ANTIGEN-ANTIBODY

REACTIONS

Presenter name:
Bharathi Gudapati
 Contents

• Introduction
• Immunity
• Antigen
• Antibody
• Antigen antibody reactions
• Conclusion
• References
BASICs
 ANTIGEN
 An antigen is a substance which when introduced into a body
evokes an immune response to produce a specific antibody with
which it reacts .

 It can be classified as-

1. Complete antigen
2. Incomplete antigen (Haptens)
• Epitope is the smallest unit of antigenicity.

• The combining site on the antibody molecule,

corresponding to the epitope is called the
Paratope.
 Antibody

 These are substances which are formed in the serum and

tissue fluids in response to an antigen and react with that antigen
specifically and in some observable manner.

 Chemically they are globulins, hence they are named

immunoglobulins.

 They constitute about 20 – 25% of the total serum proteins and

are mainly synthesized by plasma cells.
Structure of Immunoglobulin
Immunoglobulin classes
7
Abnormal Immunoglobulins
16
• Multiple myeloma
• Urine by coagulation
Bence jones protein • Light chain
• Ig G,A,D or E

Waldenstrom’s • Ig M producing cells

macroglobulinemia • M protein and their chains
• Form of paraproteinemia
Heavy chain disease causing lymphoid neoplasia
G, A, M Fc portion

• Formation of gel on
cooling and redissolves
cryoglobulinemia on warming
• IgG ,M or mixed
 MISCELLANEOUS

ADAPTIVE

LOCAL

HERD
Humoral immunity
 Primary and Secondary Immune Response

◆Primary immune response

Immune response that occurs after 1st exposure to an antigen.

◆Secondary immune response

Immune response that occurs after 2nd or subsequent exposure to

the same antigen.
 Antigen Elimination

Antibodies eliminate antigens by:

1. Neutralization of toxins and Viruses.

2. Complement activation followed by

a. Cytolysis.
b. Opsonization and phagocytosis.
c. Immune complex clearance

3. Antibody dependent cell mediated cytotoxicity(ADCC).

Monoclonal vs Polyclonal Antibodies Comparison
Area Monoclonal Polyclonal

Specificity for Varying specificities

Epitope
a single epitope. to multiple epitopes.

Identifies the entire

target protein via
binding at multiple
Identifies whether a
sites . Since multiple
Targeting particular region of a
epitopes are targeted,
protein is present
there is a higher
likelihood of detection
of the target
 Uses of Monoclonal Antibodies

• Diagnostic tests
– detect small amounts of drugs, toxins or hormones
– e.g. monoclonal antibodies to human chorionic
gonadotropin (HCG) - pregnancy test kits
– AIDS by the ELISA test
• Treat viral diseases
• Detection and immunotherapy of cancer
• Classify strains of a single pathogen, e.g. Neisseria
gonorrhoeae
 Factors Affecting the Antibody
Production
• Immunosupressive agents
– Inhibit the immune response
– Essential to prevent graft rejection in transplantation
X rays: no antibody is produced till 24 hrs after exposure
Corticosteroids: depletion of lymphocyte from blood and
lymphoid organ
Anti-metabolites: interfere the synthesis of DNA, RNA or
both. Ex: methotrexate, 6-mercaptopurine, azathioprine.
Cytotoxic Chemicals: cyclosporine – cyclic polypeptide,
most commonly used
Antilymphocytic serum (ALS): antiserum against
lypmhocytes
Radiomimetic drugs: cyclophosphamide, nitrogen
mustard
• Multiple Antigens: when 2 or more antigens are given
simultaneously the effect may vary
– No effect: antibody will be produced separately as if they
were given alone.
eg:Typhoid and cholera vaccine
– Enhanced: triple vaccine DPT
– Diminished: when only diptheria and tetanus vaccine given
together – the one in excess shows antibody production.
• person already immunised with diptheria, will not show immune
reponse to triple vaccine other than dipteria

• Effect of Antibody
– Immune response gets suppressed by passive administration
of homologous antibody.
– Rh(-ve) mother having Rh(+ve) fetus – early sensitization by
administration of anti-Rh globullin (antibody).
Super antigens

