Blood Revised

8/4/2019 Blood Revised

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BLOOD ANDIMMUNOLOGY

REBECCA A. VILLANUEVA, M.D.Associate Professor, Diplomate, Fellow, PAAB

Department of Biochemistry & Nutrition


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RULE OF 2/3

2/3/ICF

2/3 INTERSTITIAL 1/3 INTRAVASCULAR

(BLOOD)

~7% BW

1/3 ICF

2/3 WATER 1/3 SOLIDS

TOTAL BODY WEIGHT

1/3 ECF


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BLOODviscous fluid with a cellular

portion and fluid portion

I. Cellular PortionplateletsWBCRBC

II. Fluid PortionPlasma

Serum without clotting factors


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Properties of Blood 7 10 % of total body weight

To = 36.5 C to 37.5 C

pH = 7.35 7.45

Osmotic pressure = 7 8 atm

Freezing point = -0.537C

Sp. Gravity = 1.035 1.075

Viscosity = 4 5 x greater than water


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Blood: Functions

I. Transport

A. Respiration

B. Nutrition

C. ExcretionD.Fluids, electrolytes,

minerals, vitamins


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Blood: Functions

II. Regulatory

A. Blood pH

B. Body Temperature

C. Body HormonesD.Body Fluids &

Electrolytes


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III. Protective

A. Infections (WBC)BacterialParasiticViralFungal

Blood: Functions


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III. Protective

B. Immune Diseases

AllergiesImmunitySelf / Non-self

C. Bleeding (Hemorrhage)

D. Excessive clotting

Blood: Functions


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PLASMAPROTEINS


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Major Fractions of Plasma Proteins

1. Fibrinogen

2. Gammaglobulin

3. Prothrombin

4. Blood grouping globulins

5. Pure Albumina. 100 mL of 25% solution = 1 unit o

blood


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Major Fractions of Plasma Proteins

small amount ofalbumin and globulin

large amount of salts

many non-proteinsubstances of low MW


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GENERALIZATIONS

Most plasma proteins aresynthesized in the liver

Plasma proteins are generallysynthesized on membrane

bound polyribosomes Almost all plasma proteins are

glycoproteins


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GENERALIZATIONS Many plasma proteins exhibit

polymorphism

Each plasma protein has acharacteristic half-life

Levels increase during acuteinflammatory states orsecondary to certain types oftissue damage


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PLASMA PROTEINS

comprise the major parts

of solids of plasma separation by Salting Out

Method

Na+ ORAmmonium Sulfate


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PLASMA PROTEINS

Plasma CHON = 7.75 g/dl (6-8)

Albumin = 4.7 5.7 gm%

Globulin = 1.3 2.5 gm%

Fibrinogen = 200 400 mg%


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ALBUMIN


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ALBUMIN

most abundant, most homogenousin electrophoresis

MW - 69,000

Ellipsoidal in shape150 Amstrom x 38 Amstrom

small & compact

Single polypeptide chain (585 a.a.)

with 17 disulfide bonds


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ALBUMIN

compact, small in size fast to travel inelectrophoresis

high solubility

reactive groups in the molecule causealbumin to combine reversibly with agreat diversity of anions and cations

responsible for most of oncotic pressureof blood


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ALBUMIN

N-terminal a.a. Asp C-terminal a.a.

Leu human,dog & rabbit Ala other species

IpH = 4.7 More phototrophic group

(Proton binding groups)


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ALBUMIN Titration studies show molecule of

albumin at ph 7.4

100 COOH groups

58 NH2 groups

16 Imidazole

19 phenolic

22 guanidine groups


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ALBUMIN

Major protein of human plasma 4.5g/dl

Produced in the Liver 12 g/day

60% of Total Plasma Protein

75 80% of Plasma Oncotic

Pressure

Initially synthesized as PRE-PRO

PROTEIN with a signal peptide


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ALBUMIN

Decreased A:G RatioLiver and Kidney Diseases

Malnutrition

Binds various LIGANDSFFA, Calcium, Steroid Hormones

Bilirubin, Tryptophan, CopperDrugs:Sulfonamides, Penicillin G

Dicumarol, Aspirin


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GLOBULIN


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GLOBULIN

Fractions:

A. Pseudoglobulin

B. Euglobulin

glob IpH=5.06 transport (LP) globIpH=5.12 transport (LP) globulin IpH=6.0 Immune system-Ab


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FIBRINOGEN


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FIBRINOGEN: Factor 1

soluble plasma glycoprotein

46 nm in length

MW: 340,000

6 polypeptide chain

(A)2: (B)2;2 A2B2;2


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FIBRINOGEN: Factor 1

end highly negatively charged(Asp & Glu residues A, B)

B - (-) tyrosine O SO4 residue Hence:

Contribute to its H2O solubility

Repulsion of termini of otherfibrinogen molecule preventingaggregation


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Diagram Of Fibrinogen


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Fibrin Polymerization

Major Protein Component


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Major Protein ComponentOf Human Plasma

Concentration

mg/100 ml

Approximate

M.W.

