Donor Criteria

Temperature?

Pulse?

Blood pressure?

Hb and Hct?

Skin?

Donor Criteria

Temperature? No more than 37.5 (99.5)

Pulse? 50-100 (unless athlete)

Blood pressure? 180/110 max

Hb and Hct? 12.5,38% allogenic; 11,33% autologous

Skin?Free of lesions

Deferral Times

1.Viral hepatitis after 11th bday?

2.Pregnancy?

3.Parenteral Drug use stigmata?

4.FH of Creutzfeld Jacob disease?

5.Hx syphilis or gonorrhea, tx, or + syphilis screening test?

6.Recept of blood prodcuts, human tissue, or plasma derived clostting fx?

Deferral

1.Viral hepatitis after 11th bday? Indefinite

2.Pregnancy? 6 weeks postpartum

3.Parenteral Drug use stigmata? Indefinite

4.FH of Creutzfeld Jacob disease? Indefinite

5.Hx syphilis or gonorrhea, tx, or + syphilis screening test? 12 months after tx completion

6.Receipt of blood prodcuts, human tissue, or plasma derived clotting fx? 12 months

Deferral times

1. Receipt of dura mater or pituitatry GH human origin?

2.Aspirin?

3.Recent blood donation

4.Toxoids, synthetic or killed vacinnes: anthrax, cholera, diphtheria, hep A, hep B, influencza, lyme, paratypohiod, pertussis, plague, pneumococcal polysaccharide, polio -Salk, rabies, RMSF, tetatnus, typhoid (injxn)?

Deferral times

1. Receipt of dura mater or pituitatry GH human origin? Indefinite

2.Aspirin? 36 hours

3.Recent blood donation? 8 wks whole blood; 16 wks for 2 unit red cell apheresis; 72 h autologous, >48 for plasma, platelet, leukopheresis

4.Toxoids, synthetic or killed vacinnes: anthrax, cholera, diphtheria, hep A, hep B, influencza, lyme, paratypohiod, pertussis, plague, pneumococcal polysaccharide, polio -Salk, rabies, RMSF, tetatnus, typhoid (injxn)? None (sx free)

Deferral times

1. Live attenuated viral and bacterial vacc (german measles and chicken pox (varicella)?
2. Live attentuated viral and bacterial vacc: measles (Sabin oral), mumps, typhoid (oral), yellow fever?
3. Smallpox vaccine (vaccinia)?
4. Hepatitis B immune globulin?
5. Confirmed + for HbsAg or repeatedly for anti HBc?
6. + HCV infection?

1. Live attenuated viral and bacterial vacc (german measles and chicken pox (varicella)? 4 weeks
2. Live attentuated viral and bacterial vacc: measles (Sabin oral), mumps, typhoid (oral), yellow fever? 2 weeks
3. Smallpox vaccine (vaccinia)? No complications - 21 days or till scab comes off; with, 14 d after resolution
4. Hepatitis B immune globulin? 12 months
5. Confirmed + for HbsAg or repeatedly for anti HBc? Indefnite
6. + HCV infection? Indefinite

Deferral times?

1. HTLV 1 + infection?
2. Previous donation where recipient got HIV or HTLV?
3. Tattoo?
4. Mucous membrane blood exposure?
5. Nonsterile skin penetration?
6. Sexual contact with person w/ viral hepatitis?

1. HTLV 1 + infection? Indefinite
2. Previous donation where recipient got HIV or HTLV? Indefinite
3. Tattoo? 12 months
4. Mucous membrane blood exposure? 12 months
5. Nonsterile skin penetration? 12 months
6. Residing with or Sexual contact with person w/ viral hepatitis? 12 months

Deferral times

1. In jail > 72 hours?
2. Malaria? 
3. Immigrants from malaria areas?
4. Travelers to malaria areas?
5. Travelers to variant Creutzfeld Jacob areas?
6. Use of bovine insuline from UK?

1. In jail > 72 hours? 12 months
2. Malaria? 3 years after being asymptomatic
3. Immigrants from malaria areas? 3 years
4. Travelers to malaria areas? 12 months
5. Travelers to variant Creutzfeld Jacob areas? Indefinite
6. Use of bovine insuline from UK? Indefinite

Deferral times

1. Hx of babesiosis or Chagas?
2. West nile virus?
3. Stigmata of alcohol abuse?
4. Finasteride (proscar, propecia?
5. Dutasterid (avodart)?
6. Isoretinoin (accutane)?

1. Hx of babesiosis or Chagas? Indefinite
2. West nile virus? 14 d after resolved, 28 after onset
3. Stigmata of alcohol abuse? Exclude, no specific time
4. Finasteride (proscar, propecia? 1 month
5. Dutasterid (avodart)? 6 months
6. Isoretinoin (accutane)? 1 month

Deferral times

1. Acitretin (soriatane)?
2. Etretinate (tegison)?
3. Receipt of $ or drugs for sex?
4. Paying for sex?

1. Acitretin (soriatane)? 3 years
2. Etretinate (tegison)? Indefinite
3. Receipt of $ or drugs for sex? Indefinite
4. Paying for sex? 12 months

Kidd Phenotype Frequencies

White             Black

1. JK a+b-
2. Jk a+b+
3. Jk a-b+
4. Jk a-b-

Kidd Phenotype Frequencies

White             Black

1. JK a+b-           28                 57
2. Jk a+b+          49                 34
3. Jk a-b+           23                  9
4. Jk a-b-             Rare              Rare

Blood Group A Immunodominant Sugar Molecule?

