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Mercy CLS 4: Blood Bank > 11. Applications of BB > Flashcards

11. Applications of BB Flashcards

1
Q

3 general functions of HLA

A
  1. immune regulation
  2. recognition of self vs nonself
  3. coordination of cellular and humoral immunity
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2
Q

Class I HLA
nomenclature
location

A

HLA-A, B, C
platelets and most nucleated cells
Bg Ag on RBCs

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3
Q

Class II HLA
nomenclature
location

A

HLA-D-related
immune cells, interstitial epithelium, early hematopoietic cells

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4
Q

HLA on chromosome…

A

6

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5
Q

HLA gives a 3D configuration to each molecule to form a unique …
function?

A

peptide binding groove
holds processed peptides for presentation to T-cells

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6
Q

MHC I function

A
  • present peptides from intracellular proteins (self, cancer, viral)
  • present to CD8 T-cells
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7
Q

MHC II function

A
  • present peptides from extracellular/cytosolic proteins (self, bacterial)
  • present to CD4 T-cells
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8
Q

immune cells are only activated when…

A

antigenic peptide is displayed in context with the self HLA antigens

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9
Q

2 types of HLA nomenclature

A

conventional (serological)
DNA sequencing

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10
Q

HLA

capital letter denotes…
number denotes…

A

locus
antigenic specificity

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11
Q

HLA

“w” indicates… (3)

A
  • C locus specificities (distinguish from complement)
  • DP and D specificities (cellular methods)
  • Bw4 and Bw6, public antigens found on multiple B specificities
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12
Q

HLA

—- shows allele is defined by DNA techniques

A

asterisk

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13
Q

HLA

set of two digit numbers separated by colons describe…

A

gene variants

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14
Q

HLA

low resolution
high resolution

A

serologic results
genetic variants

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15
Q

2 HLA genes inherited as a ———

A

haplotype

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16
Q

HLA

parents and children always share…

A

one haplotype match

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17
Q

best source of HLA-identical matches, 25% probability

A

siblings

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18
Q

HLA phenotype represents…

A

the combined expression of both haplotypes

HLA genes codominant

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19
Q

HLA

dash represents…

A

blank where an antigen is not detected

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20
Q

required to determine haplotypes

A

family studies

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21
Q

testing used to find HLA matches

A

histocompatibility

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22
Q

historically HLA Abs for testing were from…

A

multiparous women
transplant/tranfusion recipients

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23
Q

Ab recognizing a single HLA gene product

A

private epitopes/splits

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24
Q

Ab recognizing multiple HLA gene products

A

public epitopes/crossreactive

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25
Q

CREG

A

cross-reactive group
HLA antigens that consistently demonstrate cross-reactivity

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26
Q

———- antigens are less immunogenic when transplanted, allowing mismatch

A

within CREG

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27
Q

source for HLA DNA typing

A

any nucleated cell

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28
Q

3 ways to analyze amplified DNA for HLA

A
  • sequence-specific oligonucleotides
  • sequence-specific primers
  • DNA sequence-based typing
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29
Q

HLA Ab testing indications

A
  • determine presence of Ab in a transplant or platelet transfusion recipient
  • find HLA Ab in donor plasma, which could cause TRALI
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30
Q

HLA Ab testing techniques (3)

A
  • complement dependent lymphocytotoxicity (CDC)
  • ELISA
  • microbead array assay
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31
Q

standard serological HLA Ab typing method

A

CDC

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32
Q

CDC reagent cells

A
  • T cells for class I
  • B cells for class II
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33
Q

CDC principle

A

utilizes biological function of Ab to activate complement and disrupt cell membrane

dead cells are stained for visualization

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34
Q

HLA ELISA principle

A

HLA Ag bound to wells
HLA Ab binds, visualized with enzyme-conjugated substrate that changes color

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35
Q

microbead assay principle

A

beads labelled with ratio of 2 different fluorescent markers

beads of specific HLA type assigned to different color ratio

anti-IgG used to ID Ab binding to certain beads

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36
Q

explain HLA XM

A

recipient serum + donor purified WBCs

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37
Q

HLA XM incompatibility is a contraindication for….

