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Dermatology Basics for
Small Animal Clinicians :
The Skin Immune System
Drh. Herisman Hernadi,
Resident of AiCVD
Veterinary Dermatology Practitioner
 References

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 Agenda
• Introduction
• Innate Immune Respons
• Adaptive Immune Respons
• Components of the Skin Immune System

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Introduction
 The Skin Immune System (SIS)
• An active component of the overall immune system

• Helps defend the host against many environmental insults :

microbial and parasite
• Animal skin is bombarded by parasites, bacteria, fungi, viruses,
and allergens trigger undesirable immune responses.
• Major site for autoimmunity  autoantigens in the epidermis
or BMZ are attacked by autoantibodies or autoreactive T cells

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 Introduction
The immune system is divided into :
1. Innate immunity (non-specific)
• No prior recognition of antigen
• First line of defence
• No reinforcement upon repeated exposure to antigen
2. Adaptive immunity (specific)
• Antigen recognition and a directed response

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 Innate immunity
Includes physical barriers such as the skin or gut
1. Protective substances such as mucus, enzymes, or peptide
antibiotics
2. Soluble proteins such as complement

3. Phagocytic cells such as macrophages, neutrophils, and

eosinophils

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 Innate immunity
Many cells located near body surfaces possess pattern-recognition
receptors termed :
• Toll-like receptors (TLRs) that recognize molecular patterns
(PAMPs [pathogen-associated molecular patterns]) common to
invading organisms.
• PAMPs include peptidoglycans, lipopolysaccharides (LPS),
glycolipids, and mannan-rich carbohydrates.
• TLRs are found on macrophages, eosinophils, mast cells,
dendritic cells, and epithelial cells.

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 Adaptive immunity
• Highly specific and shows phenomenal memory

• Why?....Because :
1. Many diseases require an adaptive immune response to resolve
2. Some diseases are caused by abnormalities or failures of it
3. Clinicians spend a great deal of time modifying it in one way or
another.

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 The basic features of innate and
adaptive immune responses

• Innate immune responses tend

to rely on strategies for either
blocking entry of injurious
substances or attacking them if
they breach the skin’s barrier.

• Adaptive immune responses

usually commence in response
to an antigen and eventuate in
the production of antibodies and
a population of antigen-specific
lymphocytes

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Innate Immune Responses
 Innate Immune Responses

• The epidermal barrier

• Innate attack mechanisms

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 The epidermal barrier
• The epidermis is the first
defense mechanism against
external injurious substances,
whether they be :
1. Parasites

2. Infectious agents
(Staphylococci and Malassezia)
3. Allergens

• The main protective layer is the

stratum corneum

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 The stratum corneum
• Overlapping layers of flattened
corneocytes separated by lipid,
provides a physical barrier that
is difficult to penetrate
• The continuous desquamation
of corneocytes makes it difficult
for microbes and allergens to
gain access to the body
• Within the stratum corneum,
the intercellular emulsion
contains sodium chloride, albumin,
complement, transferrin, interferons, lipids, and
antibodies, and these molecules can
provide further protection

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 The stratum corneum
• Transferrin is an iron binding
protein, and its main effect is to
prevent the buildup of free iron,
which can predispose to
bacterial infection.
• Transferrin also has other effects
that include reducing the
binding of gram-positive and
gram-negative bacteria to
surfaces.
• It is currently not known if this
mechanism is relevant in canine
or feline skin infections, but
transferrin has been shown to
inhibit the growth of Malassezia
pachydermatis in vitro

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 The epidermal barrier
• The epidermis can also produce peptides with antibiotic functions.
• The best known of these are defensins, cysteine-rich proteins containing 29 to 34
amino acids.
• Defensins are similar to broad-spectrum antibiotics that can kill a variety of
bacteria and fungi
• Synthesis of defensins is upregulated in response to inflammatory cytokines such
as IL-1 and TNF
• The role of defensins in small animal skin infections is currently uncertain, but
they have been isolated from canine skin and testes
• Human β-defensin 3 inhibits in vitro growth of Staphylococcus pseudintermedius
isolates from atopic and healthy dogs
• Expression of canine β-defensin 103 was found to be downregulated in skin of
atopic dogs and may therefore be a link to the increased incidence of bacterial
infections in atopic dermatitis.

