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Immunology > Module 3 > Flashcards

Module 3 Flashcards

1
Q

What is humoral immunity?

A

Immunity mediated by the humors, of body fluids

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2
Q

After centrifuging blood to remove RBCs what is the remaining liquid?

A

Plasma

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3
Q

What do you call plasma without the clotting agents?

A

Serum

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4
Q

What are two other names for antibodies?

A

Immunoglobulins (Ig) and gamma globulins

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5
Q

What is the structure of an antibody?

A

Two heavy chains and two light chains in the form of a Y

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6
Q

What are the two types of antibody light chains?

A

Kappa and Lambda

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7
Q

What connects the heavy chains of an amino acid as well as the light chains to the heavy chains?

A

Disulfide bridges

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8
Q

Because the heavy chains and light chains are identical, antibodies are structurally ____

A

Symmetrical

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9
Q

What is the variable region of an antibody?

A

The highly variable first 100 amino acids that make up the specificity characteristics of the antibody

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10
Q

Where is the variable region located on an antibody?

A

At the amino terminus of the light and heavy chains, at the tips of the Y

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11
Q

Where is the constant region of the amino acid?

A

The carboxy terminus, the Y except the tips

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12
Q

What are the two functional domains of an antibody?

A

Fab and Fc

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13
Q

What is the Fab (fragment antigen binding) functional domain of an antibody?

A

It is located at the split of the Y and accounts for the specificity of the single antigen binding site

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14
Q

What is the Fc (fragment crystallizable) functional domain of the antibody?

A

It is located at the bottom of the two heavy chain constant regions or the stem of the Y and serves as effector function mediation instead of binding

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15
Q

What is an epitope?

A

The sub-portion of antigen that binds to the Fab domain of an antibody

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16
Q

What determines the strength of the affinity between an antibody and an antigen?

A

The sum total of its non-covalent bonds

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17
Q

How many hyper variable regions are there in the antibody variable region?

A

Three

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18
Q

What else is the hyper variable region of an antibody known as?

A

Complementarity Determining Regions (CDR1, CDR2 and CDR3)

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19
Q

Where is the greatest amino acid variability in an antibody?

A

The hyper variable regions

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20
Q

What are the five human isotypes?

A

IgA, IgD, IgE, IgG and IgM

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21
Q

Where do isotypes differ from each other?

A

The heavy chain constant region (where the Y splits)

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22
Q

How can you tell the difference between the four subtypes of IgG?

A

They differ in length at flexible hinge region of the heavy chains (where the Y splits)

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23
Q

What is the most abundant isotype and predominates in the secondary response?

A

IgG

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24
Q

Which isotype is secreted as a pentameter with 10 Ag binding sites and is most predominant in the primary response?

A

IgM

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25
Q

Which isotype is secreted as a diner with 4 Ag binding sites and is most predominant in bodily secretions?

A

IgA

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26
Q

How is passive immunity passed through breast milk?

A

IgA is readily available in breast milk, though with no memory B cells the immunity doesn’t last

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27
Q

What are the five antibody isotypes?

A

IgM, IgG, IgA, IgE and IgD

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28
Q

What is the defining feature of IgM?

A

It is the first Ab formed in the primary response and is a pentamer

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29
Q

What are the distinctive features of IgG?

A

It is the most abundant Ag in serum, has four types and is a major part of the secondary response

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30
Q

What are the defining features of IgA?

A

It is a dimer Ag that is present in external secretions including breast milk

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31
Q

What are the distinctive features of IgE?

A

They sensitize mast cells and basophils resulting in allergy symptoms and helminth protection

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32
Q

What are the distinctive features of IgD?

A

It is expressed on naive B cells and activity is rarely demonstrated

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33
Q

What accounts for the limitless antibody varieties that can by synthesized by an individual?

A

Gene rearrangement events that occur during B cell development in the bone marrow and T cell development in the thymus

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34
Q

Where are the genes that encode for the heavy and light chains of an antibody?

A

Across three separate chromosomes

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35
Q

Immunoglobulin locusts organized into coding cadets made up of what?

A

Multiple V, J and D regions

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36
Q

What are the three gene segments of the antibody heavy chain?

A

Variable (V) segment, Diversity (D) segment and Joining (J) segment

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37
Q

What are the two light chain antibody gene segments?

A

Variable (V) segment and Joining (J) segment

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38
Q

Where do Rag1 and Rag2 (recombination activator genes) bind for VDJ gene recombination?

A

The recombination signal sequence motifs (RSS) located where genes can be recombined

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39
Q

What do RAG1 and RAG2 do as a part of VDJ gene recombination?

A

They bring together the gene segments together to recombine and cleave gene at RSS motif

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40
Q

What is the process of P additions after the VDJ recombination?

