Separation of various B-cell subpopulations from mouse spleen
1979; Elsevier BV; Volume: 46; Issue: 1 Linguagem: Inglês
10.1016/0008-8749(79)90253-3
ISSN1090-2163
AutoresSharyn M. Walker, Geraldine C. Meinke, William O. Weigle,
Tópico(s)Immune Cell Function and Interaction
ResumoMouse spleen cells were depleted of immunoglobulin (Ig)-bearing B cells by rosetting with glutaraldehyde-fixed, tannic acid-treated RBC coupled with antibody to mouse Ig (anti-Ig) and removing the rosetted cells by density gradient centrifugation. The method was routinely greater than 90% effective in removing B cells as assayed by the failure of anti-Ig rosette-depleted primed spleen cells to generate antibody-producing cells in vitro in response to specific antigen or of anti-Ig rosette-depleted nonprimed spleen cells to generate a polyclonal antibody response. T cells were not removed by the rosetting procedure as measured by helper T-cell activity. The greater effectiveness of the rosetting procedure in removing potential IgG-secreting, non-IgM-bearing B cells is shown relative to other commonly used B-cell depletion procedures. Because the RBC in the rosetting reagent are fixed with glutaraldehyde, the rosetting reagent is stable for many months. Such stability makes constantly available a convenient means for B-cell removal, as well as reducing consumption of antisera.
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