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Immunology > Lecture #3 > Flashcards

Lecture #3 Flashcards

1
Q

Immunoglobulins can be _______ or ______ receptors

A

Membrane Bound or Soluble

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2
Q

What causes a resting B cell to generate antibody?

A

Encounter with antigen turns it into a plasma cell

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3
Q

What is an epitope?

A

An antigenic determinant The part of an antigen to which an antibody binds

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4
Q

T or F. Most antigens have a single epitope.

A

False. Most antigens ahve multiple epitopes

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5
Q

What are epitopes usually made of?

A

Carbohydrates of pepride

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6
Q

Difference between DTwP and DTaP

A

DTwP = Whole Cell Pertussis VaccineDTaP = Acellular Pertussis Vaccine

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7
Q

An immune response to _____ is triggered in an HPV vaccination.

A

Caspid

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8
Q

Where are epitopes recognized by antibodies typically located?

A

At the antigen surface. There they can be directly bound

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9
Q

Two different types of epitopes an Ab can bind to.

A

Linear and Discontinuous

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10
Q

Difference between linear and confirmational/discontinuous epitopes.How is this different from T Cells

A
  • Linear epitope binding happens to series of peptide in a row. - Discontinuous epitope binds to peptides adjacent to one another because they are disrupted and unfolded. - Ts Only Do Linear
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11
Q

What lymphocytes recognize confirmational epitope?

A

B Cells.NO T Cells

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12
Q

What lymphocytes recognize linear epitopes?

A

B Cells and T Cells

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13
Q

What are Haptens?

A

Small molecules that are not immunogenic alone. They can bind Igs and TCRs.

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14
Q

What do Haptens do?

A

They can induce immune responses when linked to a lager carrier.

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15
Q

Example of a Hapten?

A

Penicillin

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16
Q

Effect of Penicilin on bacteria

A

Binds bacterial transpeptidase and inactivates it

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17
Q

Effect of Penicillin on RBC

A

Modifies RBC Proteins to generate a foreign epitope

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18
Q

What happens to RBCs after they receive their hapten epitope?

A

B Cells are activated by antigen+TH2Ab binding to RBCsRBC destruction

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19
Q

What holds together the light and heavy chains?

A

disulfide bonds

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20
Q

Where can disulfide bonds be found?

A

Between light and heavy In the Hinge Region

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21
Q

Names of the two light chains

A

Lambda and Kappa

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22
Q

Name the five heavy chains

A

Mu, Gamma, Alpha, Epsilon, Delta

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23
Q

The specificity of binding comes from the…

A

Antigen binding sites/Variable Region

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24
Q

What do you find when Ab is cleaved by Papain?How many total fragments are generated?

A

Antigen-binding FragmentCrystallizable FragmentThree

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25
Q

What do you find when Ab is cleaved by pepsin?How many total fragments are generated?

A

F(ab’)2 fragmentChopped up base2 Fragments

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26
Q

What function might F(ab’)2 have?

A

Naturally, none. In labs, it can be used in binding studies.

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27
Q

Use of the hinge region?

A

Gives flexibility at antigen binding sites

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28
Q

Downside of the hinge region?

A

Susceptible to proteolysis by bacterial and host proteases

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29
Q

How many domains in heavy chain? Light Chain?What are they?

A
  1. 3C, 1 V2. 1C, 1V
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30
Q

Significance of V and C Regions?

A

V=Epitope BindingC = Biological Fxn of Molecule

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31
Q

Describe the molecular structure of the Ab domains.

A

Two beta sheets held together by a disulfide bond”Sandwich with a Toothpick”

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32
Q

Another name for hypervariable region?

A

Complementarity-Determining Region

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33
Q

What do hypervariable regions do?

A

They provide the actual antigen binding specificity.

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34
Q

How many HV regions are in an antigen binding site?

A

6 (3 on light and 3 on heavy)

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35
Q

Are the HV regions adjacent genetically?

A

No, hydrophobic regions come together from different parts of the genome in the process of protein folding.

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36
Q

Name the 5 types of Immunoglobulins

A

Ig G, M, D, A, E

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37
Q

Which immunoglobulins have no Hinge region?What else is special about them?