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 The epidermal barrier-Parasites & Infections
• The canine or feline epidermis is unable to protect against a
number of parasites and microorganisms.
• Fleas and mites are able to survive on the skin surface and can
penetrate it to feed.
• Two common commensal microorganisms, S. pseudintermedius
and M. pachydermatis, can also lead to skin infections without
having to breach epidermal defenses.
• Epidermal physiology is influenced by many disease processes
that can alter its permeability and defense mechanisms, thus
resulting in the common phenomenon of secondary pyoderma

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 The epidermal barrier-Staphylococci
• Staphylococci have evolved efficient mechanisms for attaching
to the skin, especially if it has already been damaged.
• This process is known as adherence, and it is the first step in any
infection
• Bacteria have specific molecules on their surface known as
adhesins that allow them to do this.
• Adhesins on staphylococci include teichoic acid, lipoteichoic
acid, fibronectin-binding proteins, and laminin-binding proteins.
• These molecules bind to various proteins on the host, such as
fibronectin, laminin, and collagen

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 The epidermal barrier-Staphylococci
• Fibronectin is a protein normally found in the dermis, but in
humans it can appear on the skin surface following damage or
allergic reactions.
• Adherence of Staphylococcus intermedius to canine
corneocytes is higher in atopic dogs, suggesting that similar
mechanisms occur both in humans and dogs.
• The binding of S. intermedius and Staphylococcus aureus to
canine and human corneocytes, respectively, is very specific,
suggesting that adhesins or binding proteins are different
between species

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 The epidermal barrier-Malassezia
• Malassezia overgrowth can lead
to dermatitis, presumably
because the organism has
evolved as a skin commensal,
and the innate defense
mechanisms of the epidermis
are not equipped to eradicate
• M. pachydermatis has evolved
mechanisms that allow it to
adhere to canine corneocytes

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 The epidermal barrier-Malassezia
• Clinical problems tend to arise when the organism overgrows,
probably in response to changes in skin temperature, humidity,
or lipid profiles.
• When this happens, skin inflammation arises, most likely due to
initiation of an adaptive immune response.
• Other fungi such as dermatophytes, and viruses such as
papillomavirus, have also evolved mechanisms to overcome the
innate defenses of the epidermis to establish infection

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 The epidermal barrier-Allergens
• The skin surface may also be contacted by allergens, which in
genetically predisposed and sensitized individuals can lead to
atopic dermatitis
• Innate defense mechanisms designed to deal with infectious
agents are ineffective at dealing with allergens, and the
epidermis appears unable to prevent access of these proteins to
the underlying skin immune system
• There is also some preliminary evidence that epidermal barrier
function is reduced in canine atopic dermatitis, thus increasing
the chance of allergen entry

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 Innate attack mechanisms
Other defense mechanisms that can be used to provide protection
against parasites or infectious agents, include :
1. Activation of complement

2. Recruitment and activation of phagocytic cells (neutrophils and

macrophages)
3. Recruitment of eosinophils

4. Activation of intraepithelial lymphocytes and natural killer (NK)

cells
5. Activation of mast cells

6. Production of cytokines and chemokines

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 Complement activation
• The complement system is a series of plasma proteins that exist
in a quiescent state until they are activated.
• The role of complement is to assist the immune system in
eliminating pathogenic bacteria
• The complement cascade can be activated by three distinct
pathways:
1. The classical pathway

2. The alternative pathway

3. The lectin pathway

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 1. The classical pathway
• The classical pathway is activated by antibody-antigen complexes,
which bind to the first complement component, C1
• In a subsequent complex series of steps, the fourth (C4) and second
(C2) components of complement bind to C1, forming the active
enzyme C3 convertase.
• This enzyme catalyses conversion of the third complement
component (C3) into two parts, C3a and C3b.
1. C3a is a chemoattractant for neutrophils and can activate mast cells