• Certain protein Molecules that activate large number of

T cells, irrespective of Antigenic specificity.
Ex: Staphylococcal Enterotoxin

• Usually only few T-cells (0.001%) are stimulated –

up to 20 % of circulating T-cells
• This exaggerated T-cell activation – outpouring of
T-cell cytokines : multisystem dysfunction
(Staphylococcal Shock syndrome).
 Types of Hypersensitivity Reactions

The response of the host to the presence of foreign substances can trigger four types of
hypersensitivity reactions:
1.Immediate
2.Cytotoxic
3.Immune complex
4.Cell-mediated

Type I: Immediate Hypersensitivity (Anaphylactic reaction):

Type II: Cytotoxic Reaction (antibody dependent)
TYPE III:IMMUNE COMPLEX REACTION:
TYPE IV : DELAYED HYPERSENSITIVITY:
The complement system
 Antigen Antibody Reactions
• The antigens and antibodies combine specifically with each other.
This interaction between them is called Antigen – Antibody
reaction.

• It may be abbreviated as
Ag – Ab reaction.

• The first correct description of the antigen-antibody reaction was

given by Richard J. Goldberg at the University of Wisconsin in
1952.
• These reactions form the basis for detection of infectious disease
causing agents and also some non specific antigens like enzymes.

• The reactions between Ag and Ab occur in three stages.

 In first or primary stage the reaction involves formation

of Ag-Ab complex.

 The secondary stage leads to visible events like precipitation,

agglutination etc.

 The tertiary stage includes destruction of Ag or its

neutralization.
Classification of antigen-antibody interactions

1.Primary serological tests: (Marker techniques) e.g.

–Enzyme linked immuono sorbent assay (ELISA)
–Immuno florescent antibody technique (IFAT)
–Radio immuno assay (RIA)

2.Secondary serological tests: e.g.

–Agglutination tests
–Complement fixation tests (CFT)
–Precipitation tests
–Serum neutralization tests (SNT)
–Toxin-antitoxin test

3.Tertiary serological test: e.g.

–Determination of the protective value of an anti serum in an animal.
Its USES are
1. In vivo
• Forms basis of immunity against infectious diseases.

2. In vitro
• For diagnosis of infections
• Helpful in epidemiological studies
• For identification of enzymes
• Detection and quantification of antigens or antibodies.
 Characteristics
• Reaction is specific, an antigen combines only with its homologous
antibody and vice-versa. However, cross reactions may occur due
to antigenic similarity.

• Entire molecules of antigen and antibody react and not the

fragments.

• Only the surface antigens participate in the antigen antibody

reaction.

• The reaction is firm but reversible.

 Properties of Antigen – Antibody Reaction:

•The properties of antigen and antibody can be explained with the

help of three points. They are:

• Antibody Affinity.
• Antibody Avidity
• Cross reaction.
 Affinity of Antibody:

Antibody affinity is the strength of

the reaction between a single
antigenic determinant and a single
combining site on the antibody.

•It is the sum of the attractive and

repulsive forces operating between
the antigenic determinant and the
combining site .
 Avidity of Antibody:

•It is the strength of the bond after the formation of Ag-Ab

complexes.
•It is used to denote the overall capacity of antibodies to
combine with the multivalent antigen.
•A multivalent Ag has many types of antigenic determinants.
•When injected into the blood, each antigenic determinant
stimulates the production of a particular antibody.
 Cross Reaction:

An antiserum raised against an Ag, can also react with a similar

Ag of another type. This is called cross reaction and the Ag
which produces the cross reaction is called Cross reactive Ag.
But the strength of Ab raised against its own Ag is strong.
 Types

 Precipitation reactions

 Agglutination

 Neutralization test

 Immunofluorescence

 Radioimmunoassay

 Enzyme linked immunosorbent assay

 Immunoelectronmicroscopic test
 Precipitation Reaction:

When a soluble Ag combines with its Ab in the presence of

an electrolyte (NaCl) at a particular temperature and pH, it
forms an insoluble precipitate of Ag-Ab complex. The Ab
causing precipitation is called Precipitin and the reaction is
called as precipitation reaction.