N-acetyl-D-galactosamine

(GalNAc)

Which cells agglutinate most strongly with Ulex europaeus lectin?

A2 and O

(The main blood groups agglutinate with the following relative strength with anti-H or Ulex lectin: O > A2 > B > A2B > A1 > A1B.)

What ABH substances would be found in the saliva of a group B secretor?

H and B

( if you are a secretor, you will have both H and B substance in secretions if you are group B.)

Approximately what percentage of group A individuals could be further classified as subgroup A1?

80%

Major difference between the A1 and A2 subgroups?

Quantitative :  A1 RBCs have more A antigen than A2 RBCs

Some qualitative: A2 individuals can make anti-A1.  u clinically insignificant.

- The lectin of Dolichos biflorus only agglutinates RBCs containing A1 specificity,

-lectin of Ulex europaeusagglutinates RBCs with increased H antigen, like most A2 RBCs.

Dosage most common with which blood groups?

Kidd, Duffy, Rh, MNS

[Some antibodies react more strongly with RBC antigens 

that have double-dose (homozygous) gene expression.]

Enhanced Enzyme Classfication Blood Groups?

ABO related (ABO, H, Lewis, I system, P system)

Decreased Enzyme Classfication Blood Groups?

MNS

Duffy

Lutheran

Unaffected Enzyme Classfication Blood Groups?

Kell

Diego

Colton

Substances that neutralize red cell antibody?

ABO

Lewis

P1

Sda

Chido, Rodgers

Substances that neutralize red cell antibody?

ABO       Saliva (secetor)

Lewis     Saliva (secretor for Leb)

P1         Hydatid cyst fluid, Pidgeon egg whites

Sda       Human urine

Chido, Rodgers       Serum

Lectin Specificity

Dolichos biflorus

Ulex Europaeus

Vicia gaminea

Arachis hypogea

Glycine Max

Salivea

Lectin Specificity

Dolichos biflorus  A1

Ulex Europaeus     H

Vicia gaminea        N

Arachis hypogea     T

Glycine Max          T, Tn

Saliva                Tn

Group B sugar?

Infectious tests needed to be done on blood?

HbsAg, anti-HBc

Anti-HCV, HCV nucleic acid

anti-HTLV1, anti-HTLV 2

anti-HIV-1, anti-HIV-2, HIV nucleic acid

RPR

West nile virus (serology)

Blood group carbohydrate antigens

Blood types

Naturally occuring?

Ig class?

React at room temp?

Immediate spin or AHG reaction?

Blood group carbohydrate antigens?

Blood types-ABO, Le, I, M, N, P

Naturally occuring?-Naturally occurring

Ig class?-IgM

React at room temp?-Yes

Immediate spin or AHG reaction?-  Immediate spin

[Agglutinating antibodes]

Blood group protein antigens

Blood types?

Naturally occuring?

Ig class?

React at room temp?

Immediate spin or AHG reaction?

Blood group protein antigens

Blood types? Rh, Kidd,l Kell, S, s, Duffy (all others than carbohydrate Ag)

Naturally occuring? After exposure

Ig class? IgG

React at room temp?  37 C

Immediate spin or AHG reaction? AHG phase

H antigen (notes)

Made from type 1 and 2 precursor carbohydrate chains

In secretions, type I covertedto H antigen by enzymatic action (fucosylation) of the Se gene product.

In blood, type 2 chains converted to H antigen by fucosylation of the H gene product.

ABO blood types in US

                        Whites                   Blacks

O

A

B

AB

ABO blood types in US

                        Whites                     Blacks

O                         45                          50

A                         40                          25

B                         10                          20

AB                        5                           5

Rh Nomenclature and Incidence

             Antigens            Whites             Blacks

R1         

R2         

R0          

Rz          

Rh Nomenclature and Incidence

             Antigens            Whites             Blacks

R1            DCe                   40                   15

R2            DcE                    10                   10

R0            Dce                    5                     45

Rz            DCE              rare(6% nat am)      rare

Rh Nomenclature and Incidence

             Antigens            Whites             Blacks

r'             

r''         

r              

r^y             

Rh Nomenclature and Incidence

             Antigens            Whites             Blacks

r'              dCe                   3                    3

r''             dcE                    2                   rare

r               dce                  40                    30

r^y              dCE                  rare                 rare

-weak D occurs when someone who is D + has a form of the D antigen that doesn’t lead to a visible reaction when anti-D is mixed with his red cells. This happens, MCly, due to mutations altering (but not necessarily eliminating) portions of the D antigen within the red cell membrane.

-Weak D red cells are proven to be D-+ when an IAT is performed. The IAT is done by incubating then washing the red cell:anti-D mixture and adding AHG to detect red cells that have been coated with antibody but not agglutinated.

-Weak D can also happen when a particular RHCE allele for the C antigen (specifically, RHCe) is present on the opposite chromosome to an RHD allele (as in the combination R₀r’, also written as "Dce/dCe"). This has historically been known as the "C in trans" mechanism for weak D.

A blood group: what % are A1 vs A2?

How can they distinguished?

Significant anti-A1 in what percentage of A2 people? of A2B people?

