Shift of C3 deposition from localization in the glomerulus into the tubulo-interstitial compartment in the absence of secreted IgM in immune complex glomerulonephritis
Christine VaculikBeate M. RügerGenya YanagidaDavid HollemannA. SoleimanUdo LosertJ. ChenMichael B. Fischer
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Summary The role of secretory IgM in protecting kidney tissue from immune complex glomerulonephritis induced by 4 mg horse spleen apoferritin and 0·05 mg lipopolysaccharide has been investigated in mutant mice in which B cells do not secrete IgM, but are capable of expressing surface IgM and IgD and secreting other Ig isotypes. Glomerular size, number of glomeruli per cross-section, glomerular cellularity and urine content of protein and creatinine was comparable in treated secreted IgM (sIgM)-deficient and wild-type mice. Assessment of urinary proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed a 30 kDa low molecular weight protein in treated sIgM-deficient animals only, reflecting dysfunction of proximal tubules. A shift of bound C3 from glomeruli to the tubulo-interstitial compartment in sIgM-deficient mice also suggests tubulo-interstitial damage. In contrast, local C3 synthesis within the kidney tissue did not differ between the two treated groups. Apoptosis physiologically present to maintain kidney cell homeostasis was increased slightly in treated wild-type mice. These results indicate that secretory IgM can protect the tubulo-interstitial compartment from immune complex-induced damage without having an effect on the glomerulus.Keywords:
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Three populations of murine splenic B lymphocytes have been characterized previously (6, 7, 9) as those bearing only IgM, those bearing only IgD, and a population bearing both isotopes. These studies were designed to test the response of the IgM+ cells (IgM-only or IgM plus IgD) vs. the IgD-only cells to the B-cell mitogen, lipopolysaccharide. Results that after 1-4 days of culture, in the presence of mitogen, the IgM+ cells enlarge and elaborate an IgM polyclonal response. The IgD-only cells, in contrast, do not exhibit an IgM polyclonal response, but instead undergo blastogenesis and proliferation.
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125I-membrane IgM, 125I-membrane IgD-like molecules, and their constituent chains from iodinated murine splenocytes were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The apparent m.w. of the heavy chains decreased as the acrylamide concentration was raised. The membrane mu-chain had a slower mobility than did mu-chain from secreted IgM. Unreduced IgM and IgD-like molecules had mobilities consistent with an H2L2 structure. Intact IgD-like molecules were replaced after overnight dialysis by molecules with the properties of HL. Unreduced surface IgM had a slower mobility than that of monomeric IgM obtained by partial reduction of secreted IgM.
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A proportion of lymphocytes in blood, spleen and lymph nodes of nonhuman primates had immunoglobulin on their surfaces detectable by fluorescent antibody to human IgM and IgD. The majority of the individual lymphocytes having either IgM or IgD on their surfaces possessed both classes of immunoglobulin. Lymphocyte surface IgD was capped independently of surface IgM on the same cell when incubated at 37 degrees with anti-IgD. Lymphocytes with surface IgM and/or IgD were present in blood at birth and the percentages over the first 6 months of life were increased compared to older monkeys. A corona of cells faintly positive for both IgM and IgD was observed around germinal centres of both lymph nodes and spleen.
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An analysis was made of the immunoglobulin surface markers of the cells of patients with chronic lymphatic leukemia (CLL) in view of previous evidence of their monoclonal B-cell character. The simultaneous presence of IgM and IgD on the surface of the majority of lymphocytes was demonstrated by both immunofluorescence and hemagglutination inhibition in most cases. However, cases were observed with surface IgM without IgD as well as cases with IgD without IgM. IgG and IgA were absent. Studies of the light chains indicated only a single class in a given case. In addition to bound light chains, free light chains were readily demonstrated in most cases through the use of antisera specific for "free chain" determinants. It thus appeared that there are three major types of surface Ig on CLL lymphocytes, IgM, IgD, and free light chains.
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Abstract Human peripheral blood mononuclear cells were depleted of surface IgM + or IgD + cells and assayed for mitogen‐induced differentiation to immunoglobulin‐secreting cells (ISC) of IgM, IgG and IgA classes. Stimulatory agents included T cell‐dependent poke weed mitogen, B cell mitogen Staphylococcus aureus bacteria strain Cowan I, and a combination of the two which gives uniform, high levels of ISC from all normal donors. Depletion of either IgM‐ or IgD‐bearing B lymphocytes resulted in loss of cells bearing the other Ig class and blocked most of the mitogenic reactivity to anti‐IgM and anti‐IgD. Proliferative responses to Cowan I in these depleted populations were about 20% that of unfractionated mononuclear cells. Depletion of T cells increased the mitogenic response to Cowan I and to the two antibody preparations, showing that they are T‐independent mitogens. Depletion of IgD + cells caused partial loss of mitogen‐induced IgM ISC (22%‐60% of unseparated controls) but no loss of IgG or IgA ISC. Depletion of IgM‐bearing cells caused complete loss of IgM ISC, but no loss of IgG or IgA ISC. We previously demonstrated that anti‐IgM antibody blocked mitogen induction of Ig secretion of these three classes in spleen cells, but only IgM secretion in blood mononuclear cells. Together, the results suggest that the majority of cells in normal blood responding to mitogens to mature to IgG or IgA production belong to IgM − , IgD − B cell subsets, in contrast to precursors of secreting cells for these isotypes in the spleen. Thus, these blood precursors appear to be more mature than the corresponding spleen cells.
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Summary The distribution among murine spleen cells of a newly described class of surface immunoglobulin (Ig) with properties similar to human IgD was studied. Splenocytes were separated on the basis of size and the surface Ig on large cells (sedimenting faster than 6 mm/hr in a 1 × G velocity gradient) and small cells (sedimenting between 2.5 and 3.0 mm/hr) was analyzed. Spleen cells from young animals had virtually only IgM on the large cells but had substantial amounts of IgM and the IgD-like molecule (IgD) on small cells. Spleen cells from older animals, which have larger amounts of IgD, had IgM and IgD on both cell types; however, the amount of IgD relative to IgM on the large cells was always substantially less than that on the small ones. These observations taken together with those of other investigators support the hypothesis that a large lymphocyte with surface IgM is the precursor of a small lymphocyte with both surface IgM and IgD.
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Abstract Monoclonal B lymphocyte populations from the bone marrow of seven cases of Waldenström's macroglobulinemia were studied for membrane IgD and IgM. In all cases lymphocytes with both classes on the membrane were found as well as cells with IgD or IgM only; the relative percentages of these three groups of lymphocytes were different in different cases. The IgM‐containing plasma cells always had membrane IgM and some of these also had IgD. Of particular interest was one patient whose macroglobulin had an anti‐IgG reactivity. The peripheral blood B lymphocytes of this patient had both membrane IgM and membrane IgD: it was possible to show, by capping with anti‐IgG or with aggregated IgG, that both classes of immunoglobulin receptors shared reactivity with human IgG.
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