Basic Immunololgy

      

Introduction to Immunology Innate immunity Cell Biology Complement system Antibody dependent responses Cell mediated immune responses Immunoregulatory mechanisms

INTRODUCTION TO IMMUNOLOGY
Immunology  Biochemical study of body defenses against parasites and microbes (bacteria, virus, fungi and parasites). Immunology important in  Biomedical, clinical, dentistry, pharmacy and basic sciences  Evolutionary trends from innate to adaptive immunity  Immunodiagnostics in the treatment and management of patients including underlying mechanisms in the disease process.

Importance cont
Clinical transplantation offers life saving benefits in diseased organs or tissues  Immunotherapeutic vaccines in the control of many diseases (viral and bacterial infections)  Anthropological studies to determine migration patterns  Paternity identity and criminal identification (employment of DNA and HLA profiles)


Historical Perspectives
First studies (1796) by Edward Jenner  Showed body could respond to foreign substances and generate the ability to defend itself in subsequent infections.  Exposure to cowpox led to resistance against smallpox.

Scientist(s) Edward Jenner (1749 1823) Lowis Pasteur (1822) Pasteur (1881 1885)

Contribution Experimented with cowpox leading to small pox vaccine in 1796 Introduced vaccine with the first attenuated virus vaccine. Germ theory led to production of attenuated vaccines against anthrax, cholera and rabies. Proposed cellular theory of immunity involving phagocytosis of wondering cells (1884). Believed humoral immunity involving antibodies and not cells. Demonstrated enhancement of phagocytosis by serum opsonins indicating a linkage between humoral and cellular immunity. First described diphtheria antitoxin (antibody) (1890).

Ilya Metchnikoff (1845 1916)

Paul Ehrlich (1854 1915 Wright and Douglas (1903)

Von Behring (1854-1917) and Kitasato (1870) Robert Koch (1843-1910)

Described delayed hypersensitivity reaction to tuberculin (Kochs phenomenon). Described a heat labile serum factor (complement) Demonstrated bacteriolysis of Cholera vibrio by antibody and complement.

Buchner (1893) Bordet (1895)

Ehrlich (1897)

Proposed receptor theory of antibody synthesis and first described neutrophils and eosinophils. Discovered human ABO blood groups system Described serum sickness Identified lysozymes Demonstrated that antibody activity resided in the gamma globulin portion of serum proteins. Showed maternal transfer of species specific immunity to infant through milk. Developed avirulent BCG vaccine. Found agammaglobulinaema in a male child. Observed that antibody production in chickens dependent on the presence of the Bursa of Fabricius in birds

Landsteiner (1900) Von Pirquet (1874-1929) Fleming (1922) Tiselius and Kobet (1938)

Paul Ehrlich (1892)

Calmette Guerrin (1920) Colonel (1952) Ogden Bruton

Glick and Change (1956)

Henson (1739-1774)

Described lymphocytes and were stained and studied by Paul Ehrlich (1879). Described phagocytic defect, chronic granulomatow disease, in children. Demonstrated that T cell recognition of antigen was self MHC restricted (1996).

Holmes et al (1966) Doherty (1974) and Zinkernagel

Susumu Tonegawa (1976)

Discovered that single immunoglobulin proteins were encoded by separate rearranging genes (1987). Discovered the principle for production of monoclonal antibodies

Kohler and Milstein (1975)

Porter (1950s and 1960s)

Showed immunoglobulins were composed of two heavy and two light chains covalently bonded (1950). Contributed to development of yellow fever vaccine

Max Theiler (1938)

BRANCHES OF IMMUNOLOGY
Basic Immunology  Immunochemistry(Physiochemical Properties)  Cellular (Cells) and Molecular (Molecules) Immunology Clinical Immunology: Immunodeficiency; Allergic and Hypersensitivity; Autoimmune Diseases; Reproductive Immunology; Immunohaematology; Tumour Immunology and ClinicalTransplantation

Classification of Immune Responses

Body defence mechanisms  Innate and adaptive immunity. Humoral responses involve  Soluble components including immunoglobulins (antibodies) and complement proteins Cellular (cells) responses  Lymphocytes, macrophages and natural killer (NK) cells (principal components)

Outstanding Features
Adaptive immunity  Recognition, narrow specificity and memory. Innate immunity  Constituted by inborn defense mechanisms


Broad specificity, no memory

Characteristic Features of Immunity

Innate Immunity Adaptive Immunity

Inborn pattern recognition receptors (PRRs) recognize pathogens PRRs possess a broad specificity for various Pathogen Associated Molecular Patterns (PAMP) Response immediate

Randomly generated receptors recognize pathogens Receptors specific for particular epitopes : B cell receptor (BCR) and T cell receptor (TCR) Response slow (3 - 5days) because of immune system stimulation

No memory of prior exposure

Memory of prior exposure exists

Innate Mediators
Innate immune responses involve several determinants (factors)  Genetics, anatomical barriers, bacterial antagonisms  Pattern-recognition receptors (PRR) Pattern Nonspecific defense chemicals  Inflammation and fever.