A blood group: what % are A1 vs A2?

80% vs 20%

How can they distinguished?

By anti-A1 reagant (from B individuals), or w/ Dolichos biflorus lectin (has anti-A1 activity)

Significant anti-A1 in what percentage of A2 people? of A2B people?

5% ; 35%

RHD and RHCE gene loci on what xsome?

RHAG geen on what xome?

Most common white genotype? black genotype?

RHD and RHCE gene loci on what xsome?

1

RHAG gene on what xome?

6

Most common white genotype? black genotype?

R1/R1 or R1/r white

R0/R0 or R0/r in blacks

Sources of weak D phenotype?

Cepelli effect?

Sources of weak D phenotype?

Quantitative - gene alteration to reduced expression. Does not result in loss of D antigen epitopes

(Alteration common in blacks, often w/ Dce haplotype)

Partial D: RBCs lack D antigen components (epitopes)

Cepelli effect: People w/ Dce/Ce genotype have weakened D expression due to the presence of the C allele situated trans (on the other homologous xsome) to D

Rh null people lack _ and _ , and have weakened expression of _, _, and _ antigens.

Rh null people lack LW and Fy5 , and have weakened expression of S, s, and U antigens.

How are Rh antigens affected by enzymes?

Intravascular vs Extravascular hemolysis?

How are Rh antigens affected by enzymes?

Enhanced

Intravascular vs Extravascular hemolysis?

Extravascular

anti-c

Most with anti-E have R1R1 phenotype (DCe,DCe) and got R2 blood (DcE). As a result, develop anti-E and too weak to detect anti-c (which is a common cause of delayed hemolytic transfusion reactions)

Common practice to give c-E- blood to those with anti-E

Kidd antigens- enzyme enhanced?

Class?

Dosage?

Which phase do they react?

Kidd antigens- enzyme enhanced?

Yes

Class?

Warm reacting IgG

Dosage?

Yes

Which phase do they react?

AHG phase

Thrombotic microangiopathies (TTP, HUS, HELLP)

Massive transfusion (MTP)

DIC

PT>1.7 prior to invasive procedure or multiple factor deficiences

Warfarin- rapid reversal w/ active bleeding or imminent procedure

Congenital or acquired (inhibitors) factor deficiency

Protein C, S deficiencies, or AT

Lewis system: two precursor oligosaccharides

Type 1 found where?

Type 2?

Difference betwee i and I?

Type 1: Predominantly in secretions but also free in blood

Type 2: Only on the RBC surface

Branched type 1 and 2 are I antigens. In neonate, unbranched i antigen oligosacc predominate. Get progressively branched with age, and become I antigens. Adults mostly with I antigen.

Fucosyl transferase

Adds fucose to type 1 precursors both in secretions and free in serum. To make Lea antigen. 

Though Lewis antigens are synthesized on free type 1 precursor substance, it becomes passively adsorbed onto RBC surfaces. 

What factors are considered the most "labile" (rapidly degrading) coagulation factors in stored plasma?

Factors V and VIII

(factor VII, while it degrades very rapidly in the body, is more stable in storage conditions)

Which platelet product is commonly referred to as “random-donor” platelets?

Whole blood-derived platelets (WBD-platelets)

[AD-platelets, which are commonly known as “single donor” platelets.]

Do platelets manufacture ATP?

What is the body's platelet growth factor?

Shape of unstimulated vs stimulated platelets?

What fraction of platelets sequestered in spleen?

Do platelets manufacture ATP?

Manufacture ATP via oxidative metabolism.

What is the body's platelet growth factor?

Thrombopoietin, a hepatic-derived substance

Shape of unstimulated vs stimulated platelets?

Simple, discoid shape, while stimulated platelets develop cytoplasmic projections known as filopods

What fraction of platelets sequestered in spleen?

1/3

What HLA class present on platelets?

Group A or B more abundant on group AB platelet?

Are Rh antigens present on platelets?

What HLA class present on platelets?

Class I

Group A or B more abundant on group AB platelet?

Group A

Are Rh antigens present on platelets?
No

Platelet alpha granules contain?

Dense granules?

Platelet alpha granules contain?

vWF, Factor V (cascade substrate),

Fibrinogen,

Protein S (inhibit coagulation),

PDGF (wound healing)

Dense granules?

ADP, serotonin, calcium

Lewis phenotypes

                       White                        Black

Le a-b+

Le a+b-

Le a-b-

Le a+b+

Lewis phenotypes

                       White                        Black

Le a-b+              72                              55

Le a+b-              22                              22

Le a-b-                6                               23

Le a+b+             0                                 0

Encodes a fucosyl transferase that adds fucose to type 1 precursor ONLY if Le gene product already acted on it (thus Le^b made on Le^a)

To make Leb, must have both Le and Se gene.

% in Caucasians of K, Jk^a, E, s ?

K: 9%

Jk^a: 77%

E: 29%

s: 89%

Ficin enzymes destroys which antigens?

Fy^a

Fy^b

M, N, S, s

Xg^a

What does a negative autocontrol test indicate?

What does a negative autocontrol test indicate?

No autoantibodies present

The last phases of testing serum and red cells for compatibility, most commonly in test tubes (the first two are the immediate spin and the 37C phases).