Other Function

Albumin 3 4.5 x 103 68,000 Osm regtransport ofFA, bilirubin

Globulin

glo 100 40 35,000 CHO Not known lipo 350

450

200,000

450,000

4070%lipid

Lipid transport

2 Glo 400900

200,000

2 glyco 200,000 CHO Unknown


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Concentrationmg/100 ml

ApproximateM.W.

Other Function

Ceruloplasmin

30 150,000 CHO Coppertransport

Prothrombin 63,000 CHO Blood clotting

Glo 6001,200

lipoprotein

350

450

3 20 M 8090%lipid

Lipid transport



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Concentrationmg/100 ml

Approx.M.W.

Other Function

lipo protein 350450

200,000

450,000

40 70%lipid

Lipidtransport

TransferrinPlasminogen

40 85,000

90,000

CHO Irontransport

Precursor ofFibrinolysin

Globulins 700 1,500 150 th CHO Antibody

Fibrinogen 300 340 th CHO Blood clottingfactors



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HAPTOGLOBIN


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HAPTOGLOBIN

Plasma glycoprotein

MW = 100,000 daltons

Alpha-phoretic mobility

Irreversibly bindsextracorpuscular Hgb in a tightnon-covalent complex (Hb-Hp)


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HAPTOGLOBIN

KidneyHb Excreted in urine orprecipitates in

tubules (iron is lost)

Hb+Hp KidneyHb-HpComplex

Catabolized By Liver Cells(Iron is Conserved)


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HAPTOGLOBIN


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CERULOPLASMIN


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CERULOPLASMIN

2-Globulin Serum Glycoprotein MW = 160,000

Carries 90% of Cu present in plasma(60% in RBC Superoxide Dismutase)

Deep blue color

Single subunit contains 6 atoms ofcopper

Displays ferroxidase activity


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Physiological functions of copper:1. Oxidation of iron

2. Erythropoiesis and Leucopoiesis

3. Bone Mineralization

4. Elastin & Collagen cross-linking

5. Oxidative phosporylation

CERULOPLASMIN


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Physiological functions of copper:

6. Catecholamine metabolism

7. Melanin formation

8. Myelin Formation

9. Glucose homeostasis

10.Antioxidant protection of cells

CERULOPLASMIN


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-1 ANTITRYPSIN(-1 Antiproteinase)

1 ANTITRYPSIN


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polymorphic single chain plasmaprotein (394 AA); MW 52,000

90% of-1 fraction of humanplasma

synthesized by hepatocytes andmactophages

1 ANTITRYPSIN


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principal serine proteaseinhibition (serpin or Pl) of human

plasma

inhibits trypsin, elastase andother proteases

genetic defect which leads todeficiency causes emphysema

and liver diseases


1 ANTITRYPSIN


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smoking

oxidizes methionine residue

inactivates -1 antitrypsinincrease proteolytic

destruction of lung tissue

emphysema



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HEMOSTASIS


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VASOCONSTRICTION

HEMOSTASIS


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HEMOSTASIS

PLATELET PLUG


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HEMOSTASIS

FIBRIN CLOT


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PLASMIN - FIBRINOLYSIS

HEMOSTASIS


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10x

Inactive

Enzyme A

ActiveEnzyme A

InactiveEnzyme B

ActiveEnzyme B

10x

10xInactiveEnzyme C

Active

Enzyme C

Amplification 10x

Amplification 100x

Amplification 1000x


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PLATELET

ADHESION &

ACTIVATION


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ACTION OF PLATELETS


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ACTION OF PLATELETS

SEROTONIN GROWTH

FACTOR

LIPOPROTEINS FACTOR IV

FACTOR V

20%

FACTOR XIII

(ALPHA S.U.)