A

Hairpin DNA loops created during recombination are opened and refilled by DNA repair enzymes

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41
Q

What is the process of N additions after VDJ recombination?

A

TdT enzyme (terminal deoxynucleotide transferase) adds random nucleotides to exposes chromosome ends

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42
Q

Which is the first isotype expressed by naive B cells?

A

IgM

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43
Q

What is the process by which a DNA rearrangement occurs between constant regions in exchange of Cmu coding region for constant regions of a different isotype?

A

Isotype switching

44
Q

What is the antibody isotype found during primary response?

A

IgM

45
Q

What is the antibody isotype primarily found in secondary response?

A

IgG

46
Q

What is responsible for altering the affinity strength between an antibody and antigen?

A

Single point mutations of the somatic hypervariable region of immunoglobulin DNA

47
Q

During clonal selection, which B cell receptors are selected?

A

Those with a higher Ag affinity

48
Q

What is affinity maturation?

A

When secondary and later responses produce antibodies with increasing affinity

49
Q

What are the four steps at occur during B cell development in the bone marrow?

A
  1. V-region assembly from gene fragments
  2. Generation of junction diversity
  3. Assembly of transcriptional controlling elements
  4. Transcription activation with coexpression of surface IgM and IgD
50
Q

What are three changes that occur post activation to B cells in secondary lymph tissue?

A
  1. Membrane Ig changes to secreted antibody
  2. Somatic hypermutation
  3. Isotype switch
51
Q

What are the five effector functions of the different antibody isotypes?

A
  1. Neutralization of pathogens
  2. Opsonization
  3. NK cell sensitization
  4. Mast cell sensitization
  5. Complement activation
52
Q

What binds to viruses to prevent its attachment to a host cell receptor?

A

Fab (the arms of the antibody)

53
Q

Which antibody is the most important neutralizer for the mucosal tract?

A

IgA

54
Q

Which antibody is found in breast milk?

A

IgA

55
Q

Which antibody isotype diffuses across the placenta for passive immunity during fetal development?

A

IgG

56
Q

Which part of the antibody binds to bacterial pathogens blocking its ability to colonize?

A

Fab (the arms of the antibody)

57
Q

How are secreted exotoxins neutralized?

A

Fab binds to toxins blocking its ability to cause cellular damage

58
Q

Phagocytes (neutrophils and macrophages) have Fc receptors for which antibody isotypes?

A

IgG1 and IgG3

59
Q

What happens during opsonization?

A

Phagocytes bind to Fc portion of antibodies that coat a pathogen for enhanced uptake

60
Q

Which antibody isotypes trigger NK cells to attack?

A

IgG1 and IgG3 which coat the target cell such as a virus infected host cell or cancer cell

61
Q

Which antibody isotype coat helminthic parasite activated eosinophils to secrete toxins that damage worms?

A

IgE

62
Q

Which antibody isotype bind to Fc receptors on mast cells and basophils?

A

IgE

63
Q

Which three antibody isotypes activate the complement cascade by the classical pathway?

A

IgM, IgA and IgG

64
Q

What is the Western Blot test?

A

The process to identify protein/a set of proteins within a mixture

65
Q

What are the four steps of a Western Blot test?

A
  1. Proteins separated with electrophoresis through polyacrylamide denaturing gel
  2. Proteins transferred onto filter paper and exposed to antibody that binds to protein of interest
  3. Excess antibody washed off and secondary antibody added that is specific for the bound antibody
  4. Secondary enzyme conjugated to an enzyme that reacts and makes a color to show protein location
66
Q

What is the Enzyme Liked Immunosorbent Assay (ELISA) test?

A

A sensitive quantitative technique for detecting antigens or antibodies that can be used for diagnostics

67
Q

What are the seven steps of the ELISA test?

A
  1. Each of the 96 wells on the microtiter plate filled with a protein sample
  2. Primary antibody added to each well
  3. Antibody binds if antigen is present
  4. Excess antibody removed
  5. Secondary specific antibody added
  6. Secondary antibody changes color
  7. Amount of color measured by spectrophotometer
68
Q

What does Immunoaffinity Chromatography accomplish?

A

It is a process that purifies a specific macromolecule

69
Q

What are the four steps of immunoaffinity chromatography?

A
  1. Antibody specific for macromolecule of interest sent through chromatography beads in a column
  2. Mixture of macromolecules pass over the column
  3. Macromolecules of interest bind to the antibody and the other pass along
  4. Change in salt or pH of Washington buffer removes the bound macromolecules
70
Q

What is the process of immunofluorescence microscopy?

A

Antibodies conjugated to a fluorescent molecule can be detected under a fluorescent microscope

71
Q

What are the four steps of fluorescent activated cell sorting (FACS)?

