Basic Immunologic Procedures
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1
Examples of active immunotherapy vaccines include
Cancer vaccines e.g. HPV, HBV
2
Cytokines classified as active immunotherapy examples include
HGFs/CSFs (G-CSF, GM-CSF, EPO, IL-11), IFN-α, IL-2
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Monoclonal antibodies given as examples of passive immunotherapy include
Rituximab, Trastuzumab
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Adoptive immunotherapy examples utilizing immune cells include
Tumor-infiltrating lymphocytes (TILs), Genetically engineered T cells e.g. chimeric antigen receptors (CAR T cells)
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Force of attraction between a single Fab site on an antibody molecule and a single epitope on the corresponding antigen is defined as
Affinity
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Overall strength of antigen-antibody binding representing total binding strength is defined as
Avidity
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Ability of a particular antibody to combine with one antigen instead of another is defined as
Specificity
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Phenomenon occurring if two different antigens share an identical or very similar epitope is defined as
Cross-reactivity
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Primary level of antigen-antibody interaction involves
Combination of antigen and antibody (sensitization)
10
Comment and test requirements associated with primary level interactions are
Requires use of labels and special equipment; most sensitive and specific
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Test examples categorized under primary level of interaction include
Labeled immunoassay (RIA, EIA, FIA, CLIA, CLIA)
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Secondary level of antigen-antibody interaction involves
Lattice formation and other demonstrable consequences of ag-ab reaction
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Comment regarding sensitivity and specificity of secondary level interactions is
Easily detectable; less sensitive and specific
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Test examples categorized under secondary level of interaction include
Precipitation, agglutination, complement fixation, neutralization
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Tertiary level of antigen-antibody interaction involves
Immunologically active in vivo reactions e.g. inflammation, chemotaxis, phagocytosis, immune complex deposition
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Comment and example test associated with tertiary level interactions are
Detected by the effect the reaction has on tissues or cells; least sensitive and specific; e.g. Skin test
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Temperature and duration parameters for serum preservation via refrigeration are
2-8°C for 72 hours
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Temperature parameter for serum preservation via freezing is
-20°C for longer period
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Temperature and time required for heat inactivation of complement are
56°C for 30 mins
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Temperature and time required for reinactivation of complement if tested after 4 hours are
56°C for 10 mins
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Expression definition of simple dilution is
Volume of serum divided by total volume of solution
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Definition of compound dilution is
Involves two or more steps
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Definition of serial dilution is
Compound dilution in which the dilution factor is the same in each step; used to obtain a titer
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Definition of antibody titer is
Reciprocal of the highest serum dilution that gives a positive reaction with the antigen
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Titer increase between acute and convalescent sera indicative of a current infection is
Fourfold rise (two doubling dilutions)
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Antibody titer result given agglutination positives at 1:10, 1:20, 1:40, 1:80 and negative at 1:160 is
Titer = 80
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Zone of antibody excess where antibody not bound to antigen can be detected in the supernatant is
Prozone
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Zone of antigen excess where antigen not bound to antibody can be detected in the supernatant is
Postzone
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Zone of optimal antigen-antibody ratio where large multimolecular lattice is formed is
Zone of equivalence
30
Measurement principle of turbidimetry is
Measures the reduction in light intensity due to particles in a solution
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Measurement principle of nephelometry is
Measures the amount of light scattered at a particular angle from the incident beam as it passes through a suspension
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Diffusion setup for the Oudin test is
Single diffusion in a single dimension; antibody is incorporated in agar gel in a test tube and antigen solution is layered over it
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Diffusion setup for Oakley and Fulthorpe is
Double diffusion in a single dimension; antigen and antibody move towards each other in a tube through an intervening column of plain agar
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Plate setup and diffusion type used in Radial Immunodiffusion (RID) are
Single diffusion in two dimensions using a plate containing agar with known antibody
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Measurement timing and relationship for the Fahey method in RID are
Kinetic method; read at 18 hours; diameter of precipitin ring is proportional to log of ag conc
36
Measurement timing and relationship for the Mancini method in RID are
End-point method; read after equivalence (24-48 hours); square of diameter is proportional to ag conc
37
Diffusion setup for the Ouchterlony technique is
Double diffusion in two dimensions; both antigen and antibody diffuse horizontally and vertically through a medium
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Precipitin line pattern representing identity in Ouchterlony technique is
Smooth continuous arc (antigen in the sample is the same as known antigen)
39
Precipitin line pattern representing partial identity in Ouchterlony technique is
Spur formation (antigen in the sample has some similarities with known antigen)
40
Precipitin line pattern representing non-identity in Ouchterlony technique is
Crossed lines (antigen in the sample is different from known antigen)
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