Nature of Antigens and the

Major Histocompatibility
Complex
Immunology and Serology (IMSE 311)

adaptive immune response

- requires prior exposure to
lymphocytes --> cells
the antigen
responsible for adaptive
Rochelle D. Darlucio-Yabut, RMT, MPH immunity
- B cells
Our Lady of Fatima University - T cells
College of Medical Laboratory Science - NK cellls
 molecules capable of binding to the product of that immune response (to
antibody) --> they can bind but they CANNOT STIMULATE antibody
ANTIGEN production
• a substance that is specifically recognized by
the adaptive immune system
- it can be either: immunogenic or non-immunogenic

•IMMUNOGEN can bind to antibody and CAN STIMULATE

antibody production
• a substance capable of causing an adaptive
response.
- able of eliciting immune response by the immune system
- can trigger immune response
- IMMUNOGENIC --> can stimulate immune response
 (antigen) hapten binded to a carrier molecule

once they bind, they can

haptens alone stimulate/induce immune
cannot induce response
an immune response
 - some antigens are too
small/too difficult to bind to
antibody --> cannot stimulate
good example of immunogens immune response
are ANTIGENS

•“All immunogens are antigens

•but not all antigens are

immunogens.”
Factors Influencing the Immune Response
1.Age --> older individuals (humihina ang immune system as we grow older)
--> neonates (immune system not fully developed)

2.Dose --> the larger the dose of an immunogen, the greater the immune response
3.Route of Inoculation subcutaneous, or via oral administration
--> it can be: intravenous, intradermal,

4.Health status of the host --> malnourished, stress, fatigue

5.Genetics --> links to Major Histocompatibility Complex (MHC)
IMMUNOGENICITY
• the ability of an immunogen to stimulate a host response
Traits of Immunogen --> what they need to elicit immune response
--> MW of at least 10,000 Daltons
1.Macromolecular size --> Best immunogens: 100,000 Daltons
2.Chemical composition and molecular complexity
3.Foreignness --> non-self antigen - seen as immunogenic
4.Ability to be processed and presented with MHC
molecules.
Proteins & Polysaccharides: BEST IMMUNOGENS
(most especially protein)
- protein is made up of amino acids
 Main Parts of Antigen
where antibody binds
•1. Antigenic Deteminants / Epitope - molecular shapes or
configurations that are recognized by B or T cells
• - sequential or linear (i.e., amino acids following one
another on a single chain) or conformational.
•Haptens- are nonimmunogenic materials that, when combined with a
•carrier, create new antigenic determinants - y-shaped

becomes immunogenic where antibody binds

•2. Carrier- responsible to give the

antigen its required size.
- usually made up of protein

•VALENCE- number of combining sites

 where antibody binds
EPITOPES / Antigenic Determinant
• precise molecular shapes or configurations recognized by B cells,
or the peptide sequences detected by T cells

• linear epitopes: consist of sequential amino acids on a single

polypeptide chain

• Conformational epitopes: result from the folding of one or

more polypeptide chains, bringing together amino acids that may
be distant from each other so that they are recognized together
B cells: able to bind to those antigens with LINEAR & CONFORMATIONAL (BOTH)
T cells: only able to response to LINEAR EPITOPES

- consists of sequential amino acid

- folding of
polypeptide chains
 HAPTENS distinguished
as IMMUNOGENIC
- non-immunogenic
- when combined with
a carrier: immunogenic

Non-immunogenic
- they can bind to antibody
but no activation of immune basis for
response precipitation
or agglutination
reaction
Adjuvants
a substance administered with an immunogen that increases the
immune response (hinahalo sa vaccines)
Aluminum Salts, Complete Freund’s adjuvant
- increase size & prevent - not usually administered for humans
rapid escape from tissues
(injected in muscles) --> Hepatitis B vaccination

• Adjuvants are thought to enhance the immune response by:

1.Prolonging the existence of immunogen in the area
2.Increasing the effective size of the immunogen
3.Increasing the number of macrophages involved in antigen
processing
 Types of Antigens
"self" - usually do not evoke immune response bcs they are inside of the body (will not harm the host)
1.Autoantigens- are those antigens that belong to the host.
Self-antigens: (HLA antigens - Human Leukocyte Antigen)
2.Alloantigens- are from other members of the host’s species, and these
are capable of eliciting an immune response. - from one person to another person
- Foreign antigens, non-compatibility to tissues/organs --> e.g., during blood transfusion
3.Heteroantigens- are from other species, such as other animals, plants,
or microorganisms.

4.Heterophile antigens- are heteroantigens that exist in unrelated

plants or animals but are either identical or closely related in structure so
that antibody to one will cross-react with antigen of the other
cross reactivity or
cross immunity
 MAJOR HISTOCOMPATIBILITY COMPLEX
- genes can contribute to immune response (good example is MHC)

Large multi-gene locus consisting of several thousand kilobase

pair of DNA on a single chromosome

HUMAN: HUMAN LEUKOCYTE ANTIGENS (HLA), complex

located at chromosome # 6
- capable to mount immune response

Main function of MHC: antigen-presentation --> bring antigen to surface of T-cells

will only be activated in
presence of antigen
The Major histocompatibility complex is found on human
chromosome 6

• 1. Have a role in intercellular recognition and discrimination of self/nonself.

