43099 BASES BIOQUIMICAS DE LA INMUNOLOGIA

FUNDAMENTO Y APLICACIONES

TEMA 2b
Evolución del Sistema Inmunitario
 Evolution: The Tree-of-life

So you've been where I've just

come
LUCA:
The Last Universal Common Ancestor
The Kinks
Strangers
Evolution of the Immune system: A very simplistic view
Evolution of the Immune system: Still, a simplistic view
 Evolution of the Immune system: A less simplistic view

500-600 My ago: Appearance of Adaptive Immunity or the Big Bang of Immunity

Adaptive Immunity
Simple Immunity

Innate Immunity
Invertebrates represent 95 % of all existing
species in the Animal Kingdom. After all, they
seem to have a successful immunity!
Evolution of the Immune system: A much less simplistic view

• Immunity is ubiquitous in nature and the

frontier between adaptive and innate immunity
is not as clear cut as claimed for decades.

• Nonvertebrate immunity no longer appears to

be unspecific, and many forms of immune
specificity seem to exist in animals and plants.

• Immune memory, a characteristic ascribed to

adaptive immunity and therefore to higher
vertebrates, exist to some extent in
invertebrates.

• The adaptive immune system is not self-

sufficient. An antigen recognized only by the
adaptive but not by the innate immune system
will not trigger an immune response.
 The evolutionary advantage of Adaptive Immunity

Innate Immunity Adaptive Immunity

Innate immunity depends on Higher vertebrates have evolved
phagocytic cells, antimicrobial an adaptive immune system
molecules and germline–encoded mediated primarily by
receptors that have evolved to lymphocytes. By means of
recognize highly conserved rearrangeable immunoglobulin V,
pathogen associated molecular D, and J gene segments, a
patterns (PAMPs). These receptors lymphocyte receptor repertoire of
have therefore been termed sufficient diversity may recognize
pattern recognition receptors the antigenic component of any
(PRRs) and activate specific signal potential pathogen or toxin.
transduction pathways, and
initiate the events that will lead
eventually to antigen presentation
to lymphocytes

Anticipation
Regulation
Potentiation
Diversification
Focusing
Memory
 Landmarks of Immune evolution

Ig-based clonal, combinatorial anticipatory immunity

LRR-based, anticipatory immunity

Immunoglobulin Superfamily
Leucine-Rich Repeat proteins
Polymorphic proteins for self recognition

Large repertoir of diverse PAMP receptors

Natural cytotoxicity

Toll-Like Receptors

Immunocytes; Self recognition; Cytokines; Antimicrobial proteins

The Evolution of self versus not-self
discrimination
 The Evolution of self versus not-self discrimination

Self versus not-self discrimination is one of the most ancient features

of immune system
 The Evolution of self versus not-self discrimination

Adhesion molecules may

likely be the precursors of
self- and not-self
recognizing receptors at the
cell surface
The Evolution of antimicrobial molecules
and complement system
 Primitive Innate Immunity: Antimicrobial Peptides

Antimicrobial peptides include diverse structural types:

• α-Helices
• β-Sheet and small proteins
• Peptides with thio-ether rings
• Peptides with overrepresentation of one or two amino acids
• Lipopeptides
• Macrocyclic cystine knot peptides

Gram-positive bacteria Gram-negative bacteria Fungi

Defensins Cecropins Drosomycin
Drosocin metchnikowin
Attacins
Diptericin
MPAC
 The mammalian complement system

Time of Appearance
The final addition of the lytic pathway:
Evolution of the Complement System
C4 and C5 are created from gene duplications of C3. A
molecule with a perforin domain appeared and evolved
into C6, C7 and C8 to lyse enucleated cells. C9 was
added to form a pore capable of lysing nucleated cells.

The appearance of the Classical pathway: The amplification power of the protease cascade:
Gene duplications of MBP and the MASPs created C1q, Genome-wide duplication creates C4 and C2 to improve
C1r, and C1s. C1q evolves to bind to immunoglobulins lectin pathway.

The appearance of the Lectin pathway:

MASPs and mannose-binding protein (MBP).

