The response to lipopolysaccharide of mouse spleen lymphocytes fractionated on the basis of surface immunoglobulin and complement receptor using fluorescence-activated cell sorting and rosetting techniques

Abstract

Mouse spleen lymphocytes were stained with rabbit antisera specific for either μ chain or δ chain, followed by fluorescein-conjugated goat anti-rabbit immunoglobulin. The cells were analysed and fractionated using a fluorescence-activated cell sorter. Fifty-five per cent of the lymphocytes stained with a polyspecific anti-Ig reagent or with a combination of anti-μ and anti-δ reagents, while about 40% of the lymphocytes were stained when either the anti-μ reagent or the anti-δ reagent was used alone. Three per cent of the lymphocytes stained with the anti-μ reagent, but not with the anti-δ reagent, and eight per cent stained only with the anti-δ reagent. Unfractionated spleen cells and populations depleted of μ- or δ-bearing cells were cultured in the presence of lipopolysaccharide. All three populations responded by incorporating [³H]-thymidine and secreting IgM and IgG.

Spleen cells were fractionated by a rosetting technique into complement receptor-positive and negative populations. Both populations were able to respond to lipopolysaccharide and to synthesize Ig of both the IgM and IgG classes. Unfractionated cells and complement receptor-negative populations were stained for surface μ or δ chain and analysed on the fluorescence-activated cell sorter. The distribution of staining intensity suggested that the complement receptor-bearing population was enriched in cells which stain weakly for μ and cells which stain with a low to intermediate intensity for δ chain.

It is concluded that the precursors of IgM- and IgG-secreting cells are not limited to any one of the three populations of cells defined on the basis of surface immunoglobulin or to either of the populations defined on the basis of the complement receptor.