8a.
Specific Aims The theme of the work proposed in this program project application is to elucidate the key controlmechanisms that regulate clonal expansion (cell cycle progression, Project 1) and differentiation (IgMsecretion, Project 2) in a population of B cells, namely, B-1 cells, that, though occupying a critical rolein anti-bacterial and anti-viral immunity, constitutes just a small proportion of the total complement ofB cells in both human and murine systems. In the murine system, B-1 cells are, in some strains, thepredominant B cell population in the peritoneal cavity, and much of what is known about B-1 cellsderives from studies of B cells obtained from this location by peritoneal lavage (peritoneal 'washout').However, B-1 cells obtained by peritoneal washout are highly contaminated with many other cellularelements. Thus, studies of primary B-1 cells are inherently plagued by the twin difficulties of isolatingcells a) in sufficient numbers, and b) with sufficient purity, to allow studies to go forward in aproductive fashion. Multi-project programs have the additional responsibility to insure that allexperiments utilize similar populations of B-1 cells that do not differ in terms of isolation strategy andresultant purity. To address these issues, the investigators of this program have formulated a singlesite core to provide a steady stream of highly purified B-1 (and B-2) cells on a regular and scheduledbasis. These goals will be met through the following specific aims:1. Develop and maintain expertise in isolation and purification of murine peritoneal B-1 cells.2. Isolate and purify murine peritoneal B-1 cells and splenic B-2 cells on a regular and scheduledbasis.3. Provide purified B-1 and B-2 cells, or lysates of treated B-1 and B-2 cells, to projects 1 and 2located at Boston College and Boston Medical Center, respectively.
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