- In classic tube testing, the AHG phase occurs after the serum and red cells have undergone an incubation at 37C followed by a washing step. During this phase, anti-human globulin (yes, that's where "AHG" comes from) is added to the serum-red cell mix, and it agglutinates red cells that are coated with IgG antibody. This process is simply an indirect antiglobulin (Coombs) test, and it shows that there is an antibody present that is incompatible with the red cells present in the test tube. Most clinically significant antibodies will react at the AHG phase.

The first of the three  "phases" of testing serum and red cells for compatibility, most commonly in test tubes (the last two are the 37C and AHG phases).

-The term "immediate spin" refers to the fact that diluted red cells and patient serum or plasma are added to a test tube at room temperature, immediately centrifuged ("spun"), and then examined visually for agglutination. A positive immediate spin reaction can indicate ABO incompatibility between serum and RBCs (and this principle is used in the immediate spin crossmatch) or the presence of a "cold" or room-temperature reactive antibody.

What does chloroquine do to RBCs?

FFP left frozen fro 24 hours, then thawed in fridge till slush staged

Then centrifuged (4200 rpm) for 10 min

Supernatant plasma removed

Cryo then frozen at stored at -18 C (1 year)

Stable and labile coagulation factors (1 unit of each clotting factor and 2-4 mg of fibrinogen/mL plasma)

-Other plasma proteins (inc albumin)

-20% is CPD anticoagulant solution

Fibrinogen (200-250 mg; min 150 mg)

Factor VIII (80-120 units; min 80 units)

vWF

Factor XIII

Fibronectic

-Decreased risk of CMV, HTLV, EBV

-HLA alloimmunization

-Platelet refractoriness

-Febrile nonhemolytic reactions

-TRIM (transfusion related immunomodulation)

Duffy antigens?

Fy(a+b-) more/less common than Fy(a-b+)?

Fy(a-b-) % in blacks?

Affected by enzymes?

Duffy antigens? Fya and Fyb

Fy(a+b-) more/less common than Fy(a-b+)?

More common

Fy(a-b-) % in blacks?

68%

Affected by enzymes?

Destroyed

Duffy antibodies are what class?

Show dosage?

Duffy antibodies are what class?

IgG warm reacting

Show dosage?

Yes

MNS found on what on the RBC surface?

S,s,U ?

MNS found on what on the RBC surface?

Glycophorin A

S,s,U ? 

Glycophorin B

MNS antibodies - display dosage?

What class are they?

What class are S,s, and U?

MNS antibodies - display dosage? - Yes

What class are they?

IgM, cold-reacting; naturally occuring; clinically insignificant

What class are S,s, and U?

IgG, warm reacting, after expsoure, significant

Kell antigens affected by enzymes?

What destroy them?

McLeod phenotype? Disease associations

Kell antigens affected by enzymes? 

Unaffected

What destroy them?

ZZAP

DDT

McLeod phenotype? Lack of Kx protein depresses expression of Kell antigens - assw shortened RBC survival and reduced deformabilitiy 

Disease associations: Xlinked CGD, acanthocytosis, late-onset type muscular dystrophy 

Kell antibodies

Class? Exposure?

Kell antibodies

Class? Exposure?

Warm reacting IgG ; after exposure, significant

Most are anti-K 

(k antigen very common - 99%, K antigen - 9%)

RBC diameter?

Microaggregate filter diameter?

Standard blood filter diameter?

RBC diameter?

8 microns

Microaggregate filter diameter?

40 microns

Standard blood filter diameter?

170 microns

Factors that can decrease response to platelets?

Fever

ABO incompatibility

Splenogmegaly

Recipient HLA antibodies

Drugs: Amphotericin B

When giving blood products to a IgA deficient individual, why is it impt to remove the plasma?

IgA deficient patients can develop antibodies to transfused IgA - thus can have severe anaphylactic reaction on subsequent transfusions.

P antigens: protein or carbohydrate?

What is the P₁ phenotype?

White %? Black %? 

P antigens: protein or carbohydrate?

Carbohydrate

What is the P₁ phenotype?

anti-P_1, P+, PP₁P^k+, P^k_

White %? 80%

Black %? 95%

P2 phenotype?

Black %?

White%?

P2 phenotype

anti-P_1-, P+, PP₁P^k+, P^k_

Black % - 5%

White% - 20%

What is the rare p phenotype?

 Absence of P antigens (anti-P_1-, P-, PP₁P^k_, P^k_)

Make a potent anti PP₁P^k (can be assw delayed hemolytic transfusion reaction and HDN)

Assw between anti-P₁ and anti PP₁P^k and 1st Trimester abortion

P2 phenotype

IgM anti P₁, 4 C, not significant

In Paroxysmal Cold Hemoglobinuria, what antigens are the target?

What test used to pick it up?

In Paroxysmal Cold Hemoglobinuria, what antigens are the target?

P antigens

Auto-anti P IgG Ab act as a biphasic hemolysin. (As blood passes thru cold parts of body, it binds and activate complement, hemolysis occurs at warm temperatures)

What test used to pick it up?

Donath Landsteiner

What antigen serves as receptor for parvovirus B19?

P antigen

HLA class I antigens found on what cells?

3 loci?

HLA class I antigens found on what cells?

