Transcript Immuno assays

Principles of Immunology
by
S.S.Eghbali ,MD
5/10/1380
IMMUNE SYSTEM
Non
specific IMMUNITY
Specific
Immunity
Specific Immunity
Humoral
Cellular
Immunitiy
Immunity
Humoral Immunity
Cellular Immunity
T
CD4/TCD8 ratio
Cytokines IL-2, G-IFN, IL-4,IL-5
CTL
NK-Cells
CELL IMMUNITY IS AFFECTED
BY
AGE
Malnutrition
Cancers
Chronic
diseases
Immuno assays
Precipitation
Immunoassay
Particle Immunoassay
Radio Immunoassay
Enzyme Immunoassay
Chemiluminescent Immunoassay
Precipitation Immunoassay
Precipitation Immunoassay
A)Qualitative
Single Immunodiffusion
 Double Immuno diffusion
 Immuno electro phoresis
 Electro Immuno Fixation

B)Semi-Quantitative
Single radial
immunodiffusion
 Single dimension electro
immunodiffusion
(Rocket electrophoresis)

Particle Immunoassay
Hemaglutination
e.g
Treponema pal.
Gelatin particle agglutination
HBV,HCV,HIV
Latex agglutination HCG
Radio Immuno assay
Competitive
Non
RIA
competitive RIA
(Sandwich or IRMA)
Enzyme Immunoassay
Colorimetric
Fluorescent
Chemiluminscent
EIA
EIA
EIA
PRESENTATION OF
POSSIBLE IMMUNODEFICIENCY
* Frequent bacterial infection
* Unusually severe systemic reaction to a
virus
* Development of infection with an unusual
organism such as fungus or protozoan
* Systemic reaction following live virus
vaccination
* Family history of recurrent infections
* Exposure to the human immunodeficiency
virus.
BASIC SCANNING IMMUNOLOGY
STUDIES
Complete blood count / differential
Lymphocyte subpopulation analysis
(numbers and percentages of T and B
cells) by flow cytometry
Lymphocyte activation in vitro to
mitogens and microbial activators
Serum immunoglobulins, including
immunoglobulin subclasses if
evidence of clinical infections with
encapsulated bacteria. In some
cases, immunoglobulin levels are
normal but heterogeneous
nonbinding antibodies are
produced; thus, additional studies
are needed.
CONFIRMING AND FIRST-STAGE
ANALYTICAL STUDIES
* Radiograph for thymic shadow
* Skin test
* Natural Killer cell activity ( if child is 6
months or older)
* Cytokine production in response to
activation T-helper 1, T-helper 2 (IL-2,
interferon-y, IL-4, and so on)
* Mixed lymphocyte culture reaction with
patient as stimulator and patient as
responder
* Response to immunization
* Test for presence of ageappropriate specific antibodies
* Naturally occurring antibody
*Response to isohemagglutinins
(anti- A and –B blood group
substances) if patient has A, B, or O
blood type
* Test for adenosine deaminase and
purine nucleoside phosphorylase
enzyme deficiency
ANALYTICAL AND
IMMUNOREGULATORY STUDIES
* Development of activation antigens
during ersponse to stimulation,
such as Tac antigen, transferrin
receptor, up-regulation for MHC
class II on T cells, soluble
receptor, and so on
* Early activation response (e.g.,
calcium channels)
Immunoregulation
Response to IL-1, IL-2, interferons
Development of effector functions
Immunoglobulin synthesis in vitro
Cytotoxic T-cell activity
Suppressor cell/factor analysis
Gene activation, cell cycle analysis
Response to immunization: de novo
immunization