ADP THROMBOXANE A2

PLATELETS

vWF

COLLAGEN


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Arachidonic Acid

Cyclooxygenase

Endoperoxidase

Thromboxanesynthetase

ThromboxaneA2

Thromboxane B2(platelets)

Prostacyclinsynthetase

Prostacyclin(PGI2)

6 Keto PGF1(endothelial

cells)

PROPERTY HEPARIN WARFARIN


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PROPERTY HEPARIN WARFARIN

STRUCTURE Large Acidic

polymer

Small lipid-

soluble mol.ROUTE Parenteral Oral

ONSET OF

ACTION

Rapid (Seconds) Slow

M.O.A. ActivatesAntithrombin III

InhibitsVitamin K

ANTIDOTE Protamine Vitamin K /Plasma

USE Acute (Days) Chronic

(Wks. Mos.)


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ANTIPLATELET DRUGS

Inhibits thromboxane synthesisby blocking cycloxygenase

Used in prophylaxis and

treatment of MI, TIA and otherthrombotic events

ASPIRIN / NSAIDS


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Inhibits ADP Formation

Prevents TIA, for patients whocant tolerate aspirin

TICLOPIDINE (Ticlide 500mg)

ANTIPLATELET DRUGS


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ANTIPLATELET DRUGS

CLOPIDOGREL (PLAVIX 75 mg)

A. Inhibits the ADP pathway

B. Both are used for people with stent


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ANTIPLATELET DRUGS

CILOSTAZOL (PLETAAL)

A. Inhibits Phosphodiesterase useespecially with people withintermittent claudication


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SERINE PROTEASE INHIBITORS

Aprotinin

* Risk of kidney failure

A A GS


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Inhibits phophodiesteraseincreasing cAMP

Used in prevention of

thrombosis in artificial heartvalves

DIPYRIDAMOLE

ANTIPLATELET DRUGS

ANTIPLATELET DRUGS


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ANTIPLATELET DRUGS

SOLUDEXINE(VESSEL DUE-F)

ANTIPLATELET DRUGS


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Mechanism unknown

Rarely Used

SULFINPYRAZONE

ANTIPLATELET DRUGS


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COAGULATION

PATHWAY


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Numerical System


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Numerical SystemFor Nomenclature of

Blood Clotting Factors

Factor I Fibrinogen

Factor II Prothrombin

Factor III Tissue FactorFactor IV Ca++

Numerical System


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Factor V Proaccelerin, labile

factor, acceleratorglobulin

Factor VII Proconvertin, serum

prothrombin conversionaccelerator (SPCA),cothromboplastin

Numerical SystemFor Nomenclature of


Numerical System


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Factor VIII Antihemophilic Factor

A, Antihemophilicglobulin

Factor IX Antihemophilic Factor

B, Christmas Factor,plasma thromboplastincomponent (PTC)

u e ca SysteFor Nomenclature of


Numerical System


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Factor X Stuart-Power Factor

Factor XI Plasma thromboplastinantecedent (PTA)

Factor XII Hageman Factor

Factor XIII Fibrin Stabilizing factor(FSF), Fibrinoligase

yFor Nomenclature of


ZYMOGENS OF SERINE


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ZYMOGENS OF SERINEPROTEASES

Factor XIIBinds to exposed collagen at

site of vessel wall injuryactivated by HMWK &kallikrein

Factor XIActivated by Factor XIIa

ZYMOGENS OF SERINE


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Factor IXActivated by Factor XIa in

the presence of Ca++

Factor VIIActivated by thrombin inthe presence of calcium

ZYMOGENS OF SERINE


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Factor XActivated on surface of

activated platelets by tenasecomplex (Ca++, FactorsVIIa and IXa) and by Factor

VIIa in the presence ofFactor III & IV


ZYMOGENS OF SERINE


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Factor II

Activated on surface ofactivated platelets byprothrombinase complex

(Ca++, Factors Vaand Xa)



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COFACTORS

Factor VIII

- Activated by thrombin- Cofactor in the activation

of Factor X by Factor IXa


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Factor V

- Activated by thrombin- Cofactor in the activation

of Prothrombin byFactor Xa

COFACTORS


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Factor III (Tissue Factor)

- Lipoprotein found on thesurfaces of extravascularcells that acts as acofactor for factor VII

COFACTORS

DiagrammaticR i f P h bi


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Representation of ProthrombinNH2

(Gla-XI)P

R

O

T

H

R

O

M

B

I

N

ArgThr

ArgIIe

COO-


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THROMBIN ACTIVATES

FACTOR VII

FACTOR V

PROTEINC

FACTOR

XIIIFACTOR

VIII

REGULATORY AND OTHER


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REGULATORY AND OTHERPROTEINS