2. C3b can opsonize bacteria and promote phagocytosis and also

activate the membrane attack complex, a cylindrical structure
composed of complement components 4, 5, 6, 7, 8, and 9

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 2. The alternative pathway
• The alternative pathway is activated by direct interaction
between the surface of a microbe and C3b, which is generated
in the serum by slow, spontaneous hydrolysis of C3.
• Once bound, the C3b catalyses further conversion of C3 into
C3b, resulting in the downstream changes

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 3. The lectin pathway
• A plasma lectin binds to a mannose residue on the surface of a
microbe and activates C1 in the absence of antibodies.
• The pathway is subsequently the same as the classical pathway

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 Recruitment and activation of
phagocytic cells
• Phagocytic cells play a major role in protecting the
host against pathogenic microorganisms

• To eliminate bacteria, circulating neutrophils and

monocytes must first be recruited to sites of
infection.

• The initial response is triggered by resident tissue

macrophages that detect microorganisms and
secrete the proinflammatory cytokines IL-1 and
TNF-α

• These molecules lead to upregulation of adhesion

molecules on blood vessel walls, including
selectins and integrins, which can bind to ligands
on the inflammatory cells.

• E-selectin binds to carbohydrate ligands on the

phagocytic cell surface with low affinity, causing
the cell to slow down and roll along the
endothelium

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 Recruitment and activation of
phagocytic cells
• After arriving at a site of infection, neutrophils and macrophages
must become activated by recognizing extracellular microbes using
receptors on their surface.
• There are a number of classes of receptors that can bind
microorganisms and mediate phagocytosis or cell activation :
1. Mannose and other C-type lectin-like receptors.

2. Scavenger receptors

3. Receptors for opsonins

4. Toll-like receptors

5. Seven transmembrane receptors

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 Recruitment of eosinophils
• Eosinophils are involved in cutaneous immune responses against
ectoparasites, but also contribute to hypersensitivity and
autoimmune reactions.
• Eosinophils are attracted to sites of inflammation by specific
chemotactic factors such as histamine, complement component 5a,
leukotriene B4, parasite extracts, and various chemokines (e.g.,
eotaxins 1 and 2, RANTES [regulated upon activation, normal T-cell
expressed and secreted]).
• As with neutrophils, eosinophils possess a number of cell surface
receptors for these substances, in addition to Fc receptors for IgG and
low-affinity IgE receptors.
• At sites of inflammation, eosinophils show both phagocytic and
secretory activity

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 Recruitment of eosinophils
• They have some ability to ingest bacteria, immune complexes,
and yeasts, but much less than neutrophils
• More important is their secretory function.

• Eosinophil granules contain a number of important proteins

that include major basic protein, eosinophil cationic protein,
eosinophil peroxidase, and lysosomal enzymes.
• These proteins are toxic to helminths, cells, and bacteria and
can degranulate mast cells, but they can also cause significant
damage to host tissues in hypersensitivity disorders.

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 Recruitment of eosinophils
• Eosinophils are commonly found in parasitic skin diseases (e.g.,
flea allergy dermatitis, mosquito bite hypersensitivity, sarcoptic
mange)
• Low numbers in allergic dermatoses (e.g., atopic dermatitis).

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 Activation of Intraepithelial Lymphocytes
and Natural Killer (NK) cells
• The epithelial surfaces of mammals contain a special group of lymphocytes
known as intraepithelial lymphocytes.
• Like normal T cells, these are T cells that contain antigen receptors.
• Some intraepithelial lymphocytes act as NK cells and can recognize and kill
virus-infected cells or microbes directly.
• Intraepithelial lymphocytes act as sentinels at epithelial surfaces and
function in host defense by secreting cytokines, activating phagocytic cells,
and killing infected cells.
• Although NK cells are known to occur in dogs there is no evidence that they
are beneficial in the response to staphylococci or Malassezia organisms.
• They are probably more relevant in the defense against cutaneous viral
pathogens

