IMMUNOLOGY & SEROLOGY cells making them more susceptible to the action of

phagocytes)
IMMUNOLOGY – Study of the reactions of a host when foreign - Assists in the activation of the classical pathway of the
substances are introduced into the body. complement system.

Cardinal Signs of Inflammation

TYPES OF IMMUNITY a. Rubor – redness
1. Natural/Innate/Non-specific Immunity b. Calor – heat
- Ability of the individual to resist infection by means of c. Tumor – swelling
normally present body functions. d. Dolor – pain
2. Acquired/Adaptive/Specific Immunity e. Function laesa – loss of function
- Characterized by specificity for each individual pathogen
and the ability to remember a prior exposure, which result b. Interferon
in an increase response upon repeated exposure. - Glycoproteins produced by virally-infected cells

LINE OF DEFENSE
1. 1st Line of Defense – Skin and mucous membrane; secretions;
acidity of GIT and vagina; cilia lining the respiratory tract.
2. 2nd Line of Defense – Phagocytosis; inflammation and fever;
antimicrobial substances (complement, properdin, interferon,
tissue necrosis factor, beta-lysin)
3. 3rd Line of Defense – Specialized lymphocytes (T cells, B cells);
antibodies secreted by plasma cells
FIRST LINE OF DEFENSE
a. Physical/Anatomical/Mechanical Barriers
 Skin
 Mucous coating of epithelial and mucous cells
 Shredding off of cells that carry microbes
 Beating off of the cilia
 Physiologic actions (coughing, sneezing, vomiting,
diarrhea/defecation, urination, lacrimation, body temp,
oxygen tension)
b. Chemical Barriers/Biochemical Factors
 Acid pH
 Lysozyme
 Lactoferritin
 Lactoperoxidase
 Fibronectin
 Interleukins - Polypeptides that are secreted by antigen-
stimulated macrophages and lymphocytes that enhances T-
cell activation, proliferation, and activity during the
immune response.
 Interferons – Inhibits viral replication by inducing
neighboring cells to produce antiviral proteins that
interferes with the translation of viral mRNA.
SECOND LINE OF DEFENSE
a. Inflammatory response
C-Reactive Protein (CRP)
- Increased in inflammatory response such as infections,
burns, injuries and malignancies.
- Binds to surface of microorganisms allowing it to function
as an opsonin (a protein that bind to foreign particles and
Types:
 Alpha Interferon – Secreted by leukocytes and induced
by viruses or synthetic polynucleotides
 Beta Interferon – Secreted by fibroblasts and induced
by viruses or synthetic polynucleotides
 Gamma Interferon (Immune interferon) – Secreted by
lymphocytes following stimulation with specific
antigen
c. Phagocytosis
- Process of engulfment of foreign particles
Process:
 Initiation – Phagocytosis is initiated as a result of tissue
damage, either trauma or as a result of microorganism
multiplication.
 Chemotaxis – Migration to certain direction under
stimulation of a chemical substance.
*Positive – Towards site of stimulation
*Negative – Away from site of stimulation
 Engulfment
 Digestion
THIRD LINE OF DEFENSE
a. Active
- Antibody production is done by the body
 Natural – Infection
 Artificial – Vaccination
b. Passive
- Antibody production is not done by the body
 Natural – Transfer in-vivo-IgG (antibody); Colostrum-
IgA (Mother’s milk)
 Artificial – Administration of immune serum IgG –
injected
TYPES OF IMMUNOGLOBULINS
1. IgG
- Most predominant immunoglobulin in humans
- Function: Provides immunity for newborn; Fixation of the
complement; Opsonization; Neutralization of toxins and
viruses; Participation in agglutination and precipitation
reaction
ouens aipaq
2. IgM 4. Chemiluminescent Immunoassay
- Known as macroglobulin - The emission of light caused by a chemical reaction
- Most primitive producing an excited molecule that decays back to its
- Antibody most often formed in response to stimulus by original state.
gram negative bacteria
- Function: Complement fixation; Agglutination;
Opsonization; Neutralization of toxins and viruses SYPHILIS
3. IgA - Caused by Treponema pallidum subspecies (originally called
- Secretory component: Mucosal immunity; Prevents Spirochaeta pallida)
attachment of pathogen to mucosal surfaces; Produced by - “Great pox”, “Evil pox”, “French /Italian/Spanish Disease”
epithelial cells near IgA producing plasma cells - Transmitted by sexual contact, direct blood transmission or
4. IgD transplacental route
- Function: Immunoregulation; Second to appear on B cell
surface; Found on unstimulated but immunocompetent B Serologic Test
cells 1. Wasserman Test – Complement fixation test
5. IgE 2. Non-Treponemal Serologic Test – Non-specific ; Subject to
- “Reagin antibody” biologic false positive; Determine for the presence of
- Least abundant immunoglobulin in the serum Reagin (substance that appears in the serum of infected
- Binds strongly to receptor on mast cells and basophils and individuals that has the properties of an Ab); Based on
together with antigen, mediates the release of histamine Flocculation (special type of precipitation that involves fine
and heparin from these cells particles)
- Plays a role in allergic reactions c. VDRL (Venereal Disease Research Laboratories Test)
d. RPR (Rapid Plasma Reagin) – Charcoal agglutination
3. Treponemal Serologic Test – More specific
SUMMARY OF SOME TUMORS a. Treponema pallidum Immobilization Test (TPI)
1. AFP – Multiple Myeloma b. Fluorescent Treponemal Antibody Absorption Test
2. hCG – Nonseminomatous testicular cancer, Primary hepatoma (FTA-ABS)
3. PSA – Prostate cancer
4. CA-125 – Ovarian adenocarcinoma
5. CEA – Gastrointestinal tract HEPATITIS
6. CA 19-9 – Colonic and pancreatic adenocarcinoma 1. Primary Hepatitis Virus
7. CA 15-3 – Breast adenocarcinoma a. Hepatitis A Virus (HAV)
8. ALP – Lung cancer - Picornaviridae
9. Amylase – Pancreatic cancer - Fecal-oral
- “Infectious Hepatitis”
- HAV Ag:
SEROLOGY – Study of non-cellular portion of blood  Shed in feces of infected individuals during the
incubation period and early acute stage of
LABELED IMMUNOASSAYS infection, decline to low levels by the time
1. Radioimmunoassay (RIA) symptoms appear
- Uses radioactive substances - HAV Ab:
- Sample + (labelled analyte or antibody)  IgM Anti-HAV-marker of Acute Hepatitis A – Peak
- Immunoreaction with washing steps for separation during 1st month of illness and decline to
2. Enzyme Immunoassay (EIA) undetectable levels within 6-12 months;
- Uses enzymes as labels which can convert substrate Routinely detected by a solid-phase antibody
molecules to millions of products capture ELISA
- Very sensitive  IgG Anti-HAV – Produced as a result of natural
a. Enzyme-Linked Immunosorbent Assay (ELISA) - Specific infection or immunization; Indicates immunity to
antibody is coupled to particles such as dextran beads or HAV; Detected by competitive inhibition ELISA
to the bottom of plastic tubes forming a solid phase tests
immunosorbent.. b. Hepatitis B Virus (HBV)
b. Enzyme-Multiplied Immunoassay Technique (EMIT) – - Hepadnaviridae
The conjugation of enzymes to haptens does not disrupt - Sexual intercourse; Parenteral: blood, needles;
enzyme activity; however, the binding of hapten-specific Perinatal: mother to child
antibodies to haptens results in inhibition of enzyme - “Serum Hepatitis”
activity. - HBV Ag:
3. Fluorescent Immunoassay  Hepatitis B Surface Antigen (HBsAg) – “Australia
- Uses fluorescent compounds known as fluorophores or Ag” 1st marker to appear; Indicator of active
fluorochromes to detect antigen in tissue sections infection; Imp marker in screening blood donor

