Analysis of mouse LMIR5/CLM-7 as an activating receptor: differential regulation of LMIR5/CLM-7 in mouse versus human cells

Blood ◽  
2008 ◽  
Vol 111 (2) ◽  
pp. 688-698 ◽  
Author(s):  
Yoshinori Yamanishi ◽  
Jiro Kitaura ◽  
Kumi Izawa ◽  
Takayuki Matsuoka ◽  
Toshihiko Oki ◽  
...  
Keyword(s):  
Mast Cells ◽  
Cytokine Production ◽  
Fetal Liver ◽  
Human Cells ◽  
Cross Linking ◽  
Wild Type ◽  
Predominant Role ◽  
Innate Immunity Cells ◽  
Production Cell ◽  
Activating Receptor

We have analyzed leukocyte mono-Ig–like receptor 5 (LMIR5) as an activating receptor among paired LMIRs. Mouse LMIR5 (mLMIR5) is expressed in myeloid cells such as mast cells, granulocytes, macrophages, and dendritic cells. Cross-linking of transduced mLMIR5 in bone marrow–derived mast cells (BMMCs) caused activation events, including cytokine production, cell survival, degranulation, and adhesion to the extracellular matrix. mLMIR5 associated with DAP12 and to a lesser extent with DAP10, and mLMIR5-mediated functions of BMMCs were strongly inhibited by DAP12 deficiency. Importantly, cross-linking of endogenous mLMIR5 induced Syk-dependent activation of fetal liver–derived mast cells. Unlike mLMIR5, cross-linking of human LMIR5 (hLMIR5) induced cytokine production of BMMCs even in the absence of both DAP12 and DAP10, suggesting the existence of unidentified adaptors. Interestingly, hLMIR5 possessed a tyrosine residue (Y188) in the cytoplasmic region. Signaling via Y188 phosphorylation played a predominant role in hLMIR5-mediated cytokine production in DAP12-deficient, but not wild-type BMMCs. In addition, experiments using DAP10/DAP12 double-deficient BMMCs suggested the existence of Y188 phoshorylation-dependent and -independent signals from unidentified adaptors. Collectively, although both mouse and human LMIR5 play activatory roles in innate immunity cells, the functions