BLOOD

& LYMPH

Grace Mendoza-Bardelosa, MD, FPDS

Department of Human Structural
Biology
 Intended Learning Outcomes
•  Describe blood and its composiDon.
•  DifferenDate the formed elements of blood.
•  Discuss the clinical significance of derangements
in the composiDon of blood.
•  Discuss crenaDon and hemolysis.
•  Explain the role of formed elements of blood in
common clinical condiDons.
•  Discuss lymph formaDon.
•  Discuss common clinical condiDons that affect
lymph formaDon and flow.
 Blood
•  Specialized connecDve Dssue
•  Cells suspended in PLASMA
•  5 L move unidirecDonally in a closed
circulatory system
 Formed Elements
•  Erythrocytes
•  Leukocytes
•  Platelets
 Plasma
•  Straw-colored, translucent
•  Slightly viscous supernatant
•  55% at top of a centrifuged tube

 Hematocrit
•  UncloTed blood
•  Erythrocytes that are sedimented
•  45% in healthy adults
 Buffy Coat
•  Thin gray-white layer between plasma and
hematocrit
•  1%
•  Leukocytes and platelets
ComposiDon of Whole Blood
 Serum
•  Liquid porDon of cloTed blood
•  Contains growth factors and other proteins
released from platelets
•  Biologically different from plasma
 FuncDons of Blood
•  A distribuDng vehicle (O2, CO2, metabolites,
hormones and other substances)
•  ParDcipates in heat distribuDon
•  RegulaDon of body temperature
•  Maintenance of acid-base and osmoDc
balance
 ComposiDon of Plasma
•  An aqueous soluDon
•  pH 7.4
•  Dissolved compounds are mostly proteins
•  Nutrients
•  Respiratory gases
•  Nitrogenous waste products
•  Hormones
•  Inorganic ions aka electrolytes
 ComposiDon of Plasma
•  Albumin
–  Most abundant; maintains osmoDc pressure of blood
•  α-Globulins and β-globulins
–  Transport lipids, metal ions, iron ions in bloodstream
•  γ-Globulins
–  Immunoglobulins
•  Fibrinogen
–  ParDcipates in blood coagulaDon, precursor of fibrin
•  Complement proteins
–  Important in inflammaDon & destrucDon of organisms
Preparing a Blood Smear
ERYTHROCYTES
 Erythrocytes
•  Non nucleated
•  Completely filled with hemoglobin
•  Biconcave discs – provides large surface-to-
volume raDo, facilitates gas exchange
•  7.5 μm diameter
•  2.6 μm thick at rim
•  0.75 μm thick at center
Normal Erythrocytes
 Erythrocytes
•  NV females: 3.9 to 5.5 million/ μL
•  NV males: 4.1 to 6.0 million/ μL
•  Flexible
•  Cuplike shape
•  Rouleaux: RBC’s adhere to one another
loosely in stacks
 Erythrocytes
•  Plasmalemma, best known membrane
•  40% lipid, 10% carbohydrate, 50% protein
•  Mostly integral membrane proteins
–  Ion channels
–  Band 3 protein (anion transporter)
–  Glycophorin A
•  Glycosylated extracellular domains include anDgenic
sites
•  Basis for ABO blood typing system
 Erythrocytes
•  Peripheral proteins
•  Spectrin: forms lakce with acDn filaments
•  Ankyrin: anchors the lakce to glycophorins
and band 3 proteins
•  Stabilizes the membrane
•  Maintains cell shape
•  Provides cell elasDcity
 Erythrocytes
•  Lacks all organelles
•  Filled with hemoglobin
•  Tetrameric O2 carrying protein
•  Oxyhemoglobin (O2)
•  Carbaminohemoglobin (CO2)
•  Hemoglobin + CO
 Erythrocytes
•  No nucleus and organelles
•  No mitochondria
•  Rely on anaerobic glycolysis for minimal
energy needs
•  Cannot replace defecDve proteins
•  120 days normal life span
•  Removed in circulaDon by macrophages
 Anemia
•  CondiDon where concentraDon of RBC’s are
below the normal range
•  S/Sx: lethargy, shortness of breath, faDgue,
skin pallor, cardiac palpitaDons
•  Causes:
–  Insufficient RBC producDon
–  Blood loss
 Erythrocytosis
•  Polycythemia
•  Increased concentraDon of RBC’s
•  Physiologic
–  Areas with high alDtudes
–  O2 tension is low
–  Increase in hematocrit
Sickle Cell Erythrocyte
LEUKOCYTES
 Leukocytes
•  Granulocytes
•  Azurophilic granules or lysosomes
•  Specific granules
–  Bind neutral, basic, or acidic stains
–  Have specific funcDons
•  Polymorphic nuclei
•  Neutrophils, Eosinophils, & Basophils
 Leukocytes
•  Agranulocytes
•  No specific granules
•  Azurophillic granules
•  Nuclei indented but not lobulated
•  Lymphocytes & Monocytes
 Leukocytes
•  Defense against invading microorganisms
•  Repair of injured Dssues
Normal Leukocytes
 Neutrophils
•  12-15 μm
•  54% to 62%
•  Nuclei: 2 to 5 lobes linked by thin nuclear
extensions
•  Females: drumsDck-like appendage
•  InacDve and spherical when inacDve
•  Amoeboid during diapedesis
 Neutrophils
•  AcDve phagocytes of bacteria
•  Usually first to arrive at sites of infecDon
•  Azurophilic primary granules
•  Role in killing and degrading engulfed
microorganisms
–  Myeloperoxidase
–  Lysozyme
–  Defensins
 Neutrophils
•  Myeloperoxidase
–  Generates hypochlorite & other agents toxic to
