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IMMUNOLOGY-SEROLOGY_______________________________________________________________

-demfestin,AHSE,RN,RMT

IMMUNOLOGY:
_____________________________________________________________________________________

 Study of the reactions of a host when foreign substances are introduced into the body.
 Study of all aspects of body defences, such as antigens and antibodies, allergy and
hypersensitivity.

NOBLE PRIZE WINNERS IN IMMUNOLOGY

YEAR SCIENTIST RESEARCH

1901 Emil von Behring Serum antitoxins
1905 Robert Koch Cellular immunity in TB
1908 Elie Metchnikoff Phagocytosis:
Paul Erlich Immunity
1913 Charles Richet Anaphylaxis
1919 Jules Bordet Complement
1930 Karl Landsteiner: Human blood group antigens
1960 Macfarlane Burnet, Peter Medawar Discovery of immunologic tolerance
1972 Gerald Edelman, Rodney Porter Structure of antibodies
1977 Rosalyn Yalow Radioimmunoassay
1980 George Snell, Jean Dausset, Baruj Benaceraf Major histocompatibility complex
1984 Niels Jerne Immunoregulation
George Koehler, Cesar Milstein Monoclonal antibody
1987 Susumu Tonegawa Antibody diversity
1991 Edward Donnall Thomas, Joseph Murray Transplantation
1996 Peter Doherty, Rolf Zinkemagel Cytotoxic T cell recognition of virally
infected cells
2008 Francoise Barre-Sinoussi, Luc Montagnier Human immunodeficiency virus

SIGNIFICANT MILESTONE IN IMMUNOLOGY

DATE SCIENTIST DISCOVERY

1798 Jenner Smallpox vaccination
1862 Haeckel Phagocytosis
1880-1881 Pasteur Live, attenuated chicken, cholera and
anthrax vaccines
1883-1905 Metchnikoff Cellular theory of immunity through
phagocytosis
1885 Pasteur Therapeutic vaccination
First report of live “attenuated” vaccine
for rabies
1890 Von Behring Humoral theory of immunity proposed
1891 Koch Demonstration of cutaneous delayed-type
I hypersensitivity
1900 Ehrlich Antibody formation theory
1902 Portier, Richet Immediate-hypersensitivity anaphylaxis
1903 Arthus Arthus reaction of intermediate
hypersensitivity
1938 Merrack Hypothesis of antigen-antibody binding
1949 Salk, Sabin Development of polio vaccine
1951 Reed Vaccine against yellow fever
1957 Burnet Clonal selection theory
1975 Kohler First monoclonal antibodies
2005 Frazer Development of human papillomavirus
vaccine

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HISTORICAL PERSPECTIVES (Stanley)

1798 Edward Jenner, an English countryside physician demonstrated that protection from
cowpox could be generated by the transfer of postural material from a cowpox lesion
instead of the more hazardous smallpox lesion.
1880 Louis Pasteur demonstrated that injection of killed microbes provided protection upon
subsequent exposure to live counterpart.
1888 Elie Metchnikoff demonstrated that certain blood cells ingest foreign materials
1894 Jules Bordet discovered complement
1897 Robert Kaus discovered precipitins
1901 Emil von Behring had the distinction of being awarded as the first immunology-related
Nobel Prize for his works on serum therapy
1984 Discovery of the T cell receptor gene
1987 Susumu Tonegawa was awarded the Noble Prize for his 1987 discovery of the genetic
principles underlying the generation of antibodies with different specificities

PROPERTIES OF NATURAL AND ACQUIRED IMMUNE SYSTEM

 Natural immunity is the ability of the individual to resist infection by means of normally present
body functions.
 Acquired immunity, in contrast, is a type of resistance that is characterized by specificity for
each individual pathogen and the ability to remember a prior exposure, which result in an
increase response upon repeated exposure.

