immunology
Cytokines made by macrophages
IL-1
Costimulator of TH2 cells
Stimulates acute phase response
IL-6
Promotes B cell differentiation
Stimulates acute phase response
IL-12
Costimulator of TH1 cells
IL-18
Promotes IFN-y production by TH1 cells
TNF-a
Cytotoxic
Stimulates T cell growth
Stimulates acute phase response
Triggers inflammation
What are the proinflammatory cytokines
IL-1b
IL-6
TNF-a
CD8
Kill virus infected cells
Target viruses and intracellular bacteria
TH1 CD4
Activate infected macrophages
Help B cells produce antibodies
Target microbes in macrophage vesicles and extracellular bacteria
TH2 CD4
Help B cell produce antibodies
Help antibody switching to IgE
Targets helminths
TH17 CD4
Enhance neutrophil response
Promote barrier integrity
Target fungi
TFH
Antibody production
Isotype switching
Target all types
T Reg CD4
Suppress T cell responses
5 important cytokines
IL-1 beta
Local
Activates vascular endothelium (leaky, adhesion molecules, cytokines)
Activates lymphocytes
Activates local tissue destruction
Increases access of effector cells
Systemic
Fever
Production of IL-6
IL-6
Local
Lymphocyte activation
Antibody production
Systemic
Fever
Acute phase protein production
TNF-a
Local
Activates vascular endothelium
Increases vascular permiability
Increases entry of IgG, complement and cells to tissue
Fluid drainage to lymph nodes
Systemic
Fever
Mobilisation of metabolites
Shock
CXCL8
Local
Chemotactic factors recruit neutrophils, basophils and T cells to site of inflammation
IL-12
Local
Activates NK cells
Induces differentiation of CD4 T cells into TH1 cells
Lectin Pathway
Mannose binding lectin and ficolin's bind to carbohydrates on pathogen surface
Complement proteins assemble together to make C3 convertase
Classical
C1q component interacts with pathogen surface or antibodies from adaptive immunity that are bound to the surface of the pathogen
Complement proteins assemble on the surface of pathogen or in an infected cell to make C3 convertase
Alternative
C3 undergoes spontaneous hydrolysis at low levels in the body
Deposition of enzyme C3 convertase on microbial surface
If no infection, no deposition of enzyme on surface of pathogen
Production of C3 convertase
C3 convertase cleaves C3 molecule of complement into C3a and C3 b
C3b deposits itself on the surface of pathogen/cells
C3a is released
Steps of leukocyte migration
Interactions between activated vascular endothelium and molecules on leukocyte
Rolling adhesion
Tight binding: caused by cytokines and chemokines
Diapedesis: crossing of cell wall
Migration: follows gradient of cytokines/chemokines
Phagocytic cells
Neutrophils and macrophages
Attracted to site of infection by dead/dying cells
Phagocytosis steps
Chemotaxis
Adherence
ingestion
Destruction
Fever
Microbial components engage PAMPs on macrophages
Release inflammatory cytokines
IL1
IL6
TNFa
These cytokines especially IL-1 act on hypothalamus and increase body temperature
Systemic outcome of fever
Inflammatory response can be localised or systemic
IL-1
Acts on brain causing fever, anorexia, somnolence
Reduce energy consumption
IL-6
Acts on liver
Release of acute phase proteins = recognition and destruction of invading pathogen
Acute phase protein = c reactive protein
Elevated CRP = inflammation
Cytokines
Cell to cell communication
Immunoregulating/modulating agents
Interleukins and interferons
Can effect brain and liver and lead to acute phase response
Acute phase proteins
Bacteria induce macrophages to produce IL-6
IL-6 acts on hepatocytes in liver to induce synthesis of acute phase proteins
C reactive proteins
Binds phosphocholine on bacterial surfaces
Acts as an opsonin: coats bacteria and enhances uptake of bacterium by phagocytes
Can also help activate complement
Mannose binding lectin
Bind to carbs on surface of bacteria
Acts as opsonin increasing uptake by phagocytes
Initiate lectin pathway
Acute inflammation
Happens quickly
Major cell type is neutrophils, followed by monocytes - > macrophages
Redness (vasodilation), swelling (plasma proteins and fluid in tissue, vascular leakage, histamines, leukocytes), pain (kinins), heat (blood flow)
