Immunology : Lecture 1 : Bolded Terms & Concepts 3 steps to a successful infection / points to defend: (1) penetrate the host

(2) attach / adhere to the host

(3) thrive in the host

Can think of the 3 steps above as STAGES as well. 4 rounds to every stage: bug attacks host fights back (defends) host counter attacks bug defends Layers: (1) physical barriers (2) physiological barriers (3) inflammation (4) adaptive immunity Scenario: Imagine that you are getting out of your tub, and as you step onto the deck, you get a large splinter in your big toe. On that splinter are many bacteria, and within a few hours, youll notice that the area around where the splinter entered is red and swollen. STAGE I. EARLY in the Infection (Step 1: penetrate host) Physical Barriers first line of defense that are physical walls between us and the outside; the three most important barriers are skin, mucous membranes, fluids. Mucous membranes are like doorways from the skin into the lungs (respiratory), the genitourinary, oral/gut areas. These membranes prevent lateral movement (penetration). The mucous these membranes make can be thought of as fluids such as urination, salivation. Physical Barrier Strategies defense mechanisms of the HOST involve either pushing the pathogen out of the body, sloughing cells layers, or washing out the pathogen with fluid flow Bug Attacks (BA) + Host Response (HR): 1. Skin: BA: lesions, hair follicles, sweat glands (can remain local or spread systematically) HR: intact epidermal layer, exfoliation, sweat 2. Oral/Gut: BA: contaminated food / water / fingers (think: you digest these things!) HR: salivary flow, intact mucosa and mucous flow, peristalsis (wave-like contractions that move food down the digestive tract!) 3. Respiratory: BA: inhaled airborne or aspirated pathogens (nose/throat to airways to lungs) HR: vibrissae in nasal passage (specialized hairs), intact mucous & flow, abrupt directional changes, cough reflex, cilia 4. Genitourinary: BA: urethra, bladder, kidneys HR: urine and mucous flow, sloughing epithelium If the bug has defended itself correctly, then it will be able to bypass the physical barriers by using invasive and tissue-targeted strategies: a. digestive enzymes = breaks down tissue constituents + destroys integrity b. adhesins = surface structures that allow pathogens to stick to host tissues, can be highly specific c. motility = various means to move through host tissues and fluids, some are sophisticated organelles

Breaking down the 1st line of defense, means the bug is inside and now needs to hang on. Leads to Step 2: Adhesion or STAGE II

Immunology : Lecture 1 : Bolded Terms & Concepts STAGE II: EARLY TO MIDWAY in the Infection (Step 2: Adhere to host) Bug Attack / Defense Colonizes the establishment of the microbial pathogen at the site of entry; one bug will start to grow and multiply from 1 cell (genetically identical). 1 bug = 1 colony Virulence Factors characteristics that the microbial pathogen expresses to promote its pathogenic potential = molecules/factors that are able to cause disease + (most) influence their hosts function to allow/promote the pathogens survival. virulent = potent pathogen Use of genetic strategies which lead to phenotypic changes Adaptation: relatively reversible; involves turning on and off genes (i.e. lac operon) Mutation: relatively irreversible alteration of genes via nucleotide changes; random mutate event so could take > time Host Response Physiological Barrier Strategies defense barriers of the HOST are designed to deoptimize the growth conditions of the pathogen, thereby preventing further growth and expansion of potential microbial pathogens (i.e. affects survival!) Strategies include changing body temperature (i.e. fever), pH, oxygen content, nutrient requirements. Skin: Dryness, low pH (5.5), low T, high O2, high [salt], resident bacteria (native flora in gut) Oral/Gut: low pH, antibacterial saliva/mucous, -lytic enzymes, resident bacteria, bile Respiratory: T gradient (increasing upper to lower), high O2, antibacterial mucous Genitourinary: low pH, antibacterial mucous

