BIOCHEMISTRY

IMMUNOLOGY
Dra. Agnes A. Alba, MD || 08/30/2017
LE1

OUTLINE A. 2 TYPES OF IMMUNITY

 Innate Immunity
I. Immunity
 Natural / Native; Non- Specific
A. 2 Types of Immunity
 Consist of physical and chemical barriers, cells and
B. Difference between Innate and Adaptive molecules that recognizes a limited number of pathogen –
II. Innate Immunity associated molecular patterns and effector systems that
A. Cells involved in Inflammation facilitate destruction of pathogen.
III. Adaptive Immunity  Immediate response and it is the body‘s first line of defense
A. Cells of Adaptive Immunity  Adaptive Immunity
B. Two Types of Adaptive Immunity  Centered on lymphocytes:
1. Cell Mediated Immunity  B cells - responsible for Humoral responses
2. Humoral Immunity  T cells - responsible for cellular responses
IV. Cell Communication  Acquired; Highly specific
A. Cytokines  Specificity of the response is achieved through receptors that
B. Pattern Recognition Receptors recognize antigen
C. Antigen Presentation  Has immunologic memory which makes it remember a specific
V. Complement System encounter, thus yielding to a specific response
A. Classical  Acts when innate defenses are unsuccessful due to the
B. Lectin persistence of the triggering agent.
C. Alternative
D. MAC
VI. Immune Diseases and Clinical Examples
VII. Review Questions
VIII. References
IX. Appendix

OBJECTIVES
At the end of the lecture, students must be able to:
● Explain the role of the immune system in the maintenance of
health
● Compare and contrast between innate and adaptive immunity in
terms of the cells and mechanisms
● Compare and contrast the characteristics and mechanisms
involved in humoral and cell-mediated immunity
● Relate the role of the complement system in innate immunity
● Discuss the cells involved in the adaptive immune response – T
cells, B cells and antigen presenting cells

I. IMMUNITY
 The human immune system has developed to protect the
individual from many pathogen constantly present in the
environment
 A state of relative resistance to disease
 Defined as resistance to disease, specifically infectious
disease (Abbas, 2011)
 Results from the combined activities of many different cells
concentrated in lymphoid organs, such as the bone marrow,
thymus, spleen, and lymph nodes (Karp, 2010).
 Engages in a type of molecular screening by which they
recognize “foreign” macromolecules (Karp, 2006).
 It is good to have pathogens in our environment to help Figure 1. Mechanisms of Innate and Adaptive Immunity Response. (A) Time
response (B) Steps of infectious agent removal
improve our immune system, but it should not be too
overwhelming that we would not be able to resist the
Kinetics of immunity
problems.
The mechanisms of innate immunity provide the initial
Immunology: branch of biomedical science that deals defense against infections.
with the immune system Innate Immunity: provides intermediate protection against
Immune System: the defense system of the host microbial invasion
comprising many biological structures within an organism Epithelial barriers – prevent infections
that acts as a protection against the diseases Phagocytes, NK, ILCs, complement – eliminate microbes
Immune Repertoire: the number of different sub-types an Adaptive Immunity: develops more slowly and provides
organism's immune system makes more specialized defense against infections
The diversity of antigen receptors of T and B lymphocytes. Antibodies – block infections and eliminate microbes
T lymphocytes – eradicate intracellular microbes

Trans #8 Group #18 : Castillo, Castillo, Castillo, Castro, Castro TRANS HEAD: ACUYONG 1 of 13
Table 1. Importance of Immune System in health and disease ● Phylogenetic studies have indicated that genes for PRRs and
other components have been gradually modified over
generations by natural selection

A. CELLS OF INVOLVED IN INFLAMMATION

● When activated, innate cells can synthesize and secrete a
wide variety of inflammatory mediators.
● Some may be toxic to pathogens while others (cytokines)
may be released to activate other immune cells.
● Liver produces mediators present in the blood.

