MCB 415

NOTE

PROF. E. A. OPHORI

IMMUNOLOGY
TOPICS
1. HYPERSENSITIVITY REACTION
2. MHC
3. IMMUNOMODULATION & IMMUNOSUPPRESSION
4. IMMUNOTRANSPLANTATION

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HYPERSENSITIVITY REACTIONS

This is an abnormal state of immune reactivity which had deleterious

effectts for the host

Ag+Ab+Mastcells →Allergy→release of vasoactive

Gell α Combs classified hypersensitivity states into 4 categories

Type Manifestation Lag between Mechanism

s exp. And
symptoms
I IMHS Minutes 1gE possibly 1gG4
II Cytotoxic Variable IgG IgM
III Ag+Ab complex 6hr igG mainly
IV DHS (Arthus rxn) 12- Sensitized
48hr lymphocytes

Type 1 Systemic Anaphylactic reactions

Cutaneous

The vasoactive amines include histamine, heparin, leukotrienes C4, D4

and E4 (SRS-A-) which lead to smooth muscle contraction and most
reaction are species specific.

Systemic anaphylactic in humans present with itching, erythema,

vomitng and abdorminal cramps, diarrhea, respiratory distress and in

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severe case edema and vascular collapse leading to shock that may be
irreversible.

Examples of type 1-

(a) Asthma
(b) Hay fever
(c) Urticaria

In most cases, there is inflammation characterized by redness, swelling,

heat, pain and loss of function

Chemical mediators are signaling molecules that act on smooth muscle

cells, the endothelial cells, or WBCs to induce, maintain or limit
inflammation. The early phose is modified by histamine and serotonin,
whereas several other medicators contribute to the later phase

TERMS USED IN INFLAMMATORY REACTIONS

1. Hyperemia – increased blood in tissue, caused by vasodilation

2. Edema – excess fluid in tissue – increase vasopermeability
3. Exudate-pathologic, high protein, edema containing Ig

Types of exudate

i. Serous – thin fluid

ii. Fibrinous – containing fibrin
iii. Supurative – nuetrophils and necrotic tissues

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 iv. Hemorrhagic – containing blood
v. Fibrous – healed exudate

MAST CELL MEDIATORS

MEDIATORS STRUCTURE/C SOURCE EFFECTS

HAIN
Histamine B-imidazoleyl Mast cells, basophil Vasodilator increase
ethylamine vascular
permeability
Serotonin 5- Mast cells, platelets Vasodilation,
hydroxytryptamin increase vascular
e permeability
Neutrophil - Mast cells Chemotaxis of
chemo. Factor neutrophils
Easinophil Tetrepeptide Mast cells Chemotactic for
factor eosinophil
Thromboxane Machidonic acid Vasoconstriction
A2 bonchiel anstrikn
plateletaggregation
Prostagladin Arachidonic acid Vasodilation,
E2 (cycloxygenase increase per-
pathy) (read up) meability of
way histamine and
bradykinin
Leukotrienes Arachidonic acid Chemotaxis of
B4 clipoxygenease neutrophils
pathway
PAF (platelet Acetylated Basophils, Release of
activating glycerol ether neutrophis, mediators from
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factor) phosphocholine monocyte and superoxide,
macrophages production, platelets

Mast cell degranulation

1. Ag+mast cell+21gE
2. Dimerization of IgE receptors
3. Activation of phospholipase C on membrane phosphatidylinositol
-4,5 biphosphate- inositoltri P04 and diacylglycerol
4. Activation of protein kinase C
5. Mobilization of instracellula Ca2+
6. Protein phosphorylation
7. Enlargement of granules by protein kinase
8. Activation of phospholipase A2 with formation of lysolecithin and
arachidonic acid
9. Lysolecythin acts as a fusogen causeing granules to fuse with
membrane and release content
10. Activation of granules dependent on cAMP and cGMP
11. Release of vasoactive amines

