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Blood Bank > Immunology & Genetics > Flashcards

Immunology & Genetics Flashcards

1
Q

Immunity

A

Body’s defense against pathogens, foreign materials, and malfunctioning self-cells

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2
Q

Type 1 Hypersensitivity

A

Allergic reactions mediated by IgE antibodies and mast cells or basophils, causing histamine release

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3
Q

Type 2 Hypersensitivity

A

Antibody-dependent reactions involving IgG, IgM, complement, and phagocytic cells
- Antibodies attack own self cells

Ex: Hemolytic disease of newborn (HDN), autoimmune hemolytic anemia, transfusion reaction

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4
Q

Type 3 Hypersensitivity

A

Tissue damaging reactions due to immune complex deposition

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5
Q

Type 4 Hypersensitivity

A

Mediated by T-cells without antibody involvement, causing graft vs. host responses

  • Th1 cells activated and release cytokines —> recruit macrophages and neutrophils —> inflammatory response
  • Tc cells —> tissue destruction
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6
Q

Immune mediated diseases

A
  • Immune deficiencies: can lead to false-negative reactions
  • Monoclonal gammopathies/polyclonal gammopathies (multiple myloma) and autoimmune diseases: increase serum proteins and interfere with alloantibody detection.
    > Ex: Rouleaux formation and panagglutination
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7
Q

Major Histocompatibility Complex (MHC) aka Human Leukocyte Antigen (HLA)

A

Surface markers that allow immune cells/WBCs to distinguish “self” vs “nonself”

Found on membrane of nucleated cells

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8
Q

MHC Class I coded by what genes?

A

HLA-A
HLA-B
HLA-C

*Remember ABC

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9
Q

MHC Class II coded by what genes?

A

HLA-DR
HLA-DQ
HLA-DP

*Remember: PQR in alphabet

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10
Q

MHC Class III coded by what gene?

A

Not associated with surface membrane

Codes for complement and cytokines like TNF (Tumor Necrosis Factor)

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11
Q

Antigen Binding Groove

A

On MHC Class I and II

So endogenous and exogenous antigens can bind for WBCs to make antibodies or cytokines

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12
Q

Exogenous vs Endogenous

A

Exogenous: outside of the cell

Endogenous: inside a cell that has already invaded or tumor cells

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13
Q

MHC Class I

A

Binds endogenous antigens (tumors, viruses, intracellular bacteria - produced within the cell)

When NK cell finds a cell that lacks MHC Class I, it releases perforin and granzymes to induce cell death/apoptosis

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14
Q

MHC Class II

A

On APCs (monocytes, macrophages, dendritic cells, B lymphocytes)

Bind exogenous antigens (fragments of bacteria, virus or parasites ingested by APCs)

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15
Q

Opsonization

A

Promotes phagocytosis

C4b or C3b

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16
Q

Anaphylatoxins

A

Increase vascular permeability, contract smooth muscle, release histamine from basophils and mast cells

C3a, C4a, C5a

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17
Q

C5a

A

Chemotaxin - signals WBCs to migrate to affected area

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18
Q

C5b67

A

Promotes monocyte and neutrophil adherence to blood vessel and WBC migration to tissue

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19
Q

Classical Pathway

A

C1 = recognition unit, stabilized by Ca2+. Binds to IgM. Recruits C1q, C1r, C1s
- Cleaves C4 + C2 —> C4b2a —> C3 convertase
- C4b2a3b —> C5 convertase

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20
Q

Membrane Attack Complex (MAC)

A

C5-C9

Allows influx of water, causing cell to swell and lyse

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21
Q

Alternative Pathway

A

C3 —> C3a + C3b

C3b and Factor B require Mg+ to combine
Factor D unbinds Factor B from C3b

Properdin (Factor P) stabilizes the C3bBb complex

C3bBbP = C5 convertase

Formation of MAC

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22
Q

Mannose Binding Lectin Pathway

A

MBL = equivalent to C1q in Classical Pathway

MASP-1 and MASP-2 = function similarly to C1r and C1s

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23
Q

Innate Humoral Immunity

A

Complement Proteins

Components formed in the liver
C1 formed in intestinal epithelial cells
Factor D forms in adipose tissues

