The questions on the papers are intended to guide your thinking and approach the papers,

as well as provide a basis for discussion. Although answers to the questions will be
collected, the answers themselves will be graded on effort and thought, not correctness.
Please limit your response to 1 page, and use simple diagrams whenever you feel they
would be helpful. Please be prepared to turn in your answers to the assigned questions
for each week's article at the beginning of section.

Nov. 7 –9: Schreiber et al, 2012, Nature.

Cancer immunoediting is the process by which the immune system controls tumor
outgrowth and shapes tumor immunogenicity.

What knockout mice did the authors use to demonstrate that immunoediting of d42m1
occurs via a T-cell dependent immunoselection process? Based on past lectures,
describe the role of the knock-out gene in T-cell development.

The mice were Rag2 -/- immune deficient. The Rag gene encodes for the protein of the
same name that is crucial to VDJ recombination.

Using exome sequencing, the authors identify somatic, non-synonymous mutations in

d42m1 and H31m1 cells.

i) How were the d42m1 and H31m1 cells generated?

D42m1 and h31m1 cells were generated from Rag -/- mice after transplantation.

ii) What is exome sequencing? How did the authors modify exome sequencing for their
studies? How was this difference in experimental approach important for finding tumor
rejection antigens?

Exome sequencing is a technique for sequencing all protein coding regions of the
genome. It involves first selecting exons and then sequencing them with high
throughput DNA technology. The version of exome sequencing used in this paper
differs from regular exome sequencing in that the authors used cDNA.

iii) What dominant rejection antigen did they find in the d42m1 cell line?

Mutant spectrin beta-2