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CLS 790 > Quiz 2 - Autoimmunity & Serological Techniques > Flashcards

Quiz 2 - Autoimmunity & Serological Techniques Flashcards

1
Q

When autoimmunity is suspected, we perform this test…

A

Anti-nuclear antibodies (ANA)

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2
Q

Are men or women more predisposed to autoimmune disorders?

A

Women

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3
Q

When antigen-antibody complexes deposit in tissue, they attract…

A

Complement

  • rupture/lyse cells
  • autoimmune diseases cause tissue injuries
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4
Q

3 steps of process of disease

A
  1. Autoantibodies complex with patient’s own DNA
  2. Immune complexes deposited in vascular systems - organs & tissues
  3. Immune-mediated tissue injury due to complement
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5
Q

ANA vs Crithidia luciliae - which is the screening test and which is the confirmatory test?

A

ANA = screening

Crithidia luciliae = confirmatory

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6
Q

ANA test uses these cells as the substrate

A

HEP-2

  • epithelial cell line
  • ANA reacts with nuclear DNA/RNA
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7
Q

Why do different ANA patterns appear?

A

Due to different stages of nuclear DNA mitosis

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8
Q

How is the ANA test performed?

A

Patient sera placed on slide with HEP-2 cells and labeled with conjugate antibody with fluorescence tag

  • ie FITC
  • “sandwich” assay
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9
Q

The ANA test is an example of a “sandwich” assay. What is being sandwiched?

A

Patient’s antinuclear antibodies

-between HEP-2 DNA and conjugate antibody with fluorescence tag

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10
Q

An ANA test result showing a homogeneous pattern indicates this autoimmune disease

A

SLE = Systemic lupus erythematosus

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11
Q

An ANA test result showing a centromere pattern indicates this autoimmune disease

A

CREST syndrome

-form of scleroderma

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12
Q

An ANA test result showing a speckled pattern indicates this autoimmune disease

A

Sjogren syndrome

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13
Q

An ANA test result showing a nucleolar pattern indicates this autoimmune disease

A

Scleroderma

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14
Q

SLE - shows this ANA test pattern. What do the autoantibodies target (3)?

A

Homogeneous

anti-dsDNA
anti-ssDNA
Histones

-includes anti-Sm abs

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15
Q

Sjogren - shows this ANA test pattern. What do the autoantibodies target?

A

Speckled

anti-RNP (ribonuclear protein)

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16
Q

Scleroderma - shows this ANA test pattern. What do the autoantibodies target?

A

Nucleolar

anti-nucleolar

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17
Q

CREST - shows this ANA test pattern. What do the autoantibodies target?

A

Centromere

anti-centromere

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18
Q

After performing an ANA test for SLE, we confirm our results with this test. What does the confirmatory test target in the organism?

A

Crithidia luciliae

Kinetoplast

  • mitochondria rich in dsDNA
  • highly sensitive to HEP-2 substrate
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19
Q

The hereditary component sometimes involved in autoimmune disease involves the…

A

HLA

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20
Q

SLE - laboratory findings

A

Anemia - 50% leukopenia

Thrombocytopenia - 25-50%

Elevated erythrocyte sedimentation rate (ESR)

ANA positive - anti-dsDNA and anti-Sm. Targets nucleoprotein antigens

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21
Q

Overuse of this causes a SLE-like syndrome. The autoantibodies developed target…

A

Drugs

Histones
-milder SLE

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22
Q

SLE vs drug-induced SLE - what would the Crithidia luciliae test results be?

A

SLE = positive

Drug-induced SLE = negative
-absence of anti-dsDNA antibodies

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23
Q

SLE - clinical manifestations

A

Arthritis - chronic inflammation - skin, joints, kidneys, lungs, CNS

Butterfly rash across nose/upper cheeks

Raynaud’s syndrome

Renal disorders

Pulmonary manifestations

CNS disorders - depression, psychosis, convulsions

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24
Q

What is Raynaud’s syndrome. Which autoimmune disorders is it associated with?

