VBS 2032. EXAM 3
Define:
- Neutrophils
- Lymphocytes
- Monocytes
- Eosinophils
- Basophils
- Mast cells
- Granulocytes
- Agranulocytes
- Differential white blood cell count - answer Types of White Blood Cells (5):
1. Neutrophils: phagocytizes bacteria
- Granulocyte (granules in cytoplasm)
- most frequent type of WBC
- pale cytoplasm
- 2-3 regularly shaped nuclei
- Can be segmented or unsegmented
2. Lymphocytes: Group of WBC involved in adaptive immunity
- Involve in immune response
- Agranulocyte (no granules in cytoplasm)
- Small
- Large, round, dark nucleus
- Cytoplasm appears as a rim around nucleus
EX: B and T cells
- B cell: programmed to make antibodies
- T cell: lymphocyte that matures in the thymus
3. Monocytes:
- Garbage trucks of the immune response
- Agranulocyte (no granules in cytoplasm)
- Huge (much bigger than lymphocytes)
- Large, kidney-bean shaped dark nucleus
- Cytoplasm appears foamy
- They differentiate into either macrophages or dendritic cells when they migrate into
tissues
4. Eosinophils: Kills parasitic worms
- Granulocyte (granules in cytoplasm)
- Cytoplasm appears pink or red
- Nucleus is bilobed, evenly rounded lobes (more uniform than neutrophils)
,- Are involved in allergic reactions, causing some of the symptoms associated with
allergies, but reducing others.
5. Basophils: Releases histamine and other inflammation-inducing chemicals
- Least common WBC
- Granulocyte (granules in cytoplasm)
- Cytoplasm crowded by very dark granules
- Cannot see bilobed nucleus because granules are so dense
- Receptors on its surface bind IgE
*Are involved in allergic reactions and inflammation
Mast Cells: similar in appearance and function of basophils but found in the tissue
- Receptors of Fc portion of IgE
*Important in the inflammatory response and are responsible for allergic reactions
Granulocytes: Neutrophils, eosinophils and basophils that contain cytoplasmic granules
of proteins and other compounds used to kill bacteria.
Agranulocytes: Lymphocytes and monocytes that lac
The most common granulocyte in the blood is:
A. lymphocyte
B. neutrophil
C. leukocyte
D. RBC - answerB. neutrophil
*This multi-lobed cell is also called a polymorphonuclear leukocyte (PMN)
- What do leukocytes look like: lymphocyte, monocyte, neutrophil, basophil, eosinophil?
- What is the difference between neutrophilia and neutropenia? - answerNeutrophilia:
increased proportion of neutrophils in the blood
- Common with acute bacterial infections
Neutropenia: decreased proportion of neutrophils
- May be seen with viral infections
Define:
- Antigen
- Adaptive immune system
- Antibodies
- Enzyme-linked immunosorbent assay (ELISA)
- Primary antibodies
- Secondary antibodies
- Specificity
- Conjugated - answerAntigen: something your body perceives as foreign
EX: bacteria, viruses or fungi
,Adaptive immune system: When you are exposed to an antigen, your body responds
first with components of the innate immune system and begins to make antibodies and
flags them for destruction by other cells of the immune system.
Antibodies: proteins that recognize and bind to specific antigens
- Contains 2 light and 2 heavy chains each with a constant and variable region
Enzyme-linked immunosorbent assay (ELISA): uses antibodies to detect the presence
of a disease agent (for example, viruses, bacteria, or parasites) in your blood or other
body fluid.
*Can use antibodies to detect the presence of antibodies
Primary antibodies: In an immunoassay, the antibodies used to recognize antigens like
disease agents
Secondary antibodies: recognize primary antibodies and can be used in many different
tests. An enzyme is conjugated or covalently linked to the heavy chain.
Specificity: binding to a specific antigen of interest
Conjugated:
- Describe the steps in an ELISA assay. (Make a drawing)
- Why are the washes necessary?
- What results would you get if you forgot to add the primary or secondary antibody? -
answerELISA assay Steps:
1. Load student samples (unknown antigens) in triplicate into wells of the microplate
strips.
