INFECTION AND IMMUNITY

Immunity is the resistance exhibited by the host towards injury caused by microorganisms and their products. Protection against the infectious diseases is only one arm of the immune response, since the main purpose is the reaction of the body against foreign antigen. Immune cells and the responses they generate. There are 2 types of immunity: 1. INNATE - this is also called the NATURAL IMMUNITY since it is present even at birth. This response will not increase upon repeated exposure to the antigen. It is composed of skin, mucous membranes, secretions (saliva, tears), phagocytic cells and NK cells. Innate immunity also allows elimination of foreign substance like a bacteria, without previous exposure. It is enhanced by natural antibodies or natural cytotoxic cells like macrophages, neutrophils, eosinophils and NK cells. It is nonspecific since it is effective against a wide range of infectious agents. Factors which affect innate immunity: Age (infants are affected more by infections since they have an immature immune system); Sex(death rate is higher in males, but there is no marked differences in susceptibility between males and females);Hormonal influence (there is decreased resistance to infection in DM);Nutrition (poor nutrition leads to increased susceptibility to infections). MECHANISMS OF INNATE IMMUNITY A. MECHANICAL BARRIERS AND SURFACE SECRETIONS The intact skin and mucous membranes of the body give avery good protecton against pathogens. The skin is a resistant barrier because of its outer cornified layer consisting mostly of keratin which microorganisms cannot digest. Also, the dry condition of the skin plus the high concentration of salt in the sweat is lethal. Sebaceous secretions and sweat also contain bactericidal and fungicidal fatty acids. The sticky mucus of the respiratory tract acts as a trapping mechanism for inhaled pathogens. Nasal secretions and saliva also contain mucoploysaccharides which can block some viruses. The washing of tears and flushing of urine are effective in invasion or organisms.

There are also normal bacterial flora in the epithelial surfaces which are beneficial: they occupy the space which cannot be occupied anymore by pathogens, they have used up nutrients; they produce byrproducts which are harmful to pathogens.

B. HUMORAL DEFENSE MECHANISMS A number of microbicidal substancesare present in the tissues and body fluids. Lysozyme is a protein found in high concentrations in neutrophils and in body fluids, except for the CSF, sweat and urine. It functions as a mucolytic enzyme against gram (+) cel wall, causing their lysis.it may calso cause the intracellular destruction of gram (-) bacteria. Basic polypeptides like spermine and spermidine can kill tubercle bacilli and staphylococci. Acute-phase proteins rise highly during an infection.Endotoxin can cause the release of Interleukin-1, which in turn, stimulates the liver to produce the acute-phase proteins. The most commonly known is C-reactive protein(CRP). This bins to the bacterial cell wall. It also activates the classical complement pathway. Alpha 1-antitrypsin, alpha 2- macroglobulin, fibrinogen and amyloid are examples. Interferon are factors released by cells in response to a viral infection. It protected other cells of the same species from being infected by that virus. C. COMPLEMENT These are about 30 proteins found in the serum in their inactive forms, but can be activated to form an enzyme cascade with the following end result: Opsonization Cellular activation Cell lysis There are two pathways: the classical and the alternative pathway The classical pathway is the main antibody-directed mechanism for complement activation; it is rapid and most efficient. The complement recognizes the antigen-antibody complexes (either IgM or IgG), then bind to C1, which then activates the cascade.

The alternative pthway protects the body from pathogens in the absence of antibody; it is activated by the bacterial surface itself. The final result for both pathways is the formation of the membrane attack complex, which is formed from the reactions among C5, C6, C7, C8 and C9. D.CELLS Phagocytes are cells which engulf micro-organisms as they enter tissues, fluids or the bloodstream. Mononuclear phagocytes in the blood are called monocytes, macrophages in tissues, histiocytes in connective tissue, mesangial cells in the kidney, osteoclasts in bone, microglia in the brain, and sinus-lining macrophages in the spleen, lymph node and thymus. Phagocytes are: Actively phagocytic Contain digestive enzymes to degrade ingested material Process and present antigens Produce molecules that stimulate lymphocyte differentiation into effector cells. Phagocytosis involves recognition, binding, ingestion and digestion of the cell.