A

Cyanosis on cold temperatures. Stress-induced vasoconstriction on extremities

SLE, CREST (the “R”)

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25
Q

Sjogren’s syndrome - 90% of autoantibodies are called…

A

Rheumatoid factor

-anti-RNP associated with ANA stain

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26
Q

Sjogren’s syndrome - laboratory findings

A

ANA - anti-La

Labial salivary gland biopsy for definite diagnosis

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27
Q

Dry mouth and dry eyes are symptoms associated with…

A

Sjogren’s syndrome

-affects lacrimal, salivary & excretory glands

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28
Q

Scleroderma has 2 forms

A

Progressive diffuse

Systemic = CREST

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29
Q

What are the acronyms of CREST?

A 
Calcinosis - bone formation
Raynaud - vasoconstriction of hands/feet
Esophageal involvement
Sclerodactyly - harden finger skin
Telangiectasia - spider veins
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30
Q

Scleroderma - lab findings

A

ANA - centromere pattern (CREST) or nucleolar pattern

31
Q

Insulin-dependent diabetes mellitus - insulin production is deficient due to…

A

Destruction of cells of pancreas

  • congential rubella
  • HLA
32
Q

This autoimmune disease has symptoms that overlap to SLE. How are lab results different from SLE?

A

Mixed connective tissue disease

Absence of multiple anti-SM and anti-dsDNA

33
Q

Ankylosing spondylitis - males or females more prone? What is a common symptom?

A

Males

Fibrosis of bone synovial capsule

34
Q

Rheumatoid arthritis - primarily affects…

A

Synovial joints

-swelling, morning stiffness, weight loss, fatigue

35
Q

Rheumatoid arthritis - lab findings

A

RF
-latex agglutination detects IgM RF

Circulating immune complexes

ANA - 14-28% of patients

36
Q

What antibodies are expected for a hepatitis infection?

A

Anti-smooth muscle

37
Q

What antibodies are expected for biliary cirrhosis (PBC)?

A

Anti-mitochondrial

38
Q

What antibodies are expected for Goodpasture’s?

A

Anti-glomerular basement membrane

-kidney’s glomerulus damaged

39
Q

What antibodies are expected for pernicious anemia?

A

Anti-parietal cells

40
Q

IgG subclasses - which ones are autoimmune and which ones are infectious antibodies?

A

IgG 1 & 3 = infectious

IgG 2 &4 = autoimmune

41
Q

Multiple sclerosis and myasthenia gravis are similar in that they both affect the…

A

Nervous system

  • MS - brain, spinal cord, myelin shealth
  • MG - neuromuscular
  • do an MRI scan
42
Q

Hashimoto’s thyroiditis vs Graves disease - both have what in common? How are they different

A

Enlarged thyroid

Hashimoto’s - hypothyroidism
-weight gain, bradycardia, slow metabolism

Graves - hyperthyroidism
-weight loss, tachycardia, fast metabolism

43
Q

Pernicious anemia - this is prevented from being absorbed… due to the lack of this…

A

Vitamin B12

Intrinsic factor

44
Q

Difference between infectious liver disease and non-infectious liver disease?

A

Non-infectious (autoimmune) = ANA towards smooth muscle proteins

Infectious = no ANA
-see anti-viral, anti-bacterial antibodies

45
Q

Autoimmune liver disease - lab findings

A

anti-liver antibodies:
anti-liver soluble protein (anti-LSP)
anti-liver membrane antibody (anti-LMA)
anti-acidoglycoprotein receptor (anti-ASCP)

No anti-dsDNA

46
Q

The highest peak on a normal SPE graph corresponds to…

A

Albumin

47
Q

Multiple myeloma patients show a large spike in the SPE corresponding to…

A

gamma-globulins

48
Q

What does IFE stand for? What is the test used to determine?