- Incubate for 5 mins and rinse
*lets the antigen bind to the wells
2. Add primary antibody to all wells.
- Incubate for 5 mins and rinse
*Primary antibody is very specific and binds to antigen of interest
3. Add enzyme-linked secondary antibody to all wells.
- Incubate for 5 mins and rinse
*Secondary binds to primary antibody
*Purpose of secondary: amplify the reaction because it usually combines with multiple
primary antibodies at once and more than one site on each primary antibody
*The secondary antibody is used to conjugate the enzyme horseradish peroxide (BRP)
4. Add enzyme substrate to all wells.
- Incubate for 5 minutes
, *The HRP catalyzes the conversion of the chroma genic substrate to a color we can
measure
- (When HRP cleaves the substrate, a color in formed. Within a certain range, the
absorbance is proportional to the amount of antigen present, making this assay
quantitative.)
5. Add stop solution
- An acid that will denature the enzyme and stop its activity
Washes are necessary:
- To ensure the wash buffer is washing the unbound primary antibody out of the wells
Results if forgetting to add the primary or secondary antibody:
- Primary: there would be no attachment to the antigen
-Secondary: cannot bind to the primary antibody since it is not present and enzyme will
not conjugate or covalently link to the heavy chain.
In an ELISA, an enzyme is conjugated to:
A. the antigen
B. the primary antibody
C. the secondary antibody
D. substrate - answerC. the secondary antibody
*Secondary antibodies recognize primary antibodies and can be used in many different
tests. An enzyme is conjugated or covalently linked to the heavy chain.
What cells participate in the process of antibody production? Where are they located? -
answer- The bone marrow has stem cells to produce antibodies and other blood cells
EX: T and B cells
*Plasma cells and memory B cells produce antibodies in different situations.
- Common lymphoid progenitor
In what ways do the complement pathways and humoral immunity interact? - answer-
Both use Opsonization
- Humoral immunity antibodies can lead to complement system activation
- This is the classic pathway.
Where are lymphoid cells produced? - answerFrom stem cells in the bone marrow and
then circulated in the blood to the lymphoid tissue
Draw the structure of IgG. What part recognizes the epitope? What part interacts with
phagocytic cells? What are the parts called? - answer- Fab region contains both light
chain and heavy chain the distal end contains the variable region.
- Variable is what binds to unique epitopes.
Define:
- Neutrophils
- Lymphocytes
- Monocytes
- Eosinophils
- Basophils
- Mast cells
- Granulocytes
- Agranulocytes
- Differential white blood cell count - answer Types of White Blood Cells (5):
1. Neutrophils: phagocytizes bacteria
- Granulocyte (granules in cytoplasm)
- most frequent type of WBC
- pale cytoplasm
- 2-3 regularly shaped nuclei
- Can be segmented or unsegmented
2. Lymphocytes: Group of WBC involved in adaptive immunity
- Involve in immune response
- Agranulocyte (no granules in cytoplasm)
- Small
- Large, round, dark nucleus
- Cytoplasm appears as a rim around nucleus
EX: B and T cells
- B cell: programmed to make antibodies
- T cell: lymphocyte that matures in the thymus
3. Monocytes:
- Garbage trucks of the immune response
- Agranulocyte (no granules in cytoplasm)
- Huge (much bigger than lymphocytes)
- Large, kidney-bean shaped dark nucleus
- Cytoplasm appears foamy
- They differentiate into either macrophages or dendritic cells when they migrate into
tissues
4. Eosinophils: Kills parasitic worms
- Granulocyte (granules in cytoplasm)
- Cytoplasm appears pink or red
- Nucleus is bilobed, evenly rounded lobes (more uniform than neutrophils)
,- Are involved in allergic reactions, causing some of the symptoms associated with
allergies, but reducing others.
5. Basophils: Releases histamine and other inflammation-inducing chemicals
- Least common WBC
- Granulocyte (granules in cytoplasm)
- Cytoplasm crowded by very dark granules
- Cannot see bilobed nucleus because granules are so dense
- Receptors on its surface bind IgE
*Are involved in allergic reactions and inflammation
Mast Cells: similar in appearance and function of basophils but found in the tissue
- Receptors of Fc portion of IgE
*Important in the inflammatory response and are responsible for allergic reactions
Granulocytes: Neutrophils, eosinophils and basophils that contain cytoplasmic granules
of proteins and other compounds used to kill bacteria.