A

IgM and IgEAlso have a 4th constant domain

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38
Q

Problems with the unique structure of IgM and IgE?

A

Less Flexibility of antigen binding.

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39
Q

What are isotypic differences between immunoglobulins?

A

Like IgG vs IgA

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40
Q

What are allotypic differences between immunoglobulins?

A

Variations of immunoglobulins seen because they come from the two different genes. Usually only very small differences.

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41
Q

What are idiotypic differences between immunoglobulins?

A

Like IgGs with different epitope binding sites

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42
Q

Why care about allotypical differences (3)

A

May alter half lifeMay Affect subclass distributionAssociated with susceptibility to infectious and autoim. disease

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43
Q

How are allotypes inherited?

A

Autosomal dominant

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44
Q

T or F. Binding strength can be very variable between different antibody-epitope pairs.

A

True

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45
Q

What forces drive the antibody-antigen interaction?

A

Non-Covalent:-Electrostatic- Hydrogen Bonds- Van der Waals- Hydrophobic

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46
Q

What is affinity?

A

Strength of the interaction between the epitope and one antigen binding site.

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47
Q

What is avidity?

A

Strength of the sum of interactions between antibody and antigen. Also any other rxns that might be invovled (i.e. TCR, etc.)

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48
Q

Why might avidity differ from affinity.

A

Binding with both antigen binding sites can significantly increase the strength.

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49
Q

Explain how cross reactivity works.

A

Antiserum raised against antigen A also reacts against antigen B because they share an epitope.

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50
Q

Two scenarios in which cross-reactivity is very important.

A

Impt. for vaccines and laboratory reactions

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51
Q

How many Hs and Ns in Influenza?

A

16H and 9N

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52
Q

What are monoclonal antibodies.

A

Immortilizations of single clone of antibody-secreting cells

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53
Q

How are monoclonal antibodies made in the lab? (broad concept)

A

B Cells + Neoplastic (myeloma) cells

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54
Q

Steps of monoclonal antibody generation.

A
  1. Fuse B and Myeloma Cells2. Grow in drug-containing medium to select for hybrids3. Select for antigen-specific hybridoma4. Clone the winner
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55
Q

How are polyclonal antibodies made?

A

From serum of immunized animals. (goats, rabbits)

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56
Q

Concerns regarding polyclonal antibodies

A

Multiple specificities and affinitiesVariation from batch to batch

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57
Q

Name the four types of therapeutic monoclonal antibody.

A

MouseChimericHumanizedHuman

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58
Q

Nomenclature for Chimeric

A

-ximab

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59
Q

Nomenclature for humanized

A

-zumab

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60
Q

Nomenclature for human

A

-umab

61
Q

Problems with therapeutic mouse antibodyWhen is it still useable

A

Patients develop anti-mouse antibodies Diagnostic

62
Q

What mouse regions are maintained in humanized antibodies?

A

HVR

63
Q

Can therapeutic human antibody elicit an immune response?

A

yes

64
Q

Three types of immunoassays and level of detection.

A

Precipitation – 30ug/mLAgglutination – 1 ug/mLRadioimmunoassay/ELISA – 1 pg/mL

65
Q

Whats precipitation all about?

A

Antigens form large enough complexes to see with the naked eye. Insensitive and rarely used.

66
Q

What is agglutination all about?

A

Usually hemaglutination

67
Q

What is ELISA all about?

A

Very sensitive. Good for studies of viruses and such

68
Q

To form the largest precipitates, what proportion of antibody to antigen is required?

A

Equivalence

69
Q

Aggregates are known to get stuck in _____ and _____

A

Kidneys and Joints

70
Q

Larger antigens are more prone to have _______ epitopes

A

MoreMakes large antigens better for aggregation

71
Q

Do you have to be sensitized to have anti-A or B

A

No – prbs cross reactions with gut microbes

72
Q

Ab type used in hemagglutination

A

IgM

73
Q

Why is a different AB blood type baby fine?

A

Mom’s IgMs can’t cross the placenta

74
Q

Difference between Direct and Indirect Coombs Test?

A

Direct: anti-human Ig Abs (Coomb’s reagent) + fetal RBCsIndirect: Mom’s Serum+Rh+ RBCsAfter either, add anti-human antibody and assess aglut.

