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R B Corley

Publications and source records attributed to R B Corley.

At least 19 recordsLinked to original sources
PubMed2000

The IgA/IgM receptor expressed on a murine B cell lymphoma is poly-Ig receptor.

T560, a mouse B lymphoma that originated in gut-associated lymphoid tissue, expresses receptors that bind dimeric IgA and IgM in a mutually inhibitory manner but have little affinity for monomeric IgA. Evidence presented in this paper indicates that the receptor is poly-Ig receptor (pIgR) known in humans and domestic cattle to bind both IgA and IgM. The evidence includes the demonstration that binding of IgM is J chain dependent, and that pIg-precipitated receptor has an appropriate Mr of 116-120 kDa and can be detected on immunoblots with specific rabbit anti-mouse pIgR. Overlapping RT-PCR performed using template mRNA from T560 cells and oligonucleotide primer pairs designed from the published sequence of mouse liver pIgR indicate that T560 cells express mRNA virtually identical with that of the epithelial cell pIgR throughout its external, transmembrane, and intracytoplasmic coding regions. Studies using mutant IgAs suggest that the Calpha2 domain of dimeric IgA is not involved in high-affinity binding to the T560 pIgR. Inasmuch as this mouse B cell pIgR binds IgM better than IgA, it is similar to human pIgR and differs from rat, mouse, and rabbit epithelial cell pIgRs that bind IgA but not IgM. Possible explanations for this difference are discussed. All clones of T560 contain some cells that spontaneously secrete both IgG2a and IgA, but all of the IgA recoverable from the medium and from cell lysates is monomeric; it cannot be converted to secretory IgA by T560 cells.

Amino Acid Sequence↗
PubMed1999

Constitutive nuclear translocation of NF-kappa B in B cells in the absence of I kappa B degradation.

Members of the NF-kappa B/Rel family of transcription factors are involved in many aspects of B lymphocyte development and function. NF-kappa B is constitutively active in these cells, in contrast with most other cell types. In the inactive form, NF-kappa B/Rel proteins are sequestered in the cytoplasm by members of the I kappa B family of NF-kappa B inhibitors. When activated, NF-kappa B is translocated to the nucleus, a process that involves the phosphorylation and proteasomal degradation of I kappa B proteins. Thus, NF-kappa B activation is accompanied by the rapid turnover of I kappa B proteins. We show that while this "classical" mode of NF-kappa B activation is a uniform feature of IgM+ B cell lines, all IgG+ B cells analyzed contain nuclear NF-kappa B yet have stable I kappa B alpha, I kappa B beta, and I kappa B epsilon. Furthermore, I kappa beta epsilon levels are at least 10 times lower in IgG+ B cells than in IgM+ B cells, an additional indication that the regulation of constitutive NF-kappa B activity in these two types of B cells is fundamentally different. These data imply the existence of a novel mechanism of NF-kappa B activation in IgG+ B cells that operates independently of I kappa B degradation. They further suggest that different isoforms of the B cell receptor may have distinct roles in regulating NF-kappa B activity.

Animals↗
PubMed1999

The contribution of ER quality control to the biologic functions of secretory IgM.

Secretory IgM provides a first line of defense against pathogens and is uniquely capable of enhancing the primary humoral immune response. Complement activation is especially important for these activities. Here, Padmalatha Reddy and Ronald Corley discuss how the 'quality control' mechanisms that regulate IgM assembly and secretion play important roles in the developmental progression of B cells and in B-cell function.

Agammaglobulinaemia Tyrosine Kinase↗
PubMed1998

Production of IgM hexamers by normal and autoimmune B cells: implications for the physiologic role of hexameric IgM.