A

kidney and pancreas transplant

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38
Q

types of HLA XM used most

A

virtual
flow

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39
Q

HLA XM that utilizes fluorescence

A

flow cytometry

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40
Q

advantage of HLA’s polymorphism

A

ID of individuals for parentage or forensic testing

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41
Q

GVHD occurs when ——- are transfused into a ——- patient and…

A

HLA-homozygous donor cells
heterozygous patient
patient does not recognize them as foreign

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42
Q

causes platelet refractoriness

A

pt sensitized to HLA Ags on platelets

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43
Q

leukocyte reduced product count

A

< 5 x 10^6

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44
Q

nonimmune causes of platelet refractoriness (not true refractoriness)
(3)

A
  • sequestration —splenomegaly
  • consumption —sepsis, DIC, meds, fever, bleeding
  • prothrombotic conditions —HIT, TTP, HUS
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45
Q

HIT

A

heparin-induced thrombocytopenia

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46
Q

TTP

A

Thrombotic Thrombocytopenic Purpura

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47
Q

HUS

A

hemolytic uremic syndrome

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48
Q

alloimmune causes of platelet refractoriness

A

ABO
HLA
HPA

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49
Q

ITP

A

idiopathic thrombocytopenia purpura

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50
Q

platelet transfusion contraindications (2)

A
  • prothrombotic conditions (HIT, TTP, HUS)
  • acute ITP (bleeding complications are rare)
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51
Q

essential to detect platelet refractoriness & ID successful donor-recipient matches

A

one hour post transfusion platelet ct

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52
Q

required for HLA-matched platelets

A

irradiation

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53
Q

anti-A/B titer of —– associated with incompatible platelet XM

A

> 64

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54
Q

best way to avoid platelet refractoriness

A

pt HLA ABS and ABID

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55
Q

absence of B * 27

A

ankylosing spondylitis

56
Q

presence of DQ6

A

narcolepsy

57
Q

absence of DQB1 * 02

A

celiac

58
Q

presence of DRB1 * 03/04

A

T1DM

59
Q

acute noncardiogenic bilateral pulmonary edema

A

TRALI

60
Q

cause of TRALI

A

donor HLA or HNA Ab fix complement and cause tissue damage leading to capillary leakage and pulmonary edema

61
Q

TRALI prevention (3)

A
  • use male plasma/platelets
  • use female plasma/platelets = for HLA Ab
  • donors implicated in TRALI deferred
62
Q

organization charged with development of an equitable scientifically and medically sound organ allocation system

A

united network for organ sharing

63
Q

established to meet the needs of pateints who do not have a matched, related CTP donor

A

national marrow donor program

64
Q

UNOS responsibilities

A
  • maximize use of organs
  • ensuring quality of care
  • addressing ethical issues
  • evaluates potential donors
  • obtains appropriate consent
  • prepares organs for transportation
65
Q

goal of NMDP

A

recruit a large number of HLA-typed volunteers to be listed as potential donors
establish national coordinating center for facilitating donor searches

66
Q

NMDP uses ——- to assist in targeted recruitment of volunteer CTP donors

A

geo-coding

67
Q

serologic testing to prevent disease transmission for all donors of tissue

A
  • all testing required on blood donors
  • pretransfusion specimen preferred
68
Q

ABH antigens are concerns for ——- transplantation because they are expressed on vascular endothelium (5)

A

kidneys
heart
lungs
livers
pancreases

69
Q

ABH not important in these tissue grafts (5)

A

fascia
bone
heart valve
skin
cornea

70
Q

goal of CTP transplant HLA match

A

6-antigen match
A, B, DR most important

71
Q

required before kidney & pancreas transplant

A

ABO compatibility
HLA Xm

72
Q

required before liver, heart, lung, and heart-lung transplant

A

ABO compatibility
HLA XM only if recipient has a presensitization

73
Q

shown to tolerate ABO-mismatched heart and liver

A

pediatric patients with weak isoagglutinins

74
Q

tx acute rejection of heart or kidney

A

plasmapheresis
immunosuppressive therapy

75
Q

ECMO
function

A

extracorporeal membrane oxygenation
heart-lung machine removes Co2 and oxygenates blood