NATURAL: ACQUIRED:

NATURAL ADAPTIVE
FIRST LINE OF DEFENSE SECOND LINE OF DEFENSE THIRD LINE OF DEFENSE

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COMPONENTS OF THE NATURAL IMMUNE SYSTEM

Cellular:

Humoral:

COMPONENTS OF ADAPTIVE SYSTEM

Cellular:

Humoral:

PHAGOCYTOSIS

1. Initiation

Phagocytosis is initiated as a result of tissue damage, either trauma or as a result of

microorganism multiplication.

Activated phagocyte has increased surface receptors that allow for adherence

a. _______________________________
b. _______________________________
c. _______________________________
2. Chemotaxis

Process by which cells tend to move in a certain direction under the stimulation of a chemical
substance.

a. Positive chemotaxis: __________________________________________________

b. Negative chemotaxis: _________________________________________________

Chemotactic substances: _________________________________________________

3. Engulfment

By active amoeboid motion

Final structure is known as phagosome or phagocytic vacuole

Opsonins (antibodies and complement components) interact with the surfaces of bacteria,
rendering them acceptable to the phagocyte.

4. Digestion

Minute cell particle that contain hydrolytic enzymes and peroxidase approach the phagosome,
fuse with it, rupture, and discharge contents to it.

Cells become degranulated as foreign materials are digested

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DESTRUCTION OF THE ANTIGEN

1. Formation of the phagolysosome leading to the release of lysosomal contents including

defensins, lactoferrin and lysozyme.
2. Nitric oxide produced by activated macrophages, which is toxic to microorganisms.
3. Activation of the NADPH oxidase, which is present in the phagosome membrane, leading to
the production of reactive oxygen intermediates, superoxide anion, hydrogen peroxide and
hydroxyl radicals, which are cytotoxic for microorganisms.

INFLAMMATION:

CARDINAL SIGNS

1. ______________________________
2. ______________________________
3. ______________________________
4. ______________________________
5. ______________________________

STAGES:

1. Vascular Response:

2. Cellular Response:

3. Resolution and Repair

Initiated by fibroblast proliferation, as a result:
a. Affected area may be totally repaired
b. In jury may lead to the formation of an abscess with some loss of function
c. Granuloma may be formed, typical of delayed hypersensitivity or cell-mediated
immunity.

Characteristics of Acute Phase Reactants (Steven 3 rd edition)

PROTEIN RESPONS NORMAL FUNCTION

E TIME CONCENTRATION (mg/dL)
(hr)
CRP 6-10 0.5 Opsonisation, complement
activation
Serum amyloid A 3.0 Removal of cholesterol
Alpha1-antitrypsin 24 200-400 Protease inhibitor
Fibrinogen 24 110-400 Clot formation
Haptoglobin 24 40-200 Binds hemoglobin
Ceruloplasmin 48-72 20-40 Binds copper, oxidizes iron
C3 48-72 60-140 Opsonisation, lysis
Mannose-binding protein ? 0.15-1.0 Complement activation

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INTERFERON (IFN)

Family of glycoproteins produced by all animal cells that exert a virus-nonspecific but host-specific
antiviral activity

TYPE I IFN: ALPHA AND BETA

 Non-immune, produced primarily in the initial innate response to viral infection.

IFN-ALPHA:

IFN-BETA:

TYPE II IFN: GAMMA

 Immune, primarily produced as a component of the specific immune response to viral and other
pathogens.

IFN-GAMMA:

TUMOR NECROSIS FACTOR (TNF):

TNF-ALPHA:

TNF-BETA:

COMPLEMENT:

 Series of proteins that are normally present in serum whose overall function is the mediation of
inflammation.

PROPERDIN

 Serum protein that exerts bactericidal and viricidal effects in the presence of _______ and
______.

