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Immunohematology Lec > [11] CHAPTER V LESSON 1 > Flashcards

[11] CHAPTER V LESSON 1 Flashcards

1
Q

AHG’s obtained from immunized nonhuman species bind to human globulins such as (?] either free in serum or attached to antigens on red blood cells (RBC’s).

A

IgG or complement

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2
Q

2 major types of blood group antibodies:

A

IgM
IgG

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3
Q
  • bind to corresponding antigen
A

IgM

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4
Q

directly agglutinate RBCs suspended in saline

A

IgM

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5
Q
  • “nonagglutinating” or “incomplete antibodies”
A

IgG

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6
Q

because their single monomer structure is too small to directly agglutinate sensitized RBCs

A

IgG

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7
Q

described the use of the antiglobulin test for the detection of weak and nonagglutinating Rh antibodies in serum.

A

Coombs, Mourant, and Race

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8
Q

described the use of AHG to detect in vivo sensitization of the RBCs of babies suffering from HDFN.

A

Coombs and coworkers

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9
Q

described the principle of the test

A

Moreschi

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10
Q

Kell blood group antibodies and assoc. antigen

A

Moreschi

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11
Q

Reported weeks after the Coombs had described the test

A

Moreschi

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12
Q
  • RBCs has been sensitized by an antibody
A

Hemagglutination

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13
Q

Early AHG reagents were prepared using a (?) as the immunogen.

A

crude globulin fraction

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14
Q

demonstrated that the antibody activity that detected Rh o antibodies were associated with the anti-gamma globulin fraction in the reagent.

A

Coombs and Mourant

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15
Q

: Observed that different reaction patterns were obtained when dilutions of AHG were used to test cells sensitized with warm as compared to cold antibodies.

A

Dacie

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16
Q

published data showing that the reactivity of AHG to cells sensitized with warm antibodies resulted from anti-gamma globulin activity, whereas antinongamma globulin activity was responsible for the activity of cells sensitized by cold antibodies.

A

Dacie and coworkers

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17
Q

was shown to be beta globulin and had specificity for complement

A

nongamma globulin component

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18
Q

Later studies revealed that the complement activity was a result of

A

C3 and C4

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19
Q

can be used to detect RBCs sensitized with IgG alloantibodies, IgG autoantibodies, and complement components.

A

antiglobulin test

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20
Q

can occur either in vivo or in vitro:

A

Sensitization

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21
Q

is detected by a one stage procedure

A

In vivo sensitization

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22
Q

In vivo sensitization ex.

A

Direct antiglobulin test (DAT)

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23
Q

The use of AHG to detect [?] of RBCs is a two-stage technique

A

in vitro sensitization

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24
Q

In vitro sensitization ex.

A

Indirect antiglobulin test (IAT)

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25
Q

ANTIHUMAN GLOBULIN REAGENTS
Defined by

A

Food and Drug Administration (FDA) and Center for Biologics Evaluation and Research (CBER)

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26
Q
  • contain antibody to human IgG and to the C3d component of the human complement.
A
  1. Polyspecific AHG
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27
Q

Other anticomplement antibodies, such as anti-C3b, may also be present.

A
  1. Polyspecific AHG
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28
Q
  • contain only one antibody specificity: either anti-IgG or antibody to specific complement components (C3b or C3d).
A
  1. Monospecific AHG
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29
Q

Licensed monospecific AHG reagents in common use are

A
  1. Monospecific AHG

antiIgG and anti-C3b,-C3d.

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30
Q

Contain no anticomplement activity

A

a. Anti-IgG

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31
Q

Contain antibodies specific for the Fc fragment of the gamma heavy chain of the IgG molecule

A

a. Anti-IgG

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32
Q
  • reactive against only the designated complement components and contain no activity against human immunoglobulins
A

b. Anticomplement

Anti-C3b and anti-C3d reagents

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33
Q

Monospecific anticomplement reagents: Often a blend of

A

b. Anticomplement

monoclonal anti-C3b and monoclonal C3d.