ouens aipaq
  Hepatitis B Envelope Antigen (HBeAg) – Present
during periods of active replication of the virus;
Indicates a high degree of infectivity when
present and high vertical transmission risk
(mother to child)
 Hepatitis B Core Antigen (HBcAg) – Note
detectable in serum because of the viral envelope
that masks it; Detected only through biopsy of the
infected liver; Not a serological marker
- HBV Ab:
 IgM Anti-HBc – Useful in detecting current
infection during the “core window period”
 IgG Anti-HBc – Persists for the lifetime of an
individual
 Anti-HBe – Indicates that the patient is recovering
from HBV infection
 Anti-HBs – Appears during the recovery period of
acute hepatitis B, weeks to months after the
HBsAg disappears; Persists for years and provide
protective immunity
c. Hepatitis C Virus (HCV) or Non-A/Non-B Hepatitis (NANB
Hepatitis)
- Flaviviridae (Hepacivirus)
- Sexual intercourse; Parenteral: blood, needles;
Perinatal: mother to child
d. Hepatitis D Virus (HDV)
- Incomplete virus (Viriod)
- Parenterally transmitted infection that can only occur
in the presence of Hepatitis B
- Occurs as superinfection or co-infection
e. Hepatitis E Virus (HEV)
- Calciviridae, reclassified to Hepeviridae
- Fecal-oral; Contaminated water
- Usually presents as an acute, self-limiting hepatitis
without progression to a chronic carrier state;
Associated with a high rate of mortality in pregnant
women
- HEV Ag:
 Detected in feces of most patients for about 2
weeks after the onset of illness, but may persist
longer in some cases; Identified by means of PCR
- HEV Ab:
 IgM Anti-HEV is typically present during the acute
infection but rapidly declines in the early recovery
period
- Agglutination Test – Gel or Latex particles are
used
- Dot-Blot Testing
b. Confirmatory Tests
- Western Bot Testing
- Immunofluorescence Assay
SYSTEMIC LUPUS ERYTHEMATOSUS (SLE)
- Chronic inflammatory autoimmune disease characterized by the
presence of antinuclear antibodies
- Symptoms may include swelling of the joint, erythematous rash
and deposition of immune complexes in the kidneys.
- Laboratory Diagnosis:
o Demonstration of LE cell (using buffy coat) – LE cell is a
neutrophil that has engulfed the antibody-coated nucleus
of another neutrophil
o Detection of antinuclear antibodies (ANA) – nonspecific
RHEUMATOID ARTHRITIS
- Autoimmune disease that affects the synovial membrane of
multiple joints
- Characterized by presence of an autoantibody called
Rheumatoid Factor (RF – IgM reacting against the Fc portion of
IgG)
C-REACTIVE PROTEIN
- A trace constituent of serum originally thought to be an antibody
to the c-polysaccharide of the pneumococci
- For inflammation or tissue destruction
Erythrocyte Sedimentation Rate (ESR) – For chronic inflammatory
conditions
INFECTIOUS MONONUCLEOSIS (MI)
- An infection usually caused by Epstein-Barr Virus. The Virus
spreads through saliva, thus it is called “kissing disease”.