BETALYSIN:

 Heat-stable cationic substances with bactericidal activity found in the serum of many animal
species including humans

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ACQUIRED/ADAPTIVE/SPECIFIC IMMUNITY

TYPE
ACTIVE NATURAL

ARTIFICIAL

PASSIVE NATURAL

ARTIFICIAL

LYMPHOID ORGANS

1. Primary Lymphoid Organs/:

a. Thymus :
b. Bone marrow:

2. Secondary Lymphoid Organs/:

a. Spleen
b. Lymph nodes
c. MALT
Peyer’s patches
Tonsils
Appendix

CELLS INVOLVED IN SPECIFIC IMMUNITY

T CELLS/:
Develop in the thymus
End products of activation are cytokines
Antigens include CD2, CD3, CD4 and CD8
Identified by rosette formation with sRBCs
Located in paracortical region of lymph nodes
B CELLS/:
Develop in the bone marrow
End product of activation is antibody
Antigens include CD19, CD20, CD21, CD40, MHC
class II
Identified by surface immunoglobulins
Located in cortical region of lymph nodes

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T LYMPHOCYTES

 Represent approximately 80% of the circulating lymphocytes in the peripheral blood

 Most circulating T cells express three of the following CD markers:
a. CD2: sheep red blood cell receptor, which is the classical T-cell surface marker
b. CD3: part of T-cell antigen-receptor complex
c. CD4: receptor for MHC class II molecule
d. CD8: receptor for MHC class I molecule

Helper-inducer T-cells: _____________

Suppressor-cytotoxic T-cells: ________

Normal ratio CD4+:CD8+ cells: _______

In HIV, ratio of CD4+:CD8+: __________

T CELL DEVELOPMENT

1. Double-negative thymocytes
_____________________________

2. Double-positive thymocytes

_____________________________

3. Mature T cell:

_____________________________

Activated T cell: ___________________________________

Sensitized T cell: ___________________________________

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B LYMPHOCYTES

Bone-marrow derived lymphocytes

Precursor cells in antibody production

B CELL DEVELOPMENT

1. Pro-B cell
______________________________
Rearrangements of genes on chromosome _________ coding for the __________.

2. Pre-B cell
______________________________
May have some surface immunoglobulin consisting of heavy chains with surrogate light chains
Rearrangement of genes coding for light chains
Kappa _____________ Lambda ____________

3. Immature B cell
Expression of IgM on the surface, CD21 and CD53
First Ig on the surface of B cell _______________

4. Mature B cell
IgD in expression to IgM on surface, and increased density of IgM
Cells are released from the marrow and seed in peripheral lymphoid organs

Second Ig on the surface of B cell _____________

5. Activated B cell
When activated by antigen CD25 appears, which is a receptor for interleukin-2, this enhances
proliferation

6. Plasma cell
Result of antigenic stimulation and transformation of activated B cells

Surface markers found on mature B cells disappear

Distinguished by the presence of abundant cytoplasmic immunoglobulin, which is excreted into
the bloodstream

SURFACE MARKERS ON T AND B CELLS

ANTIGEN CELL TYPE FUNCTION

CD2 Thymocytes, T cells, NK cells Involved in T cell activation
CD3 Thymocytes, T cells Associated with T cell antigen
receptor; role in TCR
transduction
CD4 Helper t cells, monocytes and Co-receptor for MHC class II,
macrophages receptor for HIV
CD5 Mature T cells, thymocytes, Positive or negative modulation
subset of B cells (B1) of T and B cell receptor
CD8 Thymocytes subsets, cytotoxic T Co-receptor for MHC class I
cells
CD10 B and T cell presursors, bone Protease; marker for pre-B
marrow stromal cells CALLA
CD16 Macrophages, NK cells, Low affinity Fc receptor;

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neutrophils mediates phagocytosis and