All nucleated cells

Platelets

3 Loci?

HLA-A, HLA-B, HLA-C

(Class I encode a single polypeptide chain that has 3 domanins ~ Ig HC + a transmembraine domain. Class I molecules embedded as a transmembrane protein, and each non covalently assw single molecule of B2 microglobulin)

HLA Class II antigens found on what cell types?

3 Loci?

HLA Class II antigens found on what cell types?

B cells

Macrophages

Activated T cells

3 Loci?

HLA-DR, HLA-DP, HLA-DQ

(The Class II genes encode 2 polypeptide chains, alpha and beta, each with 2 domains ~ Ig LC + transmembrane domain)

What are Bennet Goodspeed antigens?

Red cells have small amouts onf Class I antigens, referred to as BG antigens. Rarely involved in alloAb mediated hemolytic transfusion reactions.

 Each unit of platelets collected by apheresis (apheresis-derived platelets) should contain at least ________ platelets, while units of whole blood-derived platelets should have at least ________.

 Each unit of platelets collected by apheresis (apheresis-derived platelets) should contain at least ________ platelets, while units of whole blood-derived platelets should have at least ________.

3.0 x 10¹¹; 5.5 x 10¹⁰

 Each unit of red blood cells contains approximately how much iron?

After 1 unit plasma exchange, the % of protein removed is ? After 2 units?

1 unit - 65%

2 units - 88%

Chance siblings are HLA identical?

1 sibling?

2 siblings?

3 siblings?

Chance siblings are HLA identical? 25%

1 sibling? 25%

2 siblings? 45%

3 siblings? 60%

Reagent cells in antibody screen express which antigens?

DCEce

MNSs

P₁

Le^a,Le^b

K,k

Fy^a, Fy^b

Jk^a, Jk^b

Examples of enhancement media designed to make Ab-Ag reactions more obvious?

PEG (polyethylene glycol)

LISS (Low ionic strength saline)

Direct antiglobulin test or direct Coombs test explanation?

AHG is added to blood. If the RBCs are coated with globulins, agglutination occurs.

Indirect antiglobulin test or indirect Coombs test explanation?

Antibodies of known specificity are added to blood followed by addition of AHG. If RBCs have antigen of interest, agglutination occurs.

-Suspected when there is weak reactivity in the AHG phase to all the cells in the antibody panel. Reacts at all dilutions >1:64

-Group of antibodies that are directed against high incidence antigens (>99.9% people)

-Significance is that they mask other alloantibodies

-Some may be clinically significant: Cartwright (Yt^a), Holley (Hy), Gregory (Gy)

-Not significant : Chido/Rogers (Ch/Rg), Sda, Bg, Csa, York (Yka)

Polyagglutination?

What does T-activation mean?

Test for polyagglutination?

-Adult sera has naturally occuring IgM antibodies to T, Tn, Tk, and Cad. 

-Some antigens (all T)  only exist after being acted upon by bacterial neuraminidase (aka T-activation)

-Cad is a rare inherited antigen

-Polyaggutinable RBCs are agglutinated by adult but not cord serum. 

Hemagglutination inhibition?

-Use of substance that contains or mimics a particular antigen

-If serum sample suspected of containing an antibody with specificty for this antigen, suspicion can be confirmed by abolition of reactivity. 

-Also, additional antibodes that are masked can get picked up

Lectin                                   Binds to? 

Dolichos biflorus

Bandeiraea simplicifolia

Ulex europaeus

Lotus tetragonolobus

Arachis hypogaea

Vicea graminea

Lectin                                   Binds to? 

Dolichos biflorus                         A1

Bandeiraea simplicifolia               B

Ulex europaeus                           H (secretor status)

Lotus tetragonolobus                  H

Arachis hypogaea                         T

Vicea graminea                           N

Neutralizing substances                 Antigenic Activity?

Guinea pig urine

Hydatid cyst fluid

Saliva

Breast Mild

Pidgeon eggs

Plasma

Neutralizing substances                 Antigenic Activity?

Guinea pig urine                                      Sd_a

Hydatid cyst fluid                                     P₁

Saliva                                                    H, Le^a

Breast Milk                                               I                                             

Pidgeon eggs                                            P₁

Plasma                                             Chido, Rodgers

Le,

I/i

P

Rh

Kidd

MNSs

Fy^a, Fy^b

Lutheran

Chido

Rodgers

Yt^a

Kell

(most remaining)

Cause of positive crossmatch with positive antibody screen?

Cause of positive crossmatch with negative antibody screen?

Cause of positive crossmatch with positive antibody screen?

AlloAb, AutoAb, Antibodies to reagents

Cause of positive crossmatch with negative antibody screen?

Anti-ABO antibodies, Antibodies to low incidence antigens

Autoantibody (MC warm autoimmune hemolytic anemia)

Anti-LW (LW antigen expressed more strongly on D+ cells than D-. If anti-LW Ab present, may appear to be reacting only with D + cells, thus stimulating anti-D antibodies)

D variant

Blood group A individual RBCs get reactivity with anti-B reagent (thus look AB).

-Occurs in A1 individuals when acted upon by bacterial deacetylases (assw bacteremia [colon Ca, obstruction, gram negative sepsis]

-To confirm suspicions 1.altered A1 antigen can by re-acetylated with acetic anhydride. 2.patient's own anti-B will agglutinate these cells

(Acidified human anti-B does not react with acquired B antigen. Patient's saliva will not contain B antigen)

Drugs assw positive DAT?

