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J E Sims

Publications and source records attributed to J E Sims.

At least 55 records · Page 3Linked to original sources
PubMed1993

Comparison of IL-1 alpha effectiveness in activating murine pre-B and T cell lines.

Interleukin-1 (IL-1) is a potent agent that induces a wide range of biological effects. The action of IL-1 is mediated by surface IL-1 receptors (IL-1R). Two types of IL-1 receptors have been identified in lymphocytes. In this study we examined activity of IL-1 alpha in two murine lymphocyte lines that express different types of IL-1 receptors. The T lymphoid cell line EL-4 6.1 C10 expresses type I IL-1R that mediates IL-1 alpha-induced IL-2 gene expression and secretion of IL-2. The pre-B lymphoid cell line 70Z/3 was previously shown to express type II IL-1R and responds to IL-1 alpha by expressing immunoglobulin kappa light chain mRNA and increased levels of surface IgM. We found that IL-1 alpha was as potent in inducing IgM expression in 70Z/3 cells as it was in inducing IL-2 secretion in EL-4 6.1 C10 cells. Likewise, the IL-1 alpha concentration sufficient to trigger kappa light chain gene expression in 70Z/3 cells was similar to the concentration of IL-1 alpha sufficient to trigger IL-2 gene expression in EL-4 6.1 C10. In both cell lines, IL-1 alpha activated NF-kappa B-like DNA-binding activity but in EL-4 6.1 C10 cells the IL-1 alpha concentration sufficient to induce NF-kappa B response was 1000-fold lower than in 70Z/3 cells. Monoclonal antibody, mAb M15, to the type I IL-1R blocked IL-1-induced responses in EL-4 6.1 C10 cells. Surprisingly mAb M15 also blocked IL-1 action in 70Z/3 cells, even though these cells predominantly express type II IL-1R. 15% of the total IL-1 binding sites in 70Z/3 cells were recognized by mAb M15. Human IL-1 receptor antagonist (IL-1ra), which binds to the natural murine type I but not the type II IL-1R, blocked IL-1 alpha responses in EL-4 6.1 C10 and 70Z/3 cells. Although at low levels, Northern blot analysis confirmed that 70Z/3 cells express low levels of type I IL-1R mRNA. Taken together, these results suggest that type I IL-1R are expressed and transduce IL-1 signals in both 70Z/3 and El-4 6.1 C10 cells.

Animals↗
PubMed1993

Carney's complex in a patient with hormone-producing Sertoli cell tumor of the testicle.

Carney's complex is an unusual disorder consisting of a variety of endocrinological and urological manifestations. The characteristic gonadal and adrenal features of Carney's complex should become familiar to urologists. A patient was evaluated for obesity, cushingoid features, hyperlipidemia, glucose intolerance, coronary artery disease, a left adrenal mass, bilateral testicular masses and cardiac myxomas. Pathological evaluation revealed the testicular tumors to be of Sertoli cell origin, the adrenal mass to be an adrenocortical adenoma and intracardiac lesions consistent with myxomas. The features of Carney's complex are discussed.

Adrenal Glands↗
PubMed1992

Vaccinia and cowpox viruses encode a novel secreted interleukin-1-binding protein.

Supernatants from vaccinia virus (VV)-infected CV-1 cells were examined and found to contain a 33 kd protein capable of binding murine interleukin-1 beta (mIL-1 beta). A VV open reading frame (ORF) that exhibits 30% amino acid identity to the type II IL-1 receptor was expressed in CV-1-EBNA cells and shown specifically to bind mIL-1 beta. A similar ORF from cowpox virus was expressed and also specifically bound mIL-1 beta. A recombinant VV was constructed in which this ORF was disrupted (vB15RKO). Supernatants from vB15RKO-infected cells did not contain an IL-1-binding protein. Supernatants from VV-infected CV-1 cells were capable of inhibiting IL-1-induced murine lymphocyte proliferation in vitro while supernatants from vB15RKO infected cells did not. Intracranial inoculation of mice with vB15RKO suggests that this ORF is involved in VV virulence. The possible role of a virus-encoded IL-1-binding protein in the pathology of a poxvirus infection and its relationship to other poxvirus-encoded immune modulators is discussed.