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 Activation of Mast cells
• Mast cells are normal residents of connective tissue and found in highest
numbers in areas of the body that interface with the environment: skin, lung,
and gastrointestinal tract
• Mast cells are important in the innate defense against intestinal helminths and
bacteria, and in wound healing
• The role of mast cells is in the gastrointestinal tract, where they play an
important role in eliminating nematode parasites
• Mast cells are functionally active during expulsion of the parasites, releasing
large quantities of granule proteases
• Critical component of the innate immune response to certain bacterial infections
• Mast cells are also involved in wound healing
• Mast cell granule proteases are also important in regulating the influx of
inflammatory cells into the skin

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 Production of Cytokines and
Chemokines
• Certain cytokines and chemokines play key roles in the innate
immune system
• The general proinflammatory cytokines TNF-α and IL-1 are important
in initiating innate immune responses
• TNF-α is produced by macrophages, activated T cells, NK cells, and
mast cells
• IL-1 is produced by macrophages, endothelial cells, and keratinocytes
• Both cytokines activate endothelial cells to enable inflammatory cell
recruitment to sites of injury, activate neutrophils, and stimulate
production of acute-phase proteins

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 Production of Cytokines and
Chemokines
• IL-12 is also important in the innate immune response.
• IL-12 is produced by macrophages and dendritic cells in response to
microbial infection and promotes T-cell differentiation, IFN-γ
synthesis, and activates NK cells and T cells.
• Production of type 1 interferons (IFN-α and IFN-β) by monocytes and
fibroblasts is an important innate defense mechanism against viral
infections.
• A specialized group of cytokines known as chemokines represent a
large number of proteins that act as chemoattractants for various
inflammatory cells.

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Adaptive Immune Responses
 Adaptive Immunity
• Mediated by cells called Lymphocytes and their products

• There are 2 major populations of lymphocytes

1. B lymphocytes
2. T lymphocytes

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 Humoral and Cell-Mediated Immunity
(Two types of adaptive immunity)

• Induced by different types of lymphocytes and function to

eliminate different types of microbes
• Humoral immunity is mediated by molecules in the blood and
mucosal secretions, called Antibodies, are produced by B
lymphocytes
• Cell-mediated immunity (cellular immunity) is mediated by T
lymphocytes

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 Adaptive Immune Responses
• Antigens and antigen recognition

• Antigen presentation

• Activation of T cells

• Immunity and inflammation

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 Antigens and antigen recognition
• Start the process with antigens, usually antigens that trigger the
immune response in the first place
• Antigens that trigger an IgE response are called allergens

• Antigens those involved in autoimmune diseases are called

autoantigens
• Antigens are normally large proteins with complex three-
dimensional structures

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 To detect antigens, the body uses
two different types of receptors:
1. Antibodies—these are present on the surface of B lymphocytes and
recognize the antigen in its native form.
They usually detect a particular part of the protein molecule that is
highly immunogenic and situated on the outside of the molecule. These
sections of the antigen are known as immunodominant epitopes.
2. T-cell receptors—these are situated on the surface of T lymphocytes.

They cannot recognize the protein in its native form. The whole antigen
must first be processed by an antigen-presenting cell in which the
protein is broken down into peptides by various enzymes including
cathepsin E. These peptides are normally about 8 to 12 amino acids long.

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Antigen in Canine and Feline skin
• Canine and feline skin is
exposed to large numbers of
antigens from various sources.
• In general, three different
categories of antigen can be
identified:
1. Antigens from parasites or
infectious agents
2. Allergens that are able to trigger
allergic inflammation
3. Autoantigens that are targeted
in autoimmune diseases
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 Antigen in Canine and Feline skin
• Dogs and cats are known to react to several proteins in the
saliva of the cat flea Ctenocephalides felis, with one such
protein having been cloned, sequenced, and designated Ctef1.
• In dogs, antigens from S. intermedius that triggered an IgG
response were identified in both healthy dogs and dogs with
pyoderma.
• The proteins identified had molecular weights ranging from 20
to 198 kD, but the majority of antigens were below 75 kD
• An antigen with a molecular weight of 28 to 29 kD was the most
obvious on Western blots.

