Laboratory Tests Flashcards by Jared Darveaux

How does nephelometry determine amount of antibody?

Scatter light used to determine turbidity created by immune complexes - automated, method of choice

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How does Radial immunodiffusion (RID) determine amount of antibody?

Antiserum mixed in gel with study Ig, look for ring of precipitation - time consuming

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How does the Laurel Rocket determine amount of antibody?

Immunodiffusion + electrophoresis - takes minutes but insensitive, semiquantitative

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What test can be used to detect both antibody and antigen excess?

ELISA

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How are proteins separated for western blot?

electrophoresis

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What method is used to confirm monoclonal gammopathy and heavy or light chain class type?

Immunofixation electrophoresis

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At what age is the nadir for IgG, when maternal IgG is waning and child has not yet started producing IgG?

6 months

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Which assay is used to detect antigen specific IgE antibody using chemiluminescent ELISA?

DPC Immunlite

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What test is used to diagnose Lyme’s disease and which is used for confirmation

ELISA to screen, western blot to confirm

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Which leukotrienes are used to detect basophil activation and how are they detected?

LTC4, LTD4, LTE4 detected with ELISA

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Which marker on flow cytometry is used to determine basophil activation?

CD63

BAT high sensitivity, low specificity

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What method is used to detect interleukins?

ELISA

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What method is used to detect prostaglandins

radioimmunoassay (RIA)

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What methods are used to detect presence of TLRs?

RT-PCR; flow cytometry (intracellular)

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What method is used to determine TLR function?

incubation with panel of ligands for TLR1-9, measure release of inflammatory molecules with ELISA, Flow, or PCR

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What method is used to detect bradykinin?

radioimmunoassay (RIA)

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Where is CD1 found and what is its function?

APCs; presents bacterial lipid antigen to T cells

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Where is CD2 found and what is its function?

NK and early T cells; activates T cells, mediates adhesion between T cell and APC, inhibits apoptosis

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Where is CD3 found and what is its function?

T cells (NOT NK CELLS!); required for TCR signal transduction; defect causes SCID

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What monoclonal antibody targets CD3 and is used in solid organ transplant and ALL?

OKT3

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Where is CD14 (LPS receptor) found and what is its function?

macrophages and monocytes; PAMP - detects lipoteichoic acid, mediates IL-12 and IFNγ production

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Where is CD16 found and what is its function?

NK cells, neutrophils; most common IgG receptor for phagocytosis

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Where is CD18 found and what is its function?

neutrophils; adhesion and signaling

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Where is CD19 found and what is its function?

B cells; coreceptor with CD21; B cell activation

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Where is CD20 found and what is its function?

B cells; transmembrane receptor needed for B cell activation

Where is CD21 found and what is its function?

Mature B cells; binds to CD23 and viruses

Where is CD22 found and what is its function?

B cells; inhibits B cell signialing

Where is CD23 found and what is its function?

mature B cells; B cell activation

Where is CD25 (IL-2 Rα) found and what is its function?

Activated B and T cells; suppression of NK cells & self-reactive T cells - elevated in HLH

Where is CD27 found and what is its function?

memory B cells; marker of B cell activation (memory B cells are CD27+ and memory T cells are CD27-)

Where is CD32 found and what is its function?

WBCs, binding of FcγRII proposed mechanism for IVIG

Where is CD35 found and what is its function?

WBCs; binds immune complexes coated with C3b and C4b

Where is CD40 found and what is its function?

APCs; T cell dependent Ig switching of B cells - defective in hyper IgM3

Where is CD45 found and what is its function?

naive, memory or activated T cells; defective in T-B+NK- SCID

Which cell marker is the receptor for adenovirus?

CD46

Which cell marker is the target of alemtuzumab used to treat CLL?

CD52

Which cell marker is the receptor for rhinovirus?

CD54 (ICAM-1) and LDL receptor

Which cellular marker is deficient in paroxysmal nocturnal hemoglobinuria?

CD55

Which cellular markers are defective in LAD2 and where are they located?

E-selectin on endothelium; P-selectin on platelets

Which cellular marker is used for lymphocyte homing to HEV of lymph nodes?

L-selectin

What is the function of CD64?

HIGH affinity receptor for IgG needed in ADCC

What is the CD marker for CD40L and where is it found?

CD154 found on T cells - defective in XHIGM

Which cellular marker is defective in ALPS?

CD95 (Fas)

What is the CD marker also called FasL?

CD178

CD28 binds to what for costimulation during T cell activation?

CD80 (B71) and CD86 (B72)

An activating mutation in CXCR4 causes what?

phagocytic defect, leads to WHIM - wart, hypogam, infection and myelokathexis

CCR5 and CXCR4 are coreceptors for which virus?

HIV

This is the most potent complement receptor and binds iC3B and ICAM-1

CR3

Which PID is associated with defective CD40 and which has defective CD40L?

hyper IgM3; x-linked hyper IgM

Chemokine receptor CCR3 is implicated in allergic disease and binds to which ligands?

CCL11 (eotaxin-1), CCL26 (eotaxin-3), CCL5 (RANTES -Regulated on Activation, Normal T Expressed and Secreted)

This chemokine receptor functions in cell trafficking, is a coreceptor for HIV, and binds CCL5 (RANTES -Regulated on Activation, Normal T Expressed and Secreted)

CCR5

This chemokine receptor functions in T cell trafficking and binds to CCL17 (TARC).