76
Q

ECMO indications

A

allows heart, lungs to rest and heal
severe illness of heart, lungs
waiting for or recovering from heart transplant

77
Q

ECMO pts require…

A

a current TS
4-6 units XMed available at all times

often transfused with 1 platelet and 2-5 pRBCs per day

78
Q

reduce risk of developing HLA Ab

A

leukocyte reduction

79
Q

BB support during liver surgery

A

preparedness, supply and responsiveness during 6-12 hr surgery

80
Q

blood loss and hypocoagulability occur during liver transplant because…

A

preexisting liver disease (coag factors)
ahepatic & early reperfusion periods

81
Q

products often used during liver transplant

A

6 pRBCs, 6 FFP, 2 platelets

82
Q

why blood product use has declined during liver transplant (6)

A
  • preop iron
  • erythropoietin therapy
  • surgical technique
  • interop blood recovery
  • antifibrinolytic drugs
  • acute normovolemic hemodilution
83
Q

CTP contain…

A

hematopoietic progenitor cells (HPC)
or
stem cells

84
Q

2 stem cell populations in CTP

A
  • capable of self-renewal and differentiation into all blood cell lineages
  • committed to certain blood cell lineage
85
Q

HPC are infused after…

A

lethal doses of radiation or chemo, killing existing marrow

86
Q

does not require a 6/6 match

A

cord blood
naive

87
Q

cord blood CTP indications

A
  • tx blood diseases in pediatric pts
  • second choice for adults lacking HLA-matched donor (may need 2)
88
Q

autologous CTP transplant indication

A

malignancies without marrow involvement (ie lymphoma) and metastatic or recurrent solid tumors

89
Q

allogeneic CTP transplant indication

A

CML, AML, ALL, aplasic anemia, malignancies with marrow involvement, hemoglobinopathies

90
Q

newly developing autologous CTP method

A

chimeric antigen receptor T-cell therapy (CART)

91
Q

preventive measures for GVHD

A
  • irradiation of blood products
  • HLA matching
  • T-cell depletion (CTP)
  • immunosuppressive drug therapy
92
Q

subtype of GVHD that occurs when lymphocytes from donor make Ab to recipient RBC Ags, causing hemolysis

A

passenger lymphocyte syndrome (PLS)

93
Q

PLS onset

A

1-3 weeks post transplant
transient DAT+

94
Q

chimerism

A

persistence of donor cells in allogeneic HSCT recipients demonstrating transplant survival

95
Q

can indicate CTP graft rejection

A

lack of/reduced presence of donor cells

96
Q

allows for analysis of CTP genomes to detect graft rejection

A

short tandem repeats

97
Q

AFDC

A

aid to families with dependent children

98
Q

reasons for parentage testing

A
  • divorce/custody disputes
  • welfare laws/child support
  • inheritance questions
  • questioned parentage (switched at birth, adoption, rape/incest cases)
99
Q

necessary steps for parentage testing

A
  1. allegation
  2. sample collection
  3. chain of custody
  4. selection of genetic markers
  5. testing
100
Q

with antigen parentage testing, children must be —- old

A

6 months

101
Q

element of chain of custody (6)

A
  • positively ID, photograph, fingerprint individuals
  • proper labelling
  • documentation in writing of everyone who handles samples
  • all test performed in parallel by 2 techs using 2 lot numbers
  • samples locked up when not in use
  • reports sent only to ordering physician/court
102
Q

requirements for genetic markers chosen for parentage testing (6)

A
  • polymorphic
  • inherited by simple mendelian genetics
  • well developed in children
  • not affected by environment
  • objectively interpreted
  • remain constant through life
103
Q

4 methods of parentage testing

A
  • RBC antigens
  • HLA markers
  • enzyme techniques
  • DNA fingerprinting
104
Q

ABO grouping can exclude —–% fathers

A

73%

105
Q

BG systems used for parentage testing

A

ABO
Rh
Kell
Kidd
Duffy
MNSs

106
Q

problems with RBC parentage testing (10)