Innate mediators cont

Phagocytic cells
 

Neutrophils, monocytes and macrophages Basophils, mast cells and eosinophils release inflammatory mediators. Alternative pathway of complement activation


Soluble factors


Provides defense against gram-negative bacteria gram-

Interferons inhibit viral replication and activate inflammatory cells.

Genetic Determinants
Ir genes  Determine susceptibility or resistance and clinical expression of various infections HLA genetic markers  Linked to susceptibility rather than resistance to particular diseases (autoimmune diseases)

Genetic Factors in Malaria

Innate protection associated with
  

Negative Duffy (a-b) blood groups (Plasmodium (avivax) Sickle cell trait A/S Plasmodium parasite infected erythrocytes highly susceptible to toxic oxygen intermediates or radicals. Intracellular development of P.falciparum in GG6-PO4 dehydrogenase deficient erythrocytes inhibited or retarded.

Physical Barriers
Skin provides  Intact outer horny layer mechanical barrier against most pathogens and their products  Protective interface between internal organs and the environment. Dermis contains lymphocytes, mast cells and tissue macrophages  Epidermis equipped with immunocompetent cells (Langerhans, keratinocytes, epithelial cells, dendritic cells and epidermal T lymphocytes)

Physical Barriers Cont

Mucous, air-blood barriers and cilia lining the airrespiratory tract  Prevent adherence onto this portal entry by infectious agents.  High viscosity mucous and beating cilia (upper and lower respiratory tracts)  Trap and expel inhaled micro-organisms and micronoxious agents through mucociliary system mediated (mucociliary elevation)  Mucous gels are inherently sticky and the layer provides a barrier to parasite establishment like intestinal worms.

GIT barriers
Epithelium of the mouth and tongue protected by antimicrobial peptides  Stomach protected by pepsin digested antimicrobial peptides and by low pH of gastric juice  Colon supports commensals prevented from invasion of its lining by antimicrobial peptides.

Physical Barriers Cont

Human large intestine (colon) contains enormous populations of commensal bacteria with several benefits  Synthesize vitamins  Digest polysaccharides without human enzymes  Prime adaptive immunity.

Physical Barriers To Infection

Barrier site (first line of defense)

Activity

Skin sweat Skin and GI tract natural fauna (commensals)

Flushing, organic acids

Compete for niches Peristalsis, low pH, bile acid, flushing, antibacterial peptides

GI tract

Lung tracheal cilia

Mucociliary elevation

Nasopharynx, mucus and saliva, eye tears

Flushing, lysozymes

Recogniton
Innate immunity recognizes a few highly conserved structures present in many different microorganisms (pathogen associated molecular patterns, PAMPs) eg  Lipolysaccharide (LPS) or endotoxin from gramgram-negative bacteria cell wall  Peptidoglycan, lipotechoic acids from gramgrampositive cell wall, the sugar mannose (common microbial glycolipids and glycoproteins)  Bacterial DNA, N-formylmethionine found in Nbacterial proteins  Double stranded RNA from viruses and glucans from fungal cell walls.

TollToll-like Receptors
PatternPattern-recognition receptors (PRRs) recognize common PAMPs of microorganisms. Macrophages, dendritic cells and epithelial cells express  Transmembrane receptors that recognize different types of PAMPs designated Toll-like Tollreceptors (TLRs) eg  TLR-2 that binds to peptidoglycan of gramTLRgrampositive bacteria (streptococci and staphylococci);  TLR-3 binds to ds RNA of viruses TLR TLR-4 binds to LPS (endotoxin) of gram TLRnegative bacteria (Salmonella and E.coli). (Salmonella E.coli).


TLRs and Functions

Binding of gram-positive bacteria to TLR-2 and gramTLRgramgram-negative bacteria to TLR-4 TLR

Enhances phagocytosis and fusion of the phagosomes with lysosomes. TLR-5 binds lysosomes. TLRto flagellin of motile bacteria (Listeria). Listeria)
TLRTLR-7 and TLR-8 bind to single stranded RNA TLR(ssRNA) genomes of various viruses (influenza, measles and mumps). mumps).

Opsonization and Phagocytosis

Opsonization involves opsonins  Fibronectin (cold insoluble globulin)  Specific IgG antibodies (Fc-fragment), (Fc C3b and iC3b, C3 products that facilitate adherence of pathogens and membrane perturbation.