The appearance of the Alternative pathway:

Increase of C3 spontaneous reactivity, tolerance to low
levels of serum C3b and addition of Factor D

The key step in evolving the complement pathway:

C3 affinity for a serine protease attach C3b and addition
of Factor B allow to deposit very rapidly large amounts
of C3b onto the surface of pathogenic molecules.

The appearance of opsonins:

Ancestor to C3 and opsonins. As a consequence,
inhibitors of C3 appear on surface of host cells

Ancestor to protease inhibitor α2-Macroglobulin (α2-M)

The Evolution of immune cells
 The Evolution of immune cells
• Present in almost all metazoans
• Absent in Caenorhabditis elegans
• Present in all vertebrates
• Arising in the common ancestor
Phagocytic cells
of vertebrates

• Relatively short life-span

• Proliferation in generative hemopoetic tissues
Lymphoid
• Activated to effector cells
cells
• Express a large repertoir of surface receptors

• Derived from committed hemopoetic precursors

• Potential for self-renewal and clonal expansion
• Undergo unique VDJ rearrangements of antigen
receptor genes
• Elimination of cells bearing autorreactive receptors
• Clonal expansion and differentiation into effector
cells or memory cells
 Primitive Innate Immunity: Phagocytic cells

Amoebocytes
Coelomocytes Pathogen recognition and Phagocytosis
Haemocytes
Hyalinocytes
Secretion of antimicrobial peptides

Encapsulation/Melanization

Blood clotting

Natural killer features and responses

Primitive Innate Immunity: Phagocytic cells
 Evolution from primitive phagocytes to modern lymphocytes

Coelomocytes of Lumbricus
spontaneously kill in coculture
the erythromyeloid human
tumor cell line K562.
Cytotoxic small coelomocytes Coelomocytes of starfish
are positive for CD11a, express IL-1 receptor and
CD45RA, CD45R0, CD49b, produce IL-1 which stimulates
CD54, CD90, CD24, Beta-2- production of nitric oxide by
Microglobulin and TNF-α. coelomocytes

Phagocyte large coelomocytes Starfish IL1 stimulates murine

are negative for CD11a, thymocyte and fibroblast
CD45RA, CD45R0, CD49b, proliferation and protein
CD54, CD90, CD24, Beta-2- synthesis.
Microglobulin and TNF-α.

Starfish IL1 activity is inhibited

Graft rejection in Lumbricus is by an antibody to human IL1.
mediated by the neutrophilic Starfish coelomocytes also
granulocytic coelomocytes produce IL6 .
 Lymphoid cells in Agnathans

• Small round cells with large nucleus/cytoplasm ratio

• Generated in hemopoetic tissues:
• Larval intestine (typhlosole)
• Protovertebral arch in adults
• Thymus? (pharyngeal gutters in gill region)
• Transform into large lymphoblastoid cells upon antigen or mitogen stimulation
• Express homologs of many genes related to Gnatosthomes :
• Lymphocyte differentiation, proliferation, migration and intracellular signaling
• Antigen processing and presentation
• Do not express genes related to rearrangeable Immunoglobulin or MHC genes
• Express Variable Lymphocyte Receptors (VLRs) based on Leucine-Rich Repeats (LRR)
• Each lymphocyte expresses a VLR of unique monoallelic sequence, recombined to
create functional VLR genes
• Antigen-specific VLRs are released into the plasma upon immunization
The Evolution of pathogen-recognition
Receptors and dependent-pathways
 The Evolution of pathogen recognizing receptors and pathways

Inflammatory Cytokines
Chemokines ROS and NOS

Pattern-recognition Antimicrobial peptides

Receptors Recognition of
microbial non-self CD80 and CD86
upregulation in APCs
PAMPs UNIVERSAL STRATEGY OF INNATE IMMUNITY

Inhibitory Recognition of Connexion to Adaptive Immunity

receptors missing-self

Recognition of Triggering of
own surface proteins inhibitory signals

A MORE RECENTLY EVOLVED MECHANISM

TLR signaling and its functions in the invertebrates
 The Evolution of pathogen recognizing receptors and pathways
Toll-like receptors (TLR) are a
major class of pattern
recognition receptor, that
exists in all coelomates
including humans.
TLR signaling occurs via a TIR
domain-containing adaptor
like MyD88, TRIF, TRAM, and
results in stimulation of NF-
κB pathway
The Evolution of pathogen recognizing receptors and pathways
TLR signaling and its functions in the invertebrates
 TLR signaling and its functions in mammals