HR: IF the bug is able to get past the physiological barriers, then the 2nd line of defense, the IIS comes in: *Innate Immunity After an invader has breached the physical barriers, it is greeted by the innate immune system = second line of defense. Inflammatory Responses cellular and systemic response designed to quickly interfere with the spread of an infection; critical very early on in an infection, from 0-4 days in holding back infection. Initiated rapidly, reaches levels of intensity = current infection. Requires (1) leukocyte recruitment and (2) leukocyte response Classic Signs of Inflammation: blood vessel dilation + increased vascular permeability and RHSP = redness + heat + swelling + pain = red hands show pain Leukocytes white blood cells; T cells + B cells (1) Recruitment moving the leukocytes or designated particles to a particular site (i.e. from blood to the site of infection) by a multi-step process of interaction and transmigration involving endothelial cells. (includes leukocyte transmigration discussed on next page) (2) Phagocytosis the ability to bind and then ingest + digest foreign or damaged particulate matter. (2) Cytokines hormone-like communication molecules that promote cellular migration, growth and/or activation of additional cells of the immune system.

Immunology : Lecture 1 : Bolded Terms & Concepts

---- ORIGIN OF CELLS ---Hematopoiesis developmental process that results from the differentiation of stem cells into all of the other cells of the blood. Lymphocytes originate from stem cells in bone marrow Stem Cells cells characterized by their ability to 1) self-replicate (make more of themselves) and 2) to differentiate into other cells by receiving signals from the bone marrow to turn off their stem cell genes and turn on other genes. Megakaryocyte lineage = platelets Erythroid lineage = erthryocytes >>>>> Myeloid lineage = monocytes + granulocytes Lymphoid lineage = lymphocytes Myeloid Cells contains two families of cells: 1) monocytes 2) granulocytes. These cells are prominent in non-adaptive inflammatory response Monocyte sublineage/family of myeloid cells; make up 5% of white cells (leukocytes); can be given signals to differentiate further to do their job (immature and mature versions); function is phagocytosis. They often reside in many tissues but are involved in recruitment during inflammation. Macrophage a type of leukocyte made from monocytes; terminally differentiated = cannot be anything else, cannot differentiate into anything else; main function is phagocytosis Granulocytes sublineage/family of myeloid cells: 1) neurophils, 2) basophils, 3) eosinophils. These cells are called granulocytes because they are very granular looking; see a bit of crystalline-like shape to them. Neutrophil also called polymorphonuclear leukocytes (PMN) because they have many nuclei, which are lobe-shaped and no two really look the same. Make up 60-70% of white cells (leukocytes) in the blood = most abundant. Also recruitable; function is phagocytosis.

Immunology : Lecture 1 : Bolded Terms & Concepts

---- HR: (1) STEPS of RECRUITMENT ---Leukocyte Transmigration / Extravasation movement of white blood cells from inside the blood to outside the blood vessels and to the site of infection. Has several steps: 1) chemoattraction, 2) primary adhesion (tethering), 3) secondary adhesion (arrest), 4) diapedisis and transmigration Chemoattraction signal that the leukocyte is supposed to go somewhere; mediated by chemokines Chemokines small bioactive particles produced by a variety of cell types, including phagocytes, fibroblasts, and endothelial cells that compose the cell walls of blood vessels, and serve as chemoattractants and activators of innate and adaptive immune cells. Chemokines are often released by damaged tissue or from the endothelium of the blood vessel and convince the WBC to move where it needs to. Chemokines are divided into families based on their common structures or cellular targets (i.e. neutrophils, macrophages, lymphocytes). Inflammatory Cytokines induces leukocyte movement from the blood to infected tissues in response to chemokines; accelerates the activities of oxidative and non-oxidative phagocytosis Adhesion Molecules surface membrane structures that promote interaction = binding between cells as they collaborate during a particular function (i.e. adhesion molcules on WBC + on endothelial cells to allow them to interact). Two groups: 1) selectins and 2) integrins Selectins family of molecules that participate in the initial stages of cell adherence; allows leukocytes to bind to the inner wall of the blood vessel with low affinity; produced by WBC; expression induced by inflammatory cytokines; mediates Primary Adhesion. Primary Adhesion (Tethering) initiates leukocyte extravasation / transmigration; results in a slowing of the forward movement of leukocytes. Leukocyte rolling (to a stop) is observed during this process. Integrins family of molecules required for more stable high affinity interactions; next step to bind WBC more tightly to the wall. It probably involves altering the 3D structure to result in Secondary Adhesion. Required for cell to cross epithelial barrier: b/w cells and b/w cell and ECF. Secondary Adhesion (Arrest) firm stable binding to endothelial cells prevents further leukocyte movement through the bloodstream; is dependent on gradients of chemoattractants outside the blood vessel; this allows for translocation to take place Diapedesis the actual process of translocation through / around endothelial cells into sites of inflammation (ENTER) Cytokinesis process of moving the WBC through the endothelial wall itself