ADCC
CD19 B cells, follicular dendritic cells Part of B cell co-receptor, signal
transduction molecule that
regulates B cell development
and activation
CD21 B cells, follicular dendritic cells, Receptor for complement
subset of immature thymocytes component C3d; part of B cell
co-receptor with CD19
CD23 B cells, monocytes, follicular Regulation of IgE synthesis;
dendritic cells triggers release of GM-CSF from
monocytes
CD25 Activated T, B cells, monocytes Receptor for IL-2
CD44 Most leukocytes Adhesion molecule mediating
homing tom peripheral
lymphoid organs
CD45R Different forms on all Essential in T and B cell antigen-
hematopoietic cells stimulated activation
CD56 NK cells, subsets of T cells Not known
CD94 NK cells, subsets of T cells Subunit of NKG2-A complex
involved in inhibition of NK cell
cytotoxicity

LABORATORY IDENTIFICATION OF LYMPHOCYTES

1. Cell Flow Cytometry

Automated system for identifying cells based on scattering of light as cells flow in single file
through a laser beam

Forward LS: ______________________

Side LS: _________________________

2. Fluorescence microscopy
Both techniques rely on the use of labelled monoclonal antibodies against specific surface
antigens

3. Other methods: Rosette test:

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MAJOR HISTOCOMPATIBILITY COMPLEX

Genes that control expression of a large group of proteins

Regulate the immune response and play a role in graft rejection

Genes coding for the MHC molecules in humans are found in the short arm of chromosome
___________

CLASS I CLASS II CLASS III

GENETIC LOCI HLA HLA
A,B,C DP, DQ, DR
CHAIN STRUCTURE

CELL DISTRIBUTION

PRESENTS Ag To: To: Ex:

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IMMUNOGEN:

Substance that is capable of inducing an immune response

ANTIGEN

Substance with the ability to combine with an antibody

FACtORS AFFECTING IMMUNOGENECITY

1. Foreignness:
Autoantigen: _________________________________________________

Alloantigen: __________________________________________________

Heteroantigen: _______________________________________________

Heterophile antigens: ____________________________________

Graft

Autograft: ___________________________________________________________

Isograft/ ____________________________________________________________

Allograft: ____________________________________________________________

Heterograft/ __________________________________________________________

NOTE: TRANSPLANTATION IMMUNOLOGY

Examples of the Immunogenicity of Different Transplant Tissues

Most Immunogenic:

Bone marrow

Skin

Islets of Langerhans

Heart

Kidney

Liver

Bone

Xenogenic valve replacement

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Least Immunogenic: Cornea

Categories and Characteristics of Graft Rejection

Type Time of Tissue Damage Predominant Cause

Mechanism
Hyperacute w/in minutes humoral Preformed cytotoxic
abs to donor ags
Accelerated 2-5 days Cell-Med. Previous sensitization
to donor ags
Acute 7-21 days Cell-Med. Development of
allogenic reaction to
donor ags
Chronic >3 mos Cell-Med. Disturbance of
host/graft tolerance

2. Size
__________________________________________________

__________________________________________________

__________________________________________________

3. Chemical composition and complexity

__________________________________________________

__________________________________________________

__________________________________________________

4. Route, dosage and timing

Generally, intravenous and intraperitoneal routes are effective; the intradermal route offers
stronger stimulus than the subcutaneous or intramuscular route (although there are exemptions
to this).

Dose response may be partially dependent on the nature of immunogenic processing. Generally,
the smaller the dose, the less likely a response.

5. Adjuvants :
_________________________________________________

_________________________________________________

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ANTIBODIES

Substances produced in response to antigenic stimulation that as capable of specific interaction with
provoking immunogen.

GENERAL FUNCTIONS OF IMMUNOGLOBULINS

1. Neutralize toxic substances

2. Facilitate phagocytosis and kill microbes
3. Combine with antigens on cellular surfaces and thereby cause the destruction of these cells
either extravascular (outside of the blood vessels within the mononuclear-phagocyte system) or
intravascular (within the blood vessels through the action of complement).

THEORIES OF ANTIBODY DIVERSITY

1. Ehrlich’s Side-Chain Theory

Paul Ehrlich formulated the side –chain theory in the early 1900s. Ehrlich postulated that certain
cells had specific surface receptor for antigen that were present before contact with antigen
occurred. Once antigen was introduced, it would select the cell with the proper receptors,
combination would take place, and then receptors would break off and enter the circulation as
antibody molecules. New receptors would form in place of those broken off and enter the
circulation as antibody molecules.