Protein C

Activated to protein Ca bythrombinbound tothrombomodulin;

then degrades FactorsVIIIa and Va



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Protein SAct as a cofactor of Protein C;

both has Gla residues

THROMBOMODULIN

Protein on the surface ofendothelial which then activateProtein C cells binds thrombin

REGULATORY AND OTHERPROTEINS



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PSPC PCa

THROMBINTHROMBOMODULIN

Va

VIIIa


PROTEINS


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FIBRIN CLOT

FORMATION OF FIBRIN CLOT


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NH3+Arg Gly

FIBRINOPEPTIDE(A OR B)

Thrombin

FORMATION OF FIBRIN CLOT

STABILIZATION OF FIBRIN CLOT


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FIBRIN-CH2-CH2-CH2-CH2-NH3+ (LYSYL) NH2-CO-CH2-CH2-

FIBRIN (GLUTAMINYL)

FIBRIN-CH2-CH2-CH2-CH2-NH-C-CH2-CH2-FIBRIN

FACTOR XIIIa(TRANSGLUTAMINASE)NH+

STABILIZATION OF FIBRIN CLOT


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FIBRINOLYSIS

DISSOLUTION OF BLOOD CLOT


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FIBRIN

CLOT TPA

FIBRIN

CLOT

PLASMINOGEN PLASMINSOLUBLE

FRAGMENTSFIBRIN

t.PA

t.PA PLASMIN

RESPECTIVE INACTIVECOMPLEXES

INHIBITORSPAI-1 PAI-2

Arg Val

DISSOLUTION OF BLOOD CLOT


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THROMBOLYTIC AGENTS


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Large human protein, MW50,000 Produced in bacteria through

recombinant DNA

Directly converts Ferin-BoundPlasminogen to Plasmin

ALTEPLASE (tPA)

THROMBOLYTIC AGENTS

THROMBOLYTIC AGENTS


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APSAC (Anisoytated Plasminogen

Streptokinase Activator Component) Prodrug (ANISOYL, Group

HYDROLYZED in VITRO)

Slow Release (Long Half Life)

Human TPA from recombinant DNA

ANISTREPLASE

THROMBOLYTIC AGENTS

THROMBOLYTIC AGENTS


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Obtained from bacterialcultures

Forms a complex with

plasminogen prior toconversion

STREPTOKINASE

THROMBOLYTIC AGENTS

THROMBOLYTIC AGENTS


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UROKINASE

Extracted from cultured humankidney cells

Directly converts plasminogen

to plasmin

THROMBOLYTIC AGENTS

PLASMINOGEN

ACTIVATION INHIBITION


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PLASMIN

VARIOUS STIMULI

BLOODPROACTIVATOR

BLOODACTIVATOR

(+) (+)

tPA, UROKINASE(+)

STREPTOKINASE

ACTIVATOR(+)

PROACTIVATOR

ANISTREPLASE

(-)

ANTIACTIVATORS

(-)

AMINOCAPROIC ACIDTRANEXAMIC ACID

Arg Val

DEGRADATION

PRODUCTS FIBRINOGEN

THROMBIN

FIBRINFIBRIN SPLITPRODUCTS

(+) (+)

ACTIVITY OF HEPARIN


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HEPARIN+

ANTITHROMBIN III

PROTAMINESULFATE

( - )

HEPARIN ANTITHROMBIN COMPLEX

THROMBIN XII a XI a X a IX a

ACTIVITY OF HEPARIN

VITAMIN K CYCLE


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VITAMIN K CYCLE

DIHYDROQUINONE / VITAMIN K(H2) + O2 + CO2

PROTEINCARBOXYLASE

VITAMIN K EPOXIDE / VITAMIN K(O) + H3O+

FACTORS II, VII, IX,X PROTEIN C / S

WARFARIN

COUMARIN

DITHIOLS (THIOREDOXIN) / X-(SH)2

PLURIPOTENT

STEM CELL


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PROERYTHROBLAST ERYTHROCYTE

MEGAKARYOCYTES THROMBOCYTES

N. MYELOCYTES PMN

PROMONOCYTES MONOCYTE

E. MYELOCYTES EOSINOPHIL

B. MYELOCYTES BASOPHIL

MYELOID

STEM CELL

B LYMPHOBLAST B CELL

PRE B CELL

T LYMPHOBLAST T CELL

PROTHYMOCYTE

LYMPHOID

STEM CELL

STEM CELL


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FOR LISTENING!!

Blood Revised

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