Immunology : Lecture 1 : Bolded Terms & Concepts

---- HR (2) STEPS of ACTIVATION + RESPONSE ---Phagocytosis the ability to bind and then ingest + digest foreign or damaged particulate matter. Endocytosis the ingestion of smaller quantities Pathogen Associated Molecular Patterns (PAMPs) molecular groupings or patterns found on the surface of the (microbial) pathogen. Pattern Recognition Receptors (PRR) expressed on the surface of the phagocyte (leukocyte!) which recognize PAMPs; includes a family of PRRs called Toll-Like Receptors; certain important signals for cytokine production are also given through PRRs Toll-Like Receptors (TLRs) extremely important for recognizing molecular patterns expressed by a variety of bacterial and fungal polysaccharides. Non-Oxidative Destruction (Phagocytosis) involves mechanisms like 1) acidified environment (pH 3.5-4.0), 2) antimicrobial peptides (defensins), 3) enzymes (lysozymes), 4) competitors (lactoferrin = soaps up all the iron from the pathogen depriving it of appropriate nutrients) Oxidative Destruction = Respiratory Burst involvement of oxygen to make potent compounds like superoxide (O2-), hydrogen peroxide (H2O2), hypochlorite ions (OCl) which is extremely toxic and reactive to anything. Also nitric oxide (NO) Mechanism: O2 -----E----- > Superoxide (O2-) by E = NADPH oxidase Superoxide (O2-) -----E----- > hydrogen peroxide (H2O2) by E = superoxide dismutase Hydrogen peroxide (H2O2) -----E----- > hypochlorite ions (OCl) by E = myeloperoxidase Also: Nitric Oxide (NO)

Immunology : Lecture 1 : Bolded Terms & Concepts BA: uses a variety of anti-phagocytic strategies Capsule in the line of bug defense, successful pathogens will overcome inflammatory barriers by producing a polysaccharide capsule that repels phagocytes by preventing attachment from occurring. Phagosome-Lysosome Fusion in the pathway of destruction by a phagocytosis, the final step of destruction is obstructed by the bug by production of factors to prevent this fusion from taking place and / or neutralization of lysosomal products. HR: overall, called Acute Phase Response! Opsonization decoration = coating of particles with particular substances = opsonins that increase the efficiency of phagocytosis; key opsonins are 1) specific antibodies and 2) complement proteins Acute Phase Response elevated of levels of acute phase proteins due to regulated release by hepatocytes. This pathway is triggered as the result of hepatocyte activation by cytokines (IL-1, IL6, and TNF-alpha) related by phagocytes. Hepatocytes then release a family of proteins referred to as acute phase proteins. Acute Phase Proteins released by hepatocytes; examples are c-reactive proteins and mannan-binding lectin. These two particular acute phase proteins often function as opsonins C-Reactive Protein binds to phosphorylcholine portion of certain bacterial and fungal cell wall lipopolysaccarhides Mannan-Binding Lectin binds to mannose residues on many bacteria which are normally masked by other sugar residues on mammalian cells. Opsonins substances that coat particles and make them more attractive to phagocytic cells and thereby increase the effectiveness of phagocytic activities; they also mimic the activation of the first components of the classical complement cascade! This results in the generation of the downstream bacteriocidal complement components Cytokine Production production of mediators that enhance phagocytic activities = activate to a higher level such that say, fusion of phagosome-lysosome happens anyway despite the bugs defense. (i.e. interferon-gamma!)

