Experiment 7,8,9 Flashcards
(21 cards)
2 types of non-adherent cells present in human blood
erythrocytes (RBC)
leukocytes (WBC)
percentage of lymphocytes
20-40%
percentage of monocytes
2-8%
percentage of neutrophils
50-70%
percentage of eosinophils
1-5%
percentage of basophils
0.5-1%
Why is it important to perform a differential white blood cell count?
Infection or disease can alter the counts
High number of neutrophils?
bacteria infection
High number of eosinophils?
Parasite infection
persistent high numbers of some cell types?
leukemia
LPS
Found on the outer membrane of Gram-Negative Bacteria
Nonspecifically activates B cells
Only activates IgM B cells
Where do we get B cells?
Blood lymphocytes
Lymph nodes
Spleen
How can we activates the B cells and relate it to the difference antibodies being produces
Activate with LPS
Why do we need to activate B cells?
Most B cells are resting
To study how B cells are activated to produce antibodies
Study how T cells govern B cell activation and antibody production
To study how added agent can alter B cell activaton and antibody production
How does B cell activation occur?
- antigen binds to B cell surface Ig (BCR)
- Adding antibodies to the B cell Igs
3.Polyclonal activators (LPS)
Direct ELISA
The antigen is immobilized on the surface of the well
The enzyme linked antibody, specific to antigen is used fro detection
Indirect ELISA
Used for measuring antibodies in serum
–> immunize animals or subjects wiht antigen
–> take blood sample and prepare serum
–> measure anti-measles virus antibodies
Compare antibody response
–> determine if the vaccine worked
–> compared levels of protection
Sandwich ELISA
–> used for measuring antigen level in a solution
–> requires antibodies which bind to the antigen
–> capture antibody to bind to the antigen
–> enzyme linked secondary antibody to detect the bound antigen
Competitive ELISA
inhibition ELISA
Used for detecting small antigens that cannot be bound by 2 different antibodies
Polyclonal activators
Agents that can activate whole groups of cells, regardless of what antigen they are specific for
What activates T cells?
Concanavlin A and Phytohemagglutinin