Chronic
Lasts longer
Leukocytes, plasma cells (different cell types)
Granuloma: collection of immune cells formed when immune system is walling off but cannot eliminate foreign object
Tissue destruction due to high numbers of inflammatory cells
Consequences of activated complement system
C3a and C5a act as chemotaxins
Recruit phagocytic cells to site of infection
Promote inflammation
phagocytes that have receptors for C3b can help engulf and destroy the pathogen
Phagocytes recognize c3b on surface of pathogens, can engulf and destroy it
Formation of membrane attack complex (MAC) = pore formation in the microbe or in infected cell
Disrupts cell membrane and leads to lysis
Chemotactic factors that attract neutrophils
Complement factors
Prostaglandins
Kinins
Steps of phagocyte migration
Margination
Rolling
Tight adhesion
Diapedesis (cell crosses blood vessel wall)
B cells and macrophages present antigens to sensitized helper T cells
Dendritic cells present antigen to naïve helper T cells
T cells and APC's (DC's) paracortex
B cells follicles
GALT = Immune
Secretory antibody IgA
Cell mediated immunity
T cells
Dendritic cells, macrophages and NK cells
Inductive sites
Sites where antigens are processed and immune responses initiated
Peyer's patches, tonsils, NALT
Effector sites
Antibodies and cell mediated responses are generated
Lymphoid nodules, lymphocytes: plasma cells
IgA dominant immunoglobulin isotype at mucosal surfaces
Called common mucosal immune system
MALT can be local, systemic or can get response from other mucosal tissue
Priming of lymphocytes at 1 MALT -> protective immunity at another MALT
Adhesion molecules and chemokines bring immune cells
Peyer's patch
Mostly secondary lymphoid tissue
Induction of adaptive immunity
Peyer's patch has…
Subepithelial dome
Follicles with B cells
T cells in between
How do bacteria manipulate or escape host immune response
Escape epithelial defences
Degrade mucous/physical barrier
Make enzymes that degrade antibodies
Antigenic variation: change surface antigens to become unrecognisable to host immune system
Changes in LPS, changes to pili, flagella
Polysaccharide capsules (coat of sugars around bacterial cells)
Escape phagocytes and innate immunity
Capsules reduce antibody binding and opsonisation
interfere with phagocytosis
Interfere with phagocyte recruitment (degrade chemokines)
Kill leukocytes
Evade adaptive immunity
Resistance to antibodies, don't bind as well to polysaccharide capsules
interfering with cytokines secretions
Interfere with antigen presentation
Inhibit B and T cell functions
Virulence factors
Motility = Flagella, cilia, fimbriae
Compete for resources like iron
More resistant and more invasive
How do they resist phagocytosis
Capsules
Protein A soaks up antibody
Fibrin clot formation
Prevent phagosome from acidifying and is protected inside
Kill or escape phagocytosis
How do they evade adaptive
Capsules
Antigenic variation
Proteases
Myobacteria cause release of immune suppressive cytokine
Interfere with antigen processing and presentation
Direct host damage
Exotoxins
Endotoxins
Killing host cells
Indirect host damage
Cause immune complexes that cause inflammation/tissue damage
Molecular mimicry = autoimmunity
Aberrant immune response causes damage to own host
Most important protective mechanisms against bacteria
Complement
PAMPs recognised by TLRs
Phagocytes
Antibodies and T cells
Interstitial space like blood and lymph
Antibodies
Complement
Phagocytes
Epithelial
Antibodies
Intracellular
T cells and NK
Intracellular in vesicles
Macrophages very important
T cells need to receive multiple signals from APC before they can be activated and differentiate into effector and memory populations
Costimulation or antigen
T helper cells drive inflammatory response
Helper other cells by producing cytokines
Help B cells be activated into plasma cells and produce specific antibodies
Help CD8 T cells activate to cytotoxic T lymphocytes
Activate macrophages to enhance phagocytosis and enhanced killing ability