Antibodies Antigens Ag-Ab complex

 Applications

• Identification of bacteria. E.g., Lancefield grouping of

streptococcus.

• Detection of antibody for diagnostic purposes. E.g.,

VDRL in syphilis

• Forensic application in identification of human blood

and seminal stains

• To standardize toxins and antitoxins.

 Types of precipitation reactions

• Ring test: CRP TEST 20

FLOCCULATION TEST: VDRL TEST SYPHILIS

Non reactive and Reactive VDRL Tests

 Agglutination
 It is an antigen antibody reaction in which a particulate antigen
combines with its antibody in the presence of electrolytes at an
optimal temperature and pH resulting in visible clumping of
particles.

 It is more sensitive than precipitation for detection of

antibodies.

 The reactions take place better with IgM antibody.

 Type
s
Slide Agglutination test
Routine procedure to identify
bacterial stains.
E.g., Salmonella species Agglutination+: ve Agglutination : -ve

 Also used for blood grouping.

Tube Agglutination test

 Standard quantitative method for determination of antibodies.
 Routinely employed in diagnosis of typhoid, brucellosis and typhus
fever
 Coombs Test

• Originally devised by Coombs, Mourant and Race (1945) for

detection of incomplete Rh antibodies.

• When sera containing incomplete anti-Rh antibodies are mixed with

corresponding Rh-positive erythrocytes but no agglutination occurs.

• When such erythrocytes are treated with antiglobulin or COOMBS

serum (rabbit antiserum with human gamma globulin), the cells
are
agglutinated.
 Agglutination reactions

particulate antigen + antibody clumping

23

COOMBS TEST LATEX AGGLUTINATION TEST COAGGLUTINATION

Anti Rh antibody , incomplete ab Hepatitis B antigen , CRP
N.Gonorrhoeae,Strep.pyogen

IgG
Protein (Fc
A

F
ab
 Neutralization Test
• Bacterial exotoxins are capable of producing neutralizing
antibodies (antitoxins) which play protective role in diseases such
as diphtheria and tetanus.

• Toxin – antitoxin neutralization can be measured in vivo and in

vitro.
In vivo tests:
 Toxigenicity test. E.g., C. diphtheriae
 Shick test (similar test in human)

In vitro test:
 Antistreptolysin ‘O’ (ASO) test. E.g., Strep
pyogenes
 Virus neutralization tests.
 Immunofluorescence
• Fluorescence is the property of certain dyes which
absorb rays of one particular wavelength (ultraviolet
light) and emit rays with a different wavelength (visible
light)

• Most commonly used dyes are:

1. Fluorescin isothiocyanate – blue green
2. Lissamine rhodamine – orange red

 They are of two types:

1. Direct immunofluorescence
2. Indirect immunofluorescence
 Direct immunofluorescence
Uses:

• Commonly employed for detection of bacteria,

viruses or other antigens in blood, urine, tissues and
other specimens.

• Sensitive method to diagnosis Rabies.

Disadvantage: Separate fluorescent labelled antibody has to

be prepared for each antigen to be tested.
 Indirect immunofluorescence

• A single antihuman globulin fluorescent conjugate employed

for detection of antibody to any antigen. This has overcome the
disadvantage of direct immunofluorescence
 ELISA

• Enzyme linked immunosorbent assay is a simple and a sensitive

test.

• Requires only microlitre quantities of test reagents.

• The principle of ELISA is almost same as that of

immunofluorescence, the only difference being, an enzyme is used
instead of fluorescent dye.

• It can be used for detection of Antigen or Antibody.

• Types: Sandwich, Indirect, Competitive ELISA

 Uses:

• Detection of HIV antibodies in serum

• Detection of mycobacterial antibodies in
TB

• Detection of Hepatitis B markers in

serum.