Plasma - components in inactive form
Once activated, components split into 2 fragments:
- smaller “a” - stimulates immune system
- larger “b” - activates cascade

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24
Q

Humoral Innate Immunity

A

Plasma proteins

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25
Q

Cell Mediated Adaptive Immunity

A

APCs, T lymphocytes, B lymphocytes

Phagocytize foreign antigen

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26
Q

TH cells

A

CD4

MHC II

Stimulate TC lymphocytes and B-cell activation

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27
Q

TC cells

A

CD8

MHC I

Produce perforin to destroy virus infected cells, tumor cells, tissue grafts

TNF (tumor necrosis factor) and interferons are released to prevent spread of virus

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28
Q

Humoral Adaptive Immunity

A

B lymphocytes secrete membrane bound antibodies, react with exogenous antigens

B cells produce specific antibodies (plasma cell)

TH dependent clonal proliferation - B cell acts as an APC to the T cell: produce memory cells

TH independent clonal proliferation - BCR are IgD that recognize polysaccharides and lipopolysaccharides: does not produce memory cells

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29
Q

Immunoglobulin

A

Glycoprotein with 2 heavy chains and 2 light chains, held together by disulfide bonds

Fab region: binds to antigen

Fc region: binds to complement and receptors on macrophages, determines Ig class

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30
Q

IgG

A

Only Ig that crosses placenta

IgG1 and IgG3 interact with macrophages through Fc receptors — bind complement and promote phagocytosis

37°C - body temp

Non-agglutinating antibodies — require AHG for hemagglutination

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31
Q

IgM

A

Pentamer, first antibody produced

Indicates active infection

Binds complement and cause intravascular hemolysis

Reacts at RT - 4°C

ABO antibodies

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32
Q

Primary Humoral Response

A

First exposure to antigen

T and B lymphocytes

4-7 days of lag phase (time it takes antibody to reach detectable level)

IgM

Antibody level declines quickly

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33
Q

Secondary Humoral Response

A

Repeat exposure to same antigen

Memory cells (anamnestic response)

1-4 days of lag phase

IgG

Greater antibody titer and level remains for a while

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34
Q

Convalescent Plasma Treatment

A

Isolating plasma from infected COVID patients (with antibodies), give plasma to sick patients to try to neutralize the virus

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35
Q

Rhogam

A

Antibody against Rh D antigen for prenatal testing

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36
Q

Immunogen

A

Chemical substance capable of eliciting an immune response

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37
Q

Epitope

A

Portion of the antigen that reacts with the antibody

A single antigen can have multiple epitopes

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38
Q

Polyclonal antibody

A

Different epitopes on a single antigen, inducing B cell clones —> results in heterogeneous population of serum antibodies

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39
Q

Monoclonal antibody

A

Single clone proliferation to produce IDENTICAL antibodies

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40
Q

Hapten

A

Small molecule that requires a carrier molecule to stimulate antibody production

41
Q

Immunogenicity

A

Antigen’s ability to elicit an immune response

Factors: age, race, genetics

42
Q

Dosage

A

Quantitative expression of specific antigen on the RBC surface based on gene inheritance

43
Q

Tolerance

A

Lack of immune response or active immunosuppressive response

44
Q

Sensitivity

A

Ability to detect antigen at a low titer

45
Q

Antigenicity

A

Antigen’s ability to bind with its antibody

Affected by antigen’s: location, size, shape, charge, solubility, digestibility

46
Q

Avidity

A

Overall strength of binding between antigens and antibodies

Influenced by valence of antibody and antigen

47
Q

Affinity vs Avidity

A

Affinity: strength of a single antibody-antigen interaction
- IgG has a high affinity to antigen

Avidity: strength of ALL INTERACTIONS combined
- IgM has a low AFFINITY antigen binding sites, but since there are 10 binding sites, the AVIDITY is high