A
  • null types
  • modified types
  • allelic alterations
  • weakened Ag expression
  • other alterations (cis AB, mosaics, chimera, bombay)
  • recent transfusion
  • DAT+
  • polyagglutination
  • ABO subgroups
  • acquired B
107
Q

HLA can exclude —–% fathers

A

90

108
Q

parentage test not run in duplicate

A

HLA

109
Q

any apparent exclusions repeated from original sample

A

HLA

110
Q

problems with HLA in parentage testing (4)

A
  • large number of lymphocytes needed
  • rare antisera
  • highly technique-sensitive typing
  • unknown allele may be mistaken for homozygous state
111
Q

red cell enzymes tested for parentage (5)

A
  • phosphoglucomutase (PGM)
  • esterase (ESD)
  • glyoxylase (GLO)
  • group-specific component (GC)
  • acid phosphatase (ACP)
112
Q

serum proteins tested for parentage
function

A

transferrin and haptoglobin
can be used to distinguish siblings

113
Q

DNA methods used to test for parentage

A
  • restriction fragment length polymorphism (RFLP)
  • PCR
114
Q

RFLP

DNA is harvested from ———-
isolation and purification of DNA is through a process of…

A

WBCs
cell separation, lysis, enzyme digestion, precipitation of protein with NaCl

115
Q

RFLP

cut DNA strands at the same sequence every time

A

restriction enzymes

116
Q

RFLP

polymorphism results from individual differences in the sequence of —— and —–

A

VNTR (variable number tandem repeats)
STR (short tandem repeats)

117
Q

RFLP

separation of DNA fragments (3)

A
  • separated by size in a gel electrophoresis
  • negatively-charged DNA fragments move toward anode side
  • smaller fragments move farther along
118
Q

RFLP

interpretation of electrophoresis (5)

A
  • Southern Blot used to transfer DNA bands from gel to membrane filter
  • the DNA is denatured to ssDNA
  • membrane dried and baked to permanently fix DNA to membrane
  • radioactively labelled probes bind the bands, and unbound probe is washed
  • x-ray is used to visualize
119
Q

RFLP

if an AF isn’t exlcuded…

A

a paternity index is calculated based on gene frequency in general pop

120
Q

RFLP

required (7)

A
  • good QC of restriction enzymes
  • standardized controls
  • standardized electrophoretic separation
  • population data sufficient to accurately assess gene frequency
  • standardized probes
  • standardization of nomenclature of probes
  • publication of genetic probe data and family studies to assure genetic inheritance patterns
121
Q

must be used in conjunction with another parental testing method, usually ABO

A

RFLP

122
Q

PCR fragments can be visualized ——- on the ———- gel with a ——–

A

directly
agarose
silver stain

123
Q

standard sample containing each of alleles run with samples, for direct visual comparison

A

PCR

124
Q

RBC excludes —-% fathers

A

73

125
Q

RBC + HLA exludes ——% fathers

A

95

126
Q

RBC + HLA + enzymes excludes —–% fathers

A

99

127
Q

RBC + DNA excludes ——% fathers

A

99.9

128
Q

direct exclusion

A

the child possesses a factor not present in the mother, or the AF

129
Q

POG

A

factor the child possesses but the mother does not

130
Q

indirect exclusion

A

child lacks a factor that the AF would’ve had to pass on (homozygous)

131
Q

likelihood of paternity based on —— principle, which states:

A

Hardy-Weinberg

selection of mates is random, and genetic markers are passed to offspring in a random manner

132
Q

probabilty calculation in a 2-allele system

A

p^2 + 2pq + q^2

133
Q

basis for calculation of the likelihood of paternity

A

POG

134
Q

genetic odds ratio which reflects how much more likely it is that an included AF is the father than a random man of same race

A

paternity index
PI = x/y

x = odds of AF passing on POG
y = gene frequency of POG

135
Q

will be 0 if any PI value is 0

A

CPI

136
Q

combined paternal index

A

product of all individual system PI values

137
Q

converts PI into a percentage

A

likelihood of paternity
W = PI/(PI + 1)

Mercy CLS 4: Blood Bank (13 decks)

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