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34
Q

FDA Licensed Antihuman Globulin Reagents

Polyspecific

Monospecific

Anticomplement

A

Polyspecific
1. Rabbit Polyclonal
2. Rabbit/murine monoclonal blend

Monospecific
1. Anti-IgG (Rabbit polyclonal)
2. Anti-IgG (Gamma-done AHG)

Anticomplement
Anti-C3d

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35
Q

Contains anti-lgG and anti-C3d (may contain other anticomplement and other antiimmunoglobulin antibodies)

A

Rabbit Polyclonal

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36
Q

Rabbit/murine monoclonal blend

Contains a blend of (?) is a murine monoclonal IgM antibody

A

rabbit polyclonal antihuman IgG and Anti-C3d

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37
Q

Contains anti-IgG with no anticomplement activity

A

Anti-IgG (Rabbit polyclonal)

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38
Q

Murine monoclonal IgM antibody secreted by a hybridoma cell line

A

Anti-IgG (Gamma-done AHG)

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39
Q

The main component of this reagent is a murine monoclonal antibody to C3d.

A

Anti-C3d

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40
Q

will cause the agglutination of red blood cells coated with human C3d and/or C3b complement components.

A

Anti-C3d

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41
Q
  • Involves injecting human serum or purified globulin into laboratory animals such as rabbits.
A

Classic method of AHG production

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42
Q

The (?) behaves as foreign antigen, the rabbit’s immune response is triggered, and an antibody to (?) is produced.

A

human globulin

human globulin

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43
Q
  1. Human IgG injected into a rabbit- results in
A

anti-IgG production

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44
Q
  1. Human complement components injected into a rabbit result in
A

anticomplement

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45
Q

PREPARATION OF AHG

This type of response produces a

A

polyclonal antiglobulin serum

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46
Q

can be made using polyclonal or monoclonal antibodies.

A

Polyspecific AHG

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47
Q

The two types of antibody production processes are very different from one another, yielding two very different advantages and disadvantages in their usage.

A

Polyspecific AHG

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48
Q

is usually prepared in rabbits.

A

Polyclonal AHG

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49
Q

are produced by immunizing one colony of rabbits with human immunoglobulin (IgG) antigen and another colony with human C3 antigen.

A

Conventional polyspecific antiglobulin reagents

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50
Q

Because of the heterogeneity of IgG molecules, using serum from many donors to prepare the pooled IgG antigen to immunize the rabbits and the pooling of anti-IgG from many immunized rabbits is essential in producing polyclonal reagents for routine use that are capable of detecting the many different IgG antibodies.

A

Polyclonal AHG

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51
Q

This is an advantage of using anti-IgG of polyclonal origin for antiglobulin serum.

A

Polyclonal AHG

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52
Q

allows for a broader spectrum of reactivity, but the source of antibody is limited to the life span of the inoculated animal.

A

Preparation of polyclonal AHG reagents

Pooled donor antigen

53
Q

may be manufactured by combining polyclonal anti-IgG with either polyclonal or monoclonal anticomplement components.

A

Polyspecific antihuman globulin

54
Q

Monoclonal antibody technique:

A

Kohler and Milstein

55
Q

Monoclonal antibody technique:

A

Kohler and Milstein

56
Q

Monoclonal antibody technique:

A

Kohler and Milstein

57
Q

Useful in producing high-titer antibodies with well-defined specificities to IgG and to the fragments of C3

A

Monoclonal AHG Production

58
Q

Monoclonal AHG Production
Procedure
1. Begins with the immunization of laboratory animals usually mice with [?]
2. After a suitable immune response, mouse spleen cells containing antibodysecreting lymphocytes are fused with [?].
3. The resulting hybridomas (?) are screened for anti -bodies with the required specificity and affinity.
4. The antibody- secreting clones may then be propagated in tissue culture or by inoculation into mice, in which case the antibody is collected as [?] (fluid accumulation in the abdomen of the mouse).

A

purified human globulin

myeloma cells

lymphocyte–myeloma hybrid cells

ascites

59
Q

Because the clonal line produces a single antibody, there is no need for absorption to remove [?].

A

heterospecific antibodies

60
Q

is a hybrid cell that is used as the basis for the production of large amounts of antibodies for therapeutic, research, or diagnostic applications.

61
Q

The production of antibody is longer lasting than the polyclonal source, as the hybridoma can live indefinitely.

A

monoclonal AHG reagents

62
Q

A monoclonal blend may be manufactured by blending

A

monoclonal anti-C3b, monoclonal anti-C3d, and monoclonal anti-IgG

63
Q

can be manufactured by conventional or hybridoma technology

A

Monospecific antihuman globulin reagents

64
Q

process is similar to that described above for polyspecific AHG

A

Monospecific AHG production

65
Q

it contains only one antibody specificity

A

Monospecific AHG production

66
Q

is produced as a monoclonal, polyclonal, or blended formula.

A

Monospecific anti-IgG

67
Q

is produced as a monoclonal, polyclonal, or blended formula.