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 Malassezia pachydermatis
• Malassezia pachydermatis produces a number of antigens and
allergens that may be clinically relevant in dogs.
• Using Western immunoblotting, four proteins of 219, 110, 71, and 42
kD were shown to be recognized mainly by non-atopic dogs with
Malassezia dermatitis as compared to healthy dogs.
• In atopic dogs, an antigen of 25 kD was identified in the majority of
cases with Malassezia dermatitis, but in only a few atopic dogs
without Malassezia overgrowth and none of the normal dogs,
suggesting that this protein may have some clinical relevance.
• Proteins with molecular weights of 45, 52, 56, and 63 kD have also
been shown to be major allergens in atopic dogs, because they were
recognized by IgE in more than 50% of the sera from atopic dogs with
Malassezia overgrowth

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 Autoantigens
• A number of autoantigens have now been characterized in dogs
with cutaneous disease, including desmogleins 1 and 3,
desmocollin 1, plakin, various basement membrane proteins
and nuclear antigens

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 Antigen Presentation
• An efficient immune response is generated by stimulating T cells
• Unlike the interaction between antibodies and antigens, T cells are
unable to recognize whole proteins
• To activate the T-cell population, antigens must first be processed and
then presented to the T-cell receptor by a “professional antigen-
presenting cell.”
• In the skin, the main antigen-presenting cells (APC) are Langerhans
cells (situated in the epidermis) and dermal dendritic cells (situated in
the dermis).
• Antigens are phagocytosed by these cells and broken down into small
peptides within phagolysosomes

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 Antigen Presentation
• It is not possible for these peptides to interact independently
with the T-cell receptor.
• The peptides must first be attached to Major Histocompatibility
Complex (MHC) protein molecules produced inside the antigen-
presenting cell.
• These proteins transport the peptides back to the cell surface
and present them to adjacent T-cell receptors.

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 Antigen presentation by class I and II major
histocompatibility complex (MHC) molecules,
TCR, T-cell receptor

The MHC gene group codes for some very

important proteins:
• Class I genes code for proteins found on the
surface of most nucleated cells.
• Class II genes code for proteins usually only
found on macrophages and lymphocytes.
• Class III genes code for a variety of
inflammatory molecules including complement,
TNF, and heat shock proteins.

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 Activation of T cells
• For the T cell to be activated, a second signal is required.

• Second signals are a series of molecular interactions between

adhesion molecules on the two cells and specific receptor
ligand interactions.
• If these second signals do not occur, the T cell does not
respond, and the host becomes tolerant to the antigen, a state
known as anergy.

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 Helper T-cell responses
The difference between helper T cells is the range of cytokines they produce
following activation:
1. T helper 1 cells (TH1) produce predominantly IL-2 and IFN-γ.
2. T helper 2 cells (TH2) produce predominantly IL-4, IL-5, IL-6, IL-10, and IL-13.
• Helper T cells are initially designated as TH0 cells but can become either TH1
or TH2, depending on the signals received from the antigen-presenting cell.
• TH1 cell responses tend to promote cell-mediated immunity and IgG
production by B cells
• TH2 responses tend to promote IgE production and allergic responses.

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 B-cell activation and antibody
production
• Activated T cells can initiate inflammation on their own by
secreting proinflammatory cytokines.
• Antibody response is also required to help eliminate
microorganisms or neutralize toxins
• Activation of B lymphocytes is required for antibody synthesis,
and this can be achieved in a number of ways:
1. B cells may be activated following direct contact with antigens.
2. The B-cell response to microbes can be greatly enhanced if the antigens
are recognized in association with complement.
3. Some B cells (called B-1 cells) are pre-programmed to produce IgM
against common bacterial polysaccharides and lipids
4. B cells may be stimulated by interacting with helper T cells.

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 Antibodies can help deal with
infectious agents in a number of ways:
• They can neutralize antigens, viruses, and toxins
• They can opsonize organisms, bind to Fc receptors on phagocytic
cells, and initiate phagocytosis
• They can form immune complexes that can activate complement,
leading to phagocytosis of C3b-coated bacteria
• By activating complement, they can initiate inflammatory cell
infiltration and lysis of bacterial cell walls
• Antibodies on the surface of B cells can activate the cell and cause it
to proliferate and start producing more antibody
• Antibodies can also present antigens to T cells and stimulate a helper
response.