Phagocytosis process
Involve  Formation of phagosome, fusion of phagosome with lysosome and signal transduction leading to stimulation of calcium intracellular influx.  Activation of protein kinase C and phospholipase occurs by diacylglycerol releasing acid hydrolases and proteolytic enzymes.  Secondary granules contain lactoferrin, gelatinase; complement receptors (CR1 and CR3) and cytochrome B558 responsible for intracellular killing machinery

Events in Phagocytosis involve (1) organism (bacterium) attaches to pseudopodia (long membrane evaginations) leading to (2) ingestion occurs forming phagosome that (3) fuses with lysosome releasing lysosomal enzymes into phagosome and (4) digestion of ingested organism leading to (5) release of products from the cell. Source; http://www.whfreeman.com/COH1/phagocytosis.htm

Antimicrobial Peptides
Antimicrobial peptides include defensins and cathelicidins  Protect against bacteria and other pathogens. Defensins  Secreted by epithelial cells (skin, GIT, genitourinary tract and nasal passages and lungs) or by recruited leukocytes (neutrophils).  Punch lethal holes facilitated by their positive charges in penetrating the bacterial membranes. Cathelicidins  Secreted by epithelial and neutrophils with secondary alpha helix structures.  Synergistically with defensins confer protection against microbes.

Microbicidal mechanisms associated with



Microbicidal Mechanisms

Assembly of NADPH oxidase  Upregulation of cytochrome B558 in the activated neutrophils leading to production of ROI (superoxides, hydrogen peroxide)  Formation of chlorinated derivatives from hydrogen peroxide reaction with chlorides generating hypochloric acid, a microbicidal agent.

Macrophage Derived Factors and Activities Products Activity

Metabolites: Reactive oxygen intermediates (ROI) Reactive nitrogen intermediates (RNI) Eicosanoids, prostaglandins leukotrienes Platelet activating factor

Inflammation and intracellular killing Inflammation and intracellular killing Regulation of inflammation Recruitment and activation of platelets.

Cytokines: IL-1, TNF-E, IL-6 IFN-E IL-10 IL-12, IL-18 TGFAdhesion molecules: Fibronectin Thrombospondin Complement: C3b, C4b and C2b Enzymes: Lysozyme Collagenase, elactase

Inflammation Th1 activation Th1 suppression, Th2 activation Activation of NK and T cells Inflammation, tissue repair Opsonisation Adhesion, phagocytosis

Opsonisation

Degrades bacterial cell walls Matrix catabolism

Natural Killer (NK) Cells

 

Large granular lymphocytes Posses CD16, CD56, CD 94, lectin-like lectinand Ig-like receptors. IgProvide early MHC independent defence against intracellular infections (herpex group viruses, Leishmania and Listeria). Listeria). Activated by IFN-E and IFN-F in IFNIFNresponse to double stranded RNA viruses.

NK cont


Produce IFN-K that prime mJ to IFNmJ secrete TNF and IL-12. IL-

Primed NK cells kill viral and tumour cells by apoptosis and recognize target cells that fail to express class I MHC molecules on their surfaces.

Inflammation
Represents generalized response to infection or tissue damage  Designed to localize invading microorganisms and arrest the spread of the infections Characterized by 4 symptoms: Heat,  Pain  Redness  Swelling


Inflammation cont
Mast cells initiate inflammation mediated by granules exocytosed when gramgramnegative bacteria derived LPS bind TLRs.  Released histamine responsible for redness and swelling associated with inflammation.  Tumour necrosis factor-alpha (TNF-E) factor(TNFand chemotactic cytokines secreted by stimulated mast cells.

Inflammation cont
Reactive oxygen species and nitric oxide produced by activated phagocytes


Toxic to microorganisms and may also lead to tissue injury.

Macrophages and monocytes main source of interleukin-1 (IL-1) causing fever. interleukin- (ILArachidonic acid derivatives (leukotrienes and prostaglandins) potent mediators of inflammation

Inflammatory response
Lyses of blood cells and release  Bradykinin and prostaglandin cause the pain associated with inflammation  Lysosomal enzymes from damaged white blood cells  Serotonin from platelets  Prostaglandins from damaged cell membranes

Acute phase proteins

Acute phase respones  Leads to elevated production of protective acute phase proteins by greater than 100 fold (C-reactive proteins, serum amyloid A (Cprotein and 2 macroglobulin).  Acute phase proteins mediate restoration of tissue integrity by restricting damage to the injured site.