2. Template Theory
Felix Haurowitz put forth a second major theory, the instructive or template theory in the early
1930s. According to this theory, antibody-producing cells are capable of synthesizing a
generalized type of antibody, and when contact with an antigen occur, the antigen serves as a
mold or template and alters CHON synthesis so that antibody with a specific fit is made. This
now-specific antibody enters the circulation, while the antigen remains behind to direct further
synthesis.

3. Clonal Selection
Niels Jerne and Macfarlane Burnet independently supported the idea of clonal selection
process for antibody formation. The key premise is that individual lymphocytes are genetically
preprogrammed to produce one type of immunoglobulin, and that a specific antigen finds or
selects those particular cells capable of responding to it, causing it to proliferate.

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PROPERTIES OF IMMUNOGLOBULINS

IgG IgM IgA IgD IgE

Structure

Percent of total
immunoglobulin

MW (Daltons)

Sedimentation
coefficient

Serum half-life
(days)

C’ fixation

Crosses
placenta

TYPES OF IMMUNOGLOBULINS

IgG

 Predominant immunoglobulin in humans

 Longest half-life of any immunoglobulin class

Subclasses: ______________________________________

IgG1 67% _________

IgG2 22% _________

IgG3 7% _________

IgG4 4% _________

Major function of IgG:

1. Providing immunity for the newborn

Crosses placenta except __________

2. Fixation of complement
Best: ________________
3. Opsonisation
4. Neutralization of toxins and viruses: _________________________________________________

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5. Participation in agglutination and precipitation reactions

IgM:

 Known as the macroglobulin

 Most primitive
 First to appear in phylogeny and the last to leave in senescence
 First to appear after a primary antigenic stimulus
 Antibody most often formed in response to stimulus by gram-negative bacteria

Functions of IgM:

1. Complement fixation
Most efficient in triggering the classical complement pathway because a single molecule can
initiate the reaction due to its multiple binding sites.
2. Agglutination
3. Opsonisation
4. Neutralization of toxins
Wasserman antibodies, heterophil antibodies, RF, cold agglutinins and allohemagglutinins
characteristically occur as IgM.

IgA

 Subclasses IgA1 and IgA2

 Primarily IgA2 is found as a dimer in secretions
 Secretory component: I.________________________________________________________
II. _______________________________________________________

IgD

 Function: _____________________________________________________
 Primarily a cell membrane immunoglobulin found on the surface of B lymphocytes in association
with IgM.
 Postulated to be an anti-idiotypic antibody (antibody to antibody) and as such may be involved
in a feedback mechanism to switch off B-cells

IgE

 Least abundant immunoglobulins in the serum

 Heat-labile antibody, originally called ________________________
 Binds strongly to a receptor on mast cells and basophils and together with antigen, mediates the
release of histamine and heparin from these cells.
 Mediates some types of hypersensitivity (allergic) reactions, allergies, and anaphylaxis and is
generally responsible for an individual’s immunity to invading parasites.

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COMPLEMENT

PROTEINS OF THE COMPLEMENT SYSTEM

CLASSICAL PATHWAY
C1q Binds to Fc of IgM and IgG
C1r Activates C1s
C1s Cleaves C4 and C2
C4 Part of C3 convertase
C2 Binds to C4b-form C3 convertase
C3 Key intermediate in all pathways
C5 Initiates membrane attack complex
C6 Binds to C5b in MAC
C7 Binds to C5bC6 in MAC
C8 Starts pore formation on membrane
C9 Polymerizes to cause cell lysis
ALTERNATE PATHWAY
Factor B Binds C3b to form C3 convertase
Factor D Cleaves factor B
Properdin Stabilizes C3 convertase
MBL
MBL Binds to mannose
MASP-1 Helps cleave C4 and C2
MASP-2 Cleaves C4 and C2