Secreted IgM is predominantly found as pentameric molecules, but IgM can also be secreted as hexamers by B cell lines. Murine hexamers activate the complement cascade more efficiently than pentamers, but the physiologic significance of hexameric IgM remains unknown. Here, we report that IgM hexamers and pentamers are cleared from the circulation with similar kinetics, suggesting that the predominance of pentameric IgM in vivo reflects the regulation of polymer assembly and secretion in responding B cells. Normal IgM-secreting B cells, particularly those from the peritoneal cavity, are capable of secreting abundant hexameric IgM in vitro. The disparity between the ability of B cells to secrete IgM hexamers in vitro and the paucity of this polymer in vivo suggest that IgM hexamers might be deleterious. In support of this, we demonstrate that the autoantibodies from a number of patients with cold agglutinin (CA) disease include both IgM hexamers and pentamers. The CA IgM hexamers lyse human erythrocytes in the presence of human complement more efficiently than CA IgM pentamers, suggesting a potential role for hexameric IgM in the pathogenesis of this autoimmune syndrome.

Animals↗
PubMed1998

Assembly, sorting, and exit of oligomeric proteins from the endoplasmic reticulum.

The endoplasmic reticulum (ER) uses various mechanisms to ensure that only properly folded proteins enter the secretory pathway. For proteins that oligomerize in the ER, the proper tertiary and quaternary structures must be achieved before their release. Although some proteins fold before oligomerization, others initiate oligomerization cotranslationally. Here, we discuss these different strategies and some of the unique problems they present for the ER quality control system. One mechanism used by the ER is thiol retention. Thiol retention operates by monitoring the redox state of specific cysteine residue(s) and was discovered in studies on the assembly of IgM, a complex oligomeric glycoprotein. This system is also involved in retaining other unassembled proteins in the ER. Mutations that result in uneven numbers of cysteine residues can subject yet other proteins to thiol retention, altering their oligomerization status and function. The implications of these results on the effects of thiol retention on protein function and cell fate are discussed.

Animals↗
PubMed1997

Secretion of soluble pre-B cell receptors by pre-B cells.

Pre-B cells can express secretory mu (mu(s))- as well as membrane mu (mu(m))-chains. We evaluated the ability of mu(s)-chains to associate with surrogate light chains and assemble into a pre-B cell receptor (BCR) complex in pre-B cells, and explored whether mu(s)-chains could be exploited to generate a secreted soluble pre-BCR. We demonstrate that mu(s)-chains can associate with SLC internally. The mu(s)-containing complexes form higher order polymeric structures, but these are never assembled into completed covalent structures. Instead, the complexes are efficiently retained and rapidly degraded. Alteration of the intracellular redox state by incubation with 2-ME resulted in the secretion of mu(s)-chains, suggesting that they are retained by a thiol-mediated retention mechanism. To identify the sequences on mu(s)-chains responsible for their retention, we generated stable transfectants of a mu-negative pre-B cell line expressing either wild-type or mutant mu(s) constructs. Mutation of a single cysteine (Cys575) in the mu(s) tailpiece resulted in the release and secretion of the mu(s) H chains. These were associated with the surrogate light chain proteins lambda5 and VpreB, and thus appear to constitute an authentic secreted soluble pre-BCR. The soluble pre-BCR has a specificity distinct from Ab consisting of the same heavy chain V region paired with conventional light chains.

Animals↗
PubMed1997

Late events in assembly determine the polymeric structure and biological activity of secretory IgM.

IgM antibodies can be secreted in at least two functional polymeric forms that can be distinguished according to subunit composition. While IgM hexamers comprise six H2L2 monomeric subunits, pentamers contain an additional polypeptide, the J chain. In the presence of high abundance J chain protein, IgM pentamers are preferentially assembled at the expense of hexamers. To determine the mechanism by which J chain regulates the assembly process, we defined the point at which J chain is added to assembling polymers. We found no evidence for the presence of J chain in small IgM assembly intermediates of IgM, suggesting that it was not stably associated with these complexes. However, J chain was found associated with large polymeric IgM complexes exhibiting sedimentation properties of intracellular pentameric structures. These complexes were frequently not completely covalently assembled; however, complete covalent assembly of J chain-containing pentameric complexes did occur prior to their maturation in the Golgi. These data argue that pentameric structures are the substrate for J chain incorporation into assembling IgM and suggest that the incorporation of J chain is thermodynamically favored over the addition of a sixth monomeric subunit into an assembling polymer. We conclude that late events in IgM polymer assembly, specifically the insertion of J chain, the exclusion of an additional monomeric subunit, and the covalent closure of the pentameric IgM molecule, determine the polymeric structure and, consequently, the biological activity of secreted IgM.

