Ø The host protein that displays the antigen fragment on the cell surface is called an MHC (Major Histocompatibility

Complex).
Ø These proteins play a critical role in B cell and T cell interaction with each other as well as other host cells and pathogens.
Ø All host cells have the MHC I class marker molecule on their membranes, and this is really important as it helps the immune
system identify the “self” cells that are considered safe and do not trigger any immune response.
Ø Cells like macrophages or B cells that can display the antigens they bind to also have a MHC II class marker, which is solely
used for carrying and displaying the antigen to helper T cell .
Ø Humoral immune response
- The humoral immune response occurs in the blood and lymph, which were long ago called body humors (fluids).
- In the humoral response, antibodies help neutralize or eliminate antigens in the blood and lymph.
- Activation of the humoral immune response involves B cells and helper T cells, as well as proteins on the surface
of pathogens.
- Stimulated by both an antigen and signalling molecules called cytokines secreted from the activated helper T cell,
the B cell proliferates and differentiates into memory B cells ( and antibody-secreting effector cells called plasma
cells)
- B cell activation leads to a robust immune response: a single activated B cell gives rise to thousands of identical
plasma cells.
- These plasma cells begin producing and secreting antibodies. Each one is able to secrete approximately 2,000
antibodies every second of the cell’s 4 to 5 day life span.
- Antibodies secreted by the plasma cells enter the blood and travel to infected areas. Antibodies do not kill
pathogens, but by binding to antigens, they mark pathogens in various ways for inactivation or destruction, via
neutralisation, opsonisation or activation of the complement system (proteins from the second line of defence)
Polyclonal and monoclonal antibodies

Antibodies (whatever their class or subclass) are produced and purified in two basic forms
for use as reagents in immunoassays: polyclonal and monoclonal.

Typically, the immunological response to an antigen is heterogeneous, resulting in many

different cell lines of B lymphocytes (precursors of plasma cells) producing antibodies to the
same antigen. All of these cells originate from common stem cells, yet each develops the
individual capacity to make an antibody that recognizes a particular determinant (epitope)
on the same antigen.

Because it contains this heterogeneous collection of antigen-binding immunoglobulins, an

antibody purified from such a sample is called a polyclonal antibody.

Because an individual B lymphocyte produces and secretes only one specific antibody
molecule, clones of B lymphocytes produce monoclonal antibodies. All antibodies secreted
by a B cell clone are identical, providing a source of homogeneous antibody having a single
defined specificity.
 Humeral and Cellular Immunity
There are two main mechanisms of immunity within the adaptive immune system – humoral and cellular.

Humoral immunity is also called antibody-mediated immunity. With assistance from helper T cells, B cells will differentiate
into plasma B cells that can produce antibodies against a specific antigen. The humoral immune system deals with antigens
from pathogens that are freely circulating, or outside the infected cells.

Antibodies produced by the B cells will bind to antigens, neutralizing them, or causing lysis (dissolution or destruction of cells
by a lysin) or phagocytosis.

Cellular immunity occurs inside infected cells and is mediated by T lymphocytes.

The pathogen's antigens are expressed on the cell surface or on an antigen-presenting cell. Helper T cells release cytokines
that help activated T cells bind to the infected cells’ MHC-antigen complex and differentiate the T cell into a cytotoxic T cell.

The infected cell then undergoes lysis.

 Complement system
• The complement system, also known as complement cascade, is a part of the immune system that enhances
(complements) the ability of antibodies and phagocytic cells to clear microbes and damaged cells from an
organism, promote inflammation, and attack the pathogen's cell membrane.

• It is part of the innate immune system, which is not adaptable and does not change during an individual's lifetime.

• The complement system can, however, be recruited and brought into action by antibodies generated by the
adaptive immune system.

• The complement system consists of a number of small proteins that are synthesized by the liver, and circulate in
the blood as inactive precursors. When stimulated by one of several triggers, proteases in the system cleave
specific proteins to release cytokines and initiate an amplifying cascade of further cleavages.

• The end result of this complement activation or complement fixation cascade is stimulation of phagocytes to
clear foreign and damaged material, inflammation to attract additional phagocytes, and activation of the cell-
killing membrane attack complex.

• About 50 proteins and protein fragments make up the complement system, including serum proteins, and cell
membrane receptors. They account for about 10% of the globulin fraction of blood serum.
Complement System is

•A system of serum and cell surface proteins that interact with one another and with other molecules of the
immune system to generate important effectors of innate and adaptive immune response.

There are three pathways of complement activation that differ in how they initiated:

1. Classical pathway activated by antigen antibody complex.

2. Alternative pathway, by microbial surface.
3. Lectin pathway, by plasma lectins that binds to microbes.

Each complement pathway consists of a cascade of protolytic enzymes that generate inflammatory mediators
and opsonins that lead to formation of a lytic complex that insert in cell membrane.

