Interrogation of individual intratumoral B lymphocytes from lung cancer patients for molecular target discovery.

Intratumoral B lymphocytes are an integral part of the lung tumor microenvironment. Interrogation of the antibodies they express may improve our understanding of the host response to cancer and could be useful in elucidating novel molecular targets. We used two strategies to explore the repertoire of intratumoral B cell antibodies. First, we cloned VH and VL genes from single intratumoral B lymphocytes isolated from one lung tumor, expressed the genes as recombinant mAbs, and used the mAbs to identify the cognate tumor antigens. The Igs derived from intratumoral B cells demonstrated class switching, with a mean VH mutation frequency of 4%. Although there was no evidence for clonal expansion, these data are consistent with antigen-driven somatic hypermutation. Individual recombinant antibodies were polyreactive, although one clone demonstrated preferential immunoreactivity with tropomyosin 4 (TPM4). We found that higher levels of TPM4 antibodies were more common in cancer patients, but measurement of TPM4 antibody levels was not a sensitive test for detecting cancer. Second, in an effort to focus our recombinant antibody expression efforts on those B cells that displayed evidence of clonal expansion driven by antigen stimulation, we performed deep sequencing of the Ig genes of B cells collected from seven different tumors. Deep sequencing demonstrated somatic hypermutation but no dominant clones. These strategies may be useful for the study of B cell antibody expression, although identification of a dominant clone and unique therapeutic targets may require extensive investigation.

Chaucer and the Disconsolations of Philosophy: Boethius, Agency, and Literary Form in Late Medieval Literature

This study argues that Chaucer's poetry belongs to a far-reaching conversation about the forms of consolation (philosophical, theological, and poetic) that are available to human persons. Chaucer's entry point to this conversation was Boethius's Consolation of Philosophy, a sixth-century dialogue that tried to show how the Stoic ideals of autonomy and self-possession are not simply normative for human beings but remain within the grasp of every individual. Drawing on biblical commentary, consolation literature, and political theory, this study contends that Chaucer's interrogation of the moral and intellectual ideals of the Consolation took the form of philosophical disconsolations: scenes of profound poetic rupture in which a character, sometimes even Chaucer himself, turns to philosophy for solace and yet fails to be consoled. Indeed, philosophy itself becomes a source of despair. In staging these disconsolations, I contend that Chaucer asks his readers to consider the moral dimensions of the aspirations internal to ancient philosophy and the assumptions about the self that must be true if its insights are to console and instruct. For Chaucer, the self must be seen as a gift that flowers through reciprocity (both human and divine) and not as an object to be disciplined and regulated.

Chapter one focuses on the Consolation of Philosophy. I argue that recent attempts to characterize Chaucer's relationship to this text as skeptical fail to engage the Consolation on its own terms. The allegory of Lady Philosophy's revelation to a disconsolate Boethius enables philosophy to become both an agent and an object of inquiry. I argue that Boethius's initial skepticism about the pretentions of philosophy is in part what Philosophy's therapies are meant to respond to. The pressures that Chaucer's poetry exerts on the ideals of autonomy and self-possession sharpen one of the major absences of the Consolation: viz., the unanswered question of whether Philosophy's therapies have actually consoled Boethius. Chapter two considers one of the Consolation's fascinating and paradoxical afterlives: Robert Holcot's Postilla super librum sapientiae (1340-43). I argue that Holcot's Stoic conception of wisdom, a conception he explicitly links with Boethius's Consolation, relies on a model of agency that is strikingly similar to the powers of self-knowledge that Philosophy argues Boethius to posses. Chapter three examines Chaucer's fullest exploration of the Boethian model of selfhood and his ultimate rejection of it in Troilus and Criseyde. The poem, which Chaucer called his "tragedy," belonged to a genre of classical writing he knew of only from Philosophy's brief mention of it in the Consolation. Chaucer appropriates the genre to explore and recover mourning as a meaningful act. In Chapter four, I turn to Dante and the House of Fame to consider Chaucer's self-reflections about his ambitions as a poet and the demands of truth-telling.