Animals↗
PubMed1996

Quality control in protein biogenesis: thiol-mediated retention monitors the redox state of proteins in the endoplasmic reticulum.

There is accumulating evidence that proteins can be retained in the endoplasmic reticulum by a mechanism that is believed to monitor the oxidation status of one or more cysteines in their sequences. For example, a single cysteine residue critical for retention of secretory IgM assembly intermediates has been mapped to the C-terminal cysteine, Cys575, of the secretory mu chain. Little is known concerning the mechanism responsible for this system of quality control, which has been termed thiolmediated retention. In particular, it is not known if the mechanism monitors the redox state of the important cysteine residue in the secretory mu protein itself or within the context of higher-order IgM complexes. To address this question, we evaluated the fidelity of retention of secretory IgM and determined the redox status of cysteines in secretory mu proteins in polymers and polymer intermediates at various stages of maturation. We demonstrate that all secreting B cells and B cell lines secrete assembly intermediates in addition to completed, covalent pentameric and hexameric IgM polymers. A fraction of assembly intermediates exit the endoplasmic reticulum as individual components, mature through the Golgi without undergoing further assembly, and most, if not all, are secreted. While the majority of IgM assembly intermediates have exposed thiols and are contained within the endoplasmic reticulum where they can be utilized for oligomerization, maturing assembly intermediates found in the Golgi and extracellular space are completely oxidized. Thus, while the retention of unpolymerized IgM is highly efficient, the retention system lacks the ability to distinguish fully oxidized assembly intermediates from fully oxidized completed polymers. The molecular mechanisms that may contribute to this aspect of IgM biogenesis and their implications for the concept of thiolmediated retention are discussed.

Alkylation↗
PubMed1995

Differential susceptibility to tolerance induction in vitro of splenic B cells from several transgenic mouse lines. Role of B1 cells.

Spleen cells from transgenic mice, whose rearranged Ig receptors reflect the repertoires of B1 (CD5+ and "sister" B cells) or normal B2 cells, were examined for their ability to be rendered unresponsive. By using an anti-Ig tolerance protocol that is independent of receptor specificity, we previously reported that peritoneal B cells, containing primarily CD5+ and "sister" B cells, were not susceptible to unresponsiveness. Herein, we show that splenic B cells from two separate transgenic mouse lines, each expressing a B1-type receptor, are resistant to tolerance induction in vitro. In contrast, splenic B cells from two other transgenic mouse lines with a large representation of conventional B cells were sensitive to anti-lg-mediated unresponsiveness. This difference does not reside in the surface Ig density, cell cycle, or activation stage of these cells, but is reflected in the initial calcium mobilization and tyrosine phosphorylation after surface Ig cross-linking. Therefore, these results support the hypothesis that the antigenic specificity of B cell receptors may drive cells toward the B1 subset, as suggested by Cong et al. (Cong, Y-Z., E. Rabin, and H. H. Wortis. 1991. Int. Immunol. 3:467-476), and that B1 cell characteristics confer the ability of B cells to withstand in vitro tolerance induction, irrespective of their anatomical location. The possibility that this results from previous antigenic experience is discussed.

Animals↗
PubMed1995

Roles of heavy and light chains in IgM polymerization.

IgM antibodies are secreted as multisubunit polymers that consist of as many as three discrete polypeptides: mu heavy chains, light (L) chains, and joining (J) chains. We wished to determine whether L chains that are required to confer secretory competence on immunoglobulin molecules must be present for IgM to polymerize--that is, for intersubunit disulfide bonds to form between mu chains. Using a L-chain-loss variant of an IgM-secreting hybridoma, we demonstrated that mu chains were efficiently polymerized independent of L chains, in a manner similar to that observed for conventional microL complexes, and that the mu polymers incorporated J chain. These mu polymers were not secreted but remained associated with the endoplasmic reticulum-resident chaperone BiP (GRP78). This finding is consistent with the endoplasmic reticulum being the subcellular site of IgM polymerization. We conclude that mu chain alone has the potential to direct the polymerization of secreted IgM, a process necessary but not sufficient for IgM to attain secretory competence.