CCR4

This chemokine receptor functions in naive T cell and DC trafficking to lymph nodes and binds to CCL21 (SLC)

CCR7

This chemokine receptor functions in B cell development and is an HIV coreceptor

CXCR4

This cell marker on T cells mediates homing to skin

CLA-1

Which surface marker is present on memory B cells and absent on memory T cells?

CD27

This cell marker on T cells mediates homing to colon

A4β7

Which surface marker would differentiate a memory T cell from a Naive T cell

CD45RO

Which B cell surface marker is inhibitory?

CD22

Which surface marker would indentify a mast cell?

CD117 (c-KIT)

What is the difference in function between conventional and plasmacytoid dendritic cells?

conventional are APCs, plasmacytoid produce IFNα

A CD3+ CD25+ cell on flow would indicate what?

T reg or activated T cell

How is a lymphocyte proliferation assay stimulation index performed?

Incubate with mitogen and radioactive tritiated thymidine - this is incorporated into dividing cells' DNA, then measure radioactivity

How is flow based T lymphocyte proliferation assay performed?

PBMCs labeled with fluorescent dye, stimulate cells with mitogen, measure fluorescence

During what part of B cell development is CD20 present?

pre-B and B cell blasts

What is the mutation in X-linked agammaglobulinemia?

Bruton's tyrosine kinase

What abnormality is seen on flow cytometry in X-linked agammaglobulinemia?

absence of CD19 B cells

What type of B cell is CD27- and IgD+?

Naive B cell

What type of B cell is CD27 + and IgD+?

Activated, unswitched B cell

What type of B cell is CD27+ and IgD-?

Memory B cell

Which mitogens stiumlate T cells?

Pokeweed, phytohemagglutinin, concanavalin A

Which mitogens stimulate B cells?

Pokeweed, LPS, SAC

What is measured in determining NK cell function?

Perforin, Granzyme, CD107a expression

In patients with hemophocytic lymphohistiocytosis (HLH), expression of what is found to be deficient on flow?

perforin, granzyme

Which surface markers are not present in LAD1?

CD18 (part of Mac complex with CD11b)

What is characterized by neutropenia, recurrent skin, sinus, pulm infxns, oculocutaneous albinism, MR and neuropathy?

Chediak-Higashi syndrome

Chediak-Higashi syndrome is due to a mutation in what?

LYST 1q42

Myeloperoxidase deficiency is usually asymptomatic. What types recurrent infections are reported in diabetic?

candida of skin bones and blood

Mutations in the NADPH oxidase system leads to what?

CGD

The NBT reduction tests assesses neutrophil function. How is this accomplished?

Neutrophils mixed with yellow dye. Normal oxidative burst will reduce dye, making it blue. Qualitative measure. NOT THE GOLD STANDARD

The DHR test is a quantitative measure of neutrophil function, How does it work?

nonfluorescent dye taken up by neutrophils, converted to fluorescent compound by NADPH, assessed by flow

What is the advantage of the DHR test over other assessments of neutrophil function?

It can distinguish between X-linked, autosomal and x-linked carriers for CGD

A DHR assay with a low, broad peak would indicate what?

AUTOSOMAL gp-47 (phox) deficient CGD

A DHR assay with almost no response would indicate what type of defect?

X-LINKED gp-91 (phox) deficient CGD

Monoclonal antibodies ending in -omab are derived from what?

mouse

Monoclonal antibodies ending in -ximab are derived from what?

chimera - combines murine mAb variable region with constant region of human Ab

Monoclonal antibodies ending in -zumab are derived from what?

humanized

Monoclonal antibodies ending in -umab are derived from what?

human

Monoclonal antibodies ending in -cept are derived from what?

fusion proteins - created by joining 2 or more genes that originally coded for separate proteins

When IgG or IgM bind to normal human cells, what inhibits immune complex formation and how?

Factor H and I inhibit conversion of C3b to iC3b

What determines the size of an immune complex?

the degree of antigen or antibody excess

What can be measured to monitor disease burden from immune complexes in SLE?

C3 and C4 (complement consumption)

What is the most common cause of a low complement level?

poorly handled specimen

AH50 assesses what?

alternative complement pathway

CH50 assesses what?

classical complement pathway

How is the MBL complement pathway assessed?

ELISA - incubate serum with mannan, measure C4b and C4d

What is used as a marker for activation of the classical or lectin complement pathway?

C4a and C4d

What is a marker of activation of the alternative complement pathway?

Which type of angioedema has low C1q levels?

acquired angioedema

Which type of angioedema has normal or elevated levels of C1 inhibitor?

HAE type II

C1q autoantibody is present in which type of vasculitis?

hypocomplementemic

What complement defect would be expected in an individual with recurrent neisseria infections?

Terminal complement components (C5-9)

Mutations in genes coding for what have been associated with type III HAE?

Factor 12 (Hageman Factor) - usually gain of function

How are TRECs measured?

PCR

What happens to the amount of TRECs as someone ages?

declines

What type of infection is associated with decreases in TRECs?

HIV-1

Reverse transcriptase (RT) PCR is used to detect what?

mRNA expression levels

When is RT-PCR used?

to detect viruses with genomes composed of RNA such as HIV and influenza A

What type of assay would you use to fund specific features of DNA for use in genetic counseling, medicine and species identification?

FISH

What lab technique is used to diagnose ALL, DiGeorge and Down syndrome?

FISH

What are gene chips used for?

sequencing large numbers of genes to identify SNPs