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1 IMMUNOLOGY MICROBIOLOGY
Dr. Fontanilla| June 18, 2013 1st 2013-2014

OUTLINE II. INNATE IMMUNITY

I. IMMUNE RESPONSE IV. ADAPTIVE IMMUNITY
A. Innate Immunity A. Humoral Immunity A. Physiologic Barriers
B. Adaptive Immunity B. Antigen Recognition Molecules  Skin
II. INNATE IMMUNITY C. Antigen processing and presentation
A. Physiologic Barriers D. B cells and antibodies o intact skin – only few microorganisms can penetrate, but they
III. MECHANISMS OF INNATE E. Steps in anti body formation can enter sweat or sebaceous glands and hair follicles
IMMUNITY F. Immunoglobulin Classes o sweat glands and sebaceous glands secretions have an acidic
A. Microbial Sensors G. Primary Response pH which acts as an antimicrobial
B. Phagocytosis H. Secondary response
C. Natural Killer cells I. Protective function of Antibodies o lysozyme is present on the skin, in tears, and in respiratory and
D. Complement J. Strategies to induce antibody cervical secretions – dissolves bacterial cell walls
E. Mediators of inflammation: Production o psoriasin – antimicrobial produced by the skin
Cytokines
 Mucous Membranes
OBJECTIVES o In respiratory tract
At the end of the lecture, the student should be able to:  mucus traps bacteria and is driven outwards by ciliated cells
1. Describe the basic principles of immunology, particularly as they relate to  some bacteria have pili which acts as an attachment on
response to infection.
2. Define the following terms:
surfaces (ex. Neisseria gonorrhea)
a. Immune response  surface epithelial cells with IgA – prevent attachment of
b. Innate immunity piliated bacteria
c. Adaptive immunity  phagocytes take up microorganisms and transport them to
d. Antigen lymph nodes
e. Antibody
f. Major histocompatibility complex
 there are also other protective mechanisms in the
g. Complement respiratory system like the cough reflex
h. Cytokines o In gastrointestinal tract
i. Hypersensitivity  saliva contains hydrolytic enzymes
j. Immunodeficiency  stomach acidity kills ingested bacteria
3. Differentiate humoral and cellular immunity
4. Describe the funcions of antibodies
 small intestines contain proteolytic enzymes
5. Enumerate and describe the different hypersensitivity reactions  the presence of normal microbial flora prevent the
6. Describe the more common deficiencies of the immune response establishment of pathogenic microorganisms
o In genital tract (vagina)
References:
 acidic pH is maintained by normal lactobacilli – to inhibit
Jawetz, Melnick & Adelberg’s MEDICAL MICROBIOLOGY, 26th Edition, 2013,
Chapter 8, pp 123-148. establishment of yeasts, anaerobes and gram-negative bacilli

Legend: Italicized – quoted from the lecturer; bold – emphasis, or from references
III. MECHANISMS OF INNATE IMMUNITY
A. Microbial Sensors
I. IMMUNE RESPONSE
 A response generated against a potential pathogen  Possessed by macrophages and other phagocytic cells
o Toll-like receptors (TLRs)
A. Innate Immunity o NOD-like receptors (nucleotide-binding oligomerization
domain-like receptors) (NLRs)
 First line of defense
o RIG-1 like helicases and MDA-5
 Non-specific to the pathogen
 Toll-like receptors (TLRs)
 Rapidly mobilized at the site of infection o first line of defense against pathogens
 Lacks immunologic memory o play a role in initiating the innate immune response
o belong to a family of evolutionary conserved (PRRs) Pattern
B. Adaptive immunity Recognition Receptors that recognize (PAMPs) Pathogen
 Second line of defense Associated Molecular Patterns.
 Specific to the invading pathogen *bacteria have certain morphology & have specific molecular
 Can confer protective immunity to reinfection with that pathogen patterns.
o recognition of microbial patterns lead to a signal transduction
Advantage Disadvantage cascade that generates a rapid inflammatory response.
