Someone help me solve my homework, a biology questions. A researcher has been gi

Question

Someone help me solve my homework, a biology questions.

A researcher has been given two CD4 T-cell clones from a collaborator. T-cell clone A is thought to be a Th17 clone specific for cytochrome C, and T-cell clone B is thought to be a Treg clone specific for myoglobin.

a) Describe how you would design an assay to confirm that T-cell clone A indeed is a Th17 clone, and not a Th1 or Th2 clone.

b) Describe how you would design an assay to establish whether T-cell clone B has suppressive Treg-cell function.

Explanation / Answer

a) Describe how you would design an assay to confirm that T-cell clone A indeed is a Th17 clone, and not a Th1 or Th2 clone.

Culture T-cell clone A with cytochrome C plus APCs. The APCs will process and present to T-cell clone A cytochrome C peptide in the context of MHC-II. In parallel cultures, culture T-cell clone A with APCs in medium alone.Measure the presence of cytokinesin supernatants (e.g. by ELISA). A Th17 clone should produce IL-17A, but not IFN-g or IL-4, in response to its specific antigen (cytochrome C).

b) Describe how you would design an assay to establish whether T-cell clone B has suppressive Treg-cell function.

Culture T-cell clone A with cytochrome C plus APCsas above. In parallel cultures, mix APCs, T-cell clone A, T-cell clone B, cytochrome C, and myoglobin in the same well. Measure the level of IL-17Ain supernatants (e.g. by ELISA). If T-cell clone B is suppressive, the amount ofIL-17A should be lower in the cultures containing the two T-cell clones compared to when T-cell clone A is cultured by itself.

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