Products of injured tissues, blood factors, leukotrienes and histamine from mast cells and neutrophils, bacterial products

Chemotaxis

Phagocytes have receptors for the Fc portion of immunoglobulins and some components of the complement cascade; these act as OPSONINS on the bacterial cell surface and enhances the digestion process

The foreign particle is engulf by the cell membrane and internalized as an ENDSOME or PHAGOSOME

LYSOSOMES in the phagocyte fuse with the phagosome forming the PHAGOLYSOSOME

Enzymes are released, killing the bacterial cell:superoxide dismutase and myeloperoxidase, lyzozyme and elastase, lactoferrin

NATURAL KILLER CELLS recognize changes in virus-infected cells and destroy them by an extracellular mechanism. This is present even without prior exposure to the infectious agent. It is performed by cell large granular lymphocytes, and by cells with T cell markers. Its activity is enhanced by Interferon. They have also been implicated in the host defense against cancer cells. EOSINOPHILS are polymorphonuclear cells with a blobed nucleus and cytoplasmic granules. They are present in very low levels in the blood, but their numbers increase with parasitic infections and allergies. They are not efficient phagocytic cells, but their granules contain molecule which are toxic to microorganisms. These released molecules target large parasites such as worms.

E.TEMPERATURE Pyrexia which follows many infections can also be a source of control. It is mediated by interleukin-1, which is produced by macrophages. It is also known as the endogenous pyrogen. F.INFLAMMATION This is the reaction of the body to injury, such as invasion by a infectious agent, exposure to a toxix chemical, or physical trauma. The five signs are redness, heat, swelling, pain and loss of function. The molecular events that happen in inflammation are: Vasodilatation Increased vascular permeability Cellular infiltration. The most common mediators of infection are: Mediator Histamine permeability Main source mast cells Basophils Function vasodilatation, increased vascular

smooth muscle contraction

Kinins Prostaglandins

plasma neutrophils Eosinophils Monocytes Platelets

same as above, plus pain vasodilatation, increased permeability, pain

Leukotrienes adherence and

neutrophils

same as histamine, plus induce cell Chemotaxis

Complement cascade C5 which is for

plasma

cause the mast cells to release chemotaxis

Plasmin Cytokines factors

plasma lymphocytes Macrophages

breaks down fibrin, kinin formation chemotactic factors, colony-stimulating macrophage activation

ACQUIRED IMMUNITY develops late in fetal life, and development continues into childhood. Continued exposure to the antigen stimulates acquired immunity. The response is specific to an individual antigen because of the antigen-specific recognition by surface antibody on B cells and by T-cell receptors on T cells. Acquired immunity may have an anamnestic response, in which subsequent response to a previously recognized antigen is more magnified due to the production of memory T and B cells. There are two types of acquired immunity: active and passive. Active immunity is induced after contact with foreign antigen, such as micro organisms and their products. This contact may be subclinical or clinical infection. Immunization with live or killed infectious agents, exposure to microbial products and transplantation of foreign cells is a form of active immunity. Long term resistance , although slow in onset, occurs in acquired immunity.

Passive immunity is the prompt availability of large amounts of antibodies, and is transmitted by antibodies or lymphocytes which have be preformed in another host. There is however, a short life span,and the possibility of hypersensitivity reactions since it is formed in another species. An example is the transfer of maternal antibodies to the fetus.

An ANTIGEN is a substance that is capable of provoking the lymphoid tissues of an animal to respond by generating an immune reaction. This reaction is specifically directed against the inducing substance only and not at any other related substances. The response is just to some parts of the antigen, or individual chemical groups, and not to the entire antigen itself. This is termed the specificity to antigenic determinants or to epitopes. Thus ,since a micro-organism may have several epitopes, more than one antibody will react with that particular organism. Functional classification of ANTIGENS 1. Immunogens, substances that are able to elicit an immune response by themselves 2. Haptens, molecules that are able to react with antibodies, but are unable to stimulate their production directly; low molecular weight; become active only when attached to carriers, which are large molecules A substance that act an as antigen in one species may not necessarily produce a reaction in another species. Antigens, in general, must have a molecular weight of more than 5000. However, even lower molecular weight substances like glucagon , can function as an antigen with they are combined with an adjuvant, which in turn, causes additional stimulus to the immune response. Aspirin, penicillin and formaldehyde may also act as antigens although they have low molecular weight.

