T Cells

Tc cells are CD8+, Th cells are CD4+

CD = cluster of differentiation Expressed by various cells at particular stages of differentiation

T Cell Receptors
Similar to Abs Contain constant & variable regions Mediate Ag recognition

Found on cell surface of T cell, but is not secreted

Major Histocompatability complex

T cells cannot recognize Ag alone
Ag must be presented in conjunction with major histocompatability complex (MHC) proteins In humans called human leucocyte antigens

 MHC proteins are glycoproteins found on cell surface Vary from 1 individual to another & serve as a label 2 classes, MHC I & MHC II are involved in Ag processing Differ in structure, location & function

 MHC I found on all nucleated cells

Important in in presenting endogenous Ag, eg virus to Tc cells Endogenous Ags - produced within cytosol, eg viral proteins in an infected cell

 MHC II found only on immunocompetent celss such as macrophages & B cells

Present exogenous Ag to Th cells

Exogenous Ag taken up by endocytosis, found in vesicles within cells Eg bacteria & their products taken up by macrophages

Generation of Ab Diversity
Vast range of Ab-producing cells exist, independent of exposure to any specific Ag Presence of a given Ag stimulates production of a particular Ab or set of Abs

Vaccination
Primary & Secondary Immune Responses o Following initial stimulation of a response by particular Ag, memory cells specific to that Ag are formed

o On 2nd encounter response is faster & stronger -Memory cells are activated by Ag -Become Ab secreting plasma cells -Form further memory cells
o Basis of vaccination

Principle of Vaccination Non-pathogenic organism primes the immune system so that on encounter with pathogen, a protective response is generated

During 1o challenge, stimulated B & T cells proliferate & differentiate to give effector cells + memory cells
Memory cells are long-lived & proliferate more rapidly o Facilitate rapid control of subsequent infections

Passive Immunization Preformed Abs are transferred to recipient, eg. o Natural maternal Abs o Artificial immune serum such as horse antivenin
Protects against snake bite Short-term protection

Active Immunization Protective immunity & immunologic memory are elicited, eg. o Natural infection such as flu virus o Artificial infection such as vaccines -Attenuated organisms
-Inactivated organisms

Route of Administration of Ag Influences type if immune response 1. Systemic Response Vaccine is introduced by injection Increase of circulating Abs & specific T cells
2. Mucosal Response Vaccine is introduced orally or by respiratory route IgA predominates

Uses of Adjuvants Certain substances, which when administered simultaneously with a specific Ag, will enhance the immune response to that Ag
Eg Aluminium Salts (hydroxyl, phosphates) -1st safe & effective cpds to be used in human vaccines -Promote good Ab response, but poor CMI

Attributes of a Good Vaccine 1. Ability to elicit the appropriate immune response for a particular pathogen -Tuberculosis- CMI -Most bacterial & viral infections- Abs 2. Long-term protection -Ideally single dose-life long immunity

3. Safety -Non-pathogenic -No side effects

4. Stable -Should maintain immunogenicity o Despite adverse conditions prior to administration 5. Cheap

Vaccine Production In developing a vaccine, consider the branch of immune system to be activated

Humoral immunity o epitopes must be accessible to B-cell receptors o represent immunodominant epitopes of infectious agent

 CMI
o vaccine should be capable of maximizing presentation of Ag with Class I MHC molecules o Must undergo transient growth in recipient

Live Attenuated Organisms -Virulence artificially reduced by in vitro culture i. Under adverse conditions, eg low To ii. Selection of mutants which replicate poorly in human host
-Often single dose reqd

Replication of vaccine strain in host i. Reproduces many features of w.t infection ii Does not cause clinical disease -Induces both humoral & CMI Long-lived -May revert to virulent form Eg. Bacillus Calmette-Guerin (BCG)-an attenuated Mycobacterium bovis (Tuberculosis) -Viral particles causing measles or mumps

Inactivated (Killed) Organisms Virulent pathogen inactivated by: o Irradiation with gamma rays o Chemicals such as formaldehyde or bpropiolactone

Produces mainly humoral immunity o Poor immunogenicity Requires multiple boosters o Short-lived

 o o o o o o

More stable Organisms need not be viable Can withstand adverse storage conditions Cannot revert to virulent form Safer Cannot replicate in host & cause disease Eg. Cholera (bacterial) Influenza (viral)

HIV Vaccine?
Problems High mutation rate of HIV Need for both CMI & humoral immunity Lack of good animal model

Trial Vaccines Recombinant peptides, gp120 DNA vaccines Live attenuated virus??

Immunochemical Techniques

Techniques that use the interaction between antibody (Ab) and antigen (Ag) to detect or quantitate either Ab or Ag.

 Ab-Ag interaction is specific -Ab to HIV reacts with HIV but not with flu virus Examples Anti-insulin Abs are used to quantify the amount insulin in a serum sample
Bovine serum albumin (BSA) is used to test for the presence of anti-BSA Abs in a rabbit immunized with BSA

Enzyme linked immunosorbent assay (ELISA) Detection reagent is an enzyme, which converts a colourless substrate to coloured product
Detected by simple eye or read by a colorimeter Can be used to assay for Ab or ag, can label either Ab or Ag

Assay Design Many different variations on assay design

Solid phase for assay is usually a 96-well microtitre plate which binds protein nonspecifically
Enzymes used mainly HRP or AP, which act on a wide range of colourless substrates to give coloured products
Small, do not interfere with Ab-Ag interaction

ELISA for Ab to HIV

Coat plate with Ag Block surface Add serum sample Add E-labelled anti-human Ab Add colourless substrate

Situations where Ab detection is inappropriate Window period Newborn with HIV+ mother
Solution Detect virus rather than immune response eg PCR to detect viral RNA