Republic of the Philippines

ISABELA STATE UNIVERSITY

City of Ilagan Campus

IMMUNOLOGY (BS PSYCH 2)

LYMPHOCYTES
Prepared by: Charles Z. Ariola Jr., MSN, LPT

OBJECTIVES:

1. Describe the process of stimulation of individual B cells to divide and secrete antibody such as to generate
immunity to a particular antigen (clonal selection)
2. Briefly outline the principles of immunoglobulin (Ig) gene rearrangement in the generation of diversity
3. Outline the differences in antibody production during primary and secondary immune responses
4. Differentiate between monoclonal and polyclonal antibody

Adaptive Immune response

 B lymphocytes operate during the adaptive immune response

 Develops after encounter of antigem
 Takes 4-7 days to develop and become effective
 Elicited antibody production specific to encountered antigen
 2 types:
Humoral- B cells -- antibodies
Cell Medicated- T cells -- cytokines, lysis of pathogens

B Lymphocytes

• White blood cells

• Derived from haemopoietic stem cells
• Are effector cells of humoral immunity; they secrete antibodies and form memory cells

Where do they come from?

• Derived in the bone marrow in the absence of antigens
• Mature in the bone marrow, whereby they express specific B cell receptors (BCR)
• Migrate into the circulation (blood, lymphatic system) and into lymphoid tissues
• Antibody production requires antigen-induced B cell activation and differentiation- this occurs in peripheral
lymphoid organs

B cell Maturation
• Pro-B Cell  Pre-B Cell  Immature B Cell  Mature B Cell
• Occurs in the bone marrow in the absence of antigen
• Mature B cells are specific for a particular antigen- their specificity
resides in B cell receptor (BCR); a membrane bound immunoglobulin
B cell Receptor (BCR)
• Transmembrane protein complex composed of:
 mIg
- central larger immunoglobulin molecule
- cytoplasmic tail too short so is not involved in signalling
 Igα/Igβ
- di-sulfate linked heterodimers
- contain immunoglobulin-fold structure
- cytoplasmic tails of Igα/Igβ is long enough to interact with
intracellular signalling molecules
• has a unique binding site- binds to ANTIGENIC DETERMINANT or
EPITOPE -made before the cell ever encounters antigen
• large monoclonal population on surface of the B lymphocyte

Antigen and BCR diversity

• For the immune system to respond to the large number of antigens we are exposed to, we need to have a
large REPERTOIRE of specific BCR on different B cells that can recognise the huge array of antigens
• 1010 different antibody molecules can be generated by B cells with specific BCR
• Functional BCR genes do not exist until they are generated during lymphocyte development
• Each BCR chain (κ & λ light chains, and heavy chain) is encoded by separate MULTIGENE FAMILIES
ON DIFFERENT CHROMOSOMES
• During maturation, these gene segments are rearranged and brought together to form the BCR
– IMMUNOGLOBULIN GENE REARRANGEMENT
• There are a number of VARIABLE; V, DIVERSITY;D and JOINING;J gene segments that may be responsible for
each chain. The Diversity segment is only associated with the heavy chain. There is also a CONSTANT REGION
associated with each chain
• This generates the diversity of the lymphocyte repertoire

Prototypical Membrane Protein Synthesis

• Genomic DNA – (transcription) – Primary transcript RNA/pre-mRNA – (Splicing) – Mature mRNA
– (translation) – Membrane protein
• Intracellular; Amino terminus of protein and protein domains relating to specific exons
• Transmembrane; relates to specific exon/s
• Extracellular; cytoplasmic tail- consists of exons and carboxyl terminus

Light Chain Synthesis

• Germline DNA– (rearrangement of V and J segments involving VDJ RECOMBINASE) – B cell DNA –
(Transcription) – Primary transcript RNA/pre-mRNA – (Splicing) – Mature mRNA – (translation) – Light
chain polypeptide (Kappa or Lamda)
• During joining of gene segments the unused DNA is looped out and removed (Germline DNA – B cell DNA)

Heavy Chain Synthesis

• Germline DNA– (rearrangement of V and J segments involving VDJ RECOMBINASE) – B cell DNA –
(Transcription) – Primary transcript RNA/pre-mRNA– (Alternative Splicing) – Mature mRNA – (translation)
– Heavy chain polypeptide
• ALTERNATIVE SPLICING; results in different mature mRNA, as the mRNA express different genes (e.g. they
may have different constant region genes present)
Adaptive Immune Response

• Antibody production is a highly regulated process after activation by epitope

• If a B cell does not meet an antigen – death
MCD Immunology
• Antibodies may keep specificity but change
class
• During immune response, the first antibody
produced is IgM, but this can change
• The adaptive immune response is characterised
by:
1. Specificity
2. Diversity
3. Memory

Clonal Selection
• Basis of adaptive immunity
• Non-self reactive mature lymphocytes then
migrate to the periphery
• Our immune system is usually exposed to multiple antigens, therefore multiple cells will be activated
• Each lymphocyte (T or B) expresses an antigen receptor with a unique specificity,
• Binding of antigen to its specific receptor leads to activation of the cell, causing it to proliferate into a clone
of cells
• All of these clonally expanded cells bear receptors of the same specificity to the parental cell
• Lymphocytes expressing receptors that recognize self molecules are deleted early during lymphocyte
development and are phagocytosed/lysed
• Result: Plasma Cells, Antibodies, Memory cells

Antibody production
• Naive antigen-specific lymphocytes cannot be activated by antigen alone; they require accessory signals
either from:
- Microbial Constituents- Thymus Independent
- Helper T cells- Thymus Dependent
 Thymus Independent
- Microbial Consistuents
- Only IgM is produced
- No memory cells formed
- Antigens directly activate B cells without the
help of T cells
- This can induce antibodies in people with no
thymus and no T cells (Di-George syndrome)
- The second signal required is either
provided by the microbial constituent or by
an accessory cell
Thymus Dependent
- Helper T cells
- All Ig-classes produced
- Memory is formed
- Membrane bound BCR binds with antigen
and is internalised and delivered to
intracellular sites
- Antigen is degraded into peptides
- Peptides associated with Self- MHC Class II,
forming a complex which is expressed at
the cell surface
- T lymphocytes with a complementary T cell
receptor (TCR) recognises the complex
- T helper cells then secrete LYMPHOKINES
- B cell then enters the cell cycle, forming a
clone of cells with identical BCRs-
differentiating into plasma and memory
cells

T.B cell collaboration

• Antigen cross link with BCR induces signal 1-- ↑MHC II, ↑B7
• Antigen is internalised and degraded, and the peptide-MHC II complex is presented

• T cell recognises complex and co-stimulation by B7and CD28 interaction activation of T cells
B7(expressed by B cell)
CD28(expressed by TH cell)
• Activated T cell expresses CD40L
• The interaction between CD40L and CD40 (expressed by B cell) induces signal 2
• Activated B cells (CENTROBLAST) express cytokine receptors
• T cell derived cytokines bind to receptors on B cells
• B cells proliferate and differentiate into antibody secreting plasma cells