Animals↗
PubMed1994

Mechanism and subcellular localization of secretory IgM polymer assembly.

The predominant functions of secreted IgM, complement fixation and transcytosis across epithelial barriers via the poly(Ig) receptor, are properties of IgM polymers; thus, the mechanisms that regulate oligomerization are critical to immune function. We have developed methods to assess the mechanism and subcellular location of IgM polymer formation. Using denaturing agarose/SDS-polyacrylamide gel electrophoresis and sucrose gradient fractionation, we demonstrate that IgM polymers are assembled in a stepwise fashion in which primary intermediates, including heavy chain-light chain complexes, are sequentially incorporated. Assembly intermediates include both covalent and non-covalent components, suggesting that IgM subunit interactions precede covalent assembly. IgM polymers are assembled from primary intermediates containing immature N-linked oligosaccharides. Polymerization is insensitive to brefeldin A and occurs at temperatures that inhibit protein transport along the secretory pathway. Together, these data demonstrate that IgM polymerization occurs early in the secretory pathway, most likely in the endoplasmic reticulum. These results are consistent with a model in which secretory IgM represents an oligomeric protein that is retained in the endoplasmic reticulum and does not mature along the secretory pathway until complete assembly is achieved.

Animals↗
PubMed1994

IgM hexamers?

There has been a universal tendency to regard IgM antibodies as pentameric molecules comprising five immunoglobulin monomeric subunits joined by a single J-chain protein. Is this the only form of secreted IgM, or are the possibilities more complex? In this article, Joseph Brewer and colleagues propose that the IgM polymers secreted in primary immune responses may be more heterogeneous than previously believed and that, as a consequence, the biological activity of IgM may have considerable flexibility.

Animals↗
PubMed1993

MHC class II limits the functional expression of endogenous superantigens in B cells.

The open reading frames of the 3' long terminal repeat of mouse mammary tumor viruses (MMTV) encode superantigens. When expressed with MHC class II molecules on the surface of B cells, superantigens stimulate T cells expressing receptors with particular V beta elements. By using B cell lymphomas as model systems, we demonstrate that stimuli, including certain lymphokines (IL-2 and IL-5) and LPS, that lead to increased MMTV transcript levels in B cells did not increase functional superantigen presentation. Furthermore, stimulation of BCL1 and CH12 cells with LPS resulted in a pronounced reduction in superantigen presentation, even though MMTV transcript levels were increased. In contrast to these effects, functional superantigen expression was increased under conditions in which class II levels were increased, even when MMTV levels remained constant, indicating that the levels of newly synthesized class II are important for functional superantigen expression. These data together imply that the amount of cell surface class II limits superantigen presentation and/or that superantigens can associate only with a limited pool of class II.

Animals↗
PubMed1993

Interleukin-5 (IL-5) and IL-6 define two molecularly distinct pathways of B-cell differentiation.

Interleukin-5 (IL-5) and IL-6 have both been reported to act as B-cell differentiation factors by stimulating activated B cells to secrete antibody. However, it has not been possible to directly compare the effects of these two lymphokines because of the lack of a suitable B-cell line capable of responding to both. We have identified a clonal, inducible B-cell lymphoma, CH12, that has this property. Both IL-5 and IL-6 can independently stimulate increases in steady-state levels of immunoglobulin and J-chain mRNA and proteins, and they both induce the differentiation of CH12 into high-rate antibody-secreting cells. Nevertheless, there are significant differences in the activities of these two lymphokines. First, while IL-6 acts only as a differentiation factor, IL-5 also augments the proliferation of CH12 cells. Second, the differentiation stimulated by IL-5 but not by IL-6 is partially inhibited by IL-4. Inhibition of IL-5-induced differentiation was not at the level of IL-5 receptor expression, since IL-4 did not inhibit IL-5-induced proliferation. Third, IL-5 but not IL-6 stimulated increased mouse mammary tumor proviral gene expression in CH12 cells. These results demonstrate that while both IL-5 and IL-6 may act as differentiation factors for B cells, they induce differentiation by using at least partially distinct molecular pathways. Our results also establish that B cells characteristic of a single stage of development can independently respond to IL-4, IL-5, and IL-6.