ARACHIDONIC ACID METABOLISM

Mast cell+Ag+Ab→IMHS-Release of vasoactive amines (early phase)

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 Prolonged response phosphohpase

C00H

SRSA

CYCLO OXYGENASE

LIPOXYGENASE
PROSTAGLANDIN E
LEUCOTRIENES
THIBUMBOXANE A2

C00H

Passive transfer of anaphylactic rxns

Pranusnitz-kustner (PK)

Prostanoic acid

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Atrophy is the tendency of some individual to become sensitized to a
variety of allergens including pollens, spores, animal danders, house
dust and foods

Type II

Cytotoxic Rxns

i. Autoimmne hemolytic anemia

ii. Goodpasteur’s syndrome
iii. Nephrotoxic nephritis

Type III

i. The arthus reaction

ii. Serum sickness

Type IV

i. The Jones-mote reaction

ii. Homograft rejection
iii. Contact hypersensitivity
iv. Transfer of DHS reactions

Type II

Autoimmune henrolytic anemia- host synthesize Abs directed to their

own RBCs and cause lysis by 2 main mechanisms

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(a) If the abs are of the IgM isotype, complement is activated up to the
C9 level (MAC)
(b) If complement fails to complete activation, the RBC will be coated
with abs and possible C3b. they are then phagocytosed and lysed
intracellularly. This ingestion and destruction of RBC takes place
in the spleen and liver and when there is a marked predominance
of splenic sequestration, splenectomy can be of therapeutic value
leading to longer half- life of the patient’s red cells.

Type III

The Arthus Rxn – Reaction leads to edema, hemorhage and later

necrotic. This is caused by complement fixing IgG and is exemplified in
individuals who have reached their maximum immunity in
immunization

Nitrogen mustard or anti-neutrophil serum is used to stop inflammatory

reaction.

Inflammatory reaction will eventually lead to tissue damage

Ag
+ immune complex
Ab

Increase vascular permeability

Circulating Ag/Ab complex
Extravascular deposition

Tissue damage Neutrophil

infiltrate chemotatic Complement activation
factors

Inflammation
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Fig: sequence of events that lead to tissue damage

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DELAYED HYPERSENSITIVITY

Characterized by (1) Redness and induration (b) Perivascular infiltration

of lymphocytes and monocytes (3) lesions are red, warm, painful, soft to
the touch

Onset of immune response

Ag+APC+MHC→IR→T-lymphocytes

This is clonally restricted. T-lymphocytes are from a specific clones of

helper CD4 T cells carrying α/β TCR2 which specifically binds the
immunogenetc peptide presented in association with MACII

Ag TCR
Induced a signal

However, this signal is not enough to trigger the activation and

differentiation of T-lymphocytes. Additional signal is needed and is
provided by surface molecules on APC and T-lymphocytes. This
strengthens the affinity of the interaction between MHC II and TCR

Two pathways have been used to explain this interaction

1. The CD2 – LFA – 3

2. LFA-1-ICAM-1 interaction.

The LFA-3 molecules are expressed by most nucleated cells and RBCs
and their natural substrates are the CD2 molecules expressed by all cells

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Ag+APC+MHC II T cells

LFA-3 TCR Conformation change

CD2R CD2
(Modified CD2

LFA-3+CD2R T lymphocytes
Additional
signal

Lymphocytes then produce cytokines

Type of Lymphokines and Related cytokines

Designatio primary source Activity

n
IL-1 Macrophages Stimulation of T-cells, activates
several types of cells,
inflammatory mediators,
pyrogen
IL-2 T-Cells Activates TB cells
IL-3 T-cells Hematopoietic growth factor
IL-6 Macrophages, Tcell Bcell differentiation
IL-7 Bone marrow, therapy Lymphocyte cell growth factor
TNF Macrophage Tcell Septic shock, tumor autolysis
INFy T cells Macrophage activator

DRUG ALLERGY

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Allergy to drugs may result from (1) overdosage (2) intolerance (3) idiosynchracy

(4) Side effect (5) 2o effects or (6) allergic response to the drug.

