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. 2000 Jul 3;192(1):87–98. doi: 10.1084/jem.192.1.87

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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graphic file with name JEM000065.f3ad.jpg — The level of loss of SAg-reactive ISCs in bone marrow and spleen after neonatal or adult treatment with SpA. Level of suppression correlates with avidity of Fab-binding activity and does not require T cells. BALB/c mice treated as (A and B) neonates or as (C and D) adults, or (E and F) neonatally treated TCR-β^−/−δ^−/− or control C57BL/6 mice, received intraperitoneal injections of saline (Naive), a control antigen (HEL), a genetic mutant tetrameric form of SpA (tetmDD′), an SpA monomer (DD′), a chemically modified form of SpA (MSpA), or native SpA. The frequencies of IgM-specific ISCs in the spleen (A, C, and E) and bone marrow (B, D, and F) were quantitated 2–5 mo later by enzyme-linked immunospot assay. Replicate wells coated with anti-IgM established the total frequency of IgM-ISCs (left); with MSpA determined the frequency of ISCs that interact with the Fab-binding site of SpA (middle); or with a control antigen established that the frequency of ISCs with nonspecific binding activity was <10/10⁶ total cells (not shown). For each mouse, the relative proportion of ISCs with SAg reactivity was determined by dividing the frequency of MSpA-reactive IgM-ISCs by the frequency for total ISCs (right). The frequency of PC-specific ISCs was below the limits of detection (not shown; reference 59). +, mean values for each group. For comparisons to control protein-treated groups, significant values are indicated as follows for SpA-treated groups: *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001. Values represent the frequency of each type of IgM-secreting cell per million mononuclear cells, or the relative percentage of these IgM-secreting cells among all IgM-secreting cells (%), as indicated.

graphic file with name JEM000065.f3ef.jpg — The level of loss of SAg-reactive ISCs in bone marrow and spleen after neonatal or adult treatment with SpA. Level of suppression correlates with avidity of Fab-binding activity and does not require T cells. BALB/c mice treated as (A and B) neonates or as (C and D) adults, or (E and F) neonatally treated TCR-β^−/−δ^−/− or control C57BL/6 mice, received intraperitoneal injections of saline (Naive), a control antigen (HEL), a genetic mutant tetrameric form of SpA (tetmDD′), an SpA monomer (DD′), a chemically modified form of SpA (MSpA), or native SpA. The frequencies of IgM-specific ISCs in the spleen (A, C, and E) and bone marrow (B, D, and F) were quantitated 2–5 mo later by enzyme-linked immunospot assay. Replicate wells coated with anti-IgM established the total frequency of IgM-ISCs (left); with MSpA determined the frequency of ISCs that interact with the Fab-binding site of SpA (middle); or with a control antigen established that the frequency of ISCs with nonspecific binding activity was <10/10⁶ total cells (not shown). For each mouse, the relative proportion of ISCs with SAg reactivity was determined by dividing the frequency of MSpA-reactive IgM-ISCs by the frequency for total ISCs (right). The frequency of PC-specific ISCs was below the limits of detection (not shown; reference 59). +, mean values for each group. For comparisons to control protein-treated groups, significant values are indicated as follows for SpA-treated groups: *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001. Values represent the frequency of each type of IgM-secreting cell per million mononuclear cells, or the relative percentage of these IgM-secreting cells among all IgM-secreting cells (%), as indicated.