48
Q

Neutralization

A

Bound antibody inactivates antigen

49
Q

Precipitation

A

Antigen-antibody complex can become insoluble

50
Q

Opsonization

A

Bound antibody stimulates phagocytosis

51
Q

Hemagglutination

A

Antibodies cross-link between RBCs, RBCs clump together

Primary testing method for RBC typing, antibody screen, ID and crossmatch

52
Q

Factors that affect Antigen-Antibody binding

A
  • Ionic strength (IgM > IgG)
  • pH (6.5-7.5 optimal)
  • Temperature
  • Number of epitopes
53
Q

Stages of hemagglutination

A

Stage 1: Sensitization - antibody attaches to epitope on RBC membrane

Stage 2: Lattice Formation (agglutination) - antibodies crosslink RBCs, shows visible clumping

54
Q

Direct Antiglobulin Test (Direct Coombs Test)

A

Detects non-agglutinating antibodies or complements coating RBCs in vivo

Antihuman globulin (AHG) reacts with human globulins bound to RBCs, causing agglutination of sensitized cells (IgG)

55
Q

Possible causes of in-vivo RBC sensitization

A

Autoimmune disease: Warm AIHA, CAD (cold autoimmune disease)

Alloimmune hemolytic anemia: HTN, HDFN

Drug induced immune hemolytic anemia

56
Q

WAIHA

A

IgG, C3

57
Q

CAD

A

C3 only

IgM

Specificity: anti-I

58
Q

Indirect Antiglobulin Test (IAT)

A

Detects non-agglutinating IgG antibodies in plasma (so due to previous exposure of foreign red cell antigen)

Used for antibody screens, antibody ID, crossmatch, and prenatal antibody titration

59
Q

Alternative Testing Methods

A

Gel System
- Utilizes microtubes with gel containing anti-IgG and Dextran microbeads
- High sensitivity, no need for washing, and automation capability
- Detects low antibody titers and provides precise results
- (+) = agglutination in the gel column at some level, (-) = cell button at bottom of column

Solid Phase Testing
- Red cell antigens coated on microtiter plate wells for antibody detection
- Indicator RBCs with AHG used for positive reactions.
- Differentiates positive and negative reactions efficiently
- (+) = diffuse adherence of indicator cells, (-) = solid button

60
Q
  • Clotted specimen
  • Blood from IV line with DEXTROSE
  • Septecemia/bacteria contaminated samples
A

False (+) DATs

61
Q
  • Presence of autoantibodies
  • Drug interaction
  • Increased serum protein (fibrinogen and globulins)
  • Over centrifugation
  • Positive DAT
A

False (+) IAT

62
Q
  • Inadequate cell washing, neutralizing anti-IgG
  • Low titer IgG - postzone
  • Deteriorating reagents
A

False (-) DAT and IAT

63
Q

Antihuman globulin (AHG)

A

Reacts with human globulins bound to RBCs, causing agglutination of sensitized cells

Used in both DAT and IAT to enhance agglutination of IgG

64
Q

Check cell

A

RBC coated with antibody and is used to verify validity of negative reaction

Basically double checks negative tests

65
Q

3 levels of genetics

A

Molecular genetics: biochemistry of gene and structures

Cellular genetics: cellular organization of genetic materials

Population genetics: genetic traits in large numbers of individuals

66
Q

DNA

A

4 nitrogenous bases: Adenine, Guanine, Cytosine, Thymine

5-carbon deoxyribose sugar
- 1 nitrous base bonded to number 1 carbon of deoxyribose
- phosphate group at 3 and 5 carbon of deoxyribose

Double stranded - antiparallel

67
Q

RNA

A

Transmits genetic info from nucleus to cytoplasm for protein synthesis

Single stranded

5-carbon ribose sugar

Uracil instead of Thymine

OH instead of H (DNA structure)