The function of complement system include:

1- Triggering and amplification of inflammatory reactions.
2- Attraction of phagocytes by chemotaxis.
3- Clearance of immune complexes and apoptotic cells.
4- Cellular activation for microbial killing.
5- Direct microbial killing.
6- An important role in the efficient development of Ab response.
Stages of Complement Activation

All three pathways involves:

1. Activation of C3 which is the most abundant and most important of complement proteins.

2. Comprise protolytic cascade in which complexes of complement proteins create enzymes

that cleave other complement proteins in an order manner to create new enzymes.

3. All activation pathways converge on a common terminal pathway- formation of membrane

attack complex (MAC), causing membrane disruption and lytic killing of pathogens.

4. All the three pathways differ from each other in their initiation till formation of C3 convertase.
Then, the remaining stages are identical in all the pathways.
Complement system Functions

Complement triggers the following immune functions:

Membrane attack – by rupturing the cell wall of bacteria. (Classical

Complement Pathway)

Phagocytosis – by opsonizing antigens. C3b has most important

opsonizing activity. (Alternative Complement Pathway)

Inflammation – by attracting macrophages and neutrophils. (Lectin

pathway)

Three biochemical pathways activate the complement system: the classical complement pathway, the alternative
complement pathway, and the lectin pathway. The alternative pathway accounts for the majority of terminal pathway
activation and so therapeutic efforts in disease have revolved around its inhibition.
Ø Classical Pathway

This pathway involves complement components C1, C2 and C4.

The pathway is triggered by antibody-antigen complexes binding to Cl, which itself has three subcomponents Clq, Clr and Cls.
The pathway forms a C3 convertase, C4b2a, which splits C3 into two fragments; the large fragment, C3b, can covalently
attach to the surface of microbial pathogens and opsonise them; the small fragment, C3a, activates mast cells, causing the
release of vasoactive mediators such as histamine.
Ø Alternative Pathway

This pathway involves various factors, B, D, H & I, which interact with each other, and with C3b, to form a C3 convertase,
C3bBb, that can activate more C3, hence the pathway is sometimes called 'the amplification loop'. Activation of the loop
is promoted in the presence of bacterial and fungal cell walls, but is inhibited by molecules on the surface of normal
mammalian cells.

Ø Mannose-binding Lectin Pathway

This pathway is activated by the binding of mannose-binding lectin (MBL) to mannose residues on the pathogen surface. This in
turn activates the MBL associated serine proteases, MASP-1 and MASP-2, which activate C4 and C2, to form the C3 convertase,
C4b2a.

Ø Lytic Pathway
This pathway is initiated by the splitting of C5, and attachment of C5b to a target. C6, C7, C8 and C9 unite with C5b, and this
membrane-attack complex (MAC), when inserted into the outer membrane of some bacteria, can contribute to their death by
lysis.
Membrane Attack Complex (Terminal Complement
Complex C5b-9)
Cytokines
1- are small soluble proteins that regulate the immune system, both innate and the adaptive response to
infection.
2- These chemical messengers, produced by several different types of cells.
3- They exert activity-modulating effects on cells of hematopoietic and immune system through the activation
of cell-bound proteins
4- Cytokins are induced in response to specific stimuli such as bacterial liposacchrides, flagellin, or other
bacterial products through the ligation of cell adhesion molecules or through the recognition of foreign
antigens by host lymphocytes.

ü The major cytokine families include:

1- Interleukins (IL)
2- Tumor necrosis factors (TNF)
3- Interferons (IFN)
4- Chemokines
5- Transforming growth factors (TGF)
6- Colony stimulating factors (CSF
Ø Diseases of complement deficiency

1- systemic lupus erythematosus (SLE)

2- recurrent bacterial infection and an increased risk of cardiovascular disease. It is thought to influence the development of
atherosclerosis.
3- With properdin deficiency, there is a particular risk of overwhelming neisserial infection.
4- Leiner's disease.

Ø Opsonization

- is the mechanism by which targeting of particles for destruction through phagocytosis becomes enhanced.
- Opsonins are molecules that mark foreign particles for phagocytosis.
- the attachment of opsonins, then makes the pathogen more visible to the phagocyte, and the opsonized
pathogen is then ingested by the phagocyte before intracellular destruction through digestion
- Phagocytosis is the cellular process for removes pathogens and dead or dying cells.
- Opsonization is the second step of phagocytosis, with chemotaxis first causing the recruitment of the
phagocyte towards the site of infection or cell death.
 Human leukocyte antigen
Ø The human leukocyte antigen (HLA) system or complex is a complex of genes on chromosome 6 in humans
which encode cell-surface proteins responsible for the regulation of the immune system.

Ø HLA genes are highly polymorphic, which means that they have many different alleles, allowing them to fine-
tune the adaptive immune system.

Ø The HLA system is also known as the human version of the major histocompatibility complex (MHC) found in
many animals.