Types Definition
Overdose Too much of the drugs
Intolerance Increased sensitivity to normal dose of the drug
Idiosynchracy Qualitatively abnormal pharmacological response, not a
result of the normal pharmacological effect
Side effect Normal but undesired effect of drug
2o effect Normal undesired effect of a drug as a result of
producing the normal effect
Allergic Rxn Reaction mediated by the immune response to the drug

CLASSIFICATION OF DRUG ALLERGY

1. neutralization

2. inactivation

3. cytotoxic

4. atopic or anaphylactic

5. immune complex

6. DHS (Delayed hypersensitivity)

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ALLERGIC RXNS TO DRUGS

Mechanisms Manifestation Example

Neutralization Resistance to therapy Insulin, blood clothing,
cancer therapy, penicillin
Cytotoxic Hemolysis, Penicillin
thrumbocytopenia,
agranulocytosis
Immune complex Skin rashes, serum Serum, insulin penicillin
sickness, nephritis vaccine
Anaphylaxis Asthma, shock, urticaria Penicillin, antibiotics
cancer agents, vaccines
DHS (Delayed Dermatitis toxic Penicillin
hypersensitivity) epidermal necrosis

The most striking allergic drug reaction is anaphylactic shock which occurs within

a few seconds or minutes after exposure, less severe cases include urticaria and

wheezing.

The most common allergic drug reaction are manifested in the skin:

i. Exanthermatic
ii. Erythematous
iii. Maculopapula rashes
iv. Urticaria
v. Angioedema
vi. Contact dermatitis
vii. Erythema multiforms
viii. Eczema
ix. Fixed eruption

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 ALLERGIC AND TOXIC RXNS TO SOME ANTIBIOTICS

Drugs Diseases Contradiction Allergic Rxn

Amebicides
Chloroquine Malaria, amebiasis Pregnancy, G6PD Shock, skin
deficiency, Psoriasis reactions, nausa,
vomiting
Antibacterials
i. Nitrofurantoin UTIs, E.coli, Staph Impaired Renal fnx, Anaphylaxis, fever,
aureus pregnancy asthma, anaemia &
hepatitis
ii. Penicillin Bacterial infections Hypersensitivity Multiple skin
rashes, leucopenia
iii. Tetracycline G-ve, rickettsia, Infants, Hypersensitivity, Nausea, vomiting
enterococci pregnancy renal toxicity
Antifungal
i.Griseofulvin ? ? ?
ii. Nyastatin
Antiviral Herpes Hypersensitivity Nausea vomiting
i. Acyclovir
Helminths
Piperazine Ascaris, pinworm Liver or renal failure Neurotoxicity skin
rashes

Read Up.

1. Penicillin allergy

2. Allergic Rxn to cancer chemotherapentic agents

Drugs Type of Rxn Risk

Asparaginase Anaphylactic hemolytic IC Appreciable
Cis platin Vasoactive agents Appreciable
Bleomycin Pyrogen Infrequent
Hydroxyurea Pyrugens Very rare

MAJOR HISTOCOMPATABILITY COMPLEX

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This is an immune mechanism that enables the host to detect self from
non-self components. A major factor in immune response is to prevent
the immune system from destroying the host cell components. Its’
equivalent is the human leucocyte antigen (HLA) in man.

In many immune responses, the MHC plays a major role in the

processing of the Ag to facilitate the immune response:

Ag+Ab→AgAb complex→IR

IR cellular

Humoral

Ag→APC→MHC Type I
IR
Type II

The functions of the MHC include

(a)Detection of self from non-self

(b) Transplant tissue rejection or acceptance
(c)Involve in specificity and memory of IR

The MHC of mammalian species contain three main genes known as

class I, class II and Class III

Class I and II genes code for cell surface recognition molecules and
class III codes for some complement component

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Class I and II products controls recognition of self and non-self, T-cells
recognize Ags but also have receptors that recognize the MHC products
of cells.