Animals↗
PubMed1992

Pattern of IL-1 receptor gene expression suggests role in noninflammatory processes.

The cytokine IL-1 can be elaborated by most nucleated cell types and exerts its pleiotropic effects on a variety of tissues through binding to its cognate receptor. Two forms of IL-1R, designated types I and II, have been identified. These proteins share limited amino acid identity but both bind IL-1 alpha and IL-1 beta. A large number of cell types have been shown to possess IL-1R in vitro, but few studies have addressed the question of in vivo expression. Using in situ hybridization in normal adult BALB/c mice, we have examined the tissue distributions of both IL-1R types. The results demonstrate that although lymphoid tissues in healthy animals express low or nondetectable levels of receptor transcript, nonlymphoid tissues constitutively express readily detectable levels of IL-1R mRNA. Thymus and spleen samples were largely negative for both IL-1R types I and II, whereas half of the lymph nodes examined expressed IL-1R type I. In nonlymphoid tissues, IL-1R type I transcript was detected in the dentate gyrus of the brain, endocrine pancreas, cardiac endothelium, epidermis, dermis, hair follicle epithelium, uterine serosa, vaginal stroma, vaginal squamous epithelium, developing oocytes, and granulosa cells of ruptured follicles. In contrast, the IL-1R type II probe hybridized to epidermis, dermis, and vaginal basal epithelium. The two types of IL-1R were rarely, if ever, co-expressed by the same cell. The distributional segregation of the receptor types and the expression of IL-1R in normal nonlymphoid tissues has broad implications for the role of IL-1 in homeostatic physiology.

Animals↗
PubMed1992

Transrectal ultrasound and biopsy in diagnosis of malakoplakia of prostate.

We report our experience with the use of transrectal ultrasound and ultrasound-guided needle biopsy in the diagnosis of 4 cases of malakoplakia of the prostate. Prostatic malakoplakia is rare with a total of 25 cases reported in the literature. It may simulate carcinoma on digital rectal examination and transrectal ultrasound. Herein are 4 additional cases in which transrectal ultrasound of the prostate was compatible with carcinoma. Subsequent needle biopsy of the suspicious regions revealed the classic pathologic features of malakoplakia. Malakoplakia is a difficult pathologic diagnosis, and the prevalence of this disease may be underestimated. As the use of transrectal ultrasound becomes more common in diagnosing carcinoma of the prostate, and due to the difficulty in diagnosing malakoplakia pathologically, we recommend maintaining a high index of suspicion for malakoplakia to avoid possible unnecessary radical surgery.

Adult↗
PubMed1992

Evidence that MAP (mitogen-activated protein) kinase activation may be a necessary but not sufficient signal for a restricted subset of responses in IL-1-treated epidermoid cells.

We have investigated the activation of mitogen-activated protein kinase (MAP-kinase) in KB human epidermoid carcinoma cells treated with interleukin 1 (IL-1). MAP-kinase activity was transient; the time required for activity to reach a maximal level was dependent upon the dose of IL-1, ranging from 15 minutes to 45 minutes. The level of kinase induction correlated well with dose-response curves for two characteristic IL-1-induced responses, PGE2 and IL-6 production. MAP-kinase activity returned to basal levels within 2 hours regardless of the amount of IL-1 added to the system. Exposure of KB cells to free IL-1 was accordingly restricted to periods of 2 hours or less, by replacing IL-1 with an excess of IL-1 receptor antagonist. Even after 2 hours exposure, the ability of IL-1 to induce IL-6 or PGE2 was still IL-1ra-inhibitable by more than 80%, suggesting that events downstream of, or parallel to MAP-kinase activation, requiring the continual formation of new IL-1 receptor complexes, are needed to fully elicit these responses. Two general serine/threonine kinase inhibitors, K252a and quercetin, were found to strongly inhibit MAP kinase in vivo with ED50s of c. 100 nM and 30 microM, respectively. At these concentrations, both compounds effectively inhibited IL-1-driven PGE2 and IL-6 induction without affecting general protein synthesis or secretion. Other non-selective kinase inhibitors had less effect on MAP-kinase activation or IL-1-induced biological responses. The transient activation of MAP-kinase induction correlated strikingly with activation of the transcription factor NF-kappa B. IL-1-induced NF-kappa B activation was, however, relatively insensitive to inhibition by K252a or quercetin. We suggest that MAP-kinase is likely to be a necessary, but not sufficient, intermediate in some (IL-6, PGE2 induction) but not all (NF-kappa B activation) IL-1 responses in these cells.