Agranulocytes: Lymphocytes and monocytes that lac
The most common granulocyte in the blood is:
A. lymphocyte
B. neutrophil
C. leukocyte
D. RBC - answerB. neutrophil
*This multi-lobed cell is also called a polymorphonuclear leukocyte (PMN)
- What do leukocytes look like: lymphocyte, monocyte, neutrophil, basophil, eosinophil?
- What is the difference between neutrophilia and neutropenia? - answerNeutrophilia:
increased proportion of neutrophils in the blood
- Common with acute bacterial infections
Neutropenia: decreased proportion of neutrophils
- May be seen with viral infections
Define:
- Antigen
- Adaptive immune system
- Antibodies
- Enzyme-linked immunosorbent assay (ELISA)
- Primary antibodies
- Secondary antibodies
- Specificity
- Conjugated - answerAntigen: something your body perceives as foreign
EX: bacteria, viruses or fungi
,Adaptive immune system: When you are exposed to an antigen, your body responds
first with components of the innate immune system and begins to make antibodies and
flags them for destruction by other cells of the immune system.
Antibodies: proteins that recognize and bind to specific antigens
- Contains 2 light and 2 heavy chains each with a constant and variable region
Enzyme-linked immunosorbent assay (ELISA): uses antibodies to detect the presence
of a disease agent (for example, viruses, bacteria, or parasites) in your blood or other
body fluid.
*Can use antibodies to detect the presence of antibodies
Primary antibodies: In an immunoassay, the antibodies used to recognize antigens like
disease agents
Secondary antibodies: recognize primary antibodies and can be used in many different
tests. An enzyme is conjugated or covalently linked to the heavy chain.
Specificity: binding to a specific antigen of interest
Conjugated:
- Describe the steps in an ELISA assay. (Make a drawing)
- Why are the washes necessary?
- What results would you get if you forgot to add the primary or secondary antibody? -
answerELISA assay Steps:
1. Load student samples (unknown antigens) in triplicate into wells of the microplate
strips.
- Incubate for 5 mins and rinse
*lets the antigen bind to the wells
2. Add primary antibody to all wells.
- Incubate for 5 mins and rinse
*Primary antibody is very specific and binds to antigen of interest
3. Add enzyme-linked secondary antibody to all wells.
- Incubate for 5 mins and rinse
*Secondary binds to primary antibody
*Purpose of secondary: amplify the reaction because it usually combines with multiple
primary antibodies at once and more than one site on each primary antibody
*The secondary antibody is used to conjugate the enzyme horseradish peroxide (BRP)
4. Add enzyme substrate to all wells.
- Incubate for 5 minutes
, *The HRP catalyzes the conversion of the chroma genic substrate to a color we can
measure
- (When HRP cleaves the substrate, a color in formed. Within a certain range, the
absorbance is proportional to the amount of antigen present, making this assay
quantitative.)
5. Add stop solution
- An acid that will denature the enzyme and stop its activity
Washes are necessary:
- To ensure the wash buffer is washing the unbound primary antibody out of the wells
Results if forgetting to add the primary or secondary antibody:
- Primary: there would be no attachment to the antigen
-Secondary: cannot bind to the primary antibody since it is not present and enzyme will
not conjugate or covalently link to the heavy chain.
In an ELISA, an enzyme is conjugated to:
A. the antigen
B. the primary antibody
C. the secondary antibody
D. substrate - answerC. the secondary antibody
*Secondary antibodies recognize primary antibodies and can be used in many different
tests. An enzyme is conjugated or covalently linked to the heavy chain.
What cells participate in the process of antibody production? Where are they located? -
answer- The bone marrow has stem cells to produce antibodies and other blood cells
EX: T and B cells
*Plasma cells and memory B cells produce antibodies in different situations.
- Common lymphoid progenitor
In what ways do the complement pathways and humoral immunity interact? - answer-
Both use Opsonization
- Humoral immunity antibodies can lead to complement system activation
- This is the classic pathway.
Where are lymphoid cells produced? - answerFrom stem cells in the bone marrow and
then circulated in the blood to the lymphoid tissue
Draw the structure of IgG. What part recognizes the epitope? What part interacts with
phagocytic cells? What are the parts called? - answer- Fab region contains both light
chain and heavy chain the distal end contains the variable region.
- Variable is what binds to unique epitopes.
