Figure 3. sCD89 induces high molecular... | Rockefeller University Press

$Figure 3. sCD89 induces high molecular mass circulating IgA1 complex formation. (A) Surface staining of CD89 on mouse blood monocytes. The white layer represents CD89Tg, dotted for α1KI-CD89Tg, dashed for α1KI, and gray for WT cells. The graph shows the mean fluorescent intensity (MFI) of CD89 on CD11b+ cells. 5–30 mice per group. Error bars show mean ± SEM. (B and C) Detection of IgA1–sCD89 complexes in the sera of mice (B) and patients (C) by ELISA. For detection of these complexes, A3 anti-CD89 was used as a coating antibody and monoclonal anti–human IgA as a secondary reagent. n = 11 mice per group and 15–25 individuals per group. Bars represent the mean of OD. (D) IgA1 levels in the serum of 12-wk-old α1KI and α1KI-CD89Tg mice measured by ELISA. 11–16 mice per group. Bars represent the mean. (E) Immunoblot of monomeric (m), dimeric (d), and polymeric (p) forms of IgA1 in sera (normalized at 0.1 µg IgA1 per sample) from α1KI and α1KI-CD89Tg mice with an anti–human IgA polyclonal antibody. (F and G) Total IgA detected by ELISA and Western blot in serum HPLC fractions of one representative α1KI (F) and one α1KI-CD89Tg mouse (G). n = 3 mice per group. Detection of IgA1–sCD89 complexes in these fractions by ELISA shows that they correspond to high molecular mass IgA1 fractions. (H) Immunoblot on material immunoprecipitated from anti-CD89 serum from α1KI and α1KI-CD89Tg mice using a mixture of anti-CD89 mAbs. *, P < 0.05; and **, P < 0.01 (Mann-Whitney U test).$

Figure 3.

sCD89 induces high molecular mass circulating IgA1 complex formation. (A) Surface staining of CD89 on mouse blood monocytes. The white layer represents CD89Tg, dotted for α1KI-CD89Tg, dashed for α1KI, and gray for WT cells. The graph shows the mean fluorescent intensity (MFI) of CD89 on CD11b⁺ cells. 5–30 mice per group. Error bars show mean ± SEM. (B and C) Detection of IgA1–sCD89 complexes in the sera of mice (B) and patients (C) by ELISA. For detection of these complexes, A3 anti-CD89 was used as a coating antibody and monoclonal anti–human IgA as a secondary reagent. n = 11 mice per group and 15–25 individuals per group. Bars represent the mean of OD. (D) IgA1 levels in the serum of 12-wk-old α1KI and α1KI-CD89Tg mice measured by ELISA. 11–16 mice per group. Bars represent the mean. (E) Immunoblot of monomeric (m), dimeric (d), and polymeric (p) forms of IgA1 in sera (normalized at 0.1 µg IgA1 per sample) from α1KI and α1KI-CD89Tg mice with an anti–human IgA polyclonal antibody. (F and G) Total IgA detected by ELISA and Western blot in serum HPLC fractions of one representative α1KI (F) and one α1KI-CD89Tg mouse (G). n = 3 mice per group. Detection of IgA1–sCD89 complexes in these fractions by ELISA shows that they correspond to high molecular mass IgA1 fractions. (H) Immunoblot on material immunoprecipitated from anti-CD89 serum from α1KI and α1KI-CD89Tg mice using a mixture of anti-CD89 mAbs. *, P < 0.05; and **, P < 0.01 (Mann-Whitney U test).

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