bacteria
•  Lysozyme
–  Degrades components of bacterial cell wall
•  Defensins
–  Cysteine-rich proteins that bind and disrupt cell
membranes of bacteria
 Neutrophils
•  Specific secondary granules
•  SecreDon of ECM-degrading enzymes such as
collagenases
•  Delivery of addiDonal bactericidal proteins to
the phagolysosomes
•  InserDon of new cell membrane components
 Neutrophils
•  Role in inflammatory response
•  Restore normal Dssue microenvironment
•  First line of defense against invasion by
pathogenic bacteria
•  Short-lived
•  Half-life of 6 to 8 hours
•  Life span of 4 days in connecDve Dssue
Neutrophils
 Eosinophils
•  1% to 3%
•  Bilobed nucleus
•  Abundance of large acidiphilic specific
granules stain pink or red
 Eosinophils
•  Major basic protein (MBP)
•  50% of total granule protein
•  Kill parasiDc worms and helminths
•  Modulate inflammatory responses triggered
by allergies
•  Remove anDgen-anDbody complexes from
intersDDal fluid by phagocytosis
•  IntesDnal lining, lung Dssues
Eosinophils
 Basophils
•  12 to 15 μm
•  Less than 1%
•  Nucleus is divided into two irregular lobes
•  Large specific granules stain purple
 Basophils
•  Heparin and sulfated GAG’s
–  Histamine
–  Platelet acDvaDng factor
–  Eosinophil chemotacDc factor
–  Phospholipase A
•  Supplement funcDon of mast cells
–  Contain heparin and histamine
–  Receptors for IgE
–  Secrete components in response to certain Ag’s and
allergens
Basophils
 Lymphocytes
•  B lymphocytes
•  T lymphocytes
–  Helper (CD4+)
–  Cytotoxic (CD8+)
•  Natural killer cells (NK cells)
•  Immune defense against invading
microorganisms, parasites, abnormal cells
 Lymphocytes
•  Small – similar to RBC’s
–  Spherical nuclei
–  Highly condensed chromaDn
–  Scanty cytoplasm
•  Large (acDvated lymphocytes or NK cells)
–  9 to 18 um
–  Slightly indented nuclei
–  More cytoplasm
–  Few azurophilic granules
Lymphocytes
 Monocytes
•  Precursor cells of macrophages, osteoclasts,
microglia and other cells of the mononuclear
phagocyte system
•  AnDgen-presenDng cells
•  Immune defense
 Monocytes
•  12 to 20 μm
•  Nucleus: indented or C-shaped
•  Cytoplasm: basophilic
•  Azurophilic granules: bluish-gray distributed
through the cytoplasm
Monocytes
 Platelets
•  Thrombocytes
•  Non-nucleated cell fragments
•  2 to 4 μm
•  From megakaryocytes
•  Promote blood clokng
•  Repair of minor tears or leaks in small BV’s
 Platelets
•  150,000 to 400,000/uL (mm3)
•  Life span of 10 days
•  Clumps
•  Discoid
•  Hyalomere
–  Lightly stained peripheral zone
•  Granulomere
–  Dark-staining central zone containing granules
Thrombocytes
 Thrombocytopenia
•  Lack of circulaDng platelets
•  Leads to a bleeding tendency
 Thrombocytosis
•  Excess of platelets
•  Leads to risk of inappropriate blood clokng
(thrombosis)
 Normal Plasma
•  Normal plasma is said to be isoosmoDc or
isotonic.
•  Isotonic soluDons maybe prepared for the
study of corpuscles outside the body
•  0.85% soluDon of sodium chloride is
approximately isotonic for the blood
 CrenaDon
•  “Scalloped” or “notched”
•  The formaDon of an abnormal notching
around the edge of an erythrocyte
•  Shrinkage aser suspension in a hypertonic
soluDon
•  Echinocytes or burr cells
 CrenaDon
•  Hypertonic soluDon is added to blood
•  Erythrocytes become shrunken and irregular
in surface contour
•  Cockle-Burr appearance
•  The process is called crenaOon
 Hemolysis
•  Breakage of the RBC’s membrane
•  Release of hemoglobin and other internal
components into surrounding fluid
•  Pink to red Dnge in serum
•  May compromise laboratory’s test parameters
•  Hypotonic soluDon: RBC’s become spheroidal
•  Swelling of cells
 Hemolysis
•  Two causes
•  In vivo: Pathological causes
–  Autoimmune hemolyDc anemia
–  Transfusion reacDon
•  In vitro: Improper specimen collecDon,
processing, transport
LYMPH
 Lymph
•  Excess intersDDal fluid from Dssue spaces
•  Does not contain RBC’s
•  Lymphocytes may be present
•  Rich in proteins
 Lymph
•  Greater hydrostaDc pressure of blood at
arterial ends of capillaries
•  Opposite at venous ends
•  OsmoDc pressure of plasma
•  Increased rate of Dssue fluid formaDon at
arterial end
•  Lesser reuptake of fluid at venous end
•  Excess fluid
 Lymph
•  Chylothorax
•  A type of pleural effusion
•  LymphaDc fluid accumulaDng in the pleural
cavity
•  Causes
–  Lymphoma
–  Trauma
 Lymph
•  Lymphoma
•  NeoplasDc proliferaDon of lymphocytes
•  Failure of lymphocytes to undergo apoptosis
•  All are considered malignant
 Lymph
•  Filariasis
•  A parasiDc disease caused by microscopic,
thread-like worms
•  Adult worms only live in human lymphaDc
system
•  Spread from person to person by mosquitoes
THANK YOU