CYTOKINE SECRETION INTRACELLULAR BIOCHEMICAL MECHANISMS

APOPTOSIS PATHOGEN CELL DEATH

The Evolution of cytokines and
their signalling
 The evolution of cytokines and their signalling

Albeit with different functions, cytokine-like molecules and intracellular signaling

pathways are present since early stages of invertebrate life forms and expanded in
vertebrates
The evolution of cytokines and their signalling
The Evolution of immunological memory
 The Evolution of immunological memory

There are multiple experimental descriptions of responses suggesting of

immunological memory in invertebrates
 The Evolution of immunological memory

The features of immunological

memory in invertebrates, although
improving secondary responses, seem
short-timed, dependent on phagocytic
cells and unrelated to the
mechanisms of lymphocyte memory in
higher vertebrates.
The Evolution of antibody-based
immune systems
 The Evolution of antibody-based immune systems
Two convergent systems for anticipatory immunity based in a large repertoir of
cell bound receptors and humoral effectors evolved in Agnathans and
Gnathostomes, 500 My ago, based on two successful families of immune proteins
Agnathans Gnathostomes

Leucine-Rich Repeat Proteins (LRR) Immunoglobulins (Ig)

 The Evolution of antibody-based immune systems

Agnathans assemble diverse lymphocyte antigen receptor genes through genomic rearrangement of
leucine-rich repeat (LRR)–encoding modules. Each lymphocyte expresses a VLR of unique sequence
in monoallelic fashion. Cell surface receptors are designated variable lymphocyte receptors (VLRs).
Antigen-specific VLRs are released into the plasma upon immunization

Gnathostomes rearrange their V(D)J gene segments to assemble complete genes for the antigen
receptors expressed by T and B lymphocytes. Antigen-mediated triggering of T and B cells initiates
specific cell-mediated and humoral immune responses
The Evolution of antibody-based immune systems

In Gnathostomes the genes

encoding immunoglobulins (and
TCRs) are composed of multiple
gene segments, which are cut-and-
pasted together to form a functional
Ig gene through a process of V(D)J
recombination .

The mechanism involves two

proteins, called RAG-1 and RAG-2.
(Recombination Activating Gene),
which recognize recombination
signal sequences (RSS).

The RAG proteins bind to the RSSs of

two gene segments, then bring them
in close proximity to each other and
cleave the DNA precisely at the
junction between the gene segment
and the RSS.
The Evolution of antibody-based immune systems
Natural selection and the immune system:
A fast host vs pathogen race…
or may be not?

Those that I fight I do not hate,

Those that I guard I do not love;
W.B. Yeats
An Irish aviator foresees his death
 Does acquired immunity come from Bacteria?

• A transposon containing the RAG genes and flanked

by RSSs integrates itself into a primordial antigen-
receptor gene, splitting it into gene segments V and J.
• The locus is transcriptionally inactive in most cell
types, and prevents the expression of the RAG genes
and removal of the integrated transposon.
• The locus becomes transcriptionally active in a
lymphocyte-like cell, the RAG genes express and
remove the transposon, reuniting the two gene
segments.
 Does acquired immunity come from Bacteria?

• The imprecise joining process generates a level of

receptor diversity.
• The gene duplicates, creating multiple pairs of V and J
segments further increasing diversity.
• The entire locus duplicates, and the two lineages form
the heavy and light chains.
• Further gene duplications create additional V, and J
gene segments, and an incorrect recombination event
creates the D segment.
• Lastly, a whole genome duplication event creates the
genes which would later become the T cell receptors.
 From the Tree of Life to the Ring of Life

The possible incorporation of a

bacterial transposon into higher Strangers on this road we are on
vertebrate genome, may be another We are not two we are one
proof of the importance of horizontal The Kinks
gene transfer in evolution Strangers