‫جامعة الفرات االوسط التقنية‬

‫المعهد التقني بابل‬

‫قسم التحليالت المرضية‬

‫مناعة‬

‫‪B Cells‬‬

‫أشراف األستاذة ‪ :‬منى جابر‬

‫‪ :‬زهراء عايد عبدالشهيد‬ ‫األسم‬
‫‪ :‬الثانية‬ ‫المرحلة‬
‫‪ :‬مناعة‬ ‫المادة‬
Introduction
B cells shown in Fig 1, also known as B lymphocytes, are a type of white blood cell of
the lymphocyte subtype. They function in the humoral immunity component of
the adaptive immune system by secreting antibodies. Additionally, B cells present
antigens and secrete cytokines.

Fig 1 – Human B lymphocyte

The function of B cells was discovered in the 1960s by Max Cooper who demonstrated
that antibody production was completely abrogated in irradiated chickens after surgical
removal of the Bursa of Fabricius (the primary site of B-cell development in birds) from
which the notation ‘B’ cell was derived.

1|PA GE
B-Cell Development

Immunoglobulins consist of two identical heavy and light chains, which are
joined by disulphide bonds. During B cell development, rearrangement of
the Ig heavy chain occurs first, commencing with D-J recombination, which
takes place in the common lymphoid progenitors (CLPs) and pre-pro B cells.
This is followed by V-DJ recombination yielding a functional heavy chain
protein (Iga) in large pre-B cells. The recombined heavy chain then
associates with the surrogate light chains and the

dimer to form the pre-B cell receptor (pre-BCR) which is expressed on

the cell surface. Signalling through the pre-BCR drives intense proliferation
and differentiation into the small pre-B cell stage. Quiescent small pre-B
cells then undergo V-J rearrangement of the Ig light chain, allowing the
production of a complete functional BCR with a unique specificity that is
expressed as IgM on the surface of immature B cells. In a bid to prevent
autoreactivity, immature B cells which encounter Ag capable of cross-
linking their newly expressed BCRs are eliminated by a variety of
mechanisms.
After production in the BM, immature surface IgM+ B cells migrate to the
spleen where they differentiate through distinct transitional B cell stages
termed T1 and T2, before differentiating into long-lived mature follicular
(FO) or marginal zone (MZ) B cells. Thus, B cells experience both antigen-
dependent and -independent phases of selection, tightly regulated through
signalling events. T3 B cells do not give rise to mature B cells, but instead
represent a subset of anergic B cells which have been selected away from the
B cell developmental pathway.

2|PA GE
 Fig 2 - B-cell development and B-cell subsets
In addition to FO and MZ B cells, a third population of mature B cells exists
known as B1 cells. B1 cells are located in a number of tissues including the
spleen, intestine, the peritoneal cavity and pleural cavities. B1 cells have
distinct haematopoietic origins in the in the fetal liver, and the initial wave
of lymphopoiesis in the embryo appears to be skewed towards B1 B cell
development.

3|PAG E
B-cell surface marker

B-cell Carry many surface marker:

 CD19
 CD21
 CD22
 Membranous immunoglobulin
 Igα and Igβ heterodimer
Fig 3 – B-cell surface marker

4|PAG E
Difference between the stages of B-cell

1. Pro -B-cell
 Is the earliest recognizable B-cell stage.
 It has following surface marker:-
 CD -45
 Igα and Igᵦ heterodimer:-part of B-cell membrane receptor in later
stage.
 CD-19
 CD-24
 CD-43
 C-kit
 •The heavy chain gene rearrangement begins in this stage and its
completion signals the end of this stage.

2. Pre-B-cell
 Characterized by the beginning of the translation of heavy chain
hene.
 Essentially require bone marrow stromal cell.
 Mu chain is usually first synthesized.
 No light chains are synthesized.
 Small peptide called Surrogate light chain are synthesized( not
true light chain)
 They complex with mu chain and this complex is expressed on
the cell surface.
 The mu chain and surrogate light chain complex associate with
Igα/Igᵦ hetrodimer to form pre-B-cell receptor.

5|P AGE
3. Immature -B-cell
 Begins with light chain gene.
 Only one type of light chain is expressed(kappa or lambda)
 Light chain associates with mu chain and so a true IgM expression
occurs on the B-cell Surface.
 This also results in B-cell commited to one particular antigenic
specificity as determined by binding site of surface IgM.
 True BCR appears.
4. Mature B-Cell
 Characterized by simultaneous expression of IgM and Igd on B-cell
surface.
 The mature B-cell then leaves the bone marrow and go to the
peripheral lymphoid tissue where they get activated on encountering
the antigen and produce two types of effector cells:-
o a)Plasma cell:- produce antibody, and
o b) Memory cell: - produce secondary immune response after re-
encounter with the same antigen.
 If the mature B-cell cannot encounter antigen, they die within few
days by apoptosis.
 So the production of mature B-cell in bone marrow is an antigen
independent phase while activation and effector cell production is an
antigen dependent phase.