Base Sequence↗
PubMed1992

Molecular characterization of the interleukin-1 receptor (IL-1R) on monocytes and polymorphonuclear cells.

Primary human monocytes and monocytic cells express an interleukin 1 receptor (IL-1R) which is similar in molecular weight and IL-1 binding characteristics to the IL-1R expressed on B lymphocytes (type II). Northern blot analysis of monocytic cells using a cDNA probe from the recently isolated type II IL-1R indicates that this mRNA is detectable by 4 h and accumulates for at least 24 h following treatment with IL-1R inducing drugs. The time course of induction of this mRNA is slower than that of the type I IL-1R mRNA which is also transcribed in monocytic cells but does not appear to be translated. Sequence analysis of a monocyte-derived cDNA corresponding to the type II IL-1R mRNA shows that the monocyte and B-cell mRNAs are identical. Comparison of monocyte IL-1R peptide maps with those of the type II IL-1R suggests that the two surface IL-1R are identical. This was confirmed serologically using a polyclonal antiserum raised against the type II IL-1R. Data are presented which indicate that primary human neutrophils can also be induced to express abundant type II IL-1R.

Blotting, Northern↗
PubMed1991

A serine/threonine kinase activity is closely associated with a 65-kDa phosphoprotein specifically recognized by the kappa B enhancer element.

The immunoglobulin kappa light chain enhancer, kappa B, is an important cis-acting transcriptional element. kappa B binds a number of proteins including the members of the ubiquitous NF-kappa B family of transcription factors. Agarose beads coupled to a double-stranded oligonucleotide containing the kappa B motif were used to isolate a 65-kDa predominantly nuclear phosphoprotein. Southwestern blot analysis demonstrated that this phosphoprotein can bind the kappa B element directly and specifically. This kappa B-associated protein was phosphorylated in vivo and in vitro by a nuclear serine/threonine kinase(s) which, in a number of different cell lines, appeared to be stimulated in response to interleukin-1 alpha and lipopolysaccharide treatment. In the B cell lines 70Z/3 and CH12 LX2B, and the T cell line EL-4 6.1 C10 the activity of the kappa B-associated kinase(s) correlated with the binding activity of nuclear NF-kappa B displayed in a gel shift assay. In vitro, the 65-kDa protein was phosphorylated in the absence of exogenously added kinase. The 65-kDa phosphoprotein and the kinase activity remained associated following sequential anion-exchange and hydrophobic interaction chromatography. These results suggest that the kappa B-associated phosphoprotein is either autophosphorylated or is phosphorylated by a closely associated kinase(s). Stimulation of a nuclear protein kinase which is closely associated with a sequence-specific DNA may reflect a novel mechanism by which growth factors regulate gene expression.

Base Sequence↗
PubMed1991

Phorbol ester induces phosphorylation of the 80 kilodalton murine interleukin 1 receptor at a single threonine residue.

The cytoplasmic domains of some cell surface receptors become phosphorylated in cells treated with phorbol esters. The present study was undertaken in order to determine whether this is also true of the 80 kDa interleukin 1 receptor (IL1R). Recombinant murine IL1R, transfected into chinese hamster ovary (CHO) cells or murine fibroblasts, was immunoprecipitated from [32P]orthophosphate-labelled cells. IL1R phosphorylation was only detected in cells pretreated with phorbol 12-myristate 13-acetate (PMA) and occurred solely on phosphothreonine. In contrast to a previous report, little or no IL1R phosphorylation occurred in response to IL1. By using a truncated receptor and receptors in which threonine residues were changed to alanines, we established that Thr537, near the carboxy-terminus, is the major site of PMA-induced phosphorylation. The human IL1R has a different sequence at this locus, and is apparently not phosphorylated. Binding studies showed that PMA-induced phosphorylation had no discernible effect on ligand binding or internalization.

