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CELLS INVOLVED CD21 145 B cells, Receptor for

LYMPHOCYTES follicular complement
20 40% of dendritic cells component C3d;
part of B-cell
circulating WBC coreceptor with
w/ large, rounded CD19
nucleus, nuclear CD 175 NK cells, Not known
chromatin is dense 56 220 subsets of T
cells
no granules
2nd abundant cells T LYMPHOCYTES
Major Lymphocytes Originated in bone marrow, matures in thymus
B cells
61-80% of lymphocytes
T cells
NK cells (natural immunity) End product: cytokines (lymphokines)
Appearance of B cells and T cells is the same Cell-mediated immunity
Cluster of Differentiation (CD) Arm-to-arm combat
o Antigenic features of leukocytes that are Cytokines activates macrophages, plasma cells
differentiated by groups of monoclonal and immune system
o Identification Markers: CD2, CD3, CD4, CD8
o Receptor Longer lifespan (4 10 years)
Traditional Marker Why do we still get reinfected after prior
Traditional marker of B cell is surface exposure?
immunoglobulin (IgM and IgG) o Organisms with different strains
T cells contains CD2 o Viruses have the ability to rearrange their
What is the red cell used to differentiate T and B antigenic component
cell? Sheep RBC since it has the ability to attach to Responsible for immune response and are
the T cell because it has CD2 and it is the receptor involved in antibody regulation
of sheep RBC D e d ce a ib d
Newest marker Subsets: T helper cells, Cytotoxic T cells
B cells: CD 19, CD20, CD21 T helper cells
T cells: CD3 acti a e imm e em, chi m a
T Helper cells: CD4+, CD8- informs about the presented antigen
T Cytotoxic cells: CD8+, CD4- ADCC, the antibody coated cell can be killed by
NK cells: CD2+, CD3-, CD16, CD56+ Cytotoxic T cells
NK cells is formerly classified as T cell because it Type 1
both have CD2 marker produces interferon-gamma (IFN-gamma), tumor
Surface Markers on T, B, and NK Cells necrosis factor-beta (TNF-beta), IL-2
AG MOL CELL TYPE FUNCTION protect cells against multicellular pathogens by
WT
(KD)
activating cytotoxic lymphocytes and
CD3 20 28 Thymocytes, T Found on all T cells; macrophages
cells associated with T- Type 2
cell antigen receptor produces IL4, IL5, IL6, IL9, IL10, IL13
CD4 55 T helper cells, Identifies T helper help B cells produces antibodies against
monocytes, cells; also found on extracellular pathogens and to generally and to
macrophages most T regulatory
cells generally regulate B-cell activity
CD8 60 76 Thymocyte Identifies cytotoxic T Type 3
subsets, cells T regulatory cells
cytotoxic T CD4 and CD5
cells
CD16 50 80 Macrophages, Low affinity Fc
5% of all CD4+ cells
NK cells, receptor for Suppress immune response to self-antigens
neutrophils antibody; mediates Inhibit proliferation of other T-cell population by
phagocytosis secreting inhibitory cytokines
CD19 >120 B cells, Part of B-cell
follicular coreceptor; To prevent autoimmune disorders
dendritic cells regulates B-cell Subpopulation:
development and Th9 (Type 9)
activation produce interleukin-9 (IL-9)
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appear to have a proinflammatory effect negative selection takes place among the
they stimulate growth of hematopoietic cells surviving DP T cells if it reacts with self antigen
may promote autoimmune inflammation Negative selection takes place in the
Th17 (Type 17) corticomedullary region and the medulla of the
produce (IL-17) and (IL-22) thymus as medullary epithelial cells express a
Cytotoxic T cells wide variety of self-antigen
Kills infected cells Clonal deletion will happen to the autoreactive
CD8+ cells
kille Mature T cells
Associated with specific killing Exhibit only one type of marker
Specific CTcells vs HepaA Single positive cells
Specific CTcells vs HepaB Activated T cell
NK cells are nonspecific, it will kill any enemy T cell that is exposed to an antigen
bodies Sensitized T cell
T Cell Receptors (TCR) Capable of producing cytokines
Consists of alpha and beta chain
B-LYMPHOCYTES
CD2 sheep red blood cell antigen
10-20% of lymphocytes
CD3 part of T cell antigen-receptor complex
End product: Antibody
CD4 receptor of MHC class II molecule (Th. Tr)
Humoral immunity
CD8 receptor of MCH class 1 molecule (Tc)
Markers: Surface immunoglobulin (IgM, IgD),
appearance of the β chain also triggers the CD19, CD20, CD21, Class II MHC molecule
thymocyte to become CD4 positive (CD4+) and
Shorter life span (3-5 days)
CD8 positive (CD8+)
Precursor cell in antibody production
capable of recognizing antigens without being
presented by major histocompatibility complex Subsets: B1, B2
(MHC) proteins B2 most abundant B-cells
Light chain: Kappa and Lambda (2Kappa:
DEVELOPMENT OF T LYMPHOCYTES 1Lambda)
Double-Negative Thymocytes Disulfide bonds connect HC to LC
lack CD4 and CD8 markers Hinge region gives flexibility to the antibody,
double-negative (DN) thymocytes rich in amino acids
actively proliferate in the outer cortex under the
influence of IL-7
rearrangement of the genes development of T-cell Light chain
receptor begins at this stage
Heavy chain
T-cell receptor (TCR)
Double-Positive Stage
thymocytes express both CD4 and CD8 antigens
e ca cla if e he he he a e T-helper
cell or Cytotoxic T-cells DEVELOPMENT OF B LYMPHOCYTES
positive selection process takes place that allows Pro B Cells
only DP cells with functional TCR receptors to Lacks surface immunoglobulin
survive Require direct contact w/ bone marrow stromal
o cells are allowed to multiply cells
thymocytes bind to self-MHC antigens in the C-kit interacts w/ a cell surface molecule called
cortex by means of kinases stem cell factor bound on stromal cells
The selection of thymocytes that will only interact 1) Rearrangement of genes that code for HC and LC
with the MHC antigens found on host cells is known of an Ab molecule (Chromosome 14)
as MHC restriction. a. Kappa LC: Chromosome 2
Any thymocytes that have either a very low or a b. Lambda LC: Chromosome 22
very high affinity for self-MHC antigens die by Differentiation of pro B-cells occurs upon successful
apoptosis. rearrangement of heavy chain genes on one of the
functioning T cells must be able to recognize chromosome 14
foreign antigen along with MHC molecules.
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Pre B cells Fluorescence microscopy
When synthesis of the heavy-chain part of the Density gradient centrifugation
antibody molecule occurs Separates lymphocytes from other blood cells
accompanied by an unusual light chain molecule Regent use: Ficoll Hyphaque
called a surrogate light chain. Ficoll separates lymphocytes from other
Immature B cells components. It will produce hazy looking specimen
distinguished by the appearance of complete IgM in between two components
antibody molecules on the cell surface Hazy lymphocytes will be added with RPMI
Completion of light chain rearrangement Specific gravity: 1.077
IgM is the first developed immunoglobulin Culture medium: Roswell Park Memorial institute
CD21, CD40, and class II MHC molecules (RPMI) culture medium for lymphocyte
Mature B Cells
Addition of surface immunoglobulin (IgD)
marginal zone B cells (B cells found on spleen)
follicular B cells (found in other areas of secondary
lymphoid organs)
Activated B cells
Plasma Cells
Represent the most fully differentiated lymphocyte
Main function: Ab production Mitogen
spherical or ellipsoidal cells characterized by Increases concentration of lymphocytes by
eccentric or oval nucleus with heavily clumped promoting mitosis
chromatin that stains darkly presence of abundant T-cell/ B-cell response may be measured by
Plasma cells has major concentration of quantitating the uptake of radioactive thymidine, a
cytoplasmic immunoglobulin and little to no surface precursor of DNA
immunoglobulin Increased thymidine uptake suggests cell division
B-cells d ha e c la mic imm gl bi i h and activation
majority of surface immunoglobulin T cells
o Pokeweed antigen (PWA)
o Phytohemagglutinin (PHA)
o Concanavalene A (Con-A)
B cells
o Pokeweed antigen (PWA)
o Lipopolysaccharide
Comparison of T, B, and NK Cells
LABORATORY IDENTIFICATION OF T CELLS B CELLS NK CELLS
LYMPHOCYTES Develop in the Develop in the Develop in the
Rosette Assay thymus bone marrow bone marrow
Found in lymph Found in bone Found in spleen,
Gold standard before nodes, thoracic marrow, spleen, liver 5 15% of
Sheep RBC duct fluid 60 lymph nodes, circulating
CD2 receptor on sheep RBC 80% of 10 15% of lymphocyte pool
circulating circulating in blood
Flow cytometry
lymphocyte pool lymphocyte pool
Confirmatory test in blood in blood
Gold standard in differentiating blood cells Adaptive Adaptive Innate immunity:
is able to segregate lymphocytes into subsets using immunity: end immunity: end lysis of virally
products of product of infected cells
a technique that relies on labeled monoclonal activation are activation is and tumor cells;
antibodies against specific surface antigens cytokines antibody production of
can simultaneously measure multiple cellular or cytokines
bead properties by using several different Antigens include Antigens include Antigens include
CD2, CD3, CD4, CD19, CD20, CD16, CD56
fluorochromes
or CD8 CD21 surface
common antigens tested for include CD2, CD3, CD4, antibody
and CD8 on T cells and CD19, CD20, CD22, and
surface immunoglobulin on B cells