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Vinnie Maugeri
Biology, Senior
At Life’s first dawn,
God chose for us,
This magic blessed place.
That we might together share,
The joys of Time’s embrace
Cellular Components,
Cytokines and Growth Factors
of the
Immune Response.
Stem Cells and Stem Cell
Transplantation
Folder Title:
Cells(NoTP)
Updated: November 1, 2016
Questions About Cellular Components
of the Immune Response
How many different cell types are there?
What are the numbers of the various cell types?
What do these different cell lineages do?
Where do they come from?
How Long do they last?
What becomes of them at the end of their functional life span?
What controls their replacement? (How does the hematopoietic
system know what needs to be replaced?)
What happens if they aren’t replaced correctly?
If they are deficient in number?
If they are produced in excess to what is needed?
If they are not structurally or functionally normal?
How We Learn Things:
Memorization vs Gradual Familiarity
Immune System Make-up
From 447Intro
Morphology &
Staining of
Blood Cells
Kuby, 3rd Ed.
Figure 3-1
About 6 u diameter
About 20 u diameter
700 RBC to one
leucocyte
Leukemic
Leucocytes:
About 20u
diameter
Red Blood
Cells:
4 to 6 u
diameter.
Diluted
1:1000 from
whole blood
In this photo:
33 RBC:1
Leukemic
Leucocyte
Replacing Cells at the Correct Time
with the Correct Cell Type
Hematopoiesis
Where Do the Blood Cells Come From?
Stem Cells and Partially Differentiated Progenitor Cells
How Can We get Our Hands on Stem Cells for
Treatments?
Stem Cell Therapy in Medicine
See Figure 2-1
p. 28, 7th Editon
Hematopoiesis
(formation
of blood cells)
Fig 2-1,
Kuby 4th Ed. p. 28
Right to Left
View
Note Cytokine
Signalling:
IL2, IL3, IL4,
IL6, IL7,
IL8,IL11,EPO,
GMCSF
Hematopoiesis
What the cells look
like as colonies
growing in soft agar
Where Hematopoietic Cells
Come From:
Separating Different Kinds
of Hematopoietic Cells
Antibodies to differentiated
end-product blood cells
Separating Out Stem
Cells (S) and
Pluripotent progenitor
cells (P)
Removes
progenitor (P) Cells
Isolation of CD34+ Pluripotent Stem Cells
from Mixed Bone Marrow Mononuclear
Cells Using Monoclonal Antibody to CD34
Marker on Stem Cells
Biotin
attached
To Antibody
Fc Region
These are the Pluripotent Stem Cells that
can make all of the other blood cells
Antibody conjugated to Biotin.
Avidin coats insoluble beads
Biotin Sticks Strongly to Avidin
200,000 peripheral blood cells restore
viability
1,000 mixed stem cells and
progenitor cells restore viability
Pure Stem Cells
30 to 100 Stem cells restore viability
See Figure 2-5
Kuby, 6th Ed.
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Myeloid Lineage
Differentiation and Function Among
“Granulocytes”, Polymorpho-nuclear
Leucocytes (PMN’s) :
Neutrophils, Eosinophils, Basophils/Mast Cells,
Monocytes
Monocytes/Macrophages, Dendritic Cells
Erythrocytes
Magakaryocytes/Platelets
Cells of Myeloid Lineage
Polymorphonuclear leukocytes: (Granulocytes)
Neutrophils, Eosinophils, Basophils, Mast Cells
Antimicrobial, allergic reactions, ADCC
Monocyte Macrophages: Mononuclear phagocytes
Antimicrobial, attack virally infected cells,
Phagocytosis, Endocytosis, & Pinocytosis
Degrade and present processed antigens
Dendritic Cells
Similar functions as for macrophages
See Slides 49 - 53
Erythrocytes: Red Blood Cells, carrying oxygen
Megacaryocytes: Produce platelets for blood clotting
Myeloid Lineage
Myeloid
Lineage
(Kuby,
See Figure 2-2
Hematopoiesis
6th Edition
p. 25)
Note roles of cytokines,
Activated T-Helper
Cells,
And Activated
Macrophages
BloodCells1
BloodCells2
Histiocyte
(Connective
Tissue)
Mesangial Cell
(Kidney)
Osteoclast (Bone)
Myeloid to Monocyte
Differentiation in the
Myeloid Series to Monocyte
Macrophage Lineage
Tissue Macrophages
Differentiation and Function in the Granulocyte Lineage
Neutrophils:
Eosinophils:
Basophils and Mast Cells
Differentiation and Function in the Granulocyte Lineage
Neutrophils:
Multi-lobed Nucleus (PMN) Polymorphonuclear Leucocyte
50% of circulating leukocytes.