6|PAG E
B cell activation and the humoral immune response
B-Cell Receptors
B cell activation is initiated when the IgD and monomeric IgM surface
receptors of B cells bind to specific antigens. Upon encounter with a
microbe or antigen, either by infection or vaccination, naïve B cells (antigen
inexperienced) become activated and differentiate into antibody-producing
plasma cells and memory B cells. Some plasma cells migrate to the bone
marrow, where they persist for several years and continue to produce
antibodies even in the absence of antigen. There are two routes to B cell
activation and initiation of the humoral immune response, which depend on
the nature of the antigen. Non-protein antigens such as lipids, nucleic acids
and glycoproteins stimulate antibody production in the absence of T cells,
and are referred to as thymus independent (TI) antigens. In contrast, the
antibody response to protein antigens requires both B and T cell
involvement, and these antigens are described as thymus dependent (TD)
antigens.
Figure 1. B-cell receptors are embedded in the membranes of B cells. The variable
regions of all of the receptors on a single cell bind the same specific antigen.

7| P A G E
T Cell-Independent Activation of B cells
Activation of B cells without the cooperation of helper T cells is referred to
as T cell-independent activation and occurs when BCRs interact with T-
independent antigens. T-independent antigens (e.g., polysaccharide capsules,
lipopolysaccharide) have repetitive epitope units within their structure, and
this repetition allows for the cross-linkage of multiple BCRs, providing the
first signal for activation (Figure 5). Because T cells are not involved, the
second signal has to come from other sources, such as interactions of toll-
like receptors with PAMPs or interactions with factors from the complement
system.

Once a B cell is activated, it undergoes clonal proliferation and daughter

cells differentiate into plasma cells. Plasma cells are antibody factories that
secrete large quantities of antibodies. After differentiation, the surface BCRs
disappear and the plasma cell secretes pentameric IgM molecules that have
the same antigen specificity as the BCRs (Figure 5).

The T cell-independent response is short-lived and does not result in the

production of memory B cells. Thus it will not result in a secondary
response to subsequent exposures to T-independent antigens.

Figure 5. T-independent antigens have repeating epitopes that can induce B cell recognition and
activation without involvement from T cells. A second signal, such as interaction of TLRs with PAMPs
(not shown), is also required for activation of the B cell. Once activated, the B cell proliferates and
differentiates into antibody-secreting plasma cells.

8|PAGE
T Cell-Dependent Activation of B cells

T cell-dependent activation of B cells is more complex than T cell-

independent activation, but the resulting immune response is stronger and
develops memory. T cell-dependent activation can occur either in response
to free protein antigens or to protein antigens associated with an intact
pathogen. Interaction between the BCRs on a naïve mature B cell and a free
protein antigen stimulate internalization of the antigen, whereas interaction
with antigens associated with an intact pathogen initiates the extraction of
the antigen from the pathogen before internalization. Once internalized
inside the B cell, the protein antigen is processed and presented with MHC
II. The presented antigen is then recognized by helper T cells specific to the
same antigen. The TCR of the helper T cell recognizes the foreign antigen,
and the T cell’s CD4 molecule interacts with MHC II on the B cell. The
coordination between B cells and helper T cells that are specific to the same
antigen is referred to as linked recognition.
Once activated by linked recognition, TH2 cells produce and
secrete cytokines that activate the B cell and cause proliferation into clonal
daughter cells. After several rounds of proliferation, additional cytokines
provided by the TH2 cells stimulate the differentiation of activated B cell
clones into memory B cells, which will quickly respond to subsequent
exposures to the same protein epitope, and plasma cells that lose their
membrane BCRs and initially secrete pentameric IgM (Figure 6).

9| P A G E
Figure 6. Click for a larger image. In T cell-dependent activation of B cells, the B cell
recognizes and internalizes an antigen and presents it to a helper T cell that is specific to the
same antigen. The helper T cell interacts with the antigen presented by the B cell, which
activates the T cell and stimulates the release of cytokines that then activate the B cell.
Activation of the B cell triggers proliferation and differentiation into B cells and plasma cells

10 | P A G E
Memory B cell activation

Memory B cell activation begins with the detection and binding of their
target antigen, which is shared by their parent B cell. Some memory B cells
can be activated without T cell help, such as certain virus-specific memory B
cells, but others need T cell help. Upon antigen binding, the memory B cell
takes up the antigen through receptor-mediated endocytosis, degrades it, and
presents it to T cells as peptide pieces in complex with MHC-II molecules
on the cell membrane. Memory T helper (TH) cells, typically memory
follicular T helper (TFH) cells, that were derived from T cells activated with
the same antigen recognize and bind these MHC-II-peptide complexes
through their TCR. Following TCR-MHC-II-peptide binding and the relay
of other signals from the memory TFH cell, the memory B cell is activated
and differentiates either into plasmablasts and plasma cells via an
extrafollicular response or enter a germinal center reaction where they
generate plasma cells and more memory B cells. It is unclear whether the
memory B cells undergo further affinity maturation within these secondary
GCs.