68
Q

4 types of RNA

A
  • Ribosomal (rRNA): cytoplasmic reticulum, translation
  • Messenger (mRNA): nucleus, transported out to cytoplasm for translation
  • Transfer (tRNA): transport amino acids to ribosome bound to mRNA during translation
  • Small nuclear (snRNA): splicing for modification of mRNA, regulatory, cell development, defense
69
Q

DNA transcription

A

One DNA strand is copied into mRNA for translation

Nucleus —> cytoplasm

  • 5’ end (7-methyl guanosyl cap): recognition site
  • Poly A polymerase: stabilizes mRNA
  • Intron: part of gene that does not code for protein, RNA slicing removes it
  • Extron: remaining sequence in mRNA
70
Q

Translation

A

RNA transcripts —> proteins using codons

Codons = code for amino acids

71
Q

Stop codons

A

UAA
UAG
UGA

72
Q

Start codon

A

AUG

73
Q

Silent mutation

A

Does not change protein, functions the same

74
Q

Transition mutation

A

One purine replaced with another purine (A and G), or pyrimidine with another pyrimidine

75
Q

Transversion mutation

A

purine substituted for pyrimidine (A with C) or vice versa

76
Q

Missense point mutation

A

Change in codon, alters amino acid

UGU - Cystine
CGU - Arginine

77
Q

Nonsense mutation

A

Point change in nucleotide of a DNA sequence, causing stop codon to form

Terminates reading of DNA sequence

Stop codons: UAA, UAG, UGA

78
Q

Insertion/Deletion (Frameshift mutation)

A

Changes the triplet codon sequence by altering the reading frame of the codon, results in large change in amino acid sequence downstream of the mutation

79
Q

Recombination/crossing over

A

Takes place in meiosis

From material and paternal homologous chromosomes —> new DNA sequence different from parent strains

80
Q

Chromatin

A

Macromolecule composed of nucleic acids and structural proteins (histones)

81
Q

Heterochromatin

A

Highly condensed region where transcription is not active

82
Q

Euchromatin

A

Region where transcription is more active

83
Q

Chromosome

A

Compressed/condensed coiled chromatin

In dividing cells

84
Q

Gamete cells

A

Sex cells

Haploids, half the number of chromosomes

85
Q

Gene

A

Section on the chromosome that encodes for a particular protein

86
Q

Locus

A

Specific location of a gene

87
Q

Allele

A

Alternative forms of the gene

Can be on the same chromosome or another homologous chromosome pair

88
Q

Mitosis

A

2 identical diploid daughter cells

Asexual reproduction

89
Q

Meiosis

A

4 unique haploid cells

Sexual reproduction

90
Q

Population Genetic

A

Principle of independent segregation and independent assortment

91
Q

Mendel’s first law of inheritance

A

Independent or random segregation

Each gene is passed onto the next generation

92
Q

Mendel’s second law of independent assortment

A

Genes for different traits are inherited separately from each other

Ex: flower color gene and seed shape gene are separate

93
Q

Linked Disquilibrium Phenomenon

A

Exception to Mendel’s second law of independent assortment

Linked genes do not assort independently

94
Q

Hardy Weinberg Principle

A

Formula that explains Mendel’s inheritance

p + q = 1

p = frequency of dominant allele
q = frequency of recessive allele

p^2 + 2pq + q^2 = 1

95
Q

Autosomal Dominance

A

Expressed when the allele is present

Homozygous or heterozygous

In males and females

96
Q

Autosomal Recessive Inheritance

A

Expressed when individual is homozygous for the allele and inherited recessive allele from both parents

97
Q

Polymerase Chain Reaction

A

DNA replicated in the test tube by cyclic changes in the reaction temperature

Denature: double helix into single strands
Anneal: binding of forward and reverse primer
Extension: DNA polymerase extends primer

98
Q

4 types of NGS (new generation sequencing)

A
  • Pyrosequencing: measures light released by pyrophosphate
  • Sequencing by synthesis: fluorescence labeled dideoxynucleotide
  • Sequencing by ligation: uses oligonucleotide probes (not DNA polymerase)
  • Ion semiconductor sequencing: measures release of H+

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