LEVEL 3 – SEMESTER 5

Lab Medicine
Lecture 9
Part 2

Dr / Y.M
Lab diagnosis of autoimmune diseases & immunodeficiency
disorders
Immunodeficiency:
defined as deficiency of one or more of the 4 major components of the immune system
(B lymphocyte, T lymphocyte, phagocytic cells, complement system) that protect the individual against different viral,
bacterial, fungal & protozoal infection.
Classification of immune deficiency disease :
 Primary immunodeficiency disease
 Secondary immunodeficiency disease

Primary immunodeficiency disease

B cell immunodeficiency disorders
 Bruton s x linked agammaglobulinemia
 Common variable ID (acquired
dysgammaglobulinemia)
 Transient hypogammaglobulinemia in infancy
 Selective IgA deficiency
 Selective IgM deficiency
 Selective IgG subclasses deficiency
 Immunoglobulin deficiency with IgM
T cell immunodeficiency disorders
DiGeorge syndrome
(congenital thymic aplasia)
1. Absence or hypoplasia of both
thymus and parathyroid glands
2. Few or no circulating T cells
3. Impaired T cell function
4. Levels of circulating Igs are low,
at least to certain antigen
5. Hypocalcemia, abnormal facial
appearance, cardiac anomalies
Chronic muco-cutaneous candidiasis
1. Impaired Tcell response to
C.albicans antigen
2. Normal Tcell response toward
other antigen
3. Normal antibody response to
C.albicans

Combined B&T cell
immunodeficiency disorders
 Sever combined
immunodeficiency disease
 Wiskott-Aldrich syndrome
 Ataxia- telangiectasia
 Nezolf s syndrome
 Reticular dysgenesis
Primary immunodeficiency disease
Phagocytic dysfunction Complement deficiency
Extrinsic defect Intrinsic defect  Defects in complement component:
Secondary to Due to enzymatic  Decrease in C1,C2,C4 cause
 decrease of the deficiencies within significant pathology in
number of phagocytic the metabolic autoimmune disease
cells pathway necessary  Decrease in C3 is associated with
 decrease of the for killing bacteria pyogenic infection
number of circulating  Defects in membrane-attack
neutrophils components (C5-C9) result in
 decrease of phagocytic susceptibility to neisseria species
function
 deficiency of antibody
and  Defects in complement control
complement(opsonins) proteins:
 Chronic granulomatous disease  Deficiency in C1 esterase inhibitor
 Myeloperoxidase deficiency lead to hereditary angioedema
 Glucose-6-phosphate deficiency syndrome  Deficiency in decay-accelerating
 Chediak-Higashi syndrome factor (DAF) & CD59 results in
 Leukocyte adhesion deficiency syndrome paroxysmal nocturnal
Most of the known defects in phagocytic hemoglobinuria
cells affect their ability to kill intracellular
and/or ingested extra-cellular bacteria
 Laboratory assessment of immune competence :
1. Protein electrophoresis
2. Quantitation of immunoglobulins : using radial immunodiffusion ,nephlometry or
ELIZA
3. IgG subclasses levels : using ELIZA technique
I- Evaluation of antibody
4. B cell quantitation using flow cytometry,
(B cell) mediated
normally: B cells represent 10-20% of total circulating lymphocyte
immunity
5. In vivo specific antibody response : immunization with tetanus or pneumococcal
polysaccharide antigens
6. In vitro stimulating of B cells : by mitogen or by antigen and assessment of
immunoglobulin release
1. Total lymphocyte count : normally at any age >1200/ul
II- Evaluation of cellular 2. Total T cell count : normally T cell represent >60% of total circulating lymphocytes
(T cell) mediated 3. Monoclonal antibodies to T cell subset : using flow cytometre
immunity 4. Delayed cutaneous hypersensitivity skin test: using PPD
5. In vitro lymphocyte response to mitogen , antigen
1. Nitro-blue tetrazolium test:
used for diagnosis and screening of chronic granulomatous disease(CGD)
2. Chemiluminescence: abmormal in CGD and myeloperoxidase deficiency
3. Chemotaxis :
 measure the ability of neutrophils to move in a directed , migratory pattern toward a
III- Assessment of chemotactic substance
phagocytic function  it is abnormal in various disorders, however it is not specific
4. assessment of Superoxide production : absent in CGD and defective in other syndromes
 1. Functional (hemolytic assay–CH50)
2. Immunoassay:
for assay of C3 & C4 levels using single radial diffusion , electro-immunodiffusion , rate
nephlomery and ELISA
3. Enzyme tests : e.g. G.6.ph-dehydrogenase & myeloperoxidase
IV- Complement assay
4. membrane glycoproteins :
Deficient in leukocyte adhesion molecule deficiency (LAD) disorders and associated with
abnormal leukocyte adherence and movement
5. genetic analysis :
available for CGD , LAD , and Chediak-Higashi syndrome