Circulating Cells
a. Neutrophils

B. DIFFERENCE BETWEEN INNATE AND ADAPTIVE

RESPONSE

Table 2. Difference between Innate and Adaptive Immunity

INNATE ADAPTIVE

Cell Can discriminate Can discriminate

Discrimination foreign particles between two similar Figure 2. Diapedesis of a White Blood Cell.
molecules
● Normally found circulating in the bloodstream
Response Time Immediate Response takes a few ● Most abundant; 60-70% of the WBCs in circulation.
days (Lag Phase) ● Removes infectious agents through the process of
phagocytosis.
Specificity Non-specific, same High, the response is → Neutrophils diapedese into the tissues in response to
response to a wide directed only to the antigenic stimulation.
variety of foreign initiating agent → Chemotactic factors that attract the neutrophil to the site
agents of inflammation include complement proteins, bacterial
products, injured tissue, and hemostatic components.
Diversity Limited Extensive
→ Opsonins such as IgG and complement component C3b
help neutrophils recognize a substance as foreign
Memory Absent, subsequent Present, subsequent
Opsonization - process by which a pathogen is marked
exposures to foreign exposures to the same
agents generate the foreign agents generate for ingestion and eliminated by a phagocyte
same response amplified responses → Phagocytosis involves the attachment of the neutrophil to
the foreign object, formation of a vacuole around the
Cell Types Phagoctyes, Natural T-Cells, B-Cells, foreign object, and neutrophilic degranulation to release
Killer (NK) cells, Antigen Presenting lytic enzymes (called respiratory burst) in an effort to kill
Dendritic cells Cells (APC) the organism.
● Sensitive to the oxidants they secrete and are destroyed in
the process.
II. INNATE IMMUNITY ● Removed by macrophages and dendritic cells.
● Natural or Native immunity b. Eosinophils
● Immediate response and First line of defense. ● 1-3% of circulating WBCs; count increases with allergic
● Defends against infection in a Non-specific manner. reactions as well as in conditions of parasitism
● Defense mechanisms present even before infection and have ● Primarily thought of as terminal effectors of allergic
evolved to specifically recognize microbes and protect responses and of parasite elimination.
multicellular organisms against infections. ● Express Fc receptors (receptors for the fragment
● Conferred by all elements with which an individual is born and crystallizable or Fc region on the tail end of antibodies that
are present and available at very short notices (Coico, 2003). are used to activate the immune system) for IgE.
● Protects an organism from attack, using physiochemical ● Has histamine containing granules.
barriers, such as the skin and mucosal epithelia, and their ● Releases substances that can neutralize products released
associated secreted products, e.g. sweat, mucus and acid by basophils and mast cells. Therefore, eosinophils modulate
(Gracie & Farrell, 2014) the allergic response.
● Contains variety of cells such as phagocytes, neutrophils,
→ Release lipid mediators, including leukotriene C4, platelet-
natural killer cells, and others.
activating factor, and liposins.
● System recognizes microbes through pattern recognition
c. Basophils
receptors (PRRs), which are receptors specific for molecular
● 0-10% of circulating WBCs; least predominant.
components of microorganisms that are not made by the host.
● Some are temporarily upregulated as a result of exposure ● Has membrane receptors IgE, and histamine-releasing
to microbes, but the components of the innate immune system granules responsible for allergic reactions.
do not change permanently during an individual's lifetime. ● Shares many features with mast cells, including the secretion
of Th2 cytokines and release of histamine after activation.

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→ Once activated, degranulation releases histamine, which Eosinophils ● Allergic responses and parasite
initiates the classic signs of immediate hypersensitivity elimination
reactions.
● Release lipid mediators
● Basophils release a chemotactic factor that attracts ● Express Class II Major
eosinophils to the site. Histocompatibility (MHC-II)
● Basophils also release heparin, an anticoagulant.
Basophils ● Secretion of Th2 cytokines and
Tissue Based Cells release of histamine after
a. Macrophages activation
● In the peripheral blood, this cell is a monocyte. In the tissue,
it is a macrophage. Monocytes ● Precursors of tissue macrophages
● Express major histocompatibility complex II (MHC II), ● Recognize antigens and initiate
complement, and IgG receptors on their surface. adaptive immune responses
● Play a role in initiating and regulating the immune response.
→ They process ingested material and process antigenic Naive ● Migrate through these peripheral
information, which is presented to the T-helper (CD4) Lymphocytes lymphoid organs, where they
lymphocyte. recognize antigens and initiate
→ They arrive at the site of inflammation after neutrophils, adaptive immune responses.
but unlike the neutrophil, the phagocytic process of the
monocyte allows it to live longer. Tissue-based Macrophages ● Constitutively present in tissues
● Secrete cytokines/interleukins and tumor necrosis factor. Cells ● “dustbin of the immune
response”
● KEY FUNCTIONS include:
→ Phagocytosis of infecting
microbes
→ Antigen presentation
→ General removal of dying or
damaged host cells.

Mast Cells ● immediate hypersensitivity

● Increase in number several-fold in
association with IgE-dependent
immediate hypersensitivity
reactions
● Preformed mediators, including
histamine, serine proteases,
Figure 3. Role of the Macrophage in the Immune Response. carboxypeptidase A, and
proteoglycans are stored in
b. Mast Cells cytoplasmic granules.
● Central to the pathogenesis of diseases of immediate
hypersensitivity.
● Increase in number several-fold in association with IgE-
dependent immediate hypersensitivity reactions. III. ADAPTIVE IMMUNITY
● Release chemical mediators that cause the characteristic  Highly specific and can discriminate between two very similar
symptoms of allergy. molecules (Karp,2006).
 Responses require a lag period
NOTE:  Immunologic memory
● Extravasation
→ In the case of inflammation, it refers to the movement of white  Mechanism to remember specific encounter
blood cells from the capillaries to the tissues surrounding them  Result of memory cells that do not require further
(leukocyte extravasation), also known as diapedesis differentiation.
→ This process forms part of the innate immune response, involving  Immunoglobulin (Ig)
the recruitment of non-specific leukocytes.  Secreted by B lymphocytes
 Has specificity for foreign substances, but not all will elicit a
Table 3. Summary of Cells Involved in Inflammation. response
Classification Type of Cell Function
 Immunogen
 Foreign substances that elicit responses
Circulating Neutrophils  Most numbered leukocyte in blood  Antigen
Cells  Found circulating the bloodstream  Any antigen that can be bound by an antibody
 Recruited to sites of infection by  Epitope
the process of extravasation  Antigenic structures that gives rise to a series of effects
 Performs NETosis, which is the A. CELLS OF ADAPTIVE IMMUNITY
process of release of neutrophil T Lymphocytes
extracellular traps as a last resort  Originates from the thymus
to microbial infections.  Each cell is genetically programmed to recognize specific cell
bound antigen via antigen-specific T-cell receptor (TCR)
 Can either signal other phagocytes or directly kill the infected
cells