75
Q

What type of Ig is used in Coombs?

A

IgG

76
Q

Why don’t the IgGs agglutinate spontaneously?

A

They’re too small, so RBC without Ab can’t

77
Q

What is Coombs test even assessing?

A

Rhesus factor antibody response in mother and child (i.e. Rh- mom and Rh+ kid)

78
Q

Should a Rh- mother be worried about her first Rh+ child?

A

Probs not. Should not have Abs developed till the 2nd time

79
Q

Steps in an ELISA

A
  1. Coat Wells with Antigen2. Add serum sample3. Add enzyme-labeled anti-human IgG4. Add substrate
80
Q

Steps in a Western Blot

A
  1. Dissociate proteins in SDS2. Run SDS-Page to separate by molecular weight3. Transfer to nitrocellulose3. Overlay with anti-serum4. Detect antibody with enzyme linked anti-IgG
81
Q

Tell me about immunoflouresence studies.

A

Put an antibody on with a fluorophore attached, send exciting light, pick up emitted light on microscope. Use to identify structures (esp. in dev. bio)

82
Q

Explain Flow Cytometry.

A
  1. Incubate samples with Ab2. Each monoclonal Ab had differently colored labels3. Run through a small stream with a laser that can count based on the diff colors.
83
Q

Value of flow cytometry?

A

Qualitative, RapidCommon in Diagnostic Labs.

84
Q

Fragments of genome seen in lambda chain? kappa chain? heavy chain?

A

l – VJCk – VJCh – VDJC

85
Q

Describe the steps of kappa chain generation

A
  1. Germline DNA undergoes VJ combination2. B cell DNA undergoes transcription into RNA transcript3. VJC is made via RNA splicing4. Translation into kappa chain polypeptide
86
Q

List the order of fragments coming together in heavy chain

A

DJ –> VDJ –> VDJC

87
Q

Significance of alrge number of gene segements in Ig loci?

A

Allow for A TON of recombinatory mechanisms

88
Q

What is the purpose of recombination signal sequences?

A

They prevent bonding of regions out of order

89
Q

Significance genetically of the 12 and 23 nuc. segments

A

12 = 1 twist of DNA helix23 = 2 twists of DNA helix

90
Q

What are the nucleotide segments involved in recombination signal sequences?

A

7-23-9 binds with 9-12-7

91
Q

Enzyme complex that conducts recombination. Mentioned components.

A

VDJ RecombinaseRAGs, TdT, Artemis Nuclease

92
Q

What are RAGs

A

Recombination Activity GenesLoop out intervening DNA to delete it

93
Q

Steps from cleaving out the loop to recombined DNA (6)

A
  1. RAG leaves DNA hairpins2. RAG nicks hairpins, generates palindromic P-nucleotides3. TdT adds in nucleotides at joining region4. Strand pairing5. Unpaired nucleotides are removes by exonuclease6. Gaps caused by DNA synth and ligation
94
Q

Three ways recombination can inadvertently promote diversity.

A
  1. TdT adds in nucleotides2. Imprecise Joining3. Unpaired nucleotide removal can lead to frameshift
95
Q

Initially, what heavy chain is made

A

IgM

96
Q

Why aren’t all Abs IgMs?

A

Isotype switching can happen later

97
Q

Naive mature B cells express what isotypes?

A

IgM and IgD

98
Q

What is addedd to the Ig following the C chain being splied in -

A

AAA

99
Q

Why are there two kinds of naive generated?

A

RNA contains both IgM and IgD componentsThey can be splicaed differently

100
Q

T or F. Alternative splicing = isotype switching.

A

False.

101
Q

Since B cell receptors lack the AA for intracellular signalling, they associate with…

A

Ig alpha and beta heterodimer

102
Q

Vaugely descripe B cell signalling

A

B cell binds, conformational change, activation of heterodimer, tyrosine kinase intracellular signalling

103
Q

What happens to B cells after activation?

A

Ab SecretionSomatic Mutation (more diversity)Isotype switching

104
Q

How does the change from Membrane to secreted Ig happen?

A

Alternative RNA processing

105
Q

T or F. Somatic hypermutation will always lead to stronger antigen binding.