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 Antibody responses
• Antibody responses to various infectious agents, allergens, and
autoantigens have been frequently documented in dogs and
cats with cutaneous disease
• IgG and/or IgE antibodies have been demonstrated to
staphylococcal proteins, Malassezia antigens, proteins from D.
farinae, and canine keratinocyte proteins.
• Although these antibody responses are supposed to be
protective, they can lead to hypersensitivity reactions and
autoimmune disease.

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 Antibody responses
• In nearly all immune responses, IgG antibodies will be
produced even if other classes are also present.
• IgG responses to multiple antigens from S.
intermedius/pseudintermedius and M. pachydermatis are seen
in both healthy and infected dogs, probably because both of
these microbes are commensals that normally reside on dogs.
• IgE responses to both these organisms have been documented
by ELISA
• IgG responses are also seen to environmental allergens such as
D. farinae in both healthy and atopic dogs

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 Autoantibodies factors
• Formation of autoantibodies is likely to involve genetic,
infectious, environmental, and hormonal factors
• Genetic factors involve genes encoding for the MHC, cytokines,
and regulatory T-cell factors
• Infections may be able to trigger autoimmune disease in certain
circumstances.
• Environmental factors include UVL, which is known to be
important in the pathogenesis of lupus.
• Hormonal factors may be involved, because some diseases are
more common in one sex

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 Factors lead to autoimmune
diseases :
1. Failure of the self-tolerance mechanisms

2. Polyclonal B cell activation by certain viruses and parasites may stimulate autoreactive cells. These
cells produce antibodies to common autoantigens such as DNA, IgG, phospholipids, erythrocytes, and
lymphocytes

3. Failure of regulatory T-cell function

4. Activation of T cells by superantigens

5. Expression of class II MHC on normal cells following damage, allowing recognition by helper T cells

6. Exposure of previously hidden antigens, such as in the CNS, testes, or lens

7. Cross-reactivity between microorganisms and self antigens—this is known as molecular mimicry, but
its full significance is not known

8. Viruses or lymphoid neoplasia may interfere with normal immune function, allowing development of
autoreactive cells

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 Immunity and inflammation
• The end result of the immunologic pathways described is
inflammation
• Inflammation is characterized by recruitment of leukocytes and
plasma proteins into tissues, where they are activated
• The cardinal signs of inflammation are rubor (redness), tumor
(swelling), calor (heat), and dolor (pain)
• The aim of this inflammation is to eliminate noxious substances
or microorganisms and establish a state of immunity

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 Immunity and inflammation
• Many of these mechanisms can either be ineffective or lead to
ongoing and detrimental inflammation.
• It is because of these failings that veterinary dermatologists
(and other clinicians) spend much of time trying to eliminate
sources of antigens by treating staphylococcal and Malassezia
infections with antimicrobial drugs, or suppressing unwanted
inflammation by using glucocorticoids and other
immunomodulatory drugs to treat allergic or autoimmune skin
diseases

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Components of the Skin Immune System
 Components of the Skin Immune System
• Antigen presenting cells  Langerhans cells

• Skin homing lymphocytes

• The microenvironment of the keratinocytes

• Dermal microvascular unit

• Neural-immunologic network

• Skin draining lymph nodes

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 How does cutaneous
inflammation occur?
• The upregulation of adhesion molecules on vascular endothelia
and on inflammatory cells makes cells sticks to the vascular
endothelia, and then exit from the blood vessels into the dermis
• 4 main families :

1. Integrins
2. Immunoglobulin superfamily members
3. Selectins
4. Mucin-like adhesion molecules

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 Immune traffic in the skin
There are 3 phase :
1. Recruitment : Mast cell, TNF, PAF, P-selectin, LFA-1, E-selectin,
VCAM-1, Chemokine
2. Retention : Lymphokine, IL-1, IL-8, Expression of ICAM-1,
Interaction of T cells and dendritic cells, Migration of T cells
3. Resolution : IL-10

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 Damage resulting from the
immune system :

1. Hypersensitivity
2. Autoimmunity
3. Immunodeficiency

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