LAB # 3  End product: Destruction/Killing of Foreign

cells.
SCREENING TEST FOR  Neutrophils: are capable of moving from the
circulating blood to the tissue through a
PHAGOCYTIC process known as diapedesis.
ENGULFMENT  This begins with margination and
Immunity adherence to the vessel wall
 All those physiological mechanisms that (endothelial)
endow the animal with the capacity to  Then neutrophils forms pseudopods
recognize materials as foreign to itself and to (fake feet), which squeeze through
neutralize, eliminate or metabolize them with junctions of the endothelial cells.
or without injury to its own tissues.  They may wander randomly through
 The immune system is structured to the tissue OR attracted to a specific
recognize, respond to, and destroy a wide area by chemotactic factors.
variety of invading organism that would (HEMOTAXIS)
otherwise be capable of promoting  Chemotaxins: are chemical messengers that
infections, harmful to the body.
causes migration of cells in a particular
TYPES OF IMMUNITY direction.
 NATURAL IMMUNITY  Factors that are chemotactic for neutrophils
 Innate/Non-specific includes complement components, proteins
 ability of an individual to resist from the coagulation cascade, products from
infections by means of normally bacteria and viruses, and secretions from
present body functions. mast cells, lymphocytes, macrophages and
 Standardized response to all antigen other neutrophils.
 Lacks memory
 Present at birth STEPS IN PHAGOCYTOSIS
 ACQUIRED IMMUNITY 1. Physical contact between the WBC and the
 Adaptive/Specific foreign particle
 type of resistance that is characterized  Attachment/Initiation – enhanced by
by specificity for each individual Opsoin (CRP, C3B, C4B, C5B, Ig) in
pathogen, or microbial agent. which serum proteins attached to cell
 Diverse response for phagocytosis
 Has memory cell
 Not present at birth
2. Formation of phagosome
 Cytoplasm overflow
st
1 LINE OF DEFENSE (NATURAL) 3. Fusion with cytoplasmic granule to form
- Skin, hair, secretions phagolysosome
nd
2 LINE OF DEFENSE (NATURAL)  Release enzyme that will further be
- Phagocytosis released to outside
rd
3 LINE OF DEFENSE (ACQUIRED)
- After natural immunity 4. Digestion and release of debris to the outside
(exocytosis)
Phagocytosis Respiratory burst
 A process of engulfment of foreign cells or - Increase in oxygen consumption
foreign materials by a phagocytic cells. - Within cell as pseudopodia enclosed the
 Neutrophil – PB particle with vacuole = Phagosome
 Macrophage – Tissues
 Monocyte – PB TYPES OF PHAGOCYTOSIS
Monocyte patrols  find foreign
 Indirect
substance  if none, leaves peripheral
blood  goes to tissues   Via opsonin receptors that recognize
Macrophage opsonins such as IgG, CRP and C3b
bound to microorganisms.
  Direct  NEGATIVE – No engulfment of bacteria
 Via Pattern Recognition Receptors
that recognize lipid and carbohydrate **Non-specific
sequences on microorganisms (Toll- **Only attachment of bacteria to WBC is seen; cell
like receptor/TLR); faster damage or breakdown cannot be seen
**BOTH ARE ACTIVATED
Broth:
 Distilled (Cocci/Bacili)
 Loop  Colony
 Tip in H2o

1. Blood extraction
2. Centrifugation (density)
3. Pipetting (Pasteur)
4. Blood smear stain (Wright stain)
 Eosin – acidic stain (bright pink);
affinity to basic cell component
 Methylene blue – basic stain (bluish)
 Fixative (Methanol) = can cause
blindness
 Eosin
 Methylene blue
 Wash
**15 seconds or more**
5. Broth culture
 Distilled (Cocci/Bacili)
 Loop  Colony
 Tip in H2o
6. Microscopy

PROCEDURE:
1. Label Test tube
2. Add 4 to 8 drops of the buffy coat from either
the patient’s heparinized blood or from the
normal control to the respectively labeled test
tubes (EDTA tubes)
3. Add 2 to 3 drops of the bacterial broth culture
to each tube.
4. Incubate test tubes at room temperature or
37oC for 30 minutes.
5. Place 1 drop of the incubated specimen on a
glass slide and prepare a smear.
6. Air dry the slides and stain with Wrights
stain.
7. Place a drop of immersion on each of smear
and examine microscopically with the
immersion objective.
Scan  LPO  HPO  OIO (mineral oil)

INTERPRETATION OF RESULT
 POSITIVE – Demonstration of engulfment
of bacteria (immune system is working
properly)