- specificity
• 2. Have a role in development of humoral and cell-mediated immune
responses.
- includes B cells & T cells
• 3. One’s set of MHC genes influences the repertoire of antigens to which that
person’s TH and TC cells can respond, therefore, one’s MHC plays a role in
susceptibility to disease and autoimmunity.
Nature of HLA Antigens
1.Glycoprotein component of cell membrane
2.Present in all nucleated cells
3.Products of the genes of MHC
• Found on the short arm of chromosome 6 and are divided into three categories or
classes.
function for antigen presentation
• Class I molecules are coded for at three different locations or loci, termed A, B, and C.

• Class II genes are situated in the D region, and there are several different loci, known as
DR, DQ, and DP.

• Class III genes, which code for complement proteins and cytokines such as tumor
necrosis factor
 CLASS I MHC MOLECULES CLASS II MHC MOLECULES

CELLULAR All nucleated cells B cells, thymocytes, macrophages,

DISTRIBUTION - HIGHEST: lymphocytes & myeloid cells dendritic cells, thymic epithelial cells
- LOW/UNDETECTED: liver hepatocytes,
- found on antigen-presenting cells
neural cells, muscle cells, sperm cells
- most effective antigen-presenting cells
(expresses high MHC on their surface levels)

STRUCTURE One α chain and β2-microglobulin An α chain and a β chain

chromosome #6 chromosome #15
(located in)

CLASSES A, B, C G: plays a role in immunity DP, DQ, DR modulates antigen binding

Non-classical: HLA E, F, G Non-classical: DM, DN, DO genes involved in antigen processing
not expressed on
E & F: don't present peptide Ag to cytotoxic cells surface of cells DM: holds to load peptides onto class II molecules

SIZE OF PEPTIDES 8 to 11 amino acids 13 to 18 amino acids

BOUND
NATURE OF PEPTIDE Closed at both ends Open at both ends
BINDING CLEFT intracellular viral infection antigens outside of body
(viruses, tumors) (bacteria)
INTERACTION WITH T- Presents endogenous antigen to Presents exogenous antigen to CD4+
CELLS CD8+ T cells T cells
T-cytotoxic cells T-helper cells
• The main role of the class I and class II MHC molecules is to bind peptides within cells
and transport them to the plasma membrane, where T cells can recognize them in the
phenomemon known as Antigen Presentation

MHC Class I: present antigen to CD8+ T cells or T cytotoxic cells

MHC Class II: present antigen to CD4+ T cells or T-helper cells
 undergoes proteolysis

transporter associated proteins

present endogenous antigen in surface

of T cells (CD8+ T cells)

The Class I MHC–

Peptide
Presentation
Pathway
- referred to as "Endogenous Pathway"

TAP contains TAP 1 & TAP 2 --> very essential in this pathway
 The Class II
MHC–Peptide
Presentation
Pathway
- present exogenous antigen on the surface of CD4+ T cells
Application of HLA typing/ Matching
1. Organ transplantation --> check first for compatibility
2. Paternity Testing
3. Forensic medicine, anthropology
4. Studies of racial ancestry and migration
5. For diagnostic and genetic counselling --> certain HLA types inherited that codes for specific diseases:
HLA B27: Ankylosing spondylitis, Reiter's Syndrome
6. Basic research in immunology HLA B47: Congenital adrenal hyperplasia
HLA B5: Behcet's Disease
HLA CW6: Psoriasis vulgaris
HLA DR3: Type 1 DM
HLA DR4: Rheumatoid arthritis
HLA DR5: Gold induce nephropathy, Kaposis's sarcoma, chronic
lymphocytic leukemia
HLA B8: HIV infection
CYTOKINES
Rochelle D. Darlucio-Yabut, RMT,MPH
Our Lady of Fatima University- Valenzuela
College of Medical Laboratory Science
Cytokines
--> proteins that takes act on both innate & adaptive immune response
--> secreted by T cells in response to antigens

are small soluble proteins that regulate the immune system,

orchestrating both innate immunity and the adaptive response to
infection

A class of non antibody molecules, usually a low molecular weight

glycoproteins produces by many different cells in a highly regulated
fashion, change the behaviour and function of many different cells
(Pleiotropism)

They are regulatory and effector molecules that act as picomolar to

nanomolar concentrations on cytokine receptors expressed by target
cells

Cytokine action is mediated by binding to specific receptors on

target cells. They tend to be very potent
 Actions of Cytokines
• Autocrine Stimulation - affecting
the same cell that secreted it
e.g., T cells produces cytokines that affects T-cells