 TLR2 – recognizes various ligands (lipoteichoic acid)
Prompt availability of
Short life span of these expressed by gram-positive bacteria
Innate Immunity large amounts of
antibodies  TLR3 – engages dsRNA in viral replication
antibodies
Slow onset of protection,  TLR1 & TLR6 – recognizes multiple diacyl peptides
Adaptive Immunity Long-term protection repeated contact with (mycoplasma)
antigen is necessary  TLR4 – gram negative lipopolysaccharides
Advantage and disadvantage of the types of immunity.  TLR5 – bacterial flagellin
 TLR7 & TLR8 – interact with ssRNA in viral replication
 TLR9 – binds bacterial DNA

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Group 4 Ambatali, Amil, Andan, Ang, Angeles, Ani, Anonas, Anupol
 Microbiology 1.1
 NOD -like receptors (NLRs) D. Complement
o located in the cytoplasm
 composed of 30 proteins found in the serum or on the membrane
o intracellular sensors for microbial products
of selected cells
o activate nuclear factor kappa-light chain : enhancer of
 complement proteins target pathogens for destruction by lysis or
activated B cells (NF-κβ) pathway
for engulfment by phagocytes
o drive inflammatory responses similar to the TLRs
 3 pathways : classical, alternative and lectin
 RIG-1 like helicases and MDA-5
 The end product of the three major pathways is the production of
o cytoplasmic sensors of viral ssRNA
Membrane Attack Complex (MAC)
o trigger type 1 IFN production
 results in lysis of offending invader
o IFNs are highly effective inhibitors of viral replication

B. Phagocytosis E. Mediators of inflammation: Cytokines

 Activated macrophages release:
 Phagocytes in the immune system
o cytokines – IL-1 and TNF-α
o monocytes and macrophages
o prostaglandins
o granulocytes – PMN, eosinophils and basophils
o leukotrienes
o dendritic cells
 Effect of mediators:
 Functions of phagocytic cells
o elicit changes in blood vessels –inflammation
o chemotaxis
o induce changes in expression of various adhesion molecules
o migration
(selectins and integrins) on endothelial cells and leukocytes –
o ingestion
promote movement across vessel wall to migrate out toward
o microbial killing
the irritant
 Hallmark of phagocytic process
 Chemokines
o rapid
o stimulate leukocyte movement
o nonspecific
o recruit monocytes and neutrophils from blood into sites of
o short duration
infection
 Antimicrobial mechanisms used by phagocytes
o synthesized by macrophages and endothelial cells
o Acidification occurs within the phagosome
 Interferons (IFNs)
 pH 3.5 – 4.0 : bacteriostatic or bactericidal
o role in defense against viral infection and against other
o Toxic oxygen derived products are generated
intracellular organisms
 superoxide O2
o 3 groups:
 hydrogen peroxide H2O2
 Type I – IFN-α and IFN-β
 singlet O2
 Type II – IFN- γ
o Toxic nitrogen oxides are produced and nitric oxide is formed
 Type III – IFN-λ
o Antimicrobial peptides participate in killing
 macrophage – cathelicidin, macrophage derived peptides
 PMN – α-defensins, β-defensins, cathelicidin, lactoferrin IV. ADAPTIVE IMMUNITY
A. Humoral Immunity
C. Natural Killer Cells (NK cells)  Antigen – a substance that can provoke the production of
 large, granular lymphocytes antibodies
 morphologically related to T-cells  Features that determine immunogenicity:
 10 – 15% of leukocytes in blood o Foreignness – for immunogenicity to occur, molecules must be
 provide protection against viruses and other intracellular recognized as “non-self”
pathogens o Size – most immunogens are usually large proteins
o Chemical and structural complexity – heteropolymers
 can recognize virus infected cells and tumor cells
containing 2-3 different amino acids are more immunogenic
 Have two types of surface receptors
o Genetic constitution of the host – 2 strains of the same species
o lectin-like NK-cell receptors – bind proteins
of animal may respond differently to the same antigen
o killer immunoglobulin-like receptor (KIRs) – recognize major
o Dosage, route and timing of antigen administration – the
histocompatibility complex (MHC) class I human leukocyte
immune response can be optimized by carefully defining the
antigen B (HLA-B) or HLA-C
dosage, route of administration and timing of administration
 Can lyse target cells – malignant transformation
 Some pathogens interfere with antigen processing
 Kill virus-infected cells with altered levels of MHC class molecules
o Ex. HIV – becomes latent due to Tat proteins; individual can
 play a role in antibody-dependent cellular cytotoxicity (ADCC)
harbor virus for up to 10 years
 contains large amounts of granzyme and perforin – mediate
cytotoxic action of NK cells B. Antigen Recognition Molecules
 with the IFN system, these are integral parts of innate immunity