• Detection of enterotoxin of E.coli in

feces.
 Auto immune disease

Normally antibodies can detect and differentiate molecules from

outside of the body and those produced inside the body as a result
of cellular activities. Self molecules as ignored by the immune
system. However, in certain conditions, the antibodies recognize
self molecules as antigens and triggers unexpected immune
responses. This results in different autoimmune diseases
depending on the type of antigens and antibodies involved. Such
conditions are always harmful and sometimes deadly. The exact
nature of antibody-antigen interaction in autoimmune disease is
not yet understood.
 RECENT ADVANCES

Uses of Histone Deacetylase (HDAC) Antibodies in Research

Histone deacetylase (HDAC) antibodies are used to study the

process of histone deacetylation, an essential activity that happens
within human cells

Numerous assays currently exist that rely on the use of histone

deacetylase (HDAC) antibodies to explore the proteins present within a
biological sample and to view how they interact with other
biomolecules, which can uncover information on a wide variety of
factors. Assays including DNA pull‐down, immunohistochemistry, and
Western blot have utilized this kind of antibody. Fields that are
benefiting greatly from these methods include cancer research and
pharmacology
Applied aspects
COMPLICATIONS DUE TO AN ALTERED IMMUNE RESPONSE

An efficient immune response protects against many diseases and disorders. An

inefficient immune response allows diseases to develop. Too much, too little, or the
wrong immune response causes immune system disorders. An overactive immune
response can lead to the development of autoimmune diseases, in which antibodies
form against the body's own tissues.

Complications from altered immune responses include:

• Allergy or hypersensitivity
• Anaphylaxis, a life-threatening allergic reaction
• Autoimmune disorders
• Graft versus host disease, a complication of a bone marrow transplant
• Immunodeficiency disorders
• Serum sickness
• Transplant rejection
• 4 autoantibodies are markers of beta cell autoimmunity in type 1 diabetes: islet cell
antibodies (ICA, against cytoplasmic proteins in the beta cell), antibodies to glutamic
acid decarboxylase (GAD-65), insulin autoantibodies (IAA), and IA-2A, to protein
tyrosine phosphatase
 Undesirable Drug Reactions in Orofacial Region

Three mechanisms have been proposed for drug allergies.

First, IgE mediated reactions occur when the drug reacts with IgE
antibodies bound to mast cells.

Second,drug allergies can involve a cytotoxic reaction in which an

antibody binds to a drug that is already attached to a cell surface.

Third, drug allergy involves circulation of the antigen for extended

periods allowing sensitization of the patient's immune system and
production of a new antibody
Allergic reactions in dentistry
 Allergy to Resin Materials Composites

The dental personnel commonly complain of contact dermatitis and asthma caused by
methacrylates . HEMA, EGDMA and TEG-DMA are responsible for occupational contact
allergies.

A study was reported in which patients had lichenoid-like reactions of lips and patch testing
revealed positive reaction to composite components.

Antifungal treatment and replacement of existing restorations resulted in improvement.

Even though resin-based restorative materials are considered safe, their constituents can leach
out and cause allergic contact stomatitis as reported in a patient with mild erythema in the
gingiva and buccal mucosa.

Fisher recognized MMA monomer as the main cause of allergic dermatitis in dentists and dental
laboratory technicians.
The prevalence of contact allergy to methyl methacrylate is 1%. Different methods reduce the
leachable substances from acrylic dentures such as immersing in hot water (50°C) for one hour
prior to inserting into the oral cavity or ultraviolet light .

Allergic Reactions to Dental Materials-A Systematic Review

Meena Syed, Radhika Chopra, and Vinod Sachdev
      
 Allergy to Mercury

Delayed hypersensitivity reactions to amalgam restorations are seen as

erythematous, pruritic lesions on the oral mucosa and skin of the face and neck. The
common manifestations of these reactions are oral lichenoid lesions (OLL).
Another manifestation of allergy to mercury is burning mouth syndrome (BMS).
 Allergy to Metals Nickel-Chromium

Nickel is the common sensitizer amongst all metals.

Nickel was considered as one of the causes of allergic
contact dermatitis in women by Fisher .

In 1889, Goldman reported the first case of Nickel

dermatitis characterized by sensitivity to nickel
compounds . Overall, nickel sensitivity is more common
in women (4-10 times) than men whereas, chromium
allergy is rare (10% in males and 3% in females).