2. Secondary Hepatitis Virus

a. Epstein-Barr Virus (EBV)
b. Cytomegalovirus (CMV)

HUMAN IMMUNODEFICIENCY VIRUS (HIV)

- Etiologic agent of the Acquired Immunodeficiency Syndrome
(AIDS)
- Laboratory testing for HIV infection:
o Antibody Detection
a. Screening Tests
- ELISA – Standard

ouens aipaq
IN LABORATORY ASPECT… B-HCG (Beta - Human Chorionic Gonadotropin)
- For hormone
Alegria – Uses spectrophotometry - Unit: mIU/mL
Centaur – Uses chemiluminescence - Female:
i-Chroma – Uses immunofluorescent assay - Male: Testicular cancer
Kits – Immunochromatographic test with lateral flow
VIDAS – Uses enzyme-linked fluorescent immunoassay which reads ASO (Anti-Streptolysin O)
the color of the sample in the strip by fluorescent light - For rheumatoid arthritis

TROPONIN I
HBsAg (Hepatitis B Surface Antigen) - Tumor marker for the heart
- Detects antigen - Significance: For cardiac arrest, myocardial infarction and
- Test: An in-vitro immunochromatographic, one step assay monitoring the heart
designed for qualitative determination of HBsAg in human serum
or plasma.
- Procedure: 100 uL plasma/serum (20 mins)
- Result: Positive or Negative

HBsAb (Hepatitis B Surface Antibody)

- Detects antibody
- Important in the follow-up of patients infected with HBV.
Important in monitoring the recipients of vaccination with
recombinant HBsAg and natural anti-HBs.
- Test: A chromatographic immunoassay for qualitative detection
of antibodies against HBV in plasma or serum.
- Procedure: 100 uL plasma/serum 920 mins)
- Results: Positive or Negative

DRT (Dengue Rapid Test)

- NS1 Ag – Non-structural protein; Antigen present in the kit
- IgM – Acute, Active, Current, Present infection
- IgG – Previous infection
- If a mosquito bites a human, dengue virus flows into the
bloodstream and replicates producing antigens. If with previous
infection, IgG will start to detect the familiar antigen and react
to it. If the infection is present/active, IgM will react to the
antigen. When the reaction takes place IgG is more visible in the
kit performed, IgM is present in the reaction but is not seen
because it will be masked by the IgG.
- Procedure: 3 drops serum/plasma to Ag and 10 uL
serum/plasma/whole blood to Ab with 4 drops of assay diluent
- Interpretation:
 NS1 Ag
*Negative – Presence of only 1 color line within the result
window
*Positive – Presence of 2 color lines
 IgG/IgM
*Negative – One line “C” in result window
*Postive
- IgM Positive – Primary dengue infection
- IgG Positive – Secondary dengue infection
- IgG & IgM Positive – Late primary or early secondary
dengue infection

CRP (C-Reactive Protein)

- For infection
- Unit: mg/L

ouens aipaq