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 Includes cell mediated immunologic processes such as graft
rejection, hypersensitivity reaction, and defense against
malignant cells
 Mature naive T cells are found in the blood and T cell zones of
peripheral lymphoid organs as they are attracted by chemokines

B Lymphocytes
 Derived from the bone marrow
 Recognizes extracellular antigens and differentiate into
antibody-secreting plasma cells
 Mediators of Humoral Immunity (as Immunoglobulins)

Figure 6. Effector Functions of T-cells

Figure 4. Cells of the Adaptive Immunity

B. 2 TYPES OF ADAPTIVE IMMUNITY

Figure 7. T-cell Receptor Complex

Figure 8. Progression of Cell-mediated Response

Figure 5. Two Types of Adaptive Immunity
NOTE:
Cell-Mediated Immune Response  Figure 8: A macrophage is activated upon encounter with its matching
 T lymphocytes function to eliminate microbes ingested by antigen. It then engulfs the antigen for digestion, with its fragments
phagocytes or are living in the cytoplasm. bound to MHC molecules for presentation. This will then attract the
presence of a helper T‐cell, which will then secrete cytokines to either
 Protects the body from virus infection and cancer, killing attract macrophages, neutrophils and other lymphocytes for cell lysis,
abnormal or virus-infected body cells or stimulate differentiation of other T- cells into cytotoxic T‐cells (TC).
 T-cell receptor Complex (TCR) These TC will then destroy infected host cells.
 Consists of lymphocytes and their products, including
antibodies. Humoral Immune Response
 Have Antigen-binding site – exposed externally from the cell  Mediated by B-cells
that will recognize the microbe or the dead cell. When the
 characterized by the release of antibodies produced by
binding occurred, protein molecule will undergo a
plasma B lymphocytes.
conformational change that will allow intracellular signaling
 physiologic function: defense against extracellular microbes
mechanism which will activate other processes inside the cell
and microbial toxins
 Effector Functions of T Cells
 Antibody, or immunoglobulin (Ig)
 T cells bound to the cell
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 Serum glycoprotein produced by plasma cells that mature
from lymphocytes, called B lymphocytes (B cells), in
response to an antigen.
 Oligomeric proteins whose individual subunits have
traditionally have been classified as Heavy (H) or Light (L)
based on their migration during the SDS-PAGE
(Polyacrylamide gel electrophoresis)
 The L and H chains are polypeptides
 5 Classes: IgA, IgD, IgE, IgG, and IgM

I. FUNCTIONS OF IMMUNOGLOBULINS
1. Antigen binding
 Bind specifically to an antigen
 Results to the protection of the host
Figure 10. Immunoglobulin Fragments
 Valency of the Ab refers to the number of antigenic
determinant Nice to know:
2. Effector functions
 Fd is the heavy chain (VH, CH1) portion of Fab
 No direct biological effect but secondary effector function  Fd ' is a heavy chain (VH, CH1) portion of Fab. The prime ( ' )
 Secondary effector in the fixation of the complement system mark denotes extra amino acids due to a pepsin cleavage site.
which ends in the lysis of cells  F(ab')2 is a dimeric molecule produced by pepsin cleavage. An
 Binding to cell types ex bind to receptors on placental immunoglobulin monomer will produce a single F(ab')2
trophoblasts—which results transfer of Ig across the placenta fragment containing two (VH, CH1‘) segments joined by
disulfide bonds. An F(ab')2 contains two epitope-binding sites.
II. BASIC STRUCTURE OF IMMNUNOGLOBULIN Source: Lippincott illustrated Review of immunology pg. 61
 Built from 4 polypeptide chains: 2 identical heavy chains
and 2 identical light chains
 Stabilized by disulfide bonds
→ Inter-chain - between the heavy and light chains
→ Intra-chain - within each polypeptide bond
● Has two main regions: Variable (V) and Constant (C)
→ Variable region - basis for binding specificity
→ Constant region - has limited variation
→ Light Chain (VL100AA) (CL110AA)
▪ Variable light chain is composed of 100 Amino Acids
▪ Constant light chain is composed of 110 Amino Acids
→ Heavy Chain (VH110AA) (CH330-440AA)
▪ Variable heavy chain is composed of 110 Amino Acids
▪ Constant heavy chain is composed of 330-440 Amino
Acids
▪ Determines the IgG class
● Hinge region – region where flexibility is noted