Summary of Innate Determinants

 

 

Genetics: Ir and HLA genes Anatomical (physical barriers): skin, mucociliary elevation, cough reflex, wax Antagonism: commensal bacteria Soluble factors/chemicals: alternative complement proteins, IFNs, cytokines, lysozymes, lactoferrin, antimicrobial peptides, acids Cells: phagocytes, granulocytes, NK cells

Summary cont
Inflammation and fever  PMNL recruitment and activation  Bradykinins and prostaglandins  Histamines  Lysosomal enzymes  Serotonin from platelets  Reactive oxygen intermediates  Cytokines (IL-1, TNF) (IL Acute phase proteins

Activation of Adaptive Immunity

Innate immunity may trigger adaptive immune responses thru  Antigen processing and presentation by macrophages and dendritic cells  Induction of costimulatory molecules (CD80/86) by interaction with PAMPs and TLRs, CD28 binding and full T cell activation.  PAMPs (LPS) bind to TLR on B cells enhancing their response to antigen.  Several adjuvants contain PAMPs thus augmenting further adaptive response to vaccines.

Heat shock proteins (HSP)

 

HSP linked to activation of adaptive linked immunity. Family of substances produced when cells and various infectious agents are exposed to elevated temperatures.

Immune System Components

Major histocompatibility complex (MHC) system  Chromosome, gene, haplotype, and polymorphism and super gene family molecules. Humoral immunity consists of  B-lymphocytes, bone marrow, stem cells, plasma cells, immunoglobulins (Igs) IgG, IgM, IgA, IgE and IgD classes, class switching, antibody, gammaglobulins and complement system proteins. Cell mediated immunity comprises  Lymphocytes(CD1,CD2, CD3, CD4, CD8), monocytesmonocytes-macrophages, NK cells and cytokines

Concepts and Principles

Antigenicity and immunogenicity require definition of  Antigen, immunogen, hapten, carrier, mitogen, alloantigen and an adjuvant. AntigenAntigen-antibody reactions involve the understanding of  Immune complex (IC), epitope, agretope, affinity, avidity, cross-reactivity, crossprecipitation reactions and agglutination reactions.

Antigens
Substance which induce and react with immune responses (B or T or B+T)  Proteins  Glycoproteins and lipoproteins  Synthetic peptides  Alloantigens (transmembrane proteins)  Nucleoproteins, hormones, drugs, metals  Polysaccharides and glycolipids  Pathogen (microbial) components and vaccines

Microbial derived antigens

Entire cell (virus, bacteria and tumour cells), parasite surface coat (protozoa) or products (toxins).
Conserved proteins major antigens in infection


 

Aldolase (target of human antibody response against P. falciparum) Myosin (antigen in S. mansoni) mansoni) Superoxide dismutase (M. leprae) and (M. leprae) cyclophilin (E. granulosus). (E. granulosus).

Sperm antigens
Sperm surface antigenic determinants and the seminal plasma constituents antigenic  Sperm X-Y specific antigens induces immune Xresponses. Fluid transport of spermatozoa (seminal plasma) contains variety of potentially antigenic components eg  Decapitation factors, immunosuppressive substances,  Low molecular weight nitrogenous substances, polyamines (spermine and spermidine),  Carbohydrates (fructose, inositol) and prostaglandins.

Sperm Antigen cont

Lactoferrin  Iron-binding protein Iron Adsorbs onto the sperm and forms a major component of sperm-coating antigen. sperm-

Vaccine Ags


Live attenuated viruses (Sabin for polio, measles, small pox, hepatitis, yellow fever); attenuated bacterial vaccines (BCG) or Inactivated microorganisms (Salk for polio, typhoid, cholera, Pertussis) Subunit vaccines (influenza, meningococcal and Pertussis antigens).

Alloantigens
Transmembrane proteins eg  Major histocompatibility complex (MHC) antigens  Blood group antigens.  T cell independent antigens activate B cells directly (polysaccharides) without T cells help  T cell dependent antigens activate B cells with T cell help (mainly proteins).

Superantigens
Microbial proteins produced by bacteria or viruses capable of stimulating massive T cell functions.  Trigger diseases eg staphylococcal toxic syndrome, staphylococcal food poisoning and long-term effects including long Autoimmune diseases (rheumatic arthritis, multiple sclerosis, diabetes and rheumatic fever), Kawasahi syndrome, atopic dermatitis and periodontal disorders.

Hapten and Carrier

Hapten: substance unable to elicit an immune response and yet readily react with preformed products of the induced response eg  Drugs, cyclosporin, biotin, polysaccharides, luteinizing-hormoneluteinizing-hormone-releasing factor (LHRF) and dinitrophenyl (DNT). Carrier: substance used in the modification of the hapten renderng it immunogenic eg  Staphylococcal enteroxin B; influenza virus proteins; human serum albumin (HSA); bovine serum albumin (BSA); fibrinogen and concanavalin A (Con A).

