BCMB 406A - Lab 2 Exam Latest Update
What is the overall purpose of the first part of the lab? - ANSWER Assess MHC
class I expression on the surface of NOP breast cancer cells that have been
treated with and without IFNγ
What is the overall purpose of the second part of the lab? - ANSWER Measure
the CD8+ T cells response against NOP tumour cells that were either treated or
not with IFNγ
How do CD8+ T cells recognize cancer cells? - ANSWER They scan the surface
of cancer cells and destroy them once they find mutated proteins
What is one way for a tumour to escape the immune system? - ANSWER
Decrease expression of MHC class I to become undetectable by cytotoxic CD8+
T cells
What are the three major steps of flow cytometry? - ANSWER Staining: cells of
interest stained with mAbs coupled to fluorophores
Acquisition
Data analysis (FlowJo)
Two major components of a flow cytometer - ANSWER Optics system: measure
the fluorescent antibodies bound to cell
Fluidics system: make sure cells pass through optics system single file
How does staining cells for flow work? - ANSWER Label cells with a cocktail of
monoclonal antibodies (mAb) that are specific to a different epitope and each
have different fluorophore. Fluorescent results show what epitopes were
present on the cells
How were NOP cells engineered? - ANSWER To express the OVA peptide that
can be recognized by T cells
For OVA to be recognized by T cells what must happen? - ANSWER Must be
presented in a complex with MHC
What is IFN-γ and what is its purpose? - ANSWER A cytokine to enhance MHC
expression (can reverse downregulation of MHC but doesn't work if pathway is
non-functional)
What is the overall purpose of the first part of the lab? - ANSWER Assess MHC
class I expression on the surface of NOP breast cancer cells that have been
treated with and without IFNγ
What is the overall purpose of the second part of the lab? - ANSWER Measure
the CD8+ T cells response against NOP tumour cells that were either treated or
not with IFNγ
How do CD8+ T cells recognize cancer cells? - ANSWER They scan the surface
of cancer cells and destroy them once they find mutated proteins
What is one way for a tumour to escape the immune system? - ANSWER
Decrease expression of MHC class I to become undetectable by cytotoxic CD8+
T cells
What are the three major steps of flow cytometry? - ANSWER Staining: cells of
interest stained with mAbs coupled to fluorophores
Acquisition
Data analysis (FlowJo)
Two major components of a flow cytometer - ANSWER Optics system: measure
the fluorescent antibodies bound to cell
Fluidics system: make sure cells pass through optics system single file
How does staining cells for flow work? - ANSWER Label cells with a cocktail of
monoclonal antibodies (mAb) that are specific to a different epitope and each
have different fluorophore. Fluorescent results show what epitopes were
present on the cells
How were NOP cells engineered? - ANSWER To express the OVA peptide that
can be recognized by T cells
For OVA to be recognized by T cells what must happen? - ANSWER Must be
presented in a complex with MHC
What is IFN-γ and what is its purpose? - ANSWER A cytokine to enhance MHC
expression (can reverse downregulation of MHC but doesn't work if pathway is
non-functional)