PLASMA COMPLEMENT REGULATION

C1 Inhibitor (C1INH) Dissociates C1r and C1s from C1q

Factor I
Factor H
C4-binding protein
S protein (vitronectin)
Decay Accelerating Factor
Cleaves C3b and C4b
Cofactor with I to inactivate C3b; prevents binding of B to C3b
Acts as a cofactor with I to inactivate C4b
Prevents attachments of the C5b67 complex to cell membranes
Accelerated association of C3 convertase

DEFICIENCIES OF COMPLEMENT COMPONENTS

DEFICIENT COMPONENT ASSOCIATED DISEASE

C1 (q, r,s) Lupuslike syndrome; recurrent infections
C2 Lupuslike synfrome; recurrent infections,
atherosclerosis
C3 Severe recurrent infections; glomerulonephritis
C4 Lupuslike syndrome
C5-C8 Neisseria infections
C9 No known disease association
C1INH Hereditary angioedema
DAF Paroxysmal nocturnal hemoglobninuria
MIRL Paroxysmal nocturnal hemoglobninuria
Factor H or factor I Recurrent pyogenic infection
MBL Pneumococcal diseases, sepsis, Neisseria
infections
Properdin Neisseria infections
MASP-2 Pneumococcal diseases

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HYPERSENSITIVITY
Heightened state of immune responsiveness

COMPARISON OF HYPERSENSITIVITY REACTIONS

Type I Type II Type III Type IV
Immune mediator

C’ involvement
(classical)

Effector cells

Immune Release of mediators Cytolysis due to Deposits of ag-ab Release of cytokines

mechanism mast cells and basophils ab and C’ complexes
Examples Anaphylaxis Transfusion Serum sickness Contact dermatitis
Hay fever reactions, AIHA, Arthus reaction Tuberculin test
Food allergies HDN SLE pneumonitis
Asthma
Bee sting
Penicillin drugs

SUMMARY OF TUMOR-ASSOCIATED ANTIGENS AND THEIR AREAS OF CLINICAL USE AND THE
ASSOCIATED TUMORS

ANTIGEN *USES ASSOCIATED TUMORS

Bence Jones Protein 1 Multiple Myeloma
Alpha Feto-Protein (AFP) 1,2,3 Nonseminomatous testicular cancer, primary hepatoma
Human Chorionic Gonadotropin (hCG) 1,2,3,4 Nonseminomatous testicular cancer. choriocarcinoma
Calcitonin 1,2,3 Familial medullary thyroid carcinoma
Prostate Specific Antigen (PSA) 1,4 Prostate cancer
CA-125 1,4 Ovarian adenocarcinoma
Cytokeratins 2 Sarcomata
HMB-45 2 Malignant melanoma
Neuron-specific enolase 2 Small cell cancers of the lung, endocrine tumors
S-100 2 Neuroendocrine tumors, melanoma
Beta-2-microglobulin 3 Lymphoma
Lactate dehydrogenase 3 Lymphoma
Carcinoembryonic antigen (CEA) 2,3,4,5 Tumors of the GIT, colorectal cancer
Estrogen receptor 3 Breast adenocarcinoma
CD45 2 Hematopoietic malignancies
IL-2 receptor (CD25) 4 T cell leukemia
Monoclobal Ig 1,2,3,4 Clonality indicates malignancy
CA-19.9 4 Colonic and pancreatic adenocarcinoma
CA-15.3 4 Breast adenocarcinoma
NRLU-10 (Ab name) 5 Small cell lung cancer
B-1 5 Lymphoma
CD20 6 Lymphoma
MAGE 6 Melanoma
*1= screening 4= monitoring
2= pathologic diagnosis 5= localization of metastasis
3= staging 6= therapy