that communicate with each other  Major Histocompatibility Complex
 primary source of IFN- Υ a potent anti-viral and immuno o MHC molecules bind peptide antigens and bind them to T-cells
regulating cytokine o cluster of genes located on chromosome 6
 lytic activity of NK cells is enhanced by type 1 IFNs – IFN-α and o genes that encode the class I, class II, and class III MHC
IFN-β – induced by invading virus proteins
 acts through MHC class I molecules which are upregulated by  Class I molecules
IFNs o found on virtually all nucleated cells in the body
o recognized by CD8 cytotoxic T-lymphocytes
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 Class II molecules
o found on macrophages, dendritic cells, B cells and other APCs
o recognized by CD4 helper T cells
 Class III molecules – encodes complement proteins
C. Antigen Processing And Presentation
 MHC Class I and endogenous antigens:
o MHC Class I molecules are synthesized in the endoplasmic
reticulum of the infected cell (viral infection)
o Viral proteins in the cytoplasm that have infected the cell are
broken down by proteolytic complex known as proteasome
o Cytosolic peptides gain access to MHC Class I molecules in the
ER via peptide transporter system (TAPs)
o MHC Class I – peptide antigen complex is formed and
transported to the cell surface for display and recognition by
CD8 cytotoxic T-cells
 MHC Class II and exogenous antigens:
o Class II MHC molecules (membrane glycoprotein) are
synthesized in the RER and proceed out to the Golgi apparatus
and transported out to the cytoplasm via exocytic vesicles
o Proteins from exogenous antigens (bacteria) are taken up by
the cell (APCs like dendritic cells and macrophages) via
endocytic vesicles
o Proteins are exposed to cellular proteases in intracellular
vesicles producing peptides
o Endosomal vesicles with peptides fuse with exocytic vesicles
containing MHC Class II molecules
o MHC Class II – peptide antigen complex is transported to the
cell surface for display and recognition by a CD4 T-helper cell
 Interference with antigen processing pathways:
o HIV virus produces a Tat protein that inhibits expression of
Class I MHC molecules
o Herpes virus proteins bind to the TAPs, preventing transport of
viral peptides into the ER where Class I molecules are being
synthesized (virus infected cells are not recognized by cytotoxic
lymphocytes)
o Superantigens like staphylococcal enterotoxin, toxic shock
syndrome toxin, group A streptococcal pyrogenic exotoxin
o Superantigens bind to “outside” portion of the MHC protein-T-
cell receptor complex
o cause T-cells to be stimulated and to release large amounts of
cytokines (IL-1, TNF) – pathogenesis of disease
D. B Cells And Antibodies
 Humoral immunity is mediated by antibodies, which are
produced by B-cells
o large pool of different B lymphocytes that have a life span of
days or weeks
o B cells are found in the bone marrow, lymph nodes and gut-
associated lymphoid tissues (appendix, Peyer’s patches,
tonsils)
 B-Cell Receptor for Antigen
o display a single immunoglobulin molecule on the surface which
serve as receptors (B-cell receptor) BCR for a specific antigen
o each B-cell can respond to only one antigen or closely related
groups of antigens
o all immature B-cells carry IgM immunoglobulins on their
surface and most carry IgD
o an antigen interacts with the B lymphocytes that shows the
best fit and binds to the BCR
o there is an antigen-binding site on a B-cell to fit almost any
antigenic determinant
Microbiology 1.1
o the B cell is stimulated to divide and form a clone (clonal
selection)
o this selected B-cell differentiate to become plasma cells and
secrete antibody
o each person can make approximately 1,000,000,000,000
different antibody molecules
E. Steps in Antibody Formation
1. Binding of antigen to the surface immunoglobulin via BCR
2. The BCR with the bound antigen is internalized by the B cell
o antigen is degraded to yield peptides that are then returned to
the cell surface bound to MHC class II molecules
3. MHC class II-peptide complex on B cells is recognized by antigen-
specific helper CD4 T-cells
o these T-cells have already interacted with APC dendritic cells
and have differentiated in response to the same antigen
4. Chemokines, such as CXCL13 and its receptor CXCR5, play an
important role in this migration process
5. CD40 ligand on T-cells binds to CD40 on B cells
o T-cell produces cytokines such as IL-4, IL-5 and IL-6 which
induce B-cell proliferation
6. Finally, the activated B cells migrate into follicles and proliferate
to form germinal centers
o germinal center B cells that survive this process now
differentiate into either antibody-producing plasma cells or
memory B cells.