Figure 11. Enzyme cleavage of immunoglobulin

IV. REGIONS OF IMMUNOGLOBULINS

 Hinge Region
 Also known as flexible region
 Allows both arms to move independently, thus facilitating the
binding to antigenic sites that may have variable distances
Figure 9. Basic Structure of Immunoglobulin  Easily cleaved by pepsin or papain
 Variable Region
III. MOLECULAR STRUCTURE OF IMMUNOGLOBULINS  Antigen binding site that dictates the specificity of antibodies
● Two fragments: Fab (Fragment Antigen Binding) and Fc  No 2 variable regions from different humans share identical
(Fragment Crystallisable) AA sequences
→ Fab  All variable regions are highly unique in amino acid sequence
▪ antigen binding fragment  Hypervariable Region
▪ Valence = 1  5-10 residue island
▪ specificity is determined by VH and VL  Also called Complementarity-Determining Region
▪ each fragment has an epitope binding site  Various combinations of H and L chain CDRs can give rise to
→ Fc multiple antibodies with different specificities (combinatorial
▪ binds to cell-surface receptors diversity)
▪ functions as effector  The essence of antigen - antibody interactions is MUTUAL
● Cleaved by protein Papain COMPLEMENTARITY between surfaces of CDRs and
epitopes through noncovalent interactions

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 The short acidic glycopeptide allows the dimerization of
secretory IgA
 Half-life of 6 days
 Primary immunologic barrier against pathogenic invasions of
mucous membranes
 Found in mucus, tears, saliva, and colostrum
 Most predominant immunoglobulin produced by MALT
 Neutralization

3. IgM (Pentameric)
 Confined in the intravascular space
 Helps eliminate circulating antigens and microorganisms
 Half-life of 5 days
 First antibody synthesized after an antigenic challenge
 1st Ig made by the fetus: marker of in utero infection if it reaches
levels above 20 ng/l
Figure 12. Ig Regions and function  Most effective classic complement pathway activator due to its
 Constant Region pentameric form
 Located within the Fc fragment  Neutralization
 particularly the CH2 and CH3 (CH4 of IgM and IgE)
 Responsible for CLASS SPECIFIC EFFECTOR 4. IgD
 FUNCTIONS such as complement fixation or transplacental  Surface receptor for antigens in B lymphocytes
passage  HIGH carbohydrate content
 Has a single interconnecting sulfide bridge and elongated hinge
V. ANTIBODY DIVERSITY DEPENDS ON GENE region making it more susceptible to proteolysis
REARRANGEMENTS  Significance of circulating IgD is still undefined
 A human genome contains less than 150,000 Ig genes
 Each person is capable of synthesizing 1M different antibodies 5. IgE
specific for a unique antigen  Similar to IgM in its unit structure
 Immunoglobulin DIVERSITY that is generated by combinatorial  Least abundant immunoglobulin
mechanisms  Antibody of allergy and anti-parasitic activity
 1st mechanism: division of a coding sequence for each  High affinity for binding sites of mast cells and basophils
immunoglobulin chain among multiple genes. A light chain → is  Antigenic binding of the Fab region induces crosslinking of high
a product of 3 structural gene that codes for a variable region, affinity receptors, granulation of cells, and then its release
region and constant region  Mediates immediate hypersensitivity (allergy) by causing the
 Several combinations → multiplicity of combinations release of mediators from mast cells and basophils upon
 2nd Diversity is augmented by the action of ACTIVATION exposure to antigens
INDUCED CYSTIDINE DEAMINASE (AID) → By catalyzing the  Does not fix complement
conversion of cystidine to uracil
 AUTOIMMUNITY: the mutagenic action of AID led to the
generation of autoantibodies
 3rd mechanism: generating antibodies target novel antigens is
JUNCTIONAL DIVERSITY - refers to the addition or deletion of
random numbers of nucleotides that takes place when certain
gene segments are joined together, somatic in nature

VI. 5 CLASSES OF IMMUNOGLOBULIN (Ig)

**See Appendix for a comprehensive table
1. IgG
 Circulates in high concentration in the plasma (75-80%)
 Smallest but most numbered antibody
 Half-life of 22 days Figure 13. Function of IgE
 Present in the ECF
 During the 18-20th weeks of pregnancy, IgG is transported in
the placenta and provides humoral immunity for the fetus and
neonate until the maturation of the immune system (Passive
Immunity)
 Major isotype used in a secondary antibody response
 Dependent on antibodies (cell-mediated cytotoxicity)
 Neutralization and Opsonization
 Complement activation via the classical pathway
 Natural killer function: cell killing of antibody - bound cells
achieved by the receptors for the Fc portion

2. IgA (Dimeric)
 Found in secretions and presents an antiseptic barrier
protecting the mucosal surfaces
 The alpha chain has 18 extra amino acids at its C terminus
Figure 14. Primary and Secondary Immune Responses

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Interleukins Mode of action is
(ILs) communication between
leukocytes.