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TUMOR MARKERS (from Dean Rodriguez Clinical Chemistry Review Handbook, 2012)
TUMOR MARKERS ASSOCIATED CANCER
AFP Hepatic and testicular cancers
ALP (placental-ALP) Lung cancer
Amylase Pancreatic cancer
BRCA-1 Breast or ovarian cancer
CA-125 Ovarian cancer (treatment and recurrence)
CA-15.3 Breast cancer (treatment and recurrence)
CA-19.9 Gastric, pancreatic and colorectal cancer
CA-50 Gastric and pancreatic cancers (treatment and recurrence)
CA-27.29 Breast cancer (treatment and recurrence)
Calcitonin Medullary thyroid cancer
Cathepsin-D Breast cancer
CEA Colorectal, stomach, breast, lung cancer (treatment and
recurrence)
CK-1 Small cell lung cancer, prostate cancer
Estrogen receptor Breast cancer
GGT hepatoma
HER-2/neu Breast cancer (efficiency of trastuzumab or Herceptin
therapy)
Nuclear matrix protein (NMP) Urinary bladder cancer

AUTOIMMUNE DISEASES

General Features of Some Autoimmune Disorders That Have Systemic, Multi-Organ, or Multi-Tissue
Involvement

Autoimmune Disease Ratio F/M HLA

Rheumatoid Arthritis 3:1 HLA-DR4

*Mixture of organ specific with systemic symptoms.

*Inflammed, swollen, and painful joints.
*RF-IgM to the Fc region of IgG.
*Peak age of onset is 40-50 years of age.

Systemic Lupus Erythematosus 10:1 HLA-DR2

HLA-DR3
*Systemic and multi-organ.
*Immune complexes are formed in serum
*immune complexes are trapped in basement membrane of glomeruli,
skin/endothelium, synovial jpints, and kidney.
*ANA, anti-ENA and antu-dsDNA may be present.
*Anti-phospolipid antibodies (lupus anticoagulant) are present.
Myasthenia Gravis 4:1 Complex
Age dependent
*Tissue sp (nerve and muscle)-with systemic affects.
*Neuromuscular transmission disorder
*Antibodies are inhibitory and block acetylcholine binding
*Some evidence of prior infection with poliovirus
Scleroderma 3:1 HLA-DR3 (weak)

*Primarily in skin/ectodermal tissues, but can cause multi-organ

pathology
*Skin fibroblasts reproduce faster and produce more collagen
Sjogren’s Syndrome 10:1 HLA-DR3

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*Clinically presents as dry eyes and mouth

*it can affect ant of the body’s glands that produce sweat, saliva or oil.

IMMUNOLOGICAL AND PATHOLOGICAL FEATURES OF SOME AUTOIMMUNE DISORDERS

DISORDER PATHOLOGY ANTIBODIES

Type 1 Diabetes
Rheumatoid Arthritis
Systemic Lupus Erythematosus
Grave’s Disease
Hashimoto’s Thyroiditis
Selective destruction of the
insulin producing B-cells of the
Islets of Langerhans in the
pancreas.
Chronic systemic inflammatory
disorder in which joint cartilage,
ligaments and tendons are
destroyed.
Immune complexes are formed
and these lodge in the
basement membrane of kidney,
skin and joints.
Unregulated secretion of T3 and
T4 due to stimulation of TSH
receptor by antibody
Destruction of the thyroid gland
Anti-glutamic acid decarboxylase (GAD)
antibodies (B islet cell antigen)
Anti-insulin antibodies
RF: IgM or IgG antibodies specific for the
Fc region of IgG.
ANA
Anti-ENA
Anti-dsDNA
Anti-phospholipid (lupus anticoagulant)
Anti-TSH receptor antibodies
Anti-thyroid peroxidase antibodies
(anti-microsomal antibodies)

Anti-thyroglobulin antibodies
Pernicious anemia Destruction of the parietal cell Anti-parietal cell antibodies
of the stomach mucosa leading Anti-intrinsic factor antibodies
to intrinsic factor deficiency

AFFINITY

Initial force of attraction that exists between a single Fab sites on an antibody molecule and a single
epitope or determinant site on the corresponding antigen.