Immunology : Lecture 1 : Bolded Terms & Concepts STAGE III: LATER in the Infection (Step 3: Thrive in host) The Bug attacks again and this time does damage to the host tissue due to further microbial growth and release of toxic products. So now, the bug has really made his mark and is in for the long haul. This is when Specific Adaptive Immunity kicks in. Specific Adaptive Immunity highly specific immune response system that has three hallmarks: (1) diversity, (2) specificity and (3) memory Lymphocytes key players in specific adaptive immunity; also a type of leukocyte; B cells + T cells (1) Diversity a diverse array of cells, recognition structures (receptors) and soluble mediators to confer protection from a diverse array of biological threats = diversity in 1) recognition, 2) protective mechanisms, 3) regulation (2) Specificity each biological threat, with its specialized pathological characteristics must be recognized independently (3) Memory the immune system must remember previous exposure to antigens Vaccination based on increasing protective immunity of the host by producing immunologic memory against potential pathogens prior to infection to create memory! Sometimes multiple exposures are required = booster shots to achieve level of desired protection. Think: Hepatitis B series!! Primary Memory / Anamnestic Response associated with small numbers of responding cells due to antigen-stimulated proliferation and expansion of populations of cells in the immune system. Secondary Memory / Anamnestic Response - associated with large numbers of responding cells due to antigen-stimulated proliferation and expansion of populations of cells in the immune system. Characteristics of secondary memory response: 1) increased rates of immune response 2) increased intensity of immune responses 3) increased affinities for binding antigen demonstrated by certain immunoreactants (i.e. Ab)

--- Lymphoid Tissues --Central / Primary Lymphocyte Tissues born and mature in the Thymus or the Bone Marrow Peripheral / Secondary Lymphocyte Tissues eventually live here in either the spleen, lymph nodes, Peyers patches, tonsils or blood; entry of lymphocytes into lymphoid tissues via high endothelial venules use adhesion/interaction mechanisms similar to those described for movement of leukocytes from blood, via endothelium, to sites of inflammation. Lymphocyte Recirculation coupled with architecture, specialized localization of lymphoid tissues maximizes the possibility of detecting invading pathogens and promoting imp interactions High Endothelial Venules the channels through which lymphocytes traffic back and forth between blood and lymphoid tissues

Immunology : Lecture 1 : Bolded Terms & Concepts Antigen a substance that is capable of being specifically recognize by receptors on cells of the immune system (B and T cells); it the basic element that allows the immune system to selectively recognize and target particular pathogens. Each antigen has a level of immunogenicity Microorganisms a collection of many individual foreign molecules = antigens; the exact combination of antigens is specific to the microorganism (species, genus, strain etc). So each antigen that reacts with an antibody does so independently of the other antigens actions (i.e. independent events) but simultaneously. Immunogenicity the ability of / potential of an antigen expressed by a pathogen to induce specific and protective immune response. It depends on many factors including health and proper function of the hosts immune system + structural + biological characteristics of the antigen itself (not all antigens will be good immunogens or are immunogens!) Biochemical properties that influence Immunogenicity: Proteins > Polysaccharides >>> lipids and nucleic acids Intrinsic factors to the antigen that INCREASE immunogenicity: Multiple differences (away) from self Large in size, complex composition, particulate matter, denatured form / degradability Extrinsic factors to the antigen that INCREASE immunogenicity: Intermediate dosage (i.e. not very high or very low) Route taken: subcutaneous > intramuscular > intraperitoneal >> intravenous Adjuvant = substances, mixed with antigen, that promote slow release and nonspecific inflammation, increase immunogenicity (i.e. mineral oil / Freunds adjuvant or Alum prepcipitates) Immunogen an antigen that can induce a specific immune response when introduced into a host; not all antigens are immunogens but ALL immunogens = antigens!