Tumor Necrosis TNF Promotes inflammation,

Factor (TNF)
TNF Promotes inflammation.

Osteoprotegerin Stimulate osteoclasts and

(OPG) bone resorption.

Chemokines Involved in chemokinesis

(movement in response to
chemical stimuli)

Figure 15. Direct and Indirect Functions of Antibodies 1. Colony stimulating factors – stimulate production of
cells, increase number of cells to prevent further tissue damage
 Subclasses of Immunoglobulin: 2. Interferons (IFN)
 IgG: IgG1, IgG2, IgG3, IgG4  inhibit and interfere the replication of infecting viruses
 IgG1: best to cross the placenta  3 classes (refer to Table 1)
 IgG2: does not cross the placenta  IFNγ– regulates cell activity by activating/inhibiting cells (cell-
 IgG4: does not fix complement mediated)
 IgA: IgA1, IgA2 3. Interleukins (ILK)
 IgA1: monomer, IgA2: dimer  Produced by leukocytes and act on leukocytes
 Structures different from one another
IV. CELL COMMUNICATION 4. Tumor Necrosis Factor (TNF) - causes cell death. Alpha &
Beta are responsible for inflammatory conditions, osteoprotegerin
stimulate osteoclasts in bone absorption.
A. CYTOKINES 5. Chemokines
 Are cell signaling molecules that aid cell to cell  Small proteins secreted by activated WBC‘s
communication in immune responses and stimulate the  Set of chemokines for innate response different from that of
movement of cells towards sites of inflammation, infection, and adaptive response
trauma.  Secreted by macrophages, dendritic cells, natural killer cells,
 Are cell signaling molecules that aid cell to cell etc.
communication in immune responses and stimulate the
movement of cells towards sites of inflammation, infection, and Table 5. Cytokines Involved in the Immune Response.
trauma. IMMUNE PRODUCING CELLS CYTOKINES
 Soluble mediators of inflammatory and immune responses. RESPONSE
 Structure: Small peptides or Glycoproteins active at
concentrations between 10-15 mol/L– produced after Innate Macrophages TNFɑ, IL12
translational processes Responses Dendritic Cells IL1 IL15
Natural Killer (NK) Cells IL 6 IL 18
 Produced by various cells, including those of the innate and
IL8 IFN
adaptive responses
 May be secreted by all the cells of the immune system as well Adaptive or T cells, especially CD4+ IL2, IL4, IL5, IL10,
as fibroblasts, epithelial cells, and adipocytes. Cell-mediated T cells IL13, IL17, IL22, TGF
→ Macrophages are the main producers during innate Responses
responses, while T cells during adaptive responses.

Table 4. Classification of Cytokines.  Cytokines allow 2 cells to communicate with each other:
 Once the cytokines are secreted from the
CLASS TYPE OF CELL FUNCTION
originating/producing cells, they are transported to the
effector cells with receptors which recognize the cytokines.
Colony Involved in the
 These recepient cells contain Pattern Recognition Receptors
Stimulating development and
(PRR)
Factors (CSF) differentiation of immune
cells from bone marrow
precursors. B. PATTERN RECOGNITION RECEPTORS
 Effector cells have a set of germline-encoded cell surface and
Interferons IFN Exhibits role in inhibiting intracellular receptors for microbial recognition.
(IFNs) viral replication  Response elicited by such receptors are amnesic, responding
similarly on reinfection.
IFN� Inhibits viral replication.  Identifies structures that are shared by various microbes which
are not present on host cells.
 These patterns are also called Pathogen-Associated Molecular
IFN�  Regulates immune
Patterns (PAMPs).
responses
 Made primarily by T cells Types of Receptors
and activates
macrophages.
I. Membrane-bound Receptors
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A. Mannose Receptors C. ANTIGEN PRESENTATION
 A free-circulating plasma protein which activated the  The process wherein antigen-presenting cells capture antigens
complement cascade via the lectin pathway upon to bind them to molecules of the major histocompatibility
recognizing and binding to antigen PAMP. complex, where a T cell may recognize it.
 A complement factor known as mannose-binding lectin  Antigen-presenting cells include:
(MBL) or mannan-binding protein (MBP), binds bacterial  Dendritic Cells, which are the main type of APC. Their main
polysaccharides to generate a complex that recruits and function is process antigenic material for presentation to
activates C4 in the complement cascade. other cells.
 B cells
B. FC Receptors  Macrophages
 Stands for fragment crystallizable. Binds to antibodies which  Natural Killer Cells
are attached to infected cells.  T cells only recognize antigenic peptides when bound to MHC
 Their activity stimulates phagocytosis. and are formally presented.
 MHCs are synthesized in the ER and processed in the GA.