Factors Influencing Immunogenicity

High degree of foreignness  Capacity of the body to discriminate between self and non-self components. non Either a productive immune response or tolerance may be generated depending on the degree of foreignness of the antigen.

Molecular size and complexity

Epitopes or antigenic determinants  High molecular weight substances greater than 6kDa (albumin, tetanus toxoid) highly immunogenic  Between 1-6 kDa (insulin, adrenocorticotropic 1hormone) less immunogenic  Less than 1 kDa (penicillin, aspirin, progesterons) not immunogenic.

Adjuvants
Substances administered together with antigens to improve their immunogenicity. immunogenicity.  Facilitate activation of accessory cells and production of cytokines &provide signals for stimulation of lymphocytes. Most common inorganic adjuvants  Aluminium hydroxide (alum); aluminium phosphate; potassium ammonium sulphate and calcium phosphate.


Antigen Administration
Routes employed  Intravenous; intradermal; intraperitoneal; subcutaneous; submucosal and direct antigen placement on/or in the mucosa. Generally  High immune responses induced when antigens administered subcutaneously or intradermally  Intravenous applications tend to activate suppressor or inhibitory mechanisms  Subcutaneous inoculation superior to intramuscular antigen administration.

Age determinant
 

Immature/preterm and old age individuals associated with diminished immune activity Poor and high responders associated with immune response genes

Summary Immunogenicity Characteristics  Molecular size  Degree of foreignness  Chemical composition and texture  Dose and route of antigen administration  Genetic constitution and age of the individual

Haemopoitic System
Haemopoiesis occurs  First in the yolk sac and foetal liver at 6 weeks, spleen at 12 weeks and finally  Bone marrow at 20 weeks of gestation as well as in adult life. life. Lymphoid lineage pathway gives rise to  Thymus-derived (T) and bone marrow derived (B) Thymuslymphocytes Myeloid stem cells generate  Mononuclear phagocytes (monocytes and macrophages); polymorphonuclear lymphocytes; mast cells; megakaryocytes and platelets.

Fig 3. Haemopoetic Cell Differentiation: Multipotent stem cells differentiate through either myeloid or lymphoid pathways. Myeloid progenitor cells arte stimulated granulocyte-macrophage colony stimulating factor (GM-CSF) into granulocyte-macrophage proginators, which differentiate into basophils, neutrophils or eosinophils, enhanced by IL-3, G-CSF or IL-5 respectively. Monocyte- colony stimulating factor (M-CSF) stimulates granulocyte-macrophage proginators into monocytes while EPO, TPO and IL-11 stimulate myeloid proginator cell differentiation into eventual red blood cells and platelets. In the lymphoid pathway, B and T lymphocytes are derived from lymphoid progenitors through differentiation of stem cells enhanced by IL-6 and IL-7 cytokines. Myeloid differentiation pattern is further illustrated in Figs 3A and 3C.

Growth Factors and Biological Activities

Growth Factors PDGF (Platelet Derived Growth Factor) EGF (Epidermal Growth Factor) Source Biological Functions Promotes proliferation of connective tissue, glial and smooth muscle cells Promotes proliferation of mesenchymal, glial and epithelial cells May be important for normal wound healing Promotes proliferation of many cells; inhibits some stem cells and embryos Promotes proliferation and differentiation of erythrocytes Anti-inflammatory; promotes wound healing; inhibits macrophage and lymphocyte proliferation.

Platelets, endothelial cells, placenta

Sub-maxillary gland, Brunners gland

TGF-E (Transforming Growth Factor alpha) FGF (Fibroblast Growth Factor)

Common in transformed cells Wide range of cells

EPO (Erythropoietin)

Kidney

TGF-F (Transforming Growth Factor beta)

Activated Th1 cells and natural killer (NK) cells

Lymphoid Cells
Distinguished by expression of  Surface immunoglobulin (SmIg) on their cell membranes. Stimulation of B cells lead to the production of  Various immunoglubulins mediating humoral (antibody) immune responses. Lymphocyte differentiation antigens Most common cluster of human lymphocyte lineage differentiation antigens (CD) exist for both B cells and T cells.