Amino Acid Sequence↗
PubMed1991

IL-1 signal transduction pathways. I. Two functional IL-1 receptors are expressed in T cells.

We have recently characterized a subline of the Th2 cell line D10.G4.1, D10A, which responds to exogenous IL-1 in the absence of cofactors. In this cell line IL-1 activates two distinct transmission signal pathways, one leading to increased levels of intracellular cAMP, and the other to the phosphorylation of an 80-kDa substrate of protein kinase C (PKC). To determine whether both pathways are activated upon occupancy of a single IL-1R, we used a mAb, M15, which blocks binding of IL-1 to the 80-kDa IL-1R, known to be expressed in Th2 cells. Whereas M15 was able to inhibit the accumulation of cAMP induced by IL-1, it had no effect on the IL-1-induced phosphorylation of the 80-kDa substrate of PKC or the c-fos mRNA expression. In addition, we show that IL-1 induces the expression of the two proto-oncogenes c-myb and c-myc through the 80-kDa IL-1R, and that this induction is PKC independent. The proliferation of D10A cells in response to IL-1 is blocked by M15, but is unaffected by H-7, a potent inhibitor of PKC. These results indicate that the IL-1-induced proliferation of D10A cells is independent of PKC. In addition, IL-1 induces c-fos mRNA expression in thymocytes but not in EL-4 cells.

Animals↗
PubMed1991

Chromosomal location of murine and human IL-1 receptor genes.

The gene for the type I interleukin-1 (IL-1) receptor has been mapped in both mouse and human. In the human genome, a combination of segregation analysis of rodent-human hybrid cells and chromosomal in situ hybridization has placed the gene on the long arm of chromosome 2, at band 2q12. This is near the reported map position of the loci for IL-1 alpha and IL-1 beta (2q13----2q21). The murine gene has been mapped by analysis of restriction fragment length polymorphisms in interspecific backcrosses to the centromeric end of chromosome 1, in a region that is syntenic to a portion of human chromosome 2. The murine Il-1r1 gene has thus been separated from the IL-1 genes, which lie on murine chromosome 2.

Animals↗
PubMed1990

Induction of an interleukin-1 receptor (IL-1R) on monocytic cells. Evidence that the receptor is not encoded by a T cell-type IL-1R mRNA.

Primary human monocytes and the human monocytic cell line THP-1 were induced to express receptors for interleukin-1 alpha (IL-1 alpha) and IL-1 beta. Treatment of primary monocytes with dexamethasone resulted in a 10-fold increase in receptor number over untreated cells, to approximately 2,000 receptors/cell. Treatment of THP-1 cells with phorbol ester followed by prostaglandin E2 and dexamethasone resulted in the expression of approximately 30,000 receptors/cell. Competitive binding assays on THP-1 cells showed that both IL-1 alpha and IL-1 beta bind to the same receptor. The monocyte IL-1R is significantly smaller (63 kDa) than the T cell IL-1R (80 kDa) and is immunologically distinct. However, induction of monocytes and monocytic cell lines leads to the appearance of an abundant mRNA of approximately 5,000 bases which hybridizes to a cDNA probe from the T cell-type IL-1R. Sequence data obtained from a cDNA clone of this mRNA indicate that the message is identical to the T cell IL-1R mRNA throughout the coding region. A smaller mRNA, also homologous to the T cell IL-1R mRNA, accumulated in induced THP-1 cells and has a shorter 3'-untranslated region than the larger. Data are presented which suggest that neither form of this message encodes the 63-kDa IL-1R, but rather that this protein is the product of a separate nonhomologous mRNA.

Cell Line↗
PubMed1990

Expression cloning of a human granulocyte colony-stimulating factor receptor: a structural mosaic of hematopoietin receptor, immunoglobulin, and fibronectin domains.