Types of Major Histocompatibility Complexes

● MHC Class I
→ Leads to CD8+ T-cytotoxic responses.
→ Found in all nucleated cells.
→ Antigen binds to MHC I in the ER after being broken down
by proteasome, a proteolytic enzyme.
● MHC Class II
→ More restricted than class I, being expressed mainly on
professional APCs (e.g. macrophages).
→ Exogenous antigens are usually displayed on MHC Class
II molecules, which interact with CD4+ helper T cells.
Figure 16. An Fc receptor as an example of PRR

C. Scavenger receptors
 Allow direct microbial recognition by phagocytes; detects
negatively charged molecules and scavenger cells.

D. Toll-Like Receptors (TLR)

 Membrane-bound PRRs responsible for recognizing
pathogen associated molecular patterns.
 Launches immune and inflammatory responses to defend
from foreign bodies.
 Polypeptide moving in a serpentine manner, allowing
recognition of cytokines.
 Around 30 TLR‘s identified; expression of TLR by certain
cells depend on specific ligand molecules brought by
different pathogen species

II. Intracellular Receptors

A. NOD-like Receptor (NLR)
→ Located within the cytoplasm. They act as intracellular
sensors, triggering the N FKB pathway.
B. RIG-like Receptor (RLR) Figure 18. Mechanism of MHC Class I and II Processing.
→ Detect viral RNAs and stimulate an antiviral response
through the production of interferons.
V. COMPLEMENT SYSTEM
 COMPLEMENT
→ Immunoglobulins form the core of the adaptive immune
system
→ It can generate antibodies with new antigen binding
specificities upon encountering a novel agent
→ Plasma borne arm of the innate system is the complement
→ It acts consequent to and in support of the
immunoglobulins of the adaptive immune system
→ Has circulating zymogens that remain catalytically dormant
until activated by proteolytic cleavage
→ Synthesized by the hepatocytes, macrophages, monocytes
and intestinal endothelial cells
 COMPLEMENT SYSTEM
→ Consists of more than 30 proteins soluble in the blood or
Figure 17. Membrane-bound and Intracellular PRRs; membrane-associated.
Cytokines bind to these receptors to evoke certain responses such as the → Activation of complement leads to a SEQUENTIAL
secrete of more cytokines CASCADE of ENZYMATIC REACTIONS resulting in the
formation of the potent anaphylatoxins C3a and C5a that

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elicit a plethora of physiological responses that range from Table 6. The Activation of the 3 Different Pathway
chemoattraction to apoptosis (Sarma &Ward, 2011). Pathway Activated by:
→ Used to activate membrane attack complex (MAC) Classical  Immune complexes / antigen – antibody complex
→ MAC: made up of several factors from C5 to C9 (i.e. IgG bound to an antigen)
→ Consist of 3 Pathways:  Apoptotic cells
 Gram‐negative bacteria and viruses
▪ Classical Pathway
 C‐reactive protein bound to a ligand
▪ Lecithin Pathway or the Mannose-Binding Lecitin Lecitin  Microbes with terminal mannose group
(MBL) Pathway
 Mannose are highly selective : surface of gram
▪ Alternative Pathway (+) bacteria
Alternative  Interaction between lipid-carbohydrate
complexes on the surface of microbes (i.e. gram
negative bacterial wall made up of
lipopolysaccharide)
 Viruses
 Tumor Cells

Additional information:
*In the absence of antibody and as part of the innate response, the
alternative and lectin pathways activate complement during infection.

Figure 19: Three Biochemical Pathways That Activate the Complement System

Activation leads to a sequencial cascade of enzymatic

reactions:
1. Inflammatory effect:
a. Recruitment of inflammatory cells (e.g. neutrophils)
b. Induction of histamine release from mast cells and basophils
c. Anaphylatoxin activity – degranulation of immune cells (mast
and/or basophils) induces histamine release
2. Opsonization:
a. Complement coating of pathogenic organisms or immune
complexes for identification by the phagocyte, thereby
facilitating phagocytosis
3. Cytotoxic effect:
a. the membrane attack complex (MAC) is a multimolecular
complex that breaches the integrity of the surface of the
infecting organism by insertion into its membrane and
causing osmolysis which results in cell death

Figure 21. More detailed diagram of the 3 Pathway

A. Classical Pathway
 Activator: immune complexes (antigen‐antibody complex)
such as IgM/IgG with respective antigen
 Inhibitor: C1 inhibitor (C1‐INH)

1. The antigen‐antibody complex binds and activates C1

2. C1 catalyses decomposition of C4b2 into C2b and C4b2a
(C3 convertase)
3. C1 may catalyze the decomposition of C4 into C4a and
C4b in the MBL pathway which produces C3 convertase
as well
4. C3 is activated
5. Upon activation of C3, MBL and the classical pathway
share a common convertase,the C5 convertase
(C4b2a3b)
6. Production of C5 would proceed until the membrane
attack complex (MAC) is activated
Figure 20: The Complement System
*MBL and the Classical Pathway both lead to the synthesis of the
C3 convertase that allows C3 to be activated.