All nucleated cells express class I MHC Ags on their surface while class
II Ags are expressed on some cells such as B-Cells, macrophages but not
on others.

CLASSIFICATION AND DETECTION OF HLA GENES AND

GENE PRODUCTS

Six different loci of the HLA system have been identified and are
divided into two classes

Class 1- 3 major loci ( HLA – A, B, C)

4 minor loci (HLA – E, F, G, H)

Class II – 3 main loci – (HLA-DP,DQ,DR)

2 minor loci – (HLA-DN,DO)

The class II are the most polymorphic and the different antigenic
specificities of each is recognized by two major techniques

1. Serological technique to identify class I and II

2. The mixed lymphocyte technique assay

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GENETIC RULE OF INHERITANCE IN THE MHC

Since there are many alleles, there is only an extremely remote chance
that two unrelated humans will be found who share an individual set of
MHC Ags. This is the basis for their use as genetic markers, which
finds a major practical application in paternity testing / studies. The
expression of MHC markers is governed by two rules

1. Each antigenic specificity of any given MHC locus is determined

by one structural gene
Homozygous B27 B27
B27
Heterozygous →B8/B27 B8
B27
2. Individuals are able to respond to an MHC alloantigen they do not
express, but they cannot respond to an alloantigen which they
express

LINKAGE DISEQUILLIBRIUM

Father Mother
1 WI 5 WI 2. WI 5 W2
(A) (C) parent
(B) phentype
3 W2 8 W4 3 W3 8 W4
Loci A C B D A C B D
Parents haptotype

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(A) AI , CW1, B5 , DW1 (c) A2, CW1 B5 DW2
(B) A3 CW2, B8, DW3 (D) A3, CW3, B8, DW4
(Ac) 1 W1 % W1 (Bc) 3 W2 8 W3
2 W1 5 W2 2 W1 5 W2
(AD) 1 W1 5 W1 (BD) 3 W2 8 W3
3 W3 8 W4 3 W3 8 W4

These are 4 Possible Haplotype

Each parent → 2 haplotypes. Paternal haplotypes are designed A&B, &

maternal C&D. each offspring will receive 1 haplotype from each
parent.

THE MAIN XTICS OF MHC ANTIGENS CLASSES

Xtics Class I Class II Class III

Loci (mouse) K,D,L 1(a,-e) S (C4)
Loci (man) ABC DP, DQ, DR C4, C2
Factor B
No of alleles >100 >20 >2
Distribution of All nucleated Lymphocytes Serum RBC
the products cells macrophages membrane

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Structure
Mol. Wt=43-48k
α2 ∝1
Class 1 Formed by 2 non-identical
polypeptide chains
α1
Major chains α2
“ Bm α3
ss Rk dalton
α3 “

Hydrophobic region 24aas

The aas sequence and nucleotide sequence of the various domains of
-30-35 aas
class 1 MHC shows that the α1 and α2 domains are highly variable and
Cell membrane

most of the aa α nucleotide changes responsible for the diff between

alleles occur in the domains.

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Class II
- No B2 globulin

2 distinct polypeptide chain –

a L-chain 28k (B) which
express the greatest degree of
genetic polymorphism

∝1
Each polypeptide chainßI has 2 extracellular domains ( α 1 , α2 and β 1 , β 2 ) , a
short transmembrane domain and an intracellular tail.