Penicillin, Cephalosporins, Procainamide, Aldomet

Questions to be asked when autoantibody is suspected?

Patient hemoylzing? (bili, haptob,Hb, LDH)

Drugs? (penicillin, ceph, procain, aldomet)

Recent transfusion (mixed field reaction)

Uncommon: Post bone marrow transplant (mixed field), anti-lymphocyte globuin (ALG)

Most reliable CBC index in presence of cold agglutinins?

Titer of benign cold agglutinins usually < _ ?

Most reliable CBC index in presence of cold agglutinins?

Hemoglobin

Titer of benign cold agglutinins usually < _ ?

64 at 4 C

Pathologic cold autoagglutinins 

DAT?

C3d?

IgG?

Titer?

Pathologic cold autoagglutinins 

DAT? 2+ to 3+

C3d? Positive

IgG? Negative

Titer? >1000 at 4 C

Idiopathic cold autoimmune hemolytic anemia (CAIHA)

Antibody?

Idiopathic cold autoimmune hemolytic anemia (CAIHA)

Antibody?

IgM with anti-I, anti-i

(rarely anti-Pr)

Acrocyanosis, Raynaud, moderate hemolytic anemia

-Agglutination in extremities and complement fixation leading to intravascular lysis

Secondary cold autoimmune hemolytic anemia

Etiologies?

Secondary cold autoimmune hemolytic anemia

Etiologies?

M. Pneumonia - transient anti-I

Lymphoproliferation ~ persistent anti-I

Mono sometimes causes anti-i

(all these are IgM)

Both cold IgM and warm IgG autoantibodies

Reaction with IgG and C3 at antiglobulin phase

Presentation: Acute onset hemolytic anemia (idio or assw lupus)

Tx: Very responsive to corticosteroids

Paroxysmal Cold Hemoglobulinuria?

MC affects children with viral illness

Clinical: Paroxysmal episodes of hemoglobulinuria assw cold exposure. Fever, abdominal pain.

Anemia severe

Tx: Warm, Transfuse as necessary

Donath Landsteiner Test?

Dx Paroxysmal cold hemoglobinuria

IgG biphasic with anti-P specificity. Produces hemolysis when incubated at 2 diff temperatures

Testing: DAT postive with polyspecific AHG, negative with anti IgG, positive with anti-C3. Perform at 4 and 37 C.

Drug Induced Positive DAT

1. Drug absorption

(Penicilin prototype)

Positive DAT.

Dose dependent.

Mechanism: Drug absorbed onto RBC, and RBC then becomes coated with anti-penicillin antibodies.

Lab: Serum and eluate react with drug treated RBCs but not untreated RBCs.

Drug Induced Positive DAT

2. Non Immune protein adsorption?

(Cephalothin prototype)

Drug Induced Positive DAT

2. Non Immune protein adsorption?

(Cephalothin prototype)

Mech: Drug adsorbed onto RBC. RBC becomes nonspecifically coated with all sorts of serum proteins, including antibodies

Lab: RBCs + with IgG, IgA, C3 specific antisera, but serum and eluate are non-reactive with RBCs.

Drug Induced Positive DAT

3. Immune Complex?

(Quinidine, phenacetin, cephalosporins prototypes)

Drug Induced Positive DAT

3. Immune Complex?

(Quinidine, phenacetin, cephalosporins prototypes)

Mech: Drug elicits complement-fixing antibodes - leads to 'innocent bystander' lysis of RBCs

Lab: RBCs + for C3d only

Drug Induced Positive DAT

4. True autoimmune hemolytic anemia?

(aldomet, procainamide)

Drug Induced Positive DAT

4. True autoimmune hemolytic anemia?

(aldomet, procainamide)

Warm reacting RBC autoantibody - leads to syndrome resembling idiopathic types of warm autoimmune hemolytic anemia

1. Very little volume to begin with

2. Potential platelet loss and potential platelet functional defects

3. Potential bacterial contamination

4. Time delay

Autoantiboies agains platelets (~PL^A1)

Risk fx during pregnancy: previous splenectomy for ITP, previous infant with ITP, gestational (maternal) platelet count <100,000

Alloantibodies that cross placenta and cause destruction of fetal platelets.

MC agains PL^A1

IgG1, IgG3, IgG4

(IgA, IgG2, and IgM DO NOT CROSS)

MCC of severe HDN?

2nd MC?

MCC of severe HDN?

Kell

2nd MC?

Anti-c

Vials of RhIg = [maternal whole blood volume (mL)) x (% fetal cells in mom's blood)] / 30

blood volume = kg x 70 mL/kg (about 5000 mL)

% rbc- Kleihauer Betke test

Each full dose protects agains 30 mL whole blood OR 15 mL RBCs

Round up 1 or 2.

D+ indicator cells form rosettes around D+ fetal cells

As little as 10 cc fetal blood can be detected

Qualitative results (KB or ELAT then indicated

Since HbF is resistant to acid elution, acid elution affects adult Hemoglobin

Afterwards, stained with Wright and cells that stain are HbF containing RBCs

Cell count performed to determine % of fetal red cells

Stroke

Retinal artery occlusion

Splenic sequestration crisis

Acute chest syndrome

Aplastic crisis

Target HbS in children?

In adults?

Target HbS in children?

<30%

In adults?