Short-lived (Hours or Days).
Phagocytic
Circulates, extravasates out of vasculature into tissue.
Responds to chemotactic factors released by infection and inflammation
(e.g. from complement or blood-clotting reactions or cytokines released by
T-cells or macrophages).
Granules release peroxidase, lysozyme, hydrolases, proteases, collagenase.
Antimicrobial agents released.
Part of innate natural immune response.
Differentiation and Function in the Granulocyte Lineage
Eosinophils: Acidic Granules
Anti-parasitic immunity
1% of circulating leukocytes
Phagocytic
Basophils:
Less than 1% of circulating leukocytes
Non-phagocytic
Degranulate to release substances supporting allergic attack
Mast Cells:
Similar to Basophils, but in tissue sites
Secrete histamine in allergic attacks
Erythropoetin
ROS = Reactive
Oxygen Species
MIP-1a = Macrophage
Inflammatory protein
See Appendix III, p. C-1, 7th Edition
Note Name Changes & Complexity
See Slide Following from Science, 2015, 349, Nov. 2015
Neutrophils trails guide virus-specific CD8+ T cell migration.
(CXCL12 is a chemokine: chemical agent attracting cell movement)
Virus-specific Cytotoxic T-Cell
Blebs containing CXCL12 from Uropod
(Trailing membrane region) of Neutrophil
“Bread-crumb” trail
Kihong Lim et al. Science 2015;349:aaa4352
Cytotoxic T-Cell migration through trachea to virus-infected cells
Published by AAAS
Differentiation and Function in
Monocyte/Macrophage Dendritic
Cell Lineage
Monocyte to Macrophage
Macrophage (MPH or MO)
Monocyte
Monocyte to Macrophage
Kuby, Immunology. 6th Edition
Figure 2-7
MC&MPH
Macrophage Ingesting and Degrading Bacterial Targets
Kuby, Immunology, 6th Edition, Figure 2-8
Macrophage
bacteria
Antigen presentation
MPHIngest
Kuby, 4th Edition, p. 44
Macrophage Factors
Cytokines
Cytokine
Cytokine
Growth
Factors
MPhMake
Macrophage and PMN Killing
Kuby, 4th Edition, p. 43
Agents
MPhKill
Types of Antigen-Presenting
Dendritic Cells.
(Shown in
Lymphoid Series
in Slide 53)
Dendritic Cells from Sci Am
Dendritic and T-Cell
Dendritic Cell micrograph
Megakaryocyte – Platelet Lineage:
Blood-Clotting Function
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October 27, 2016
Differentiation and Function in
the Lymphocyte Series
Cells of Lymphocyte Lineage
B-Lymphocytes: Antibody receptors and antibody production
T-Lymphocytes (Thymus-derived lymphocytes):
T-Cell Receptors
Helper T-Cells “CD4 positive”
Cytotoxic T-Cells “CD8 Positive”
Natural Killer Cells (“Non-B-Cell, Non-T-Cell Lymphocytes)
Recognize virally infected or transformed cells
Bind to antibody labelled cells as part of antibodydependent, cell mediated cytotoxicity (ADCC)
Lymphoid Lineage
Fig 2-1 Kuby 4th Ed
p. 28
Natural Killer Cell
(Non-B, Non-T Cell)
Bone-marrow-derived
lymphocyte (Makes Antibodies)
Helper T-Cell
(Helps B-Cell Differentiation)
Promotes Cellmediated Immunity
Committed Lymphocyte Progenitor
Lymphocytes
“Educated” in Thymus
Cytotoxic T- Cell
Promotes Humoral
Immunity (Helps BCells make Ab’s &
Itself Makes
Cytokines)
TH1 and TH2
RBC?
Note size
differences
RBC????
Antigen-activated B- Cells
Differentiated B-Cell;
Makes Antibodies
Natural Killer Cells
•
•
•
•
Large Granular Lymphoctyes
Part of innate natural immune response
Usually without T-Cell Receptor or Membrane Antibody
Recognize patterns of surface molecules or unusual
expression of self-molecules (Class I MHC)
• Have anti-tumor and anti-viral activity
• CD16 Membrane Receptor for specific antibody regions
gives Antibody-dependent cell-mediated cytotoxic activity
• NKT Cells have TCR,
– Bind to MHC-like molecules CD1
– Secrete cytokines
See Appendix 1; Pages A1 to A29, Edition 7
350 CD Antigens on Leucocytes. What Cell Express Them
What they do/
Differentiation Antigen Markers (CD Antigens) on Lymphocytes
p. 34
Unique TCell Marker
Unique TCell Marker
Distinguishes
Tc from TH
Unique
NK-Cell Marker
Unique BCell Marker
Unique TCell Marker
Unique BCell
Markers
Unique NK-Cell Marker
Used to identify sub-populations of lymphocytes and
to isolate them
Blood Cell Replacement Problems
At the correct time:
When cells are damaged, aged, or no longer functional or
necessary.