The α1 and ß1 terminal domain contain the NH2 region and it’s the
H-chain (α) which is less
hypervariable region, involved in immune reactions.
polymorphic (33k)
α2 ß2

Cell membrane

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MHC-DISEASE ASSOCIATIONS
They are usually classified as autoimmune disease and are mostly linked
to MHC-II
Disease
A. Inflammatory diseases
1. Ankylosing spondylitis
2. Porter’s syndrome
3. Juvenile rheumatoid
arthritis (type II)
4. Multiple sclerosis
5. Rheumative fever
B. Endocrine disease
1. Diabetes mellitus
2. Addison’s disease
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Description of disease
Inflammation of the spine, leading
to stiffening of vertebral regions
Prostate and parts of the eye
Joint lesions and fever
Inflammatory disease of train and
spinal cord
An autoimmune disease whereas
antibodies raised during B-
heolytic streptococcal pharyngitis
cross react with heart tissues to
give rise to damaging myocarditis
Deficiency of insulin production
Deficiency of production of
adrenaglad
MHC AND IMMUNE RESPONSE

Class I - viruses and intracellular parasite

Class II – soluble antigens

In the class I, the ß2M is involved in the immune response while in the
type II the α1, α2 region that is highly variable is involved.

FACTORS /MECHANISM OF MHC ASSOCIATED DISEASES

1. The B2M protin of the class I refund determine the degree of fit
between MHC II α Ag . All the autoimmune diseases in humans
are linked to the class II MHC and the DQ protin
2. Pathogens infect the HLA B27 (intracellular parasites and cause
disease
3. Ag crocs rx+ with B27 eg yersmia predo TB

1. IMMUNOMODULATION AND IMMUNOSUPPRESSION

Immunosuppression: it is the reduction or absence of immune response

to foreign substances. It is very efficient in (i) autoimmune diseases (ii)
transplant rejection (iii) where the immune system plays a significant
pathogenic role

Immunosuppressant include: Mabs to Tcell, immune toxins, IL-2 toxins

congugete & idiotypic abs

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23
immunesuppresive drugs: pharmaecological & immunological Aspects

possible target sites are phagocytosis and AP macerphages antigen

necognitin by T.cells, Diff of lymp, proliferatin of T & B lymp,
productin of il & aetivation of immune effective mechanisms including
the production & release of cytotoxic leucocytes abs & DHS mediators

i. Harmones
ii. Alkalyting agents & antimatabolites
iii. Inhibitors of signal transduction: cyclosperin A and
ramylyinyan

Hormones: These include glucorticosteroids of the corenel complex

cortex and their synthetic analog

Prednisone and prednisolone

Corticosteriods are known to have an anti-inflammatory rxn due to their

ability to decrease the no α and proinflammatory activity of
mononuclear phafocytes at sites of inflammation.

The interaction of receptors in host animals and corticosteroids lead to

inhibition of cellular metabolism, including nucleic acid synthesis and
glucose uptake. In steroid sensitive sp, both the 1 o & 2o IR impared but
not in man (steroid resistant).

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Prednisone in a dosage of 40mg/day orally will result PPD skin test
change from ve to –ve after an average of about 2wks.

Glucocorticosteronds also inhibit chemostaxis neutrophils & have

marked effects on the distribution of lymp in peripherical blood.

Effects of predison-lymphocytoperia, monocytoparia, depletion of T-

cells.

All effects return to normal in about 24hrs after administration

2. ALKYLATING AGENTS AND ANTIMETABOLITES

These are cycle-active and non cycle-active alkalyting agents such as x-

ray causes cellular death.

Cell cycle

1. Gio-resting
2. G1 = presynthetic phase
3. S CDNA synthesizing phase
4. G2 (premitotic restivity phase)
5. M= (mitosis)

Cycle-active agents may kill by acting at different parts of the cycle.

Antimetabolites such as methotrexate, azathioprine appear to act only on

cells in cycle at the S-phase.