BIOCHEMISTRY IMMUNOLOGY 9 of 13
 Notes: *Alternative Pathway also has a different C3 convertase (C3bBb)
 C1 – trimolecular complex (C1q, C1r, C1s) held together by and a different C5 convertase (C3bBb3b) as compared to
Ca2+ ions Classical and MBL
 C1q is the largest; in order to initiate the cascade, it must
attach to 2 Fc fragments: therefore 2 IgG molecules must  Notes:
bind to adjacent sites while only 1 IgM molecule is needed  Factor B – C3 proactivator
for complement activation; attaches to the Ig initiating  Factor D – plasma serine protease
complement activation  C3bB – magnesium-ion dependent complex
 C1s – ―C1 esterase‖  When stabilized by Properdin (P) on the microbial surface,
 C4 – covalently binds to surface of microbe/cell where Ab is the C3bBb complex becomes a C3 convertase
bound  C3bBb3b becomes C5 convertase
 C3 – most abundant complement component
 C5b – 1st component of MAC; receptor for C6 and C7
 C5-C9 – membrane attack complex
 C3 convertase – C4a2b
 C5 convertase – C4a2b3b

Figure 22. Alternative Pathway Key Components

D. Membrane Attack Complex (MAC)

Figure 22. Classical Pathway Key Components

B. Lectin Pathway
 Activator: microbes with terminal mannose group

1. The MBL is first converted to a C1‐like complex by MBL‐

associated serine proteases 1 and 2
2. C1‐like complex converted to C4, decomposing to C4a
and C4b
3. C1 from the classical pathway helps in forming C4b2 with
C2 and C4b
4. C4b2 decomposes into C2b and C3 convertase (C4b2a)
5. C3 is activated
6. Upon activation of C3, MBL and the classical pathway
share a common convertase, the C5 convertase
(C4b2a3b)
7. Production of C5 would proceed until the membrane
attack complex (MAC) is activated

 Notes:
 MBL, which is akin to C1q, binds to the cell wall of bacteria Figure 23. Membrane Attack Complex
with mannose-containing surfaces (mannans) which then
associates with 2 serine proteases: MASP-1 (akin to C1r) 1. C3b from C3 splits C5
and MASP-2 (akin to C1s) which forms a trimolecular a. *C3b performs phagocytosis while C3a along with
complex C5a releases histamines and bind to mast cells
during inflammation
C. Alternative Pathway 2. C5b fragment binds C6-C9 and together form a cylinder‐
shaped membrane attack complex (inserts into the
 Activator: lipid‐carbohydrate complexes on the surface of plasma membrane)
microbes and also by viruses, tumor cells 3. MAC performs cytolysis or the bursting of the microbe
 Binding of the complement factors B, D, and Properdin due to the inflow of ECF fluid through channels formed
with the surface of the microbes immediately activates C3 by MAC

*Has a shorter pathway compared to lectin and the classical

pathway because it only takes the binding of the complement
factors with the antigen surface for C3 to be formed

1. Activation of C3
2. C3 decomposes into C3a and C3b
3. C3b binds to the Bb fragment of Factor B leading to the
synthesis of another C3 convertase, C3bBb
4. C3bBb cleaves another C3 into C3a an C3b
5. Formation of another C5 convertase C3bBb3b
6. Production of C5 would proceed until the membrane Figure 24. Membrane Attack Complex Key Components
attack complex (MAC)is activated
BIOCHEMISTRY IMMUNOLOGY 10 of 13
4.The antigen presenting cell activates the adaptive immune
system by expressing:
A. HLAI C. MHCI
B. HLAII D. MHCII
5.The antibody that is active during the first exposure in this
case is:
A. IgA B. IgD C. IgG D. IgM

CASE: SD is a 6-week-old baby girl presenting with acute onset of

swelling of the ears and extremities, an erythematous rash and
‗crabbiness‘ that could not be soothed. An hour prior to the onset
of the rash SD had been given 40 ml of cow‘s milk baby formula
which she vomited immediately. Within the hour she developed
swelling and redness of her ears, hands and feet and urticaria of
her face, limbs and body.