o some bacterial antigens can directly stimulate antibody
production
 thymus independent antigens
 induce B-cell responses with limited class switching
 do not induce memory B cells
 eg. bacterial polysaccharides and lipopolysaccharides
F. Immunoglobulin Classes
 IgG
o made up of four subclasses – based on amino acid sequence
differences in the H chain constant region
 IgG1 - 65 % of total IgG
 IgG2 - directed against polysaccharide antigens
- against encapsulated bacteria
 IgG3 - activator of complement due to rigid hinge region
 IgG4 - does not activate complement due to compact
structure
o predominant antibody in secondary immune responses
o important defense against encapsulated bacteria
o readily crosses the placenta; most abundant Ig among
newborns
o mediates opsonization of antigen through antigen-antibody
complexes to Fc receptors on macrophages and other cells
 IgM
o the main immunoglobulin produced early in the primary
immune response
o present on the surface of all uncommitted B cells
o composed of five H2L2 units and one molecule of J chain =
pentamer with 10 identical antigen-binding sites
o most efficient immunoglobulin in agglutination, complement
fixation and other Ag-Ab reactions
o has the highest binding capacity and cross linking of all Igs
o evaluation of the presence of serum IgM is useful in the
diagnosis of certain infectious diseases
o does not cross the placenta; presence of IgM in the newborn is
evidence of intrauterine infection
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 IgA
o main immunoglobulin responsible for mucosal immunity
o found in secretions such as milk, saliva and tears; respiratory,
intestinal and genital tract secretions
o each secretory IgA molecule consists of two H2L2 and one
molecule of J chain and secretory component
o a receptor binds IgA dimers and facilitates transport across
mucosal epithelial cells
o two subclasses IgA1 and IgA2
o some bacteria (Neisseria spp) can destroy IgA1 by producing a
protease
o Protects mucous membranes from attachment by bacteria
and viruses
 IgE
o Fc region of IgE binds to its high affinity receptor on the surface
of mast cells, basophils and eosinophils
o the bound IgE acts as a receptor for the antigen that stimulates
its production
o resulting Ag-Ab complex triggers allergic responses of the
immediate (anaphylactic) type
 IgD
o acts an antigen receptor when present on the surface on
certain B lymphocytes
o trace amounts in serum
o function is unclear
G. Primary Response
 an individual encounters an antigen for the first time
 antibody produced against that antigen will be detectable in the
serum within days or weeks
 serum antibody concentration continues to rise for several weeks
and then declines
 first antibodies formed are IgM, followed by IgG, IgA or both
 IgM levels then decline sooner than IgG levels
H. Secondary Response
 second encounter with the same antigen or a closely related
“cross-reacting” antigen months or years after the primary
response
 produce qualitatively similar IgM
 much more IgG is produced; persists longer than the primary
response
 antibody produced tends to bind antigen more firmly, have
higher affinity and dissociate less easily
Microbiology 1.1
I. Protective Function of Antibodies
 Enhanced phagocytosis
o antibodies coat organisms (opsonizing); more readily ingested
by phagocytes
o antibody-mediated immunity against bacteria is most effective
in bacterial infections in which virulence is related to
polysaccharide capsules
 Streptococcus pneumoniae,
 Haemophilus spp.
 Neisseria spp.
 Virus Neutralization
o viruses need to invade the cell; obligate intracellular pathogens
o antibodies directed against specific viral proteins bind to the
virus and block the ability of the virus particle to attach to cell
membrane receptor
 Neutralization of Toxins
o Corynebacterium diphtheriae – diphtheria toxin
o Clostridium tetani – tetanus toxin
o Clostridium botulinum – botulism
 Complement-Mediated Lysis
o attachment of antibodies to viral proteins on virus infected
cells or to a microbial cell can activate complement system
leading to cell lysis
 Antibody-Dependent Cell Cytotoxicity (ADCC)
o attachment of antibodies to viral proteins on virus infected
cells --- antibody-coated cells interact with killer cells = lysis
J. Strategies to Induce Antibody Production
 Active Immunity
o induced after contact with foreign antigens (microorganisms
and their products)
o clinical infection, subclinical infection (does not develop full
blown symptoms), immunization with live or killed infectious
agents or their antigens, exposure to microbial products
(toxins, toxoids) or transplantation of foreign cells
o the host actively produces antibodies
o protection is delayed until antibody production reaches an
effective level
 binding of antibodies to antigens leads to the formation of
circulating immune complexes – kidneys, heart valves –
organ dysfunction
 Passive Immunity
o generated by administration of preformed antibodies
o prompt availability of large amounts of antibody
o eg. After needle stick injury vs Hepatitis B virus
 hypersensitivity reaction if preformed antibody is produced
from other species – horse, sheep
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