HUMORAL IMMUNE RESPONSE

MICROBIOLOGY LAB

Humoral immunity to combate microbe

Be divided into innate and adaptive immunity Innate immunity is performed by complement, which have main role to lyse extracellular microbes Adaptive humoral immunity is conducted by antibodies (Ab), which have physiologic function in defense against extracellular microbes and microbial toxin

ANTIBODIES (Immunoglobulins/Ig)
are circulating protein that are produced in vertebrate in responses to exposure to foreign structures known as antigens

Antigens

Most are proteins or large polysaccharides from a foreign organism.

Microbes: Capsules, cell walls, toxins, viral capsids,

flagella, etc.
Nonmicrobes: Pollen, egg white , red blood cell

surface molecules, serum proteins, and surface molecules from transplanted tissue.

Lipids and nucleic acids are only antigenic when combined with proteins or polysaccharides. Molecular weight of 10,000 or higher. Hapten: Small foreign molecule that is not antigenic. Must be coupled to a carrier molecule to be antigenic. Once antibodies are formed they will recognize hapten.

Antigens
Epitope: Small part of an antigen that interacts with an antibody. Any given antigen may have several epitopes. Each epitope is recognized by a different antibody.

Epitopes: Antigen Regions that Interact with Antibodies

Antibody

produced by B ly in the lymphoid organ and bone marrow their effector function at sites distant from their production derived from long-lived plasma cells (in secondary immune responses by the activation of memory B cells) the effector functions are mediated by the heavy chain (Fc) constant regions of Ig molecules, e.g. phagocytosis Ab activates effector function when they are needed effector functions are triggered by binding of Ag to the variable regions (Fab)

Antibody structure

The simplest molecular structure is called monomer has four protein chains :
Two identical light chains (L-chain) Two identical heavy chains (H-chain)

The chains are joined by disulfide (S-S) bonds/links to form a Y shaped or T shaped The two arms of Y shaped is called variable (V) regions as antigen (epitope) binding sites (Fab regions) The stem of Y shaped is called Fc regions

Fab

Heavy chain

Ag binding site

Epitope

Light chain Fc
Hinge region

ANTIBODY

Antibodies can exist in two forms:

1. As membrane-bound antibodies on the surface of B lymphocytes function as receptors for Ag 2. As secreted antibodies in the circulation, tissues, and mucosal site which will bind antigens, neutralized toxins, and prevent the entry and spread of pathogens

EFFECTOR FUNCTIONS of Ab
1. Neutralization of microbes and toxins 2. Opsonization and phagocytosis of microbes 3. Mediate ADCC (Ab dependent cell-mediated cytotoxic) 4. Lysis of microbes and inflammation through the activation of complement 5. Feedback mechanism

ANTIBODY FUNCTIONS

Functions of Ab isotypes
Ab isotypes
IgG

Effector function
Opsonization of Ag for phagocytosis by Mo and neutrophils Activation of the classical pathway of complement ADCC mediated by NK cells and Mo Neonatal immunity: transfer of maternal antibosy across the placental and gut Feedback inhibition of B cell activation

IgM
IgA IgE IgD

Activation of the classical pathway of complement Ag receptor of naive B cells

Mucosal immunity: secretion of IgA into the lumens of GIT and resp tract o ADCC involving eosinophils o Mast cells degranulation (immediate hypersensitivity) Antigen receptor of naive B cells

Neutralization of microbes and microbial toxin

requires only the Fab region most neutralizing Ab in the blood are of IgG, in mucosal organs are of the IgA isotype

Neutralization of microbes and toxins

Opsonization and phagocytosis

IgG isotype coat (opsonize) microbes and promote their phagocytosis by binding to Fc receptors on phagocytes Mononuclear and neutrophils express receptor for the Fc portion of IgG (FcRI) Microbes may also be opsonized by a product of complement activation (C3b) The IgG and C3b are called opsonin

Opsonization and phagocytosis

Antibody-Dependent Cell mediated Cytotoxic (ADCC)

NK cells and other leucocytes bind to Ab coated cells by Fc receptors and destroy these cells Eosinophils mediate ADCC against helminth coated by IgE through FcRI

Engagement FcRIII (Fc receptor on NK cells) activates the NK cells to synthesize and secrete cytokine such as IFN- as well as the content of the granule (killing functions of NK cells)

Antibody-Dependent Cell mediated Cytotoxic (ADCC)

Complement functions

opsonization to enhance phagocytosis phagocyte attraction and activation through inflammation process lysis of bacteria

Complement functions

Consequences of Antibody Binding

Immunological Memory
Antibody Titer: The amount of antibody in the serum.

Pattern of Antibody Levels During Infection Primary Response: After initial exposure to antigen, no antibodies are found in serum for several days. A gradual increase in titer, first of IgM and then of IgG is observed. Most B cells become plasma cells, but some B cells become long living memory cells. Gradual decline of antibodies follows.

Immunological Memory (Continued)

Secondary Response: Subsequent exposure to the same antigen displays a faster and more intense antibody response. Increased antibody response is due to the existence of memory cells, which rapidly produce plasma cells upon antigen stimulation.

Antibody Response After Exposure to Antigen