The clinical signs and symptoms of nickel allergy

include burning sensation, gingival hyperplasia,
numbness on sides of tongue and the final diagnosis is
confirmed by patch test using 5% nickel sulphate in
petroleum jelly. In sensitized individuals, nickel
exposure leads to systemic allergic contact dermatitis
 Allergy to Latex Gloves

The frequent risk of population for latex allergy includes children with spina bifida (highest
risk), patients who underwent surgery before one year of age, latex-fruit syndrome (allergy to
various fruits) and healthcare workers (second highest risk) who have higher risk of latex
allergy due to sweating and multiple glove changing.

The allergic reactions to latex vary from stomatitis to airway compromise. There is no “gold
standard” for diagnosing latex allergy as no test is 100% accurate.

The diagnostic algorithm for latex allergy includes

obtaining medical history, skin patch testing for
type IV delayed hypersensitivity; measurement of
serum IgE for type I immediate hypersensitivity and
glove testing when patient’s history is not correlating
with IgE results
 Allergy to Local Anaesthetics

Although LA are well-tolerated drugs, they precipitate adverse reactions which are related
to LA , doses (toxic reaction or overdosage) or psychogenic factors.

Adverse reactions are reported of lignocaine , prilocaine mepivacaine or to their

components like methylparaben or metabisulphite.
 Allergy to Materials Used in Endodontics Formaldehyde

Formaldehyde is a common cause of allergic contact dermatitis .

In the dental literature 28 patients with immediate symptoms to formaldehyde

containing root canal compounds have been described. The characteristic features of
formaldehyde allergy are anaphylactic reaction or shock and generalized urticaria.

The most useful and diagnostic tool to determine formaldehyde allergy is the
assessment of specific IgE antibodies to formaldehyde
 Allergy to Impression Materials

Allergic reactions are reported to polyether impression materials which manifests as swelling,
itching and redness. It was seen on patch testing that a component of the catalyst paste caused
the allergy and on replacement of this component, no allergic reactions were observed.

There is only a single allergic case reported in which a patient developed hypersensitivity
reaction to polysulfide material in the form of redness, itching and edema following secondary
impression for upper and lower complete dentures and on treatment with topical
corticosteroids (Betamethasone valerate ointment 0.1%) she recovered.

There have been reported cases of allergic reactions to alginate impression material. Alginate
impression material is contraindicated in patients with a history of severe allergic reaction to
crystalline silica, calcium sulfate, or potassium titanium fluoride. In these cases, it is better to
use an alginate-free impression material.
Allergic reactions in implant
dentistry
 Titanium

The first case of delayed hypersensitivity reaction to titanium in the form of local
granulomatous reaction was described in patients wearing cardiac pacemakers.
Titanium allergy has a low prevalence rate of 0.6% and presents with urticaria,
eczema, redness of the mucosa.

It was observed that presence of elements in titanium alloys could cause allergic
reactions in dental implant patients such as beryllium (Be), cobalt (Co), chromium
(Cr).

Patch tests have limited use due to poor sensitivity and the test validated to detect
titanium sensitization is MELISA test . Alternate substitutes like Polytheretherketone
(PEEK) which offer mechanical properties and bone forming capacity similar to
titanium are also under investigation
 Conclusion

For establishing diagnosis, it is essential to obtain proper history related to allergy,

clinical examination and confirmatory tests like patch tests and MELISA. Thus, due to
rise in number of patients with allergies from different materials, the practicing dentists
should be aware about the allergies documented to known materials and thus prevent
allergic manifestations in the dental clinic.
 Conclusion
• Therefore we see the application of antigen antibody reactions in
the diagnosis of diseases which can help in developments of
varieties of diagnostic tests.
• In clinical practice, they help in:
 Preventing destructive diseases.
 Preventing progression of the diseases.
 Identifying high risk patients
 Target treatment of specific diseases
 Monitor the effects of the treatment.
 References

1. Textbook of Microbiology for dental students. Dir. Prof. C. P Baveja. 4th edition

2. Textbook of Microbiology for dental students.Ananthnarayan and Paniker. 8th edition

3. Essential Microbiology for Dentistry, 4th Edition. Lakshman Samaranayake.

4. https://en.wikipedia.org/wiki/Antigen antibody_interaction

5. Allergic Reactions to Dental Materials-A Systematic Review by Meena

Syed, Radhika Chopra, and Vinod Sachdev