6. Which cell of the innate immune system is expected to

migrate to the affected area?
A. basophil C. mast cell
B. macrophage D. neutrophil
Figure 25. The Multiple Activities of the Complement System 7. The effect of the cell is characterized as:
A. exhibiting memory C. discriminating
B. highly-specific D. short-lived
IV. IMMUNE DISEASES 8. Which molecule is secreted by the cell of the immune
system to combat the nonself?
 OVERREACTIVITY A. cytokine C. histamine
 Attacks and damages one self is called production of B. interferon D. thromboxane
autoantibodies 9. The antibody that is active during the first exposure in this
 Example: Rheumatoid arthritis, SLE case is:
A. A B. E C. G D. M
 DEFICIENCY 10. The precursor cell for this antibody is:
A. basophil C. mast cell
 Decrease the ability to fight invaders causing vulnerability to
B. eosinophil D. plasma cell
illness
 Example: Di George, Wiskot Aldrich, AID
CASE: A 20-year-old student was seen in the university infirmary
for fever, rhinorrhea, congestion, and sneezing. His pharynx,
A. CLINICAL EXAMPLES tonsils and buccal mucosa were red and inflamed. He was
diagnosed as having acute viral tonsillitis and advised fluid intake
 An acute onset of sepsis was noted in the newborn, to measure and antipyretic for the fever.
the acute response we have acute phase reactants as a 11. Which cell of the innate immune system is expected to
measure of the inflammatory activity. migrate to the tonsils?
 Examples are: ESR, CRP A. basophil C. mast cell
 A patient was noted to have high grade fever for the last 6 days B. macrophage D. neutrophil
and it was noted that they requested for Ig G Dengue and IgM – 12. The effect of the cell is characterized as:
IgM means current infection and IgG means past infection why A. exhibiting memory C. discriminating
is this so B. highly-specific D. short-lived
13. Which receptor in the innate immune cell will recognize
the pathogen?
REVIEW QUESTIONS A. Mannose C. RIG-like
2020 QUIZES: B. NOD- like D. Toll-like
CASE: A 5-year-old boy was brought by his mother to a 14. The antigen presenting cell activates the adaptive immune
pediatrician with a 36-hour history of acute malaise, shivering and system by expressing:
vague pains in his legs. For 12 hours he complained of a dry, sore A. HLAI C. MHCI
throat and had B. HLAII D. MHCII
vomited twice. He was febrile (T= 40.2ÅãC) with a fast heart rate 15. The antibody that is active during the first exposure in this
of 140 beats/min and tender, bilateral, cervical lymphadenopathy. case is:
His pharynx, tonsils and buccal mucosa were red and inflamed A. A C. G
and his tonsils were studded with white areas of exudate. He was B. D D. M
diagnosed as having acute bacterial tonsillitis (Gram positive).
2020 LE:
1.Which cell of the innate immune system is expected to CASE: A 12 y/o student sustained an infected wound in his R leg.
migrate to the tonsils? Staining showed Gram (+) bacteria
A. basophil C. mast cell
B. macrophage D. neutrophil 1. Which protein: function pair is observed during recruitment
2.The effect of the cell is characterized as: of an innate immune cell?
A. exhibiting memory C. discriminating a. Cytokine: Phagocytosis
B. highly-specific D. short-lived b. Integrin: Adhesion
3.Which receptor in the innate immune cell will recognize the c. L-selectin: NETosis
pathogen? d. P-selectin: Rolling
A. Mannose C. RIG-like
B. NOD- like D. Toll-like

BIOCHEMISTRY IMMUNOLOGY 11 of 13
2. Re-exposure to the same antigen activates an effector that c. Genetic recombination of receptor genes
functions to simulate d. Expression of MHCs
a. Production of MALT
b. Opsonization by macrophages 8. Which effector cell: cytokine pair is activated in the case?
c. Maturation of B cells a. Basophil: IL33
d. Activation of lectin complement b. Eosinophil: IL4
c. Macrophage: IL2
3. Which cell-mediated effector: cytokine pair inactivates the d. Mast cel: IL13
pathogen?
a. TH17: IL-17 b. TH2: IL-4 c. TH1: IFN- d. Treg: IFN- 9. Which immunoglobulin exhibits memory of the allergen?
a. A b. E c. G d. M
4. Which sequence of complement proteins is activated?
a. C1 – like - C4-C3-C5
b. C1 – C4+C2-C3-C5 Ans: 2020 QUIZES: 1d, 2d, 3d, 4d, 5d, 6c, 7d, 8c, 9b, 10d, 11b,
c. C3 – B – C3-C5 12d, 13c, 14c, 15d
2020Les: 1b, 2b, 3d, 4b, 5a, 6d, 7c, 8d, 9b
5. The reaction catalyzed by C3 convertase leads to:
a. Amplification of C5 convertase activity REFERENCES
b. Binding of patter recognition receptors ● Dr. Agnes A. Alba‘s lecture and ppt
c. Completion of membrane attack complex ● Rodwell, V.W., Bender, D.A., Botham, K.M., Kennelly, P.J., &
d. Formation of a membrane pore by C3b Weil, P.A. (2015). Harper‘s illustrated biochemistry (30th ed.).
McGraw Hill.
6. Which complement factor triggers opsonization of infected ● Lippincott‘s Illustrated Review of Immunolgy (2nd Ed.)
cells? ● 2018, 2019, 2020B Trans
a. C4a b. C4b c. C3a d. C3b

CASE: A patient with known history of hypersensitivity to peanuts

complained of swelling with itchiness (pruritis) of her upper arms
and redness of her face and upper body.

7.Diversity of adaptive immune response is accounted for by:

a. Ability to recognize microbe-specific patterns
b. Amount of pathogen associated molecular patterns

APPENDIX

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BIOCHEMISTRY IMMUNOLOGY 13 of 13