11 | P A G E
Primary and Secondary Responses
T cell-dependent activation of B cells plays an important role in both the primary
and secondary responses associated with adaptive immunity. With the first
exposure to a protein antigen, a T cell-dependent primary antibody
response occurs. The initial stage of the primary response is a lag period,
or latent period, of approximately 10 days, during which no antibody can be
detected in serum. This lag period is the time required for all of the steps of the
primary response, including naïve mature B cell binding of antigen with BCRs,
antigen processing and presentation, helper T cell activation, B cell activation,
and clonal proliferation. The end of the lag period is characterized by a rise in
IgM levels in the serum, as TH2 cells stimulate B cell differentiation into plasma
cells. IgM levels reach their peak around 14 days after primary antigen
exposure; at about this same time, TH2 stimulates antibody class switching, and
IgM levels in serum begin to decline. Meanwhile, levels of IgG increase until they
reach a peak about three weeks into the primary response (Figure 7).

During the primary response, some of the cloned B cells are differentiated
into memory B cells programmed to respond to subsequent exposures.
This secondary response occurs more quickly and forcefully than the primary
response. The lag period is decreased to only a few days and the production of
IgG is significantly higher than observed for the primary response (Figure 4). In
addition, the antibodies produced during the secondary response are more
effective and bind with higher affinity to the targeted epitopes. Plasma cells
produced during secondary responses live longer than those produced during the
primary response, so levels of specific antibody remain elevated for a longer
period of time.

Figure 7. Compared to the primary response, the secondary antibody response occurs more quickly
and produces antibody levels that are higher and more sustained. The secondary response mostly
involves IgG.

12 | P A G E
B-Cell Types
1. Plasmablast - A short-lived, proliferating antibody-secreting cell arising
from B cell differentiation.
2. Plasma cell - A long-lived, non-proliferating antibody-secreting cell
arising from B cell differentiation.
3. Lymphoplasmacytoid cell - A cell with a mixture of B lymphocyte and
plasma cell morphological features that is thought to be closely related to
or a subtype of plasma cells.
4. Memory B cell - Dormant B cell arising from B cell differentiation.
5. B-2 cell - FO B cells and MZ B cells.
 Follicular (FO) B Cell (also known as a B-2 cell) - Most common type
of B cell and, when not circulating through the blood, is found mainly
in the lymphoid follicles of secondary lymphoid organs (SLOs).
 Marginal zone (MZ) B cell - Found mainly in the marginal zone of the
spleen and serves as a first line of defense against blood-borne
pathogens, as the marginal zone receives large amounts of blood from
the general circulation.
6. B-1 cell - Arises from a developmental pathway different from FO B
cells and MZ B cells.
7. Regulatory B (Breg) cell - An immunosuppressive B cell type that stops
the expansion of pathogenic, pro-inflammatory lymphocytes through the
secretion of IL-10, IL-35, and TGF-β.[25] Also, it promotes the
generation of regulatory T (Treg) cells by directly interacting with T cells
to skew their differentiation towards Tregs.

13 | P A G E
 SUMMARY

 B lymphocytes or B cells produce antibodies involved in humoral

immunity. B cells are produced in the bone marrow, where the initial
stages of maturation occur, and travel to the spleen for final steps of
maturation into naïve mature B cells.
 B-cell receptors (BCRs) are membrane-bound monomeric forms of IgD
and IgM that bind specific antigen epitopes with their Fab antigen-
binding regions. Diversity of antigen binding specificity is created by
genetic rearrangement of V, D, and J segments similar to the mechanism
used for TCR diversity.
 Protein antigens are called T-dependent antigens because they can only
activate B cells with the cooperation of helper T cells. Other molecule
classes do not require T cell cooperation and are called T-independent
antigens.
 T cell-independent activation of B cells involves cross-linkage of BCRs
by repetitive nonprotein antigen epitopes. It is characterized by the
production of IgM by plasma cells and does not produce memory B cells.
 T cell-dependent activation of B cells involves processing and
presentation of protein antigens to helper T cells, activation of the B cells
by cytokines secreted from activated TH2 cells, and plasma cells that
produce different classes of antibodies as a result of class switching.
Memory B cells are also produced.
 Secondary exposures to T-dependent antigens result in a secondary
antibody response initiated by memory B cells. The secondary response
develops more quickly and produces higher and more sustained levels of
antibody with higher affinity for the specific antigen.

14 | P A G E
References

1. B Cell Activation(https://www.sciencedirect.com/topics/medicine-and-dentistry/b-cell-
activation)
2. An Overview of B Cells – from Discovery to Therapy(https://www.bio-rad-
antibodies.com/static/2016/b-cell/minireview/an-overview-of-b-cells---from-discovery-to-therapy-
mini-review.pdf)
3. B Cells (https://www.immunology.org/public-information/bitesized-immunology/cells/b-
cells#:~:text=The%20function%20of%20B%20cells,'B'%20cell%20was%20derived. )
4. B-Cell And Humoral Immunity
5. B Cell (https://en.wikipedia.org/wiki/B_cell#Development)