Ø Mutations in HLA genes may be linked to autoimmune disease such as type I diabetes, and celiac disease.

o HLAs corresponding to MHC class I (A, B, and C), HLA Class1 group, present peptides from inside the cell. For example, if
the cell is infected by a virus, the HLA system brings fragments of the virus to the surface of the cell so that the cell can
be destroyed by the immune system.

- These peptides are produced from digested proteins that are broken down in the proteasomes.
- In general, these particular peptides are small polymers, of about 8-10 amino acids in length.
Foreign antigens presented by MHC class I attract T-lymphocytes called killer T-cells (also referred to as CD8-positive or
cytotoxic T-cells) that destroy cells.
o HLAs corresponding to MHC class II (DP, DM, DO, DQ, and DR) present antigens from outside of the cell to T-lymphocytes.

These particular antigens stimulate the multiplication of T-helper cells (also called CD4-positive T cells), which in turn stimulate
antibody-producing B-cells to produce antibodies to that specific antigen.

ü Self-antigens are suppressed by regulatory T cells.

ü HLAs corresponding to MHC class III encode components of the complement system.

o HLAs have other roles.

1) They are important in disease defense.

2) They are the major cause of organ transplant rejections.
3) They may protect against or fail to protect (if down-regulated by an infection) against cancers.

Codominant expression of
Schematic representation of
HLA genes
MHC class I
 Role of complement and HLA in oral disease
Chronic periodontitis (CP) is an infectious inflammatory disease that affects tooth-supporting structures and
in which dental plaque bacteria, immune mechanisms and genetic predisposition play important roles.

Ø HLA-A, B, C, DR antigen frequencies and Properdin factor B (Bf) allotypes were studied in a group of 44
patients with rapidly progressive periodontitis.

Ø HLA-A9 (A24) was the only antigen with a frequency statistically significantly different from the control
population.

Ø HLA-A, B and DR antigen frequencies were determined in three groups of periodontally diagnosed subjects: 49 patients
with rapidly progressive periodontitis, 40 elderly subjects with minimal disease (considered as a resistant group) and 30
young subjects with minimal disease.

- The relative risk for HLA-A9 (previously reported to be associated with periodontal disease) was 15.5.
- HLA-A9 was present in 36.7% of the patients and 2.5% of the resistant group.
- HLA-A10 showed a significantly increased incidence in the resistant group (30.0%) compared to a non-periodontally
diagnosed control population (9.0%), and was absent from the patient group.

ü These findings provide additional evidence for the involvement of HLA-A9 in susceptibility to periodontitis, and suggest
that HLA-A10 may play a role in resistance to the disease.
Interleukin 4 (IL-4) is a key anti-inflammatory cytokine with relevant action in imbalances in inflamed periodontal
tissue.

- Individuals carrying the TCI/CCI genotype (S-haplotype) of the IL-4 gene are 5 times more susceptible to CP,
whereas the CTI/TTD genotype (P-haplotype) confers protection against CP.

- Compared with the S-haplotype, subjects with the P-haplotype produce higher levels of the IL-4 protein after
non-surgical periodontal therapy.

Peripheral blood was collected from 6 subjects carrying each haplotype, and their immune cells were challenged
with periodontopathogens to compare responses of the different haplotypes with regard to gene expression,
protein secretion and the immunophenotype of T helper responses.

- We found higher IL-4 mRNA and protein levels in the P-haplotype, which also presented higher levels of anti-
inflammatory cytokines. In contrast, cells from S-haplotype subjects responded with higher levels of pro-
inflammatory cytokines.

S-haplotype individuals exhibited significantly greater polarization toward the Th1 phenotype, whereas the P-
haplotype was associated with an attenuated response to periodontopathogens, with suggestive skewing
toward Th2/M2 phenotypes.
Individuals carrying the ATC/TTC haplotype (Hap-1) in the interleukin 8 (IL8) gene were reported as more
susceptible to chronic periodontitis (CP), an infectious disease associated with Gram-negative bacteria, in
comparison to patients with the ATT/TTC haplotype (Hap-2).

Ø This study investigated the functionality of the IL8 haplotypes in lymphocytes and monocytes of individuals
carrying the Hap-1 or Hap-2 IL8 haplotypes in the response to CP-associated Gram-negative bacteria
(periodontopathogens).

Peripheral blood was collected from 6 subjects carrying each haplotype, and their immune cells were challenged with
periodontopathogens.

Subjects carrying the Hap-1 haplotype showed increased expression of IL8 and TNFA and significantly skewing towards pro-
inflammatory Th1/M1/Th17 phenotypes. There was increased percentage of ROS-producing monocyte-derived macrophages
from individuals carrying the Hap-1 haplotype.

The Hap-1 haplotype previously associated with increased susceptibility to CP demonstrated greater skewing to
pro-inflammatory Th1/M1/Th17 phenotypes and production of ROS.

Cells from individuals presenting the Hap-2 haplotype had an overall attenuated response to periodontopathogens, with a
significant shift towards the Treg phenotype.

In conclusion, the IL8 haplotypes showed to be functional both in monocyte-derived macrophages and lymphocytes.