A

Monospecific anti-IgG

68
Q

is produced as a monoclonal, polyclonal, or blended formula.

A

Monospecific anti-IgG

69
Q

ANTIHUMAN GLOBULIN REAGENTS

A
  1. Polyspecific AHG
  2. Monospecific AHG
70
Q
  1. Polyspecific AHG
A

a. Polyclonal AHG Production

b. Monoclonal AHG Production

71
Q

ANTIBODIES REQUIRED IN AHG

A
  1. Anti-IgG
  2. Anticomplement
72
Q

AHG must contain antibody activity to non-agglutinating blood group antibodies.

73
Q

The majority of these antibodies are a mixture of IgG1 and IgG3 subclasses.

74
Q

Rarely, non-agglutinating IgM antibodies may be found; however, they have been shown to fix complement and may be detected by anti-complement.

75
Q

is the only FDA-licensed monoclonal IgG AHG in the United States

A

Gamma-clone anti-IgG (Immucor, Inc., Norcross, GA)

76
Q

detects IgG1, IgG2, and IgG3, but as described in the package insert, it fails to react with IgG4.

A

Gammaclone

77
Q

This is not regarded as a significant limitation since [?]- only antibody specificities are uncommon, have not been associated with acute hemolytic reactions, and often have high-titer low-avidity-type features associated with clinically insignificant reactivity.

78
Q

Some antibodies “fix” complement components to the RBC membrane after complexing of the antibody with its corresponding antigen.

A

Anticomplement

79
Q

The terms most, some, and rare in the table refer to antibodies that bind complement the vast majority of the time, that show variability in their ability to bind complement, and that rarely bind complement, respectively.

A

Anticomplement

80
Q

These membrane-bound complement components can be detected by the [?] activity in AHG.

A

Anticomplement

81
Q

As a result of studies published during the 1960s indicating the need for [?] activity in AHG to allow the IAT to detect antibodies, a polyspecific reagent was introduced.

A

Anticomplement

82
Q

Evidence was also presented showing that the presence of [?] activity would enhance the reactions of clinically significant antibodies (e.g., anti-Fya and anti-K).

A

Anticomplement

83
Q

Antibodies Capable of Binding Complement MOST

A

ABO
Lea
Leb
Jka
Jkb
Sc1
Co3
Ge2
Ge3
Ii
P
P1PK
Vel

84
Q

Antibodies Capable of Binding Complement SOME

85
Q

Antibodies Capable of Binding Complement RARE

A

D
P1
Lua
Lub
Kell
Fya
Fyb
Coa
Cob
Dia
S
S
Yta

86
Q

contains both anti-IgG activity and anti-C3d activity.

A

• Polyspecific AHG

87
Q

• Because most clinically significant antibodies detected during antibody screening are IgG, the most important function of [?] is the detection of IgG antibodies.

A

polyspecific AHG

88
Q

• Many blood banks have begun using [?] for routine pretransfusion testing, citing cost containment measures necessitated by the high number of repeats versus the rarity of complement-only detected antibodies such as anti-Jka.

A

monospecific anti-IgG

89
Q

• Milam states that rare clinical transfusion intolerance, when using [?]over polyspecific AHG reagents to screen for unexpected antibodies and to test for blood group compatibility, offers reliability without interference from common and clinically insignificant IgMcomplement fixing antibodies.

A

monospecific anti-IgG

90
Q

Antibody molecules and complement components are

91
Q

Injecting an animal with [?] stimulates the animal to produce antibody to the foreign protein (i.e.,AHG).

A

human globulin

92
Q

Serologic tests employ a variety of AHG reagents reactive with various human globulins, including anti-IgG antibody to the C3d component of human complement, and polyspecific reagents that contain both [?] activity.

A

antiIgG and anti-C3d

93
Q

AHG reacts with [?] molecules, either bound to RBCs or free in serum.

A

human globulin

94
Q

Washed RBCs coated with human globulin are agglutinated by [?].

95
Q

detects in vivo sensitization of RBCs with IgG or complement components

A

direct antiglobulin test (DAT)

96
Q

Clinical conditions that can result in in vivo coating of RBCs with antibody or complement are the following:

A
  1. Hemolytic disease of the fetus and newborn (HDFN)
  2. Hemolytic transfusion reaction (HTR)
  3. Autoimmune and drug-induced hemolytic anemia (AIHA)
97
Q

o Maternal antibody coating of fetal RBCs

A
  1. Hemolytic disease of the fetus and newborn (HDFN)
98
Q

o Recipient antibody coating donor RBCs

A
  1. Hemolytic transfusion reaction (HTR)
99
Q

o Autoantibody coating individual’s RBCs

A
  1. Autoimmune and drug-induced hemolytic anemia (AIHA)
100
Q

DAT Panel
Initial DATs include testing one drop of a 3% to 5% suspension of washed RBCs with [?].