<50%

MC autoantibodies that develop in sickle cell patients?

Kell, C, E, Jk^b

Dextrose?

Adenine?

Citrate?

Sodium phoshpate?

[blood storage]

Dextrose?

-Carbohydrate for glycolytic ATP production

Adenine?

-Substrate for ATP production

Citrate?

-Anticoagulant (chelates calcium)

Sodium phoshpate?

-pH buffer

Solution         Contents          Storage Time     Comment

Heparin

CPD

CPDA-1

AS-1

Solution         Contents          Storage Time     Comment

Heparin            Heparin            48 hours         Not used

CPD-Citrate, Na phosphate, dextrose; 21 days

CPDA-1 (same as CPD+adenine) ; 35 days

AS-1 (dex, aden, na phos, mannitol, Na cholide); 42 d; must be added w/in 72 hours of collection

White cells in typical RBC unit?

Leukoreduced WBC number (to prevent febrile reactions)?

Leukoreduced WBC number (to prevent HLA alloim and CMV)?

White cells in typical RBC unit?

5x10⁹

Leukoreduced WBC number (to prevent febrile reactions)?

<5x10⁸

Leukoreduced WBC number (to prevent HLA alloim and CMV)?

<5 x 10⁶

Shelf life of washed RBCs?

Purpose of washed RBCs?

Shelf life of washed RBCs?

24 hours

Purpose of washed RBCs?

Prevent allergic reactions (ex. anaphylaxis in IgA deficient recipient)

Shelf life of Irradiated RBCs?

28 days or original outdate

200 mg iron (1 mg iron/ 1 mL RBCs)

Total volume of bag 250 mL (200 mL RBCs, 50 mL plasma and anticoagulants)

Pheresis platelets equal to how many packs of regular platelets?

Average platelet lifespan?

6

 (3 x 10¹¹; vs 5.5 x 10¹⁰)

Expected increase of 1 pack: 5000/uL

1 apheresis unit: 30,000 /uL

Average platelet lifespan?

9.5 days

Granulocyte collection and administration?

Via pheresis, stored at 20-24 C up to 24 hours

Administer without a filter

FFP

Preparation?

Contains?

Indications?

Dosing?

Preparation?

Plasma placed at -18 c w/in 8 hours. Expires 1 year. Thaw to 30-37 c- expires 24 hours

Contains?

200 mL; 1 IU all coag fx (400 mg fibrinogen)

Indications?

DIC, warfarin, massive transfusion,

Plasmapheresis (TTP, ATIII def)

Dosing?

2 units (each 20% expected increase)

10-15ml/kg neonates

(make sure ABO compatible as it contains antibodies)

Cyroprecipitate preparation?

Contains?

FFP thawed to 1-6 C

Precipitate is cryoprecipitate

W/in 1 hour, store at -18 C for 1 year.

Contains: 15 mL, 150 mg fibrinogen, 80 IU fx 8, vWF, fx 13a.

DOES NOT have fx 5.

Tx- hemophilia A if fx 8 concentrate not available

Bags = [(plasma volume) x (amt fx 8 needed)]/80

Plasma volume = (kgx70mL/kg) x (1-Hct)

Fx 8 needed = (Desired activity) - (current activity)

[100% desired if going to surgery; 50% to stop spontaneous bleeding]

100% fx 8 activity = 1 IU/mL

Half life of Fx 8?

Half life of Fx 9?

Fx 8 - 12 hours

Fx 9 - 8 hours

(initial tx of choice for vwD: DDAVP)

Fibrinogen deficiency MC in what setting?

DIC

Tx: FFP and cryo (don't give alone bc doesn't have enough Fx 5)

fibrin glue factor: fx 13

3.5 hours

(indication: inhibitors against Fx8 or Fx9, trauma, 7 def, excessive coumadin, liver failure, tcytopenia)

Irradiation

Dose to midplane?

Dose to any portion?

Irradiation

Dose to midplane? 25 Gy (2500 cGy)

Dose to any portion? 15 Gy

(28 days expiration or original outdate)

(Indication: BMT, neonates, blood from 1st degree relatives, Hodgkin/NHL, congenital T-cell defects)

NOT in B cell or mphage defects

Leukoreduction

Amt to prevent febrile illness?

Amt to prevent CMV transmission?

Indications?

Leukoreduction

Amt to prevent febrile illness? <5 x10⁸

Amt to prevent CMV transmission? <5 x 10⁶

(washed RBCs and frozen,deglycerolized cells considered LR)

Washed RBCs indications?

Prevent allergic reactions

FNHTR

Neonates from parent, ABO incompatibility

Classification of hemolytic transfusion reactions?

Intravascular vs Extravascular

Acute vs Delayed

ABO, Kidd- Acute intravascular

Non-ABO alloantibodies: acute or delayed extravascular hemolysis

[NO real examples of delayed intravascular hemolysis]

Long Storage

Inappropriate temperature storage

Mechanical devices

Blood warmers

Needles

Addition of stuff other than NS

Bacterial contamination (Clostridia)

RBC unrecognized defect

Lab Associations (Intravascular or Extravascular Hemolysis?)

Postive DAT?

Pink serum?

Hyperbilirubinemia?

Hemoglobinuria?

Coagulation abnlties?

Schistocytes?

Spherocytes?

Lab Associations (Intravascular or Extravascular Hemolysis?)