A

F. Can increase or decrease specificity or have a neutral effect.

106
Q

What mediates somatic hypermutation activity of CDR 1/2?What does it do?

A

Activation-induced Cytidine deaminaseCytosine->Uracil. Changes base pairs

107
Q

Outcome of isotype switching?

A

Igs with different C regions, but identical antigen specificity.

108
Q

Steps of Isotype Switching.

A
  1. IgM and IgD are produced. 2. AID selectively targets S(mu) and S(gamma1) switch3. DNA of both switch regions is nicked4. DNA btw nicks is looped out5. IgG1 is produced
109
Q

What is AID?

A

Activation-Induced Cytosine DeaminaseImportant in Somatic hypermutation AND isotype switching

110
Q

After isotype switching, can you go back?

A

No. DNA is spliced out.

111
Q

Effector fxns of Abs?

A

Neutralization (bind to prevent receptor binding)OpsonizationComplement Activation

112
Q

Whats so cool about IgM?

A

Pentamer StructureCan be transported in mucosal secretions

113
Q

IgM have _____ Affinity and _____ Avidity

A

Low Affinity and High Avidity

114
Q

Who is the first Ig synthesized in immune response.

A

IgM

115
Q

Roles of IgM

A

Activates complementAgglutination

116
Q

___ stabilizes the IgM pentamer

A

J chain

117
Q

Which Ab has the highest conc. in the serum?

A

IgG

118
Q

How many IgG subclasses?

A

4

119
Q

Complement activation of the 4 subtypes?

A

IgG3 > IgG1 > IgG2. Ig4 doesn’t do shit.

120
Q

Roles of IgG

A

Binds Fc receptors of neutrophils, macrophages, NKsComplement Activation

121
Q

Difference between the 4 IgGs?Which is especially prevalent?

A

Hinge Region StructureIgG3 has a freakin huge hinge – very susceptible to proteases and shorter half life

122
Q

Whats unique about IgG4

A

Functionally Monovalent

123
Q

What does functionally monovalent mean?

A

Can dissociate at hinge region and recombineForms bivalent Ab molecules w/ 2 binding sites

124
Q

End effects of functional monovalence?

A

Less effective

125
Q

Secretory AgA is a….

A

Dimer with secretory component

126
Q

Subclasses of IgA?

A

IgA1 and 2

127
Q

Is IgA1 usually a monomer or dimer?

A

Monomer

128
Q

Is IgA2 usually a monomer or dimer?

A

Dimer

129
Q

Where do you tend to see IgA?

A

Mucosal Secretions

130
Q

Primary role of IgA? Response to this?

A

IgA does neutralizationMany bacteria produce IgA proteases to fuck it up

131
Q

IgA sucks at…

A

activating complementHelps not kill epithelial barriers of mucosa in infection

132
Q

Which Abs have no hinge?

A

ME

133
Q

IgE is normally at a ___ concentration

A

Low

134
Q

What patient would you expect heightened IgE in?

A

Helminth infections or Allergies

135
Q

What does IgE bind?>

A

Fc receptor of Mast/Baso

136
Q

Where is IgD found?

A

URT

137
Q

Lowest half life?

A

IgE

138
Q

Relative frequency?

A

IgGMiddle IgA, MLowest ED

139
Q

Which Igs primarily do neutralization?

A

G,A

140
Q

Which Igs primarily do opsinization?

A

G1,G3

141
Q

Which Igs primarily do NK killing?

A

G1, G3

142
Q

Which Igs primarily do Mast cells?

A

E

143
Q

Which Igs primarily do complement system?

A

M, G1, G3

144
Q

Which Igs primarily do transport across epithelium?

A

A

145
Q

Which Igs primarily do transport across placenta?

A

G134

146
Q

Which Igs primarily do extravascular diffusion

A

G,A

147
Q

List the seven events of a B cell’s life

A
  1. V region assembly2. Generation of Junctional Diversity3. Assembly og Transcriptional Controlling elements4. Trans. activated with IgM and D5. Synthesis changes to secreted6. somatic hypermutation7. Isotype Switch
148
Q

Which two (of the seven event in a B cells life) are reversible?

A

Transcription of IgM and DSynthesis changes from membrane to secreted antibdy.

Immunology (10 decks)

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