ANTIGENIC DETERMINANTS A response to antigen involves the specific interaction of components of the immune sysem, antibodies and lymphocytes, with epitopes on the antigen. The lymphocytes have receptors on their surface that function as the recognition unit:

Surface-bound immunoglobulin on B cells T cell receptor on T cells The part of the lymphocyte that attaches to the epitopes is called a PARATOPE. Antigenic determinants may be formed in two ways: 1. May be contained in a single segment of primary sequence(SEQUENTIAL EPITOPES) 2. May be assembled from residues far apart in the sequence, but are brought together in close proximity due to folding of the molecule(CONFORMATIONAL EPITOPES)

ANTIGENIC SPECIFICITY is based on: 1. Subtle chemical differences between molecules 2. Foreignness of a substance to an animal can depend on the presence of chemical groupings that are not normally found in the animal’s body. The ability of the antibody to form a high-affinity interaction with an antigen depends on the two molecules coming together in a very precise bond. The antibody may bind to another, structurally similar antigen, but the bond will be weaker. CROSS REACTIVITY occurs when the antibody to a particular antigen is found capable of combining with the with an apparently unrelated antigen. For example, Forssman antigen is found in RBC and pneumococci and salmonella. It is thus termed a HETEROPHILE ANTIGEN. The antibodies then clump the rbc, thus they are called ISOHAEMAGGLUTININS.

IMMUNOGLOBULINS 19TH CENTURY, von Behring and Kitasato in Berlin discovered that the serum of an appropriately immunized animals contained specific neutralizing substances. They were called ANTIBODIES OR IMUNOGLOBULINS. General characteristics of antibodies: 1. Glycoprotein

2. Present in the serum and body fluids 3. Induced when immunogenic molecules are introduced into the host’s lymphoid system 4. Reactive with, and are specific, to the antigen that induced their formation Antiserum contains antibodies formed in reaction to the antigen. This is then separated by electrophoresis into alpha, beta and gamma fractions. Most of the immunoglobulins will be in the gamma fractions, hence they are also called GAMMAGLOBULINS.

ANTIBODY STRUCTURE All antibodies have the same basic structure: 2 light chains and 2 heavy chains. The two light chains are one of two types: either a Kappa or a Lambda. The molecular weight is about 25000. They are composed of 2 areas, which are called DOMAINS, and these are approximately 110 amino acids. One end of the chain is constant, and is identical in all members of the isotype. This is termed the CL region. The other end shows sequence variation, and is called the VL region. The heavy chains determine the isotype, and have a molecular weight of 50000 to 70000. They also have two domains which show sequence variation ( VH); while the other domain is similar for all isotypes (CH). The tertiary structure of the VL and VH regions determines the shape of the antigencombining site or paratope. The two chains are held together by disulfide bridges and non-covalent interactions. Since the two light and two heavy chains are identical, there will be two binding sites for the antigen (2 paratopes ) at the amino- terminal or N- terminal. The carboxyl – terminal or C-terminal at the other end of the antibody will be the same for all the members of the isotype. It is responsible for the biological properties of the isotype. There is an area in heavy chains which is called the HINGE REGION. Enzymes may break up the antibody, dividing them into Fab and Fc region. The Fab region (fragment antigen binding) contains the paratope. The Fc (fragment crystallizable) is similar for all the antibodies of the same isotype. The differences from the other isotypes define the biological activity of the antibody.

The variability in the amino acid sequence in the variable domains of the light and heavy chains is not found over their entire length, but restricted to short segments only. These segments show variation, thus are called HYPERVARIABLE REGIONS. These are the areas that come in direct contact with the antigen. 5 CLASSES OR ISOTYPES 1. IgA 2. IgM 3. IgD 4. IgG 5. IgE

IgG This is the major immunoglobulin of the serum, as well as the CSF and the lymph, making up almost 75%. It has a molecular weight of 150,000 in humans. It is also the major antibody of the secondary response, and is an important defense against bacteria and viruses. It is the only antibody to cross the placenta, so it is the highest antibody in the newborn, protecting it in the first 4-6 months of life. The levels may be abnormally raised in Malaria, Kala-azar and Myeloma. Its functions are: 1. Activates the complement 2. Acts as an opsonin 3. Mediates antibody-dependent cellular immunity