Antigenic Determinants = Epitopes = regions of an antigen that can each be recognized by the immune system Linear Epitope one continuous string of molecular constituents (i.e. sequential amino acids in a protein) Conformational Epitope epitopes formed by discontinuous / noncontiguous regions brought together by the 3D folding of the antigen Haptens low molecular weight substances that can be recognized by antibodies of the appropriate specificity but cannot induce an antibody response unless coupled to an immunogenic carrier

Immunology : Lecture 1 : Bolded Terms & Concepts BUG ATTACKS Extracellular Infection when pathogen stays outside the hosts tissues Intracellular Infection when the pathogen moves to the inside of the host cells followed by parasitizing of those cells. HOST RESPONSE Typical Immune Response involves 1) recognition of an antigen sequence resulting in 2) proliferation and 3) differentiation of lymphocytes so that they can go out and 4) eliminate the pathogens1 Recognition of Antigen each lymphocyte has ONLY 1 receptor specific to antigen TWO Response types: (1) Humoral Immunity and (2) Cell-Mediated Immunity (1) Antibody-Mediated Arm Humoral Immunity one arm of the immune system that is especially good at combating extracellular infections; goal is the activation of B lymphocytes which in turn produce antibody-secreting plasma cells. Antibodies antigen-reactive glycoproteins 3 main cell types: a. Antigen-Presenting Cells often macrophages or dendritic cells, take up antigen, process/degrade that antigen, and then present antigen fragments to Helper T cells b. Helper T Lymphocytes which then become activated c. B Lymphocytes are exposed to the same antigen, which provides a FIRST signal that results in their partial activation. The interaction of the helper T cells and the B cells, providing a SECOND signal to the B cells to differentiate into plasma cells, which are high rate secretors of antigen-reactive antibody = Clonal Selection and Expansion Antigen reactive antibody can protect the host by: a. blocking of microbial attachment b. neutralization of microbial toxins c. opsonization of microbial surfaces d. collaboration with serum complement proteins leading to the destruction of microbial membranes Antibody has 2 antigen combining sites = 4 chain molecule, 2 peptides are heavy and 2 are light. Can be either: a. membrane-bound form on the surface a B cell as a receptor for antigen b. soluble secreted form product of a plasma cell Clonal Selection each lymphocyte is genetically programmed to produce on its surface membrane one homogenous family of receptor molecules with specificity for a particular epitope Monoclonal Response activation of a single lymphocyte that produces a clone of lymphocytes with homogenous specificity for a single epitope. Polyclonal Response activation of numerous clones of lymphocytes for multiple epitopes. Cross-Reactive when an antibody recognizes two antigens that arent identical but share 1+ related/identical epitopes such that one antibody can recognize two antigens

Immunology : Lecture 1 : Bolded Terms & Concepts (2) Cell-Mediated Arm/Immunity the second arm of the immune system that is especially good at combating intracellular infections. T cells are activated and confer effective resistance to intracellular infections, certain extracellular pathogens, and certain malignant host cells. Two pathways: 1. Antigen T cells inflammatory T cells -- inflammatory cytokines 2. Antigen T cells Cytotoxic T cells cytolysis: kill pathogens or kill pathogens host cell T Cells key lymphocytes in cell-mediated immunity; functional subsets 1) Cytolytic effector T cells (Killer T cells) 2) inflammatory T cells 3) regulatory T cells = regulators of both #1 and #2 as well as B cells. Inflammatory T Cells suited for amplifying innate immune responses + activities of phagocytic cells. Accessory Cells required for the processing and presentation of antigen to the T lymphocytes (i.e. macrophages, dendritic cells, B cells)