• Paracrine - affecting a target cell in

close proximity

• Endocrine – the effect is systemic

- enter into the circulation
 Features of Cytokines

• Pleiotropism --> single cytokines can have diff. effects

• Redundancy --> several cytokines can have the same effect
single

• Act in networks cytokine

• Act as hematopoietic growth factor

Cytokine Storm: massive overproduction of cytokines

--> leads to shock, multiorgan failure, death
 Cytokines and the Innate Response to
Extracellular Microbes
once they are released, they have different
• interleukin-1 (IL-1) effects:
• interleukin-6 (IL-6) 1. increased capillary permeability -->
• TNF-α antimicrobial proteins (complement, c-reactive
(Tumor Necrosis Factor-alpha) protein) will enter the tissues from the blood
plasma
2. increased platelet aggregation --> prevent
infection in the blood
3. alteration of adhesion molecule expression
on capillary endothelial cells --> enhance entry
of leukocyte into the blood
 The Interleukin-1 (IL-1) Cytokine Family
• IL-1a --> expressed in phagocytic cells, epithelial cells from skin, lungs, and GIT
--> referred to as "endogenous pyrogen" --> ability to induce fever (purpose during infection--high
• IL-1β body temperature)--inhibit growth of many bacteria and fungi, also increased microbicidal activities
of macrophages & neutrophils, also
• IL-1RA (IL-1 receptor antagonist) contribute to feelings of discomfort &
fatigue (conserve energy to combat
usually expressed in monocytes, macrophages, and dendritic cells infection)

IL-1a & IL-1B --> usually released from macrophages & monocytes
 --> before this is cytokines that kill cancer cells
Tumor Necrosis Factors
• TNF-α
• TNFβ --> called as "lymphotoxin" --> produced by lymphocytes
can cause: cell death, cytotoxicity

has 2 factors:
TNFR1 - expressed by many cells in our body
TNFR2 - seen only in hematopoietic & endothelial cells
 Interleukin-6
• IL-6 receptor (IL-6R)
--> able stimulate acute-phase reactants (produced in the liver - under innate immunity)

for adaptive immunity:

- increases activation of B & T lymphocytes; modulates immunoglobulin synthesis --> able to cause B
cells to proliferate to be differentiated in plasma cells
 - derived from cytokines
& chemotaxis
Chemokines
--> movement of cells
towards a stimulus
Transforming Growth Factor-β tumor cells
--> induce growth arrest in

• TGFβ1 - has anti-proliferative activity

- primary regulator of cell
• TGF-β2 growth, differentiation,
• TGF-β3 apoptosis, and migration
 IFN - interferon

Interferon-α and Interferon-β (Type I Interferons)

most important cytokines in response to viral infection
• Type I --> contains IFN-a & IFN-B
• Type II --> IFN-g

INTERFERONS --> substances produced by virally infected cells and

interferes with viral replication
 Cytokines Produced by Th1 Cells
• IL-2 --> T cell growth factor
• IFN-γ --> takes place in enhancement of Ag presentation by class I
and class II MHC molecules

- in adaptive immunity: T cells produced cytokines

Cytokines Produced by Th2 Cells
also expressed in lymphocytes & some non-hematopoietic stem cell types
• IL-4 --> induces the production of MHC class I
• IL-10 --> inhibitory - IL-4, IL-5, IL-13: also produced under MHC Class I
- IL-4, IL-5: codes for eosinophils (allergic reactions, parasitic infections)
effects on immune
response
Cytokines Associated With T-Regulatory Cells
• How Tregs suppress immunity:
(1) the production of suppressive cytokines, such as IL10 and TGF-
β;
(2) disruption of T-cell metabolism
(3) direct cytotoxic killing of T cells and APCs
(4) modulation of signaling between APC and T cells
Th17 Cytokines in Innate and Adaptive Immune
Responses
• IL-17
• TGF-β
• IL-6
• Interleukin-23 --> produced by macrophages & dendritic cells
Hematopoietic Growth Factors
• CSFs (Colony-Stimulating factors)
▫ IL-3 --> for bone marrow to produce B cells & T cells
▫ erythropoietin (EPO) --> RBC production
--> when activated: will became
▫ granulocyte colony-stimulating factor (G-CSF) neutrophil
▫ macrophage colony-stimulating factor (M-CSF) --> when activated: will became
macrophage
▫ granulocyte–macrophage colony-stimulating factor (GM-CSF)
--> when activated: produce other WBC types
IL-3 + GCM-CSF = devt. of basophils and mast cells
IL-3 + IL-5 + GM-CSF = devt. of eosinophils
 development of
hematopoietic stem cells

Thrombopoietin

growth of common myeloid progenitors

(cells under innate immune system)
DEPENDS ON PRODUCTION OF IL-3

<-- myeloid progenitors

Clinical Assays for Cytokines
uses diff. antibodies
• Multiplexed enzyme-linked immunosorbent assays (ELISAs)
• Microbead assays --> detection of multiple cytokines in a single tube
• ELISpot assays. --> detection of individual cytokines-secreting cells
References:
• Stevens, C and Miller, L. (2021). “Clinical Immunology and
Serology, A Laboratory Perspective”. 5th edition