1. ___________________________________
2. ___________________________________
3. ___________________________________
4. ___________________________________

AVIDITY

Sum of all attractive forces between an antigen and an antibody

_____________________________________________________

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SEROLOGY

Study of the noncellular portion of blood

Precipitation
Involves combination of soluble antigen with soluble antibody to produce insoluble complexes that are
visible.

Precipitation Reactions

I. Precipitation in a fluid medium

1. Turbidimetry:
2. Nephelometry:

II. Precipitation by Paasive Immunodiffusion

1. Radial Immunodiffusion
Antibody is uniformly distributed in support gel, and antigen is applied to a well cut into
the gel

A. Mancini/Endpoint Method:
Antigen is allowed to diffuse to completion and when equivalence is reached, there
is no further change in ring diameter.

______________________________

______________________________

B. Fahey and McKelvey/Kinetic Method

Measurements taken before the point of equivalence is reached

______________________________

______________________________

2. Ouchterlony Double Diffusion

Both antigen and antibody diffuse independently through a semisolid medium in two
dimensions.

A. Serological Identity:

B. Nonidentity:

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C. Partial identity:

III. Precipitation by Electrophoretic Techniques

1. Rocket Immunoelectrophoresis:

___________________________

___________________________

2. Immunoelectrophoresis

___________________________

___________________________

___________________________

___________________________

3. Immunofixation Electrophoresis

___________________________

___________________________

Agglutination

Process by which particulate antigens such as cells aggregate to form larger complexes when a specific
antibody is present.

READING OF AGGLUTINATION
Grade Description
Cells Supernate
0 No agglutinates Dark, turbid, homogenous
W+ Many tiny agglutinates Dark, turbid
Many free cells
May not be visible without microscope
1+ Many small agglutinates turbid
Many free cells
2+ Many medium-sized agglutinates Clear
Moderate number of free cells
3+ Several large agglutinates Clear
Few free cells
4+ One large, solid agglutinates Clear
No free cells

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Agglutination Reactions

1. Direct Agglutination
Antigens are found naturally on the surface of the particle

Hemagglutination

2. Passive Agglutination:
____________ is attached to the carrier particle
Agglutination occurs if patient ___________ is present

3. Reverse Passive Agglutination

____________ is attached to the carrier particle
Agglutination occurs if patient ________ is present.

4. Coagglutuination

Uses bacteria as the inert particles to which antibody are attached

__________________________ is most frequently used:

5. Agglutination Inhibition

Lack of agglutination is an indicator of a positive reaction

Classic example is the hCG test

6. Antiglobulin – Mediated Agglutination

Detects mon-agglutinating antibody by means of coupling with a second antibody.

A. Direct Antiglobulin Test

a. ________________________________

b. ________________________________

c. ________________________________

d. ________________________________

B. Indirect Antiglobulin Test

a. ________________________________

b. ________________________________

c. ________________________________

d. ________________________________

TYPES OF AHG REAGENTS

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a. Polyspecific AHG contains: _________________________________________________

b. Monospecific AHG contains: ________________________________________________

O CHECK CELLS

 Group O RBCs sensitized with IgG

 Added to negative antiglobulin tests to validate the negative reaction.
 Lack of agglutination after putting O CHECK CELLS means invalidated result.

FALSE – POSITIVE REACTIONS IN ANTIGLOBULIN TESTING

 Contamination of reagents
 Over centrifugation
 Direct agglutination by strong agglutinins
 Over-incubation with enzyme-treated cells
 Improper use of enhancement reagents
 Saline stored in glass or metal containers

FALSE – NEGATIVE REACTIONS IN ANTIGLOBULIN TESTING

 Reagent failure
 Improper washing
 Failure to add antiglobulin reagents
 Improper centrifugation
 Serum/cell ratio too low
 Delayed washing (elution of weakly attached antibody)

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