IgA This is the predominant antibody in seromucous secretions (saliva, tears, colostrums, respiratory, GIT and GUT secretions). Its functions are: 1. To specifically bind antigen at mucosal sites

2. Protect mucous membranes from attack by bacteria and viruses.

IgM This is the antibody produced during the immune response, especially to bacteria and viruses. Due to its large size, this isotype is mainly confined to the intravascular pool. The presence of IgM in the newborn may indicate intrauterine infection such as Syphillis, Rubella, HIV infection and Toxoplasmosis. Its functions are: 1. Most efficient in agglutination, complement fixation and other antigenantibody reactions 2. Can be produced in a fetus with an infection.

IgD This account for less than 1% of circulating antibodies. It is very susceptible to proteolytic attack , thus has a very short half-life in serum. Its functions are: 1. B cell antigen receptor 2. Marker for mature B cells 3. Occurs on cells of some lymphatic leukemias

IgE This is present in very low levels in the blood; found on the surface of mast cells and basophils, which possess a receptor specific for the Fc part of the immunoglobulin. It is also called the REAGINIC ANTIBODY since it is associated with allergic response and immediate hypersensitivity reaction. Functions: 1. It is associated primarily with allergic and parasitic infections.

ABNORMAL IMMUNOGLOBULINS Bence Jones protein is found in multiple myeloma. It can be identified in the urine by its characteristic property of coagulation when heated to 50C, but redissolving at 70C. CRYOGLOBULINEMIA is a condition where a gel or precipitate forms on coolng the serum, which redissolves on warming. This may not always be involved in diseases, but it is often found in myeloma and SLE.

There are 3 terms to describe variants among antibody molecules. 1. Isotype refers to the subclass of the immunoglobulins ( G,A,M,E,D) 2. Allotype refers to the genetically determined difference in a molecule between two members of the same species; this is also called genetic polymorphism. It is due to the substitution of only one or two amino acids in the constant regions of heavy and light chains. They have no biological significance. 3. Idiotype refers to the unique, individual differences between antibodies of different antigen binding specificities. They are determined by the structure of the variable regions of antibodies.

MAJOR HISTOCOMPATIBILITY COMPLEX This is a collection of highly polymorphic genes encoding the proteins that regulate an immune response. These genes include the class I and class II cell surface proteins, and the class III which encode for the complement proteins. In humans, the MHC is called HUMAN LEUKOCYTE ANTIGENS (HLA). And are found on the short arm of chromosome 6. The HLA proteins are present on the cell surfaces that enable the T cells to recognize and bind antigenic peptides(immune recognition). HLA CLASS I ANTIGENS are membrane glycoproteins on the surface of ALL nucleated cells and platelets. They are necessary for antigen recognition( on viruses, intracellular bacteria, parasites, tumor antigens) by CD8+ cytotoxic T lymphocytes. Pepetide of normal intracellular proteins are bound to HLA class I proteins, and displayed on the cell surface. Proteins made by intracellular viruses, bacteria, parasites or neoantigens made by tumor cells will also be displayed on

HLA class I proteins. In humans, the 3 types of class I genes are called HLA-B, HLAB, and HLA-C antigens. HLA CLASS II ANTIGENS are proteins expressed on more restricted set of cells, including the antigen-presenting cells (Langerhans cells, activated macrophages)B cells, and thymic epithelial cells involved in T-cell maturation. They are necessary for antigen recognition by Th cells. In humans, the class II genes include HLA-DR, HLA-DQ and HLA-DP. Class II proteins are found on cells that interact directly with the Th cells, including B cells, monocyte-macrophages, langerhans cells, dendritic cells and thymic epithelium. The proteins made by extracellular bacteria or parasites or injected vaccines will also be displayed on HLA class II proteins. HLA CLASS III ANTIGENS. Class III genes encode for the complement components or regulators of serum complement component complex.

Many diseases are associated with an increased frequency of the HLA antigens. Ankylosing spondylitis (HLA-B27), rheumatoid arthritis (HLA-DR4), and IDDM (HLADR3 and HLA-DR4). Tissue typing for organ transplantation involves matching the HLA Class I and HLA Class II antigens.