Postive DAT?     I or E

Pink serum?         I

Hyperbilirubinemia?   E 

Hemoglobinuria?       I

Coagulation abnlties?     I

Schistocytes?          I

Spherocytes?          E

Alloantibodies against what antigens MCly causing DHTR?

Findings in DHTR?

(Extravascular hemolysis)

Kidd, Kell, Duffy

Findings: Positive DAT (often mixed field), icteric serum, anemia, spherocytes, no free Hb

Tx: avoid further exposure

Yersinia enterocolitica

Serratia liquifaciens

Citrobacter

Pseudomonas

Two conditions needed for donor cells to survive in recipient?

Immunocompromised recipient

HLA similarity (no response mounted)

[TVHD: can occur with transfusion or solid organ transplantation)

Tetrad of TAGVHD?

Dermatitis (periauricular, palmar, plantar)

Enterocolitis (watery diarrhea)

Hepatitis (aminotransferase elevation)

BM suppresssion (pancytopenia)

Proposed pathogenesis of TRALI?

Donor HLA antibodies against reciepient WBCs.

(2 grps at risk: induction tx for heme malig, bypass surgery)

(Likelihood raised: multiparous donors, prolonged storage)

(Plasma containg products more risk: FFP, platelets)

Plasma proteins.

(IgA deficiency)

Post transfusion Purpura

Affects who?

Etiology?

Tx?

Post transfusion Purpura

Affects who?

Multiparous women

Etiology?

Plate alloantibodes (anti PL^A1) [98% positive for PLA1]

Tx?

Platelet Refractoriness Etiologies?

Infection

Splenomegaly,

Drugs (amphotericin)

DIC

HLA alloimmunization (Class I antigens found on platelets, MCly HLA-A and HLA-B)

Platelet count increment calculation?

CI = (posttransfusion plt ct) - (pretransfusion plt ct) x BSA / (platelets transfused x 10¹¹)

[expected increase 1 unit is 5000, apheresis (6 pack) = 30,000)

Transfusion Risks

HBV?

HCV?

HAV?

HIV?

Transfusion Risks

HBV?

1:100,000

HCV?

1:[800K-1.7 million]

HAV?

[1:1 million]

HIV?

1: [400K to 2.4 million]

Naturally occuring antibodies?

Antigens that display dosage?

Naturally occuring antibodies?

ABO, I/i, Le, Lu, M, N, P

Antigens that display dosage?

MNS, Kidd, C/c, E/e, Duffy

Antibodies reacting at room temperature?

Antigens that are enzyme enhanced?

Antibodies reacting at room temperature?

Anti-M, N, P¹, Le^a, Le^b

Antigens that are enzyme enhanced?

I/i, P, Le, Rh, Kidd

Antigens destroyed by enzymes?

4 most common antibodies implicated in delayed HTR?

Antigens destroyed by enzymes?

MNSs, Fy^a, Fy^b, Lutheran, Chido

4 most common antibodies implicated in delayed HTR?

anti-Jk^a, anti-E, anti-D, anti-C

Mixed field reactions expected with? 

Antibodies assw Intravascular Hemolysis?

Mixed field reactions expected with? 

-Lutheran, Sid, A₃, post BMT

Antibodies assw Intravascular Hemolysis?

-ABO, Kidd, P (Paroxysmal Cold Hemoglobinuria)

Associations

Fy (a-b-)

McLeod (Kell-null,Kx)

Anti-P

Anti-I

Associations

Fy (a-b-) : P.vivax resistance

McLeod (Kell-null,Kx): CGD, Acanthocytosis, Musc Dystrop

Anti-P: PCH, Syphilis

Anti-I : Mycoplasma Pneum

Associations

Anti-i

Anti-N

Acquired B phenotype

Rh null (Bombay)

Associations

Anti-i  : EBV

Anti-N : Renal dialysis

Acquired B phenotype: Colon Ca, Gram Neg Sepsis, Bowel                                 obstruction

Rh null (Bombay): Hereditary Stomatocytosis

Associations

Diego Negative

Gerbich Negative

Associations

Diego Negative: Diego is an epitope on band 3 protein

                       Band 3 deficiency - HS and HE

Gerbich Negative: Gerbich: epitope on Glycophorin C; A                                         cause of HE

Shelf Lives

RBC (saline washed or thawed and degycleroized)

Thawed FFP:

Thawed Plasma:

Pooled Platets:

Thawed Cryoprecipitated AHF, unpooled:

Thawed Cryoprecipitated AHF, pooled:

Shelf Lives

RBC (saline washed or thawed and degycleroized): 24h

Thawed FFP: 24h (afterwards, called plasma)

Thawed Plasma: 5 days

Pooled Platets: 4h

Thawed Cryoprecipitated AHF, unpooled: 6h

Thawed Cryoprecipitated AHF, pooled: 4h

Clinically insignificant antibodies? 

Clinically insignificant antibodies? 

M, N, P, Lewis, Lutheran, I 

Cushing syndrome (elevated serum cortisol)

Cushing disease (elevated serum cortisol due to a pituitary adenoma). 

Addison syndrome is defined as hypocortisolism

 while Addison disease is defined as hypocortisolism due to primary adrenal insufficiency. 

constellation of symptoms secondary to

hyperaldosteronism (hypernatremia, hypokalemia, hypertension).