Replace with the correct cell type.
In the correct number.
Do not propagate errors arising during cell division.
Blood Cell Survival Times and Turn-Over
Erythrocytes (Red Blood Cells)
~ 4 Months
Neutrophils
1 Day
Lymphocytes
Years
White Blood Cell Generation
3.7 x 1011/day
(50 x World Human Population per Day)
Replacing Cells at the Correct time
Getting Rid of Aged or Damaged Cells
Without generating inflammation:
Genetically Programmed Cells Death (Apoptosis)
vs Inflammatory Lysis and Necrosis
Genetically Programmed
Cell Death: Apoptosis
Cysteine – aspartate protease
Bcl-2 gene: B-cell Leukemia associated gene
Hematopoeisis and Leukemogenesis (Leukemia)
What happens if damaged cells are not destroyed?
What happens if Apoptosis is not invoked?
Bcl-2 gene up-regulation in leucocytes leads to leukemia.
(Strong inhibition of Apoptosis)
FAS Gene or Caspase Genes down-regulated or lost in
cells leads to leukemia and other cancer.
(Failure to initiate or promote Apoptosis)
“Caspase” = cysteine-aspartate proteaase
Increasing this leads to failure of cell death and to leukemia
Cysteine – aspartate protease
Shutting these down leads to failure of cell death and to leukemia
Bcl-2 gene: B-cell Leukemia associated gene
Cytokines, Cytokine Receptors,
and Normal and Pathological
Cell Signaling
How do the multiplicity of hematopoietic cells at
distant sites in the host “talk” to one another?
How do the cells “know” where to go and what to
do?
Cytokine Signaling and Cytokine Receptors
in
Normal Hematopoiesis and in Leukemia
An Exercise
In Cytokine Signaling and Cytokine
Receptors
Need persons who speak:
Spanish
Chinese
Japanese (or Other?)
Key Hematopoietic Growth Factors
and Their Targets
Relatively Multi-Specific:
Granulocyte-Macrophage Colony-Stimulating Factor
GMCSF
Interleukin III - IL3
Relatively Mono-Specific:
Granulocyte Colony Stimulating Factor - GCSF
Macrophage Colony Stimulating Factor - MCSF
Erythropoietin - EPO
GrowFact
Cytokine Table
See
Table 3-1
Kuby
3rd Ed.
Autocrine stimulation
(Self-Signaling) and the
Generation of Leukemias and
other Cancers
See Figure 3-6
Kuby, 3rd Ed.
Appendix II, Pages B1 to B6, 7th Edition
58 Cytokines from Interleukin 1
to Tumor Necrosis Factor Beta (TNF-B)
Autologous Cells in
Transplantation Medicine
Stem Cell Therapy in
Medicine
Autologous Transplant
Step 2
Step 1
Step 3
Stem Cell Transplantation in Medicine:
(See pages 42-43, 7th Edition)
Treatments with Stem Cells
In Immune deficiency diseases
In immuno-suppressed states
Autologous Transplants
Non-Self Donors
Syngeneic Donors
Allogeneic Donors
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To Get to Animations & Molecular Visualizations
Produced for Kuby Immunology
http://bcs.whfreeman.com/immunology6e/
(Or search “Kuby Immunology”,
Click on “Kuby Immunology 6e” , go to Student Resources)
Animations
Chapter 2: Cells; Cell Death
Chapter 10: Cell Death
Chapter 11: Signal Transducton
Chapter 13: Leucocyte Extravasation
Molecular Visualization
Chapter 2: Cells and Organs
Chose Other Chapters for Other Molecular Visualization
Edition 6 (2007) :Appendix 1: Pages A1 to A26
339 CD Antigens on Leucocytes
Edition 7 (2013): Apendix 1: Pages A1 to A29
350 CD Antigens on Leucocytes
What cells types express them
What they do
e.g. CD4 is a co-receptor on helper T-cells.
Confirms binding of T-Cell with its T-Cell Receptor to
an antigen-presenting cell.
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