TABLE 6: Major B Cell And T Cell Differentiation Antigens

B Cell Types Expressing the Antigen Antigen Cluster (CD) T Cell Types Expressing the Antigen

Antigen Cluster (CD)

CD 10 CD 19 CD 20 CD 21 CD 22 CD 23 CD 24 CD 25 CD 37 CD 38 CD 39 CD 40

Committed B cell progenitors; Pre-B cells All cells from early pre-B stage to plasma stage (>90% of blood B cells) Late pre-B to plasma stage cells All B cells in blood lymphoid tissues Early pre-B cell stages terminal plasma cell phase and

CD 1

Thymocytes, dendritic cells E-resetting T cells

CD 2 CD 3 CD 4 T cell (pan T) associated with TCR Helper/Inducer T cells T cell (Pan T) CD 5 T cell (Pan T): some B cells CD 6 T cell (Pan T) CD 7 Cytotoxic/suppressor T cells CD 8 Early B cells, some T cells and granulocytes Activated T and B cells CD 30 Thymocytes; lymphocytes Supp/indu/T cells; B cells CD 45R active

to

Small activated B cells in tissue B cell progenitors Activated T and B cells Late-B cell stage and early terminal plasma Germinal center B cells Mentlezone B cells CD 38 B cells; follicular dentritic cells (FDC)

CD 10

Organs of Immune System

Pluripotent stem cells in the bone marrow  Differentiate into B lymphocytes and T lymphocytes. lymphocytes. Lymphoid immune system consists of 0  Primary lymphoid (1 ) organs and 0  Secondary lymphoid (2 ) organs.)

Primary Lymphoid Organs

Three human primary lymphoid organs


Thymus, bone marrow and Peyers patches

Thymus arises from endoderm of the third and fourth pharyngeal pouches  A flat and bilobed organ above heart and below the thyroid gland  Largest relative size at birth and actual largest size at puberty.

Thymus cont
After puberty the thymus shrinks and T cell production declines  Children born without a thymus suffer from DiGeorge syndrome. Each lobule organized into  Cortex (infiltrated with rapidly dividing lymphocytes, thymocytes)  Medulla (containing visible epithelial cells). Both cortex and medulla contain stroma cells


Composed of loosely packed thymic epithelial cells, intergitating dendritic cells and macrophages

Thymus cont
Differentiation and maturation of T cells mediated by  Thymic epithelial cells and derived  Hormonal factors (E-thymosin, F4thymosin, thymopoietin, thymulin) and ILIL-7. A series of gene rearrangements select antigenic diversity of T cell receptor (TCR).

Thymus cont
Thymocytes undergo thymic education  Thymic cortical epithelial cells function in positive selection process.  T cells which bear TCR binding self-MHC selfmolecules are selected to survive and proliferate (positive selection)  Vast majority of remaining (95-99%) are (95eliminated by programmed cell death (apoptosis). (apoptosis).

Selection Process
Positive selection occurs when


Double positive T cells bind cortical epithelial cells expressing class I or II MHC molecules plus self peptides with a high affinity for survival signal. Double positive T cells bind macrophages and dendritic cells expressing class I or class II MHC molecules plus self peptides with high affinity for apoptosis signal

Negative selection occurs when



Bone Marrow
All cells of blood originate from  Pluripotent stem cells in the bone marrow (site of B lymphocyte maturation) involve stroma cells and IL-7. ILNegative selection thru apoptosis involve prepreB cells  If they do not form functional Ig molecules  Possess Ig molecules that recognize and bind to self antigens. B cells that survive negative selection (about 10%) leave the bone marrow through efferent blood vessels

Peyers patches
Located in the human wall of the intestine. intestine. PP exits with  Primary lymphoid function and another  With secondary lymphoid function


Bursa of Fabricious
BF a round, sac like structure located above the cloaca of birds  Contain epithelial cells with lymphoid cells.  Follicles divided into cortex and medulla Cortex contain macrophages, lymphocytes and plasma cells.  BF serves as site of B cell maturation and differentiation

Secondary Lymphoid Organs

Secondary lymphoid organs  Sites where lymphocytes encounter antigens  Interact with other cells and enlarge in response to antigenic stimulation.  Poorly developed in germ-free animals. germLymphoid organs morphologically divided into: cortex, paracortex and medulla

Second Lymph organ cont

Cortex (outermost layer full of B lymphocytes and macrophages arranged in primary follicles)  Paracortex (beneath the cortex, populated with T lymphocytes and dendritic cells)  Medulla (innermost with sparsely populated lymphocytes and plasma cells). Lymphocyte circulation involve


Nave T cells that bind to endothelium of venules in the paracortex T cells leave the bloodstream via conventional blood vessels and carried to the lymph nodes via tissue fluid (plasma).

A schematic presentation of secondary lymphoid organ is provided in Fig. 8. Fig.