We report the isolation from a placental library, of two cDNAs that can encode high affinity receptors for granulocyte colony-stimulating factor (G-CSF) when expressed in COS-7 cells. The cDNAs are predicted to encode integral membrane proteins of 759 and 812 amino acids in length. The predicted extracellular and membrane spanning sequences of the two clones are identical, as are the first 96 amino acids of their respective cytoplasmic regions. Different COOH termini of 34 or 87 residues are predicted for the two cDNAs, due apparently to alternate splicing. The receptor with the longer cytoplasmic domain is the closest human homologue of the murine G-CSF receptor recently described by Fukunaga et al. (Fukunaga, R., E. Ishizaka-Ikeda, Y. Seto, and S. Nagata. 1990. Cell. 61:341). A hybridization probe derived from the placental G-CSF receptor cDNA detects a approximately 3-kb transcript in RNAs isolated from placenta and a number of lymphoid and myeloid cells. The extracellular region of the G-CSF receptors is composed of four distinct types of structural domains, previously recognized in other cell surface proteins. In addition to the two domains of the HP receptor family-defining region (Patthy, L. 1990. Cell. 61:13) it incorporates one NH2-terminal Ig-like domain, and three additional repeats of fibronectin type III-like domains. The presence of both an NH2-terminal Ig-like domain and multiple membrane-proximal FN3-like domains suggests that the G-CSF receptor may be derived from an ancestral NCAM-like molecule and that the G-CSF receptor may function in some adhesion or recognition events at the cell surface in addition to the binding of G-CSF.

Amino Acid Sequence↗
PubMed1990

Regulation of alloreactivity in vivo by a soluble form of the interleukin-1 receptor.

In vitro studies have shown that cytokines are involved in the regulation of the immune response, but their role in vivo is less well defined. Specific cytokine antagonists enable the identification of particular cytokines involved in the response and offer a means for modifying it. Systemic administration of a soluble, extracellular portion of the receptor for interleukin-1 (sIL-1R) had profound inhibitory effects on the development of in vivo alloreactivity. Survival of heterotopic heart allografts was prolonged from 12 days in controls to 17 days in mice treated with sIL-1R. Lymph node hyperplasia in response to a localized injection of allogeneic cells was completely blocked by sIL-1R treatment. The inhibition was overcome by simultaneous administration of interleukin-1 (IL-1); thus, sIL-1R acts by neutralizing IL-1. These results implicate IL-1 as a regulator of allograft rejection and demonstrate the in vivo biological efficacy of a soluble cytokine receptor.

Animals↗
PubMed1990

Biology of the interleukin-1 receptor.

The biological effects of the two interleukin-1s on cells of connective tissue origin are mediated by specific cell-surface receptors. Molecular cloning studies have revealed that these receptors are identical in protein sequence to the IL-1 receptors on cells of the T-lymphocyte lineage. The functional interleukin-1 receptor on T-cells and fibroblasts is composed of a single polypeptide chain that binds both IL-1 alpha and IL-1 beta. The single chain appears to be all that is required to transduce a signal to cells. While the nature of the signal is unknown, the structure of the receptor is inconsistent with its possessing any protein tyrosine kinase activity. It is therefore not surprising that the mitogenic activity of IL-1 for fibroblasts is mediated by IL-1 induction of PDGF-A gene transcription. Finally, IL-1 is known to modulate fibroblast-matrix interactions in several ways. It is interesting therefore, that the majority of the IL-1 receptors on cultured fibroblasts are clustered into focal adhesions.

Animals↗
PubMed1990

Molecular heterogeneity of interleukin-1 receptors.

Previous studies have shown that binding of IL-1 to its receptor and intracellular processing of the IL-1/IL-1 receptor complex appear to be different in B- and T-cells. The current report summarizes recent studies from our laboratory that show that the murine and human IL-1 receptors present on T-cells and fibroblasts are identical in primary sequence within each species, and highly similar even when the two species are compared. At present no cDNA clones have been isolated for IL-1 receptors present on B-cells. However, a monoclonal antibody raised to the receptor on murine T-lineage cells did not bind to a pre-B-lymphoma cell line that displays IL-1 binding sites, nor would cDNA probes derived from a T-cell IL-1 receptor clone cross hybridize at high stringency to mRNA prepared from these cells. In addition the two receptors differ substantially in size, as determined by affinity crosslinking with radiolabeled IL-1 alpha. Taken together, these observations show that major structural differences exist between the IL-1 receptors on B and T lymphocytes, while the receptors on T-cells and fibroblasts are identical polypeptides. We propose that the T-cell/fibroblast receptor be called IL-1RI and the B-cell type IL-1RII.

Animals↗