Animals↗
PubMed1992

Direct evidence that J chain regulates the polymeric structure of IgM in antibody-secreting B cells.

IgM is secreted in two functional polymeric forms. Secreted IgM was originally thought to be exclusively a pentameric molecule containing J (joining) chain, but many B cells also secrete hexameric IgM lacking J chain. Hexameric IgM may play an important role in the immune system, since it is up to 20 times more active than pentameric IgM in initiating the complement cascade. The predominant polymeric form of IgM secreted by B cell lines, either pentameric or hexameric, correlates with the concentration of J chain present during polymerization, and cells that express high levels of J chain secrete mostly IgM pentamers. The B cell lymphoma WEHI-231 does not express J chain, and the majority of its secreted IgM is polymerized as hexamers. When a J chain-encoding cDNA was expressed in these cells, the secreted IgM was found to be almost exclusively pentameric. However, although the expression of J chain dramatically altered the phenotype of the IgM secreted by these cells, it had little effect on their secretory rate. We conclude that J chain regulates the structure and function of the IgM polymers secreted by B cells, but it is not necessary for either IgM polymerization or secretion.

Amino Acid Sequence↗
PubMed1992

Major histocompatibility complex-restricted recognition of retroviral superantigens by V beta 17+ T cells.

It has been established that at least some V beta 17+ T cells interact with an endogenous superantigen encoded by the murine retrovirus, Mtv-9. To analyze the role of major histocompatibility complex (MHC) class II molecules in presenting the Mtv-9 encoded superantigen, vSAG-9 to V beta 17+ hybridomas, a panel of nine hybridomas was tested for their ability to respond to A20/2J (H-2d) and LBK (H-2a) cells which had been transfected with the vSAG-9 gene. Whereas some of the hybridomas recognized vSAG-9 exclusively in the context of H-2a, other hybridomas recognized vSAG-9 exclusively in the context of H-2d or in the context of both H-2d and H-2a. These results suggest that: (a) the class II MHC molecule plays a direct role in the recognition of retroviral superantigen by T cells, rather than serving simply as a platform for presentation; and, (b) it is likely that components of the TCR other than V beta are involved in the vSAG-9/TCR/class II interaction.

Animals↗
PubMed1992

J chain synthesis and secretion of hexameric IgM is differentially regulated by lipopolysaccharide and interleukin 5.

Two functional polymeric forms of IgM can be produced by antibody-secreting B cells. Hexameric IgM lacks detectable J (joining) chain and activates complement 17-fold better than pentameric IgM, which usually contains one J chain per pentamer. Using the inducible B-cell lymphoma CH12, we determined if the synthesis of a particular polymeric form of IgM is a fixed property of B cells or can be altered. Lipopolysaccharide (LPS)-stimulated CH12 cells produced mixtures of IgM hexamers and pentamers, resulting in antibody with high complement-fixing activity. In contrast, interleukin-5-stimulated CH12 cells secreted predominantly pentameric IgM, with a correspondingly lower lytic activity. Differences in lytic activity were due only to the amount of hexameric IgM in the secreted antibody. Interleukin 5 stimulated higher production of J chain RNA and protein than LPS, while LPS induced the highest levels of the secretory form of mu protein. The amount of hexameric IgM secreted was therefore inversely proportional to the level of intracellular J chain protein in the responding B cells. We conclude that the biologic function of IgM produced by B cells differs depending on how they are stimulated and that this difference may be regulated by the relative availabilities of J chain and secretory mu proteins during IgM polymerization.

Animals↗