Depends on cytokines released from APC's
TH1 cytokines
IL2
IFN-Y
TNF-a
Inflammatory response
Respond to viruses and intracellular pathogens
Allograph rejection
Ig switching
TH2 cytokines
IL-4
IL-5
IL-13
IL-10
Humoral B cell responses: class switching to IgE
Helminth Immunity
Allergy
Activate mast cells and eosinophils
Cytotoxic T cells
Kill site infected with intracellular bacteria, viruses and tumours
Intrinsic pathway = Injects enzymes (granzymes and perforins) and cause death, apoptosis
Extrinsic = CD95 L, interaction outside cell = apoptosis
Function of antibodies
Enhance phagocytosis via opsonisation (taken up easier)
Enhance classical pathways of complement mediate killing initiated by antibodies = Classical pathway
Neutralize microorganisms and toxins
Prevent pathogen attachment
Antibody dependent cellular cytotoxicity
3 main types of fungal infections
Primary infections on skin and other surfaces
Primary infections by dimorphic fungi: respiration
Secondary infection by opportunistic fungi: temporary immunodeficiency
Once established fungi destroy TH1 cells
Type 4 hypersensitivity to fungal infections
Acute
Rapid onset of disease with symptoms
Small incubation period
Cytopathic
Self limiting
Examples of acute
Influenza
Smallpox
SARS
Chronic
Long incubation period
No immediate cell death so mostly non-cytopathic
Virus is not cleared by immune system: high amount of virions remain
Persists in host for long period of time
Eventually symptoms appear
Examples of chronic
Hepatitis B and C
Latent infection
Begins like acute infection
Later there is latency: no virion production, no shedding of virus and no symptoms
Virus can reactive with stress, illness
Episodic reactivation: more virus is produced and symptoms occur
Cytopathic when reactivated
Do not always have symptoms
Examples of latent
Shingles and herpes
Slow- initial infection appears acute
Infection is largely cleared but low levels of virus remain
Can ultimately defeat the host
Often target immune system specifically
Examples of slow
HIV
NK cells and interferons important for defending against virus
Interferons = family of cytokines that inhibit viral growth
Recognise viral nucleic acids by cytoplasmic and membrane sensor proteins = PRR's
Inhibit transcription and translation of viral proteins
Adaptive immunity = Increase expression of MHC molecules on surface of cells and enhances antigen presentation
IFN-a = type 1 interferon
Produced by virus infected leukocytes
IFN-b = type 1 interferon
Produced by virus infected fibroblasts and epithelial cells
IFN-Y = type 2 interferon
Released only by antigen stimulated T cells (mostly TH1 and NK cells)
NK cells
Kill infected abnormal/stressed using cytotoxic granules
Activated based on lack of MHC
MHC molecules on normal cells recognised by inhibiting receptors
Cytotoxicity of NK cells is stimulated by type 1 interferons (a and B)
Viruses can downregulate MHC 1 on surfaces of infected cells
Evades killing by cytotoxic T cells
Increases NK cell killing
CD4 TH1
Cells activate macrophages enabling them to destroy intracellular microorganisms
Help CD8 cells
Help B cells switch antibodies from IgM to IgG
CD4 TH2
Activate B cells to produce different Ig isotypes
Antigenic drift
Mutations alter epitopes in haemagglutinin so antibody no longer binds
Antigenic shift
RNA segments exchanged between 2 viruses
Occurs in secondary host
No cross protective immunity because it creates a new haemagglutinin
Protozoa
Innate
Phagocytosis and complement
TH2 = Extracellular
Antibodies, Mast cells, eosinophils
TH1 = intracellular
Cytotoxic T lymphocytes
Macrophages activated by IFN-Y
Immune evasion strats of protozoa
Antigenic variation
Intracellular localisation
Some protozoa can hide in RBC's which don't have MHC molecules so cannot be killed by cytotoxic T cells
Protozoa can lead to hypersensitivity
Type 1 (allergic)
Local irritation and inflammation involving mast cells and eosinophils
Type 2 (cytotoxic reactions)
Immune destruction of RBC's = anaemia with babesia
Type 3 (immune complex deposition)
Vasculitis, glomerulonephritis = leishmanias
Type 4 (delayed type hypersensitivity)
Granulomas
Autoimmunity
Immune responses against self = Trypasnomes
Immune strategies of Helminth
Thick cuticle: cannot be penetrated by complement, T cells and enzymes
Different immune profile
Cytokines often dampened
Type 2 immunity
Roundworms, cestodes and trematodes
CD4 TH2 cell is major player = produces range of cytokines
IL-4 is major, IL-5 and IL13
CD4 TH2 is major player in Type 2 Helminth Immunity = secretes IL-4 (IL-5 and IL-13)
Helminth Infection
IL-4 release
TH2 cells secrete other cytokines IL-4, IL-5, IL-10 (immunosuppressive blocking IL-12)
The secretion of IL-4 induces IgE and some IgG
Expanded populations of Eosinophils, Mast cells, basophils and alternatively activated macrophages (wound repair)
Cytokines released by TH2
IL-13
Epithelial cell repair = Increased cell turnover: shedding of parasitized cells
Goblet cell release more mucous = lack of adherence to GIT = loss of parasite
Increase smooth muscle contraction enhancing worm expulsion
IL-4 and IL-13
Recruit and activate M2 macrophages
Increase smooth muscle contraction
Tissue repair
IL-5
Acts on eosinophils
Eosinophils Kill helminth parasites
ADCC of parasites if They have IgE
Enzymes and toxic elements
IL-3 and IL-9
Arms Mast cells against helminth
Mast cells produce histamine and TNF-a when bound to IgE
Recruit inflammatory cells and remodel mucosa
Mast cells have receptors that bind to IgE
When allergen or antihelminth binds it degranulates: histamine, heparin, proteases, IL-4 and IL-5
Mast cells and eosinophils involved in antihelminth
Mast cell bound IgE (sensitized mast cell) triggers degranulation when bound to helminth antigens
Release products
Smooth muscle contraction
Increase vascular permeability
Epithelial cell turnover
Eosinophils
Make products that damage helminth cuticles
Make many different molecules that can damage and penetrate (ROS, phospholipases)
How do they make ADCC = IL-4 class switching to IgE, eosinophils' receptors are engaged
2 different mechanisms of expulsion of intestinal nematodes
Mast cell dependent
Mast cell degranulation
inflammation
Increase permeability and fluid secretion
Mast cell independent
TH2 response releases IL-4 and IL-13
goblet cells proliferate and Increase in mucous
Smooth muscle contractions
TH2 Often decline during chronic helminth infections = increase in regulatory cells (T-reg)
Expand numbers of T-reg = asymptomatic tolerance
Shedding of surface antigens
Protease production to neutralise anti parasite immunity and degrade Ig
Adsorbing host antigens and masking parasite antigens
Regulation of host functions
Suppress neutrophils and macrophages
Suppress TH response
Neutralise respiratory burst and antioxidants
Use of cytokines as growth factors
Hypersensitivity
Sensitisation phase
First response to antigen
Re exposure phase
Type 1 Immediate hypersensitivity
Happens within minutes
Localised or systemic
Allergy and anaphylaxis
Allergens generally encountered on mucosal surfaces
Atopic dermatitis
Presence of eosinophils at given site is hallmark of type 1
Atopy: Overactive TH2 immunity = More IL-4 and IgE genetic predisposition to type 1 immediate hypersensitivity
Itching
Redness
hairloss
Main players in type 1
Mast cells
IgE
Eosinophils
Basophils
TH2 cells and release of IL-4
Sensitisation phase
B cells make IgE against that allergen (TH2 switching to IL-4)
IgE binds to surface of mast cells creating sensitized mast cell
Re-exposure phase
Mast cells are re-exposed to allergen causing them to degranulate and release histamine, prostaglandins/leukotrienes, chemotactic factors for neutrophils and eosinophils
These can restrict smooth muscle in airways, vasodilation, acute inflammation, nerve endings causing itchiness
Hours after
Eosinophils and macrophages
Examples of Type 1
Allergy
Anaphylaxis
Asthma
Hay fever
Allergic conjunctivitis
Hives
Type 2 Antibody mediated cytotoxic
Cytotoxic: cell death (destroyed by antibodies)
Inflammation
Cell destruction
Complement
Main players in type 2
Antibodies = IgG and IgM good at binding to complement and initiating classical pathway
Macrophages phagocytose
NK cells kill
Examples
Incompatible blood transfusion (RBC's have surface antigens)
haemolytic disease of newborn
Autoimmune haemolytic anaemia
Autoimmune thrombocytopaenia (kills platelets)
Graft rejection
Incompatible blood transfusion
Lysis of RBC's in incompatible blood transfusion causes high concentration of free haemoglobin being released which can be damaging to kidney
Potassium also released
Blood clotting
Complement activation
Mast cells degranulate and vasoactive molecules produced
Haemolytic disease of newborn
Female sensitized to foetal foreign blood group antigens during pregnancy
Makes Ab against foetal antigens
Newborn absorbs colostrum with antibodies
These antibodies taken up by newborn and react with antigens on red blood cells of newborn
Rapid haemolysis (mostly in horses and cats)
Increased pregnancies can be more severe
Antibody reacts with surface of normal cells destroying them via complement, opsonisation, phagocytosis, cytotoxic cells (NK)
Lead to cell lysis and membrane attack complex
Lesions result from cell destruction
Type 3 Immune complex hypersensitivity
Antigen-antibody complexes mediate tissue damage
Also mediated by antibodies but no cytotoxicity
IgG and IgM
Immune compelxes
Immune complexes form by antigen and antibody complexing together leading to complement activation
Occurs with subsequent exposure
May activate locally or circulate and be deposited in blood vessel walls = systemic
Phagocytic cells often recruited
C3a and C5a lead to inflammation
Examples
Arthus reaction
Serum sickness
Lupus (deposition of immune complexes)
Antigens, antibodies and complement come together to form an immune complex
Antibody binding causes classical pathway
Complement activation also releases C3a and C5a, neutrophil accumulation = inflammation and tissue damage
Arthus reaction
Local tissue inflammatory reaction
Immune complexes deposited at site of penetration
Sometimes associated with vaccine = high local conc of antigen at site, high circulating antibodies
Pain, swelling, thrombosis occurs at site
Local vasculitis due to immune complexes in blood vessels = inflammation
Occurs if there are high local concentrations of antibodies
Serum sickness
Damage to kidneys
Anaemia
Thrombotyopaenia
Vasculitis
Arthritis
Depends where immune complex deposits
Phagocytic cells can remove immune compelxes
Soluble immune complexes deposited in vessel walls
Presentations
Neutrophil accumulation: arthritis
Deposited in glomeruli: inflammation/nephritis
Arterial intima: neutrophil accumulation = arteritis
Acute
Large dose of foreign antigen administered
Patient needs treatment with tetanus antiserum
Chronic
Multiple small intravenous doses
Chronic damage to basement membranes
Type 4 Delayed Type hypersensitivity
Different from other types because it is cell mediated by T cells
Delayed, prolonged onset
Can occurs days after exposure to antigen
Primarily cell mediated: antigen specific TH1 cells and its cytokines
Contact dermatitis
Main players
Cytokines
T cells (TH1)
Sensitisation phase is about generating TH1 cells and antigen specific memory TH1 cells
Subsequent exposure
Reactivation of memory T cells
Drive inflammation
Once TH1 cells are reactive usually in lymph node, go to site
Release cytokine IFN-Y
Recruit other inflammatory cell tissue damage at site of injection
Detected by presence of memory T cells at least 72 hours after exposure
Delayed type hypersensitivity
Injection
Tuberculin skin test for TB
Severity depends on number of Memory T cells present
Uptake of antigen by APC's
Phase 1
Migrate to draining lymph node
Present to naïve T cells
They respond and proliferate and secrete cytokines
Phase 2
TH1 effector cells migrate to site
Release cytokines/chemokines
Recruit inflammatory cells
Contact dermatitis
Syndromes for type 4 hypersensitivity
Antigen is often insect venom, micro bacterial proteins
Often caused by haptens penetrating skin and attaching to self proteins in epidermis and recognised as foreign
Exposure to foreign material
Skin thickening and fibrosis
Contact hypersensitivity
Absorbed through skin
Hapten
Attach to self epidermal proteins and recognised by immune system (poison ivy)
2 types of immunological disorders
Primary
Genetic
Innate and adatpive
Secondary
Acquired
Innate deficiency
Complement deficiency cause extracellular bacterial infections
Phagocytic deficiency
Defects in entry to inflamed tissues
Mutations cause white blood cells stop adhering to endothelium
Neutrophils can't exit blood vessels
Severe bacterial infections
Adhesion deficiencies
Symptom = high levels of neutrophils because they can't leave blood
Red setters and Holsteins
Cyclical or severe neutropenia
Grey colie syndrome, light coloured
Low levels of neutrophils
Effects bone marrow, so platelets and monocytes can also be affected
Cyclical function in number of blood cells, especially white
Take blood multiple times
Issue with stem cell growth factors
Bacterial infections
Defects in neutrophil granules (also NK cells and CTL's)
Neutrophil number are still there but poor function due to defect in their granules
Susceptible to bacterial infections
Decreased colour because melanin granules can also be affected
Adaptive
Cellular or antibodies
Number of cells may be normal but there could be a deficiency in the function
Often issues in progenitor cells
B and T cell deficiencies are worst
Issue in lymphoid precursors = combined immunodeficiency (B and T cells)
Thymic aplasia = T cells mostly effected
Bone marrow/Bursa = B cells don't develop properly, can't make antibody = agammaglobulinemia
IgA deficiencies if problem further down
Defects in APC's
Issues in antigen presentation to T cells
Causes Problems in other cells
B cell deficiencies
Antibody deficiency
More likely to die from extracellular bacteria disease
Selective Ig deficiencies
Sharpies and German shepherd = deficiency in IgA
T cell deficiencies
More likely to die from viral infections
Viruses replicate in cytoplasm and are killed by cytotoxic T cells
Mild defects in thymus developments
Combined T and B deficiencies
Reoccurring infections
Lack of stem cell development
Variety of mutations can cause it
CD4 both
Defects in antigen receptor genes
Defects in cytokine genes
Defects in antigen presentation
Severe combined immunodeficiencies (SCID)
Pneumonia most common in equines
Secondary
Previously had normal immune function
Acquired
Common
Cytotoxic drugs, chemotherapy, overexercise
Tumours
Old = thymic involution = thymus shrinks with age and less naïve T cells leave thymus
Immunosenescent old age
thymic involution = thymus shrinks with age and less naïve T cells leave thymus
Decrease in circulating CD4 T cells
Fewer naïve cells leave bone marrow and thymus
Old animals still have high levels of serum antibodies and mucosal IgA
Older animals have good recall for secondary immune responses
Lesser ability to mount primary immune response
Medically induced immunosuppression
Virally induced immunosuppression
Virus kills B and T cells
Can destroy lymphoid organs
Retroviruses in cats
Intense excessive exercise can cause stress and high levels of cortisol suppresses immune response
Tumours
Release molecules that suppress immune function
Impairment of T cell proliferation
Autoimmune disease
Autoreactive lymphocytes escape and enter lymphoid organs = no central tolerance
Immune system responds to self antigen via self reactive T and B lymphocytes
Central and peripheral tolerance disrupted
Mechanisms of autoimmunity
Predisposed factors
Environmental factors
MHC genes
Characterised by tissue damage = result of hypersensitivity reactions
Autoreactive T cells or self reactive antibodies
Excessive response to allergens
Organ specific = diabetes, thyroiditis, uveitis
Systemic immunological = rheumatoid arthritis
Pathogenesis of autoimmunity
Normal immune responses not around when self tolerance develops (recognised as foreign)
Molecular mimicry = microbe changes its antigens to be similar to host antigens, immune system targets own cells
Failure of normal regulatory mechanisms, reactive lymphocytes are not deleted in bone marrow or thymus
