Laboratory Tests Flashcards
How does nephelometry determine amount of antibody?
Scatter light used to determine turbidity created by immune complexes - automated, method of choice
How does Radial immunodiffusion (RID) determine amount of antibody?
Antiserum mixed in gel with study Ig, look for ring of precipitation - time consuming
How does the Laurel Rocket determine amount of antibody?
Immunodiffusion + electrophoresis - takes minutes but insensitive, semiquantitative
What test can be used to detect both antibody and antigen excess?
ELISA
How are proteins separated for western blot?
electrophoresis
What method is used to confirm monoclonal gammopathy and heavy or light chain class type?
Immunofixation electrophoresis
At what age is the nadir for IgG, when maternal IgG is waning and child has not yet started producing IgG?
6 months
Which assay is used to detect antigen specific IgE antibody using chemiluminescent ELISA?
DPC Immunlite
What test is used to diagnose Lyme’s disease and which is used for confirmation
ELISA to screen, western blot to confirm
Which leukotrienes are used to detect basophil activation and how are they detected?
LTC4, LTD4, LTE4 detected with ELISA
Which marker on flow cytometry is used to determine basophil activation?
CD63
BAT high sensitivity, low specificity
What method is used to detect interleukins?
ELISA
What method is used to detect prostaglandins
radioimmunoassay (RIA)
What methods are used to detect presence of TLRs?
RT-PCR; flow cytometry (intracellular)
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
What method is used to detect bradykinin?
radioimmunoassay (RIA)
Where is CD1 found and what is its function?
APCs; presents bacterial lipid antigen to T cells
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
Where is CD3 found and what is its function?
T cells (NOT NK CELLS!); required for TCR signal transduction; defect causes SCID
What monoclonal antibody targets CD3 and is used in solid organ transplant and ALL?
OKT3
Where is CD14 (LPS receptor) found and what is its function?
macrophages and monocytes; PAMP - detects lipoteichoic acid, mediates IL-12 and IFNγ production
Where is CD16 found and what is its function?
NK cells, neutrophils; most common IgG receptor for phagocytosis
Where is CD18 found and what is its function?
neutrophils; adhesion and signaling
Where is CD19 found and what is its function?
B cells; coreceptor with CD21; B cell activation
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?
Bb
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