A

polyspecific (anti-IgG, anti-C3d) reagent

101
Q

DAT Panel
Positive results are monitored by a DAT panel using [?] to determine the specific type of protein sensitizing the cell.

A

monospecific anti-IgG and anti-C3d

102
Q

Anti-IgG +
Anti-C3d +

A

WAIHA

Mixed-type AIHA (cold and warm)

103
Q

Anti-IgG +
Anti-C3d -

104
Q

Anti-IgG -
Anti-C3d +

A

CAS; PCH; WAIHA

105
Q

The DAT with [?] is helpful in classifying AIHAs.

A

monospecific antiglobulin reagents

106
Q

Other procedures and studies are necessary to diagnose and characterize which form of [?] is present.

A

autoimmune disease

107
Q

CAS =

A

cold agglutinin syndrome

108
Q

PCH =

A

paroxysmal cold hemoglobinuria

109
Q

WAIHA =

A

warm autoimmune hemolytic anemia

110
Q

states that “a positive DAT result alone is not diagnostic of hemolytic anemia.

A

AABB Technical Manual

111
Q

Understanding the significance of this positive result requires knowledge of the [?].”

A

patient’s diagnosis; recent drug, pregnancy, transfusion, and hematopoietic transplantation history; and the presence of acquired or unexplained hemolytic anemia

112
Q

Answering the following questions before investigating a positive DAT for patients other than neonates will help determine what further testing is appropriate:
1. Is there evidence of [?]?
2. Has the patient been [?] recently? If so, did the patient receive blood products or components containing ABOincompatible plasma?
3. Does the patient’s serum contain [?]?
4. Is the patient receiving any [?]?
5. Is the patient receiving [?]?
6. Is the patient receiving [?]?
7. Has the patient received a [?]?

A

in vivo hemolysis

transfused

unexpected antibodies

drugs

antilymphocyte globulin or antithymocyte globulin

intravenous immune globulin (IVIG) or intravenous Rh immune globulin (IV RhIG)

marrow or other organ transplant

113
Q

The [?] is performed to determine in vitro sensitization of RBCs

A

INDIRECT ANTIGLOBULIN TEST

114
Q

INDIRECT ANTIGLOBULIN TEST is used in the following situations:
a. Detection of [?] to potential donor RBCs (compatibility testing) or to screening cells (antibody screen) in serum
b. Determination of [?] using known antisera (e.g., weak D, any other antigen testing that requires IAT)
c. Titration of [?]

A

incomplete (nonagglutinating) antibodies

RBC phenotype

incomplete antibodies

115
Q

[?], with sensitization occurring in vitro, looking for unknown antibody in the patient’s serum/plasma.

A

Indirect AHG (IAT)

116
Q

(Note: If the [?] was identifying RBC antigens, the RBCs would be from the patient and the antibody would be the known reagent).

A

Indirect AHG (IAT)

117
Q

[?] with sensitization already occurring in vivo.

A

Direct AHG (DAT)

118
Q

(Note: The polyspecific AHG reagent is a mixture of anti-IgG and anti-C3d.)

A

Direct AHG (DAT)

119
Q

Detects IgG and complement coated red cells

A

Direct Antiglobulin Test

120
Q

Detects IgG and complement coated red cells

A

Indirect Antiglobulin Test

121
Q

IgG attachment to red cells has occurred within the patient’s body One stage procedure

A

Direct Antiglobulin Test

122
Q

IgG attachment to red cells has occurred during the incubation step

A

Indirect Antiglobulin Test

123
Q

One stage procedure

A

Direct Antiglobulin Test

124
Q

Two-stage procedure

A

Indirect Antiglobulin Test

125
Q

Patient’s red cells are tested with antiglobulin reagent without an incubation step

A

Direct Antiglobulin Test

126
Q

Test requires an incubation step before the addition antiglobulin reagent

A

Indirect Antiglobulin Test

127
Q

Test for certain clinical conditions: HDFN, HTR, AIHA

A

Direct Antiglobulin Test

128
Q

Used as a reaction phase of several tests immunohematology

A

Indirect Antiglobulin Test

Immunohematology Lec (35 decks)

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