None activated lymp are killed by x-rays

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S-phase inhibitor, include: methrotrexate, 6-mercaptoprine (6-mp) &
azathioprine are cell cycle specific Sphase inhibitors, kill rapidly & cells
cyclosporine has both cycle-active and non-cycle active properties

cyclophosphamide kills both dividing & nondividing cells Cytotoxic

drugs commonly used are

i. Cyclophophanode  1o α2oIR (humural) suppressors

ii. Azathepine  DHS
 Skin-graft refection
iii. Methotrexacte  Auto-immune diseases in animals

Effects of drugs with alkylating and Antimetabolite activity

Effect Acyclo Azathepine Methorecxate

phosphamide &6MP
1 Reduced IoIR + ++ ++
2 Reduced IC ++ + +
3 Antinflammatory + ++ +
effect
4 Initostatic effect ++ ++ ++
5 Reduced DHS ++ + +
6 Reduction of passive ++ + +
transfer of cellular
immunity
7 Lymphopenia ++ +
8 Facilitation of ++ +
tolerance induction

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3. USE OF IMMUNOSUPPRESSIVE DRUG IN
IMMUNOLOGICAL DISORDERS
1. Corticosteroids
- Brondinel asthma
- Autoimmune thrombocytopenis purpura
- Autoimmune hepatitis
- SLE
- Rejection of transplant cysis
2. Cytototic agents and cyclosporine A
- Lnpus glomerulonephritis
- SLE

4. ADVERSE EFFECTS OF IMMUNOSYPRESSIVE DRUGS

1. Bone marrow suppression caused by cytototin drugs
2. Neutropenia-results in pyogenic infections
Use of CSF & am –CSF provides control measure
3. Depression of cellular immunity-leading burral and candida and
aspergillums
4. Chromosomel changes and teratogenia effects
5. Malignancy eg skin cancer and lymphorecticular neoplasm

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 5. IMMUNODIAGNOSIS OF CANCER

They are less organized, less differentiated. Most cancer cells produce
underdeveloped molecules which are used as biomarkers

Some clinical application of biomarker

1. Diagnosis such as monoclonal Ig, carcino-embroyenic ag (CEA),

α-fetoprotein (AFP), ectopic hormoes, cell surface marker such as
T-cell markers
2. Prognosis – CEA, AFP, Bence-Jones protein
3. Therapy-Ab, monocline lab, radiolabel Ab, monoclonel antibodies

Types of cancer markers

Type Example
1 Deletion of blood group ABH
markers
2 Exposure of blood group Monobialoganglosides
meter
3 Isozyme Alkaline phosphatese
4 Tumor Ags Melaomeproteus
5 Ectopic hormone Human chorionic gonado thropin

Levels of expression of onco developmental Ag/markers

Example Normal Ambryogeneticall Distantl Different

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 producing y closely related y related
tissue
1 Carcinoembroyeni Colon Stomach pancres, Lung, Lymphoma
c Ag liver breast
2 Α-fetoprtein Liver, yolksac Colon, Lungs -
stomachipanciae
3 Serotonin Entervendocric Andrenal Oat cell Epidermal
e carcinoid lung lung

Cancer vaccine – Read up

Immunotherapy to human cancer

a) Specific
i. Active-autologoas tumor cells & extracts – not effective
ii. Passive (cellular) autologons lymphocyter/allogeneic lymp not
effective – immune RNA, transfer factors
iii. Passive aumone- xenogeneic antisera- not effective monoclonal
ab-effective
b) Non-specific
BCG, interferon, ILZ, TNF, mycobacterial extract

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Problems and possible solution to monoclone ab therapy

Problem Possible solution

1 Tumors not antigenic Use of immune enhancing mechanism
such as adjustment
2 Tumor ags present in - Selection of onoclonel abs that will
normal tissues select tumer cells
3 Modulation of tumor - Use of diff abs sequentially
Al
4 Selection of non - Use of diff abs sequentially
expressing clones
5 Lack of cytotoxic - Selection of cytotoxic mabs,
effect of monclonel conjugation of mabs to cytotoxc
Ab drugs

Strategy for developing mabs for use in human immunotherapy of

cancer

1. Précises identification of cancer Ags in cancer tissues or those in

low concentration
2. Production of hybridoma cells line producing Mabs to tumor cells
3. Selection of Mabs that select and kill tumer cells
4. Isolation and characterization of Mabs

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