for allergen, they were assumed
to be antibodies, but the nature of these P-K antibodies, or
reagins, was not demonstrated for many years.

• Experiments conducted by K. & T. Ishizaka in the mid- 1960s.

• Ishizakas called this new isotype IgE in reference to E Ag of

ragweed that they used to characterize it.

• Serum IgE levels in normal - 0.1–0.4 g/ml

• Most severely allergic individuals – rarely IgE levels greater
than 1 g/ml.
• Low levels made physiochemical studies of IgE difficult
• Discovery of IgE myeloma by S.G.O. Johansson & H. Bennich in
1967 - extensive chemical analysis of IgE could be done.
• IgE - found to be composed of 2 heavy & 2 light chains
(combined mol weight of 190,000). Higher mol weight wrt IgG
(150,000) - due to presence of an additional Constant-region
domain.
• Additional domain (CH4) - contributes to altered conformation
of the Fc portion - enables it to bind to glycoprotein receptors
on surface of basophils & mast cells
• Half-life of IgE in serum is 2–3 days. Once IgE bound to its
receptor on mast cells & basophils  stable in that state for a
number of weeks.
Fc Receptors
 IgE-BINDING Fc RECEPTORS

• Reaginic activity of IgE depends on its ability to bind to a

receptor specific for Fc region of the heavy chain.

• 2 classes of FcR: expressed by different cell types & differ by

1000-fold in their affinity for IgE.
– FcRI

– FcRII
 HIGH-AFFINITY RECEPTOR (FCRI)
• Mast cells & basophils express FcRI - binds IgE with high
affinity.
• High affinity - enables FcR to bind IgE despite low serum
concentration of IgE.
• Between 40,000 - 90,000 FcRI molecules - present on a
human basophil.
• STRUCTURE
• FcRI receptor - contains 4 polypeptide chains: an α & β chain
and 2 identical disulfide-linked  chains
• External region of __ chain – have 2 domains of 90 aa
-homologous with Ig-fold structure (placing FcRI in Ig
superfamily).
• FcRI interacts with CH3/CH3 & CH4/CH4 domains of IgE via
two Ig-like domains.

• __ chain spans - plasma membrane 4 times - thought to link

α chain to  homodimer.

• Disulfide-linked  chains extend considerable distance into

cytoplasm.

• Each chain have conserved sequence in its cytosolic domain

- immunoreceptor tyrosine-based activation motif (ITAM).
• ITAM motif on these receptors - interacts with protein
tyrosine kinases to transduce an activating signal to the cell.

• Allergen mediated crosslinkage of bound IgE results in

aggregation of FcRI receptors & rapid tyrosine
phosphorylation, which initiates the process of mast-cell
degranulation.

• Role of FcRI in anaphylaxis - confirmed by experiments in mice

that lack FcRI. These mice have normal levels of mast cells but
are resistant to localized & systemic anaphylaxis.
 LOW-AFFINITY RECEPTOR (FCRII)
• FcRII (or CD23) - specific for CH3/CH3 domain of IgE and has
lower affinity for IgE than does FcRI.

• FcRII - appears to play variety of roles in regulating intensity of

IgE response.

• Allergen crosslinkage of IgE bound to FcRII - shown to activate

B cells, alveolar macrophages & eosinophils.

• When this receptor is blocked with mAbs, IgE secretion by B

cells is diminished.
• Soluble form of FcRII (or sCD23) - generated by
autoproteolysis of membrane receptor - shown to enhance
IgE production by B cells.

• Atopic individuals - have higher levels of CD23 on their

lymphocytes & Ms and higher levels of sCD23 in serum.
 IgE Crosslinkage Initiates Degranulation
• Although mast-cell degranulation generally is initiated by
allergen crosslinkage of bound IgE,

• No. of other stimuli can also initiate the process, including the
anaphylatoxins (C3a, C4a, and C5a) and various drugs.
 RECEPTOR CROSSLINKAGE
• IgE-mediated degranulation begins when an allergen
crosslinks IgE that is bound to Fc receptor on mast cell or
basophil surface.
• In itself, binding of IgE to FcRI apparently has no effect on
target cell.
• Only after allergen crosslinks the fixed IgE-receptor complex -
degranulation proceeds.
• Importance of crosslinkage - indicated by inability of
monovalent allergens, which cannot crosslink the fixed IgE, to
trigger degranulation.
• Experiments have revealed -
essential step in
degranulation is crosslinkage
of 2 or more FcRI — with or
without bound IgE.
• Crosslinkage - normally
effected by interaction of
fixed IgE with divalent or
multivalent allergen
• Also can be effected by
variety of expt. means that
bypass need for allergen & in
some cases even for IgE.
 Methods to Detect Type I
Hypersensitivity Rxns

• Commonly identified & assessed by skin testing.

• Small amounts of potential allergens - introduced at specific
skin sites by intradermal injection or superficial scratching.
• Number of tests can be applied to sites on forearm or back of
an individual at one time.
• If person is allergic to allergen - local mast cells degranulate &
release histamine and other mediators – produces wheal &
flare within 30 min.
• Advantage of skin testing - relatively inexpensive and allows
screening of a large number of allergens at one time.

• Disadvantage of skin testing - it sometimes sensitizes the

allergic individual to new allergens and in some rare cases
may induce systemic anaphylactic shock.
• Few individuals also manifest late-phase rxn, which comes 4–
6 h after testing and sometimes lasts for up to 24 hr.
Eosinophils accumulate during late-phase reaction, and
release of eosinophil-granule contents contributes to tissue
damage in late-phase reaction site.
 RIST - Radioimmunosorbent test
• Method to determine serum level of total IgE Ab.
• Highly sensitive technique, based on radioimmunoassay
• Can detect nanomolar levels of total IgE.
• Patient’s serum is reacted with agarose beads or paper disks
coated with rabbit anti-IgE.
• Beads or disks are washed  125I-labeled rabbit anti- IgE is
added.
• Radioactivity of beads, measured with gamma counter, is
proportional to the level of IgE in the patient’s serum.
 RAST - Radioallergosorbent test
• Detects serum level of IgE specific for a given allergen.
• Allergen is coupled to beads or disks patient’s serum is
added  unbound Ab is washed away.
• Amount of specific IgE bound to the solid-phase allergen is
then measured by adding 125I-labeled rabbit anti-IgE,
washing the beads, and counting the bound radioactivity.