A
  1. Antibodies conjugated to a fluorescent molecule
  2. Stream of fluid passes through flow cytometer nozzle that allows only one cell at a time
  3. Laser activates fluorescent antibody
  4. Detected by a computer
72
Q

What is the difference between immunofluorescence microscopy and fluorescence activated cell sorting?

A

Both use fluorescent tagging, but only FACS allows for the cells to be separated and counted

73
Q

What is the benefit of monoclonal antibodies (MAb) over polyclonal antibodies?

A

When antibodies are used as a research tool or therapeutic drug it is best to have a pure antibody

74
Q

How are monoclonal antibodies produced?

A

Immunize mice with antigen of interest, causing an immune response. Remove the B cells and fuse with myeloma cells. Hybridomas can then be analyzed by flow cytometry and ELISA to identify antibodies

75
Q

What is a hybridoma?

A

A normal B cell fused with a cancerous B cell (myeloma) which can grow indefinitely in culture

76
Q

What is HAMA and how is it created?

A

Human anti-mouse antibody, it is created when the human immune system reacts against mouse antibodies

77
Q

What three diagnoses are treated with humanized monoclonal antibodies?

A

Breast cancer, rheumatoid arthritis and MS

78
Q

How are chimeric monoclonal antibodies made?

A

mRNA from mouse hybridomas modified using recombinant DNA techniques to replace antibody coding with human sequence.

The mouse V region (murine) is maintained

79
Q

How are humanized monoclonal antibodies produced?

A

mRNA from mouse hybridomas modified with recombinant DNA technique to replace antibody protein coding with human sequence.

Only the hypervariable murine regions (CD1, CD2 and CD3) maintained and the rest wrapped with human sequences

80
Q

What is required for a full humanized monoclonal antibody?

A

A knockout-transgenic mouse

81
Q

What process of producing monoclonal antibodies does not require mice?

A

Fully humanized phage display

82
Q

Antigens are made up of many antigenic determinants, also known as?

A

Epitopes

83
Q

Bacterial cells have many different epitopes, which of which bind to?

A

Specific antibodies

84
Q

An individual antibody is made against?

A

One epitope on a cell

85
Q

Each antigen has ___ epitope(s)?

A

One

86
Q

Each antigen has many epitopes, and a different antibody is formed against ____

A

Each epitope

87
Q

Antibodies are?

A

Immune system-related proteins also known as immunoglobulins

88
Q

The antigen binding site of immunoglobulin is formed from what?

A

Paired V chain region of a single heavy chain and single light chain

89
Q

When IgG is cleaved with a protease that targets the hinge, what is generated?

A

Two Fab fragments and two Fc fragments

90
Q

What is the part of the antibody that binds to an antigen?

A

The V domain

91
Q

What contributes 50 lads to the molecular weight of IgG?

A

The heavy chain

92
Q

What comprises the beta strands and loops not involved in antigen binding?

A

The framework region

93
Q

Which is the most conserved region of the molecule with limited variation between antibodies?

A

The constant region

94
Q

What is located within the V region and varied greatly between antibodies?

A

The hypervariable region

95
Q

What pairs with the amino-terminal part of the heavy chain to form an antibody arm?

A

A light chain

96
Q

Which epitope is formed as a result of 3D folding of a protein, and which would be destroyed if the protein denatures?

A

A conformational protein epitope

97
Q

All epic exclusion ensures that B cells do what?

A

Express only one V, J and D segment during somatic recombination

98
Q

Which IgG subclass is most efficient at activating complement?

A

IgG3, because it’s hinge is longest and makes its Fc region more accessible for C1 binding.

99
Q

Which IgG subclass is least effective at complement activation?

A

IgG4 fils to activate complement because it’s Fc region binds poorly to C1

100
Q

Which antibody plays a role in opsonization, complement activation, and is the most abundant?

A

IgA

101
Q

Which antibody sensitizes basophils?

A

IgD

102
Q

Which antibody sensitizes mast cells, basophils and is the least abundant?

A

IgE

103
Q

Which antibody plays a role in opsonization and complement activation, as well as transports across the placenta, sensitizes masts cell, NK cells and is abundant?

A

IgG

104
Q

Which antibody only plays a role in complement activation?

A

IgM

105
Q

What are humanized monoclonal antibodies?

A

Human antibodies where the CDR loops have been replaced by mouse-derived CDRs of the desired specificity

106
Q

Which IgG subclass cannot activate complement?

A

IgG4 can not activate complement because it’s Fc region binds poorly to C1.

107
Q

Why can the immune system continue to make antibodies after treatment with anti-CD20 antibody rituximab?

A

Plasma cells do not express CD20 on their surface and cells protected by these cells continue as normal.

Immunology (11 decks)

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