A

Immunofixation Electrophoresis

Determines what immunoglobulin abnormalities exists

49
Q

After band separation, IFE uses ___ to probe for immunoglobulins and visualize bands

A

Anti-antibodies

50
Q

Complement can be inactivated by ___

A

Heat

-56C for 30 mins

51
Q

The complement system is activated by…

A

Antibodies

52
Q

Name two assays to detect complement

A

ELISA

CH50

53
Q

The CH50 assay is used to detect hemolytic activity of… What does the CH50 value mean?

A

Complement

Amount of patient serum required to lyse 50% of a standard concentration of sheep RBC

54
Q

How is titer calculated from a dilution?

A

Titer is the reciprocal of the dilution

55
Q

Strep toxin can cause this substrate, ___, to change from blue to pink. What does strep toxin do? If patient has antibodies against strep toxin, what color will the test be?

A

Toluidine blue

Depolymerize DNA = pink color

Blue

  • toxin neutralized
  • no color change
56
Q

How is the dilution calculated from the dilution factor?

A

Reciprocal

-dilution factor 5 is 1:5 dilution or 1/5

57
Q

Antigen-antibodies can form 3 zones depending on their concentration. What are the 3 zones as it relates to the amounts of antibodies and antigen?

A

Prozone = too much antibody

Zone of equivalence

Postzone = too much antigen

58
Q

Precipitation test uses the concept of… using this media…

A

Immunodiffusion

Agar plate

59
Q

3 types of agglutination tests

A

Latex agglutination - mono, RA, rubella

Hemagglutination

Inhibition reactions

60
Q

Difference between turbidimetric and nephelometric

A

Turbidimetric = measure light absorbance by immune complex

Nephelometry = measure light scatter by immune complex

61
Q

Immunodiffusion assays have this weakness. However, they’re still useful for detecting ___ infections

A

Takes a long time to do

Fungal

62
Q

A double immunodiffusion assay is also called… What are the patterns that form?

A

Ouchterlony

Identity
Partial Identity
Non-identity

63
Q

Describe what is happening in each Ouchterlony assay reaction

A

Identity - both antigen/antibody and antigen/unknown have matching epitopes and will diffuse radially towards one another

Partial identity - share a few epitopes, but have more not in common. Forms small bifurcation/spur

Non-identity - precipitation lines cross, do not share epitopes

64
Q

Radial Immunodiffusion (RID) - describe assay. What does the result look like?

A

Antibody infused into agar. Inoculate patient sample into well and let diffuse out to zone of equivalence

Zone of equivalence = precipitation ring
-ring diameter proportional to concentration

65
Q

IEP vs IFE - which is more sensitive and produce sharper bands? How are they similar?

A

IFE

Both use electrophoretic immunodiffusion

66
Q

Terms for when Igs in a patient are lower than normal, higher than normal, and none

A

Hypogammaglobinemia

Hypergammaglobulinemia

Agammaglobulinemia

67
Q

Agglutination tests that use RBCs in the assay are called…

A

Hemagglutination tests

68
Q

Agglutination assays can be direct or indirect. What does each one mean?

A

Direct - antigen-antibody bind directly

Indirect - antigens are coated on a carrier molecule
-makes reaction more visible, easier to interpret

69
Q

Flocculation assay is a subset of this type of test… It is rarely used besides detection of this disease…

A

Agglutination
-precipitation of fine particles

Syphilis
-VDRL and RPR

70
Q

Describe the complement fixation test. Describe the positive and negative results

A

Complement is “fixed” with antigen-antibody so it cannot react and lyse RBCs
-uses sheep RBCs

Positive = antigen-antibody complex present, no lysis
Negative = antigen-antibody complex not present, lysis
71
Q

Immunoblot - difference between southern blot and western blot

A

Southern = DNA

Western = protein

72
Q

Define heterophile antibodies. An example of a heterophile antibody is known as…

A

Antibodies that react with antigens not responsible for their production = non-specific

Forssman antibody

  • IgM in nature
  • made against guinea pig kidney but reacts with sheep RBCs
73
Q

2 different tests to detect heterophile antibodies

A

Paul Bunnell heterophile antibody test

Davidsohn differential test

CLS 790 (13 decks)

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