Fig. 8 Structure of a Se condary Lymphoid Organ: Refer to text for the det ails. Source: http://www-immuno.path.com.oc.uk

Lymph Node
Lymph nodes  Encapsulated and packed with lymphocytes, macrophages and dendritic cells,  Filter antigens from the lymph,  Consist of cortex, paracortex and medulla. Cortex (outer most layer) contains  Mostly B lymphocytes, follicular dendritic cells and macrophages all arranged in clusters (designated primary follicles

Antigen Stimulation
Following antigen stimulation the follicles become  Secondary follicles consisting of  Concentric rings (germinal centres) of densely packed lymphocytes, macrophages and dendritic cells. Germinal centres  Sites of intense B cell activation and differentiation into plasma cells and memory cells. Paracortex (just beneath the cortex) designated T dependent region  Populated with T lymphocytes and interdigitating dendritic cells important in T cell activation.

Medulla
Medulla (inner most regions)  Sparsely populated by plasma cells, activated Th and Tc cells and high concentration of Igs.  Lymphocytes enter the lymph node between specialized capillary endothelial cells in the postcapillary venules designated high endothelial venules (HEVS)

Lymphatic vessels
Lymphatic vessels responsible for flow of lymph within the lymphoid system.  Fuid from the tissues flows from the intercellular tissue spaces into lymphatic capillaries and then into lymphatic vessels.  Lymph flows through regional lymph nodes and eventually enters the circulatory system

Other Lymphoid organs

Less organized aggregates of lymphoid organs include  Tonsils, appendix, Peyers patches in the lining of small intestine (gut associated lymphoid tissue or GALT)  Lymphoid tissue beneath the mucous membranes of the bronchi (bronchial(bronchialassociated lymphoid tissue or BALT) BALT)

Other Lymph organ cont



Aggregates of lymphatic tissue throughout the mucous membranes (mucosal associated lymphoid tissue or MALT) and Beneath the skin (skin associated lymphoid tissue or SALT).

Pathogens and other antigens enter tissues, transported by tissue fluid into lymphatic vessels.

Regional lymph nodes

In the regional lymph nodes  Microbes and antigens in the lymph encounter changing populations of BBlymphocytes  Filtered out and phagocytosed by fixed macrophages and dendritic cells.

Antigen Uptake
When microorganisms and other antigens enter blood  Transported by blood vessels to the spleen where they encounter changing population of B lymphocytes Germinal centers  Secondary follicles found in tissues containing activated cells following continuous exposure to antigen.  Sites of antigen driven activation of B cells in secondary lymphoid organs  Leads to the development of active B cells, plasma cells and macrophages.

B Cell Priming
Antigen specific B cells  Activated in the T cell zones of secondary lympoid tissue and  Migrate to B cell zones to form germinal centers within the reticulum of follicular dentritic cells (FDCs).  Primary follicles consist of a  Network of FDCs in which no antigenantigendriven process is occurring.

Lymphocyte Priming
FDCs

Depot of native unprocessed antigen, in an immune complex form  Hold Ag in a nondegraded form for long periods. Follicular B cells can  Take up antigen from FDCs and process and present in a form that activates T cells.  T cell B cell collaboration essential in the development of germinal centers


MHC System Genetics

9 MHC genes and include  HLA-A, HLA-B, HLA-C, HLA-DPA 1, HLAHLA- HLA- HLA- HLAHLADPB1, HLA-DQA1, HLA-DQB1, HLA-DRA HLAHLAHLAand HLA-DRBI. HLAMHC region divided into three regions  Class I (HLA-A,B and C) (HLA Class II (HLA-DP, DQ and DR) and (HLA Class III genes encode complement components (C2 (C2, C4 and Factor B), cytokines (TNF- ) (TNF MHC genes display high levels of allelic diversity (polymorphism).

Class 1 MHC Molecules

Class I MHC molecules consist one long heavy chain of 346 amino acids and a short one of 99 amino acids. Heavy chain consists of five regions  A transmembrane domain and a cytotoplasmic domain (with the C terminal). terminal).  Three extracellular domains including N terminal,  Protein molecule beta-2 microglobulin (F2-M) beta(F nonnon-covalently attached to the heavy chain.

Class I polymorphism
Class I MHC region encodes HLA-A, B HLAand C molecules.


HLAHLA-A, -B and -C heavy chain genes are highly polymorphic with known 60, 138, and 40 alleles, respectively.

Class II MHC Molecules

Human class II molecules are designated HLA-D and HLAclass II MHC region encodes  Molecules HLA-DP ( , ); HLA-DNA; HLA-DOB; HLA); HLA-DNA; HLA-DOB; HLAHLA-DQ ( , ) and HLA-DR ( , ). HLA). Other encoded molecules are  Transporter proteins (TAP 1 and 2) involved in antigen presentation; presentation;  Low molecular weight polypeptides (LMPs and 7) and HLAHLA-DM gene products. products. MHC Class II molecules are found only on  Professional antigen presenting cells (APCs) including macrophages/monocytes, dendritic cells, activated T cells and
B cells. cells.

Class III MHC Molecules

Class III MHC region  Located in the central part of the MHC system and  Contains genes which code for complement components


Factor B (BF), C2, C4b and C4a]; tumour necrosis a]; factor (TNF); lymphotoxin (LT); heat shock (TNF); (LT); proteins (HSP 70-1 and HSP 70-2). 7070-

Proteins produced also include 21 hydroxylase enzyme (21-Ohase), deficient in (21patients with congenital adrenal hyperplasia.

MHC System Components

Class MHC Loci Tissue distribution of MHC gene products

HLA-A,B,C

All nucleated cells, lymphoid, epithelial and mesenchymal tissues

II

HLA-DP, HLA-DNA, HLA-DOB, HLA-DQ HLA-DR

Accessory antigen presenting cells like monocyte-macrophage lineage cells, B-cells and activated T-cells

III

Central MHC region

C2,BF, C4b, C4a, and 21-hydroxylase enzyme (21- Phase), tumour necrosis factor (TNF), lymphotoxin and heat shock proteins

MHC Region Characteristics

MHC Region posses characteristic features  Multiplicity, high polymorphism and extensive heterozygosity. High polymorphism  Genes in the MHC loci exist in different or alternative multiple forms designated alleles.

HLA Specificities (Alleles)

A A1 A2 A3 A9 A10 A11 AW19 A23(9) A24(9) A25(10) A26(10) A28 A29(W19) A30(W19) A31(W19) AW34(10) AW36 AW43 B B5 B7 B8 B12 B13 B14 B15 B16 B17 B18 B21 BW22 B27 B35 B37 B38(16) B39(16) B40 BW41 BW42 C CW1 CW2 CW3 CW4 CW5 CW6 CW7 CW8 CW9(W3) CW10(W3 ) CW11 D DW1 DW2 DW3 DW4 DW5 DW6 DW7 DW8 DW9 DW10 DW11(W7) DW12 DW13 DW14 DW15 DW16 DW17(W7) DW18(W6) DW19(W6) DR DR1 DR2 DR3 DR4 DR5 DRW6 DR7 DRW8 DR9 DRW10 DRW11(W7) DRW12(5) DRW13(W6) DRW14(W6) DRW15(2) DRW16(2) DRW17(3) DRW18(3) DRW19(3) DRW52 DR21 DRW53 NB. The alleles that have been reported and provisionally given a number pending verification and official recognition are referred to by the letter W before the number such as Aw33(w19). DQ DQW1 DQW2 DQW3 DQW4 DQW5(W1) DQW6(W1) DQW7(W3) DQW8(W3) DQW9(W3) DP DPW1 DPW2 DPW3 DPW3 DPW5 DPW6

Inheritance of Haplotypes
MHC genes are co-dominant and the alleles equally coexpressed. Given a simple Mendelian pattern of inheritance,  Distribution of MHC haplotypes in a family: 25% identical; 25% non-identical and 50% partially nonidentical. Linkage disequilibrium  Certain gene combinations occur more frequently than is expected given random Mendelian combinations and permutations  HLA-B8 and HLA-DR3 alleles more closely HLAHLAlinked (97%)

Alleles in Linkage Dysequilibrium

A1, A1, A2, A2, A11, A29, B8 BW17 BW12 BW16 B5 B12 AW30 AW33, CW1, CW1, CW3, CW4, CW5, B13 BW14 B5 BW16 BW15 BW35 B12 DW1, DW, DW4, BW35 BW7 BW15

Importance of MHC Antigens

MHC antigens-a group of cell membrane antigenscomponents  Expressed on cells by certain individuals in a population referred to as alloantigens. Clinical transplantation  Matching donor HLA antigens with those of recipient to avoid induction of graft rejection reactions

Patenity identity
HLA gene products of a child compared with

Putative candidates to ascertain the biological father in a given ethnic or racial group.  Match between the HLA antigens of the child and candidates will establish patenity.


Anthropological studies
HLA closely linked and their distribution in the population markedly restricted. HLA polymorphism shows


SubSub-Saharan African populations genetically most diverse (founding human race probably of the African descent). Liberians appear more closely related to a distinct Malawian population than to their West African neighbours.

Migration patterns of peoples in populations determined through HLA characterization.



Forensic use of HLA typing

DNA probes used in resolving inconclusive blood group typing or finger print results in criminal investigations. HLA and disease association Theories include  Linkage disequilibrium,  Antigenic mimicry  Role of various infectious agents



Strong HLA disease association exists between M.tuberculosis and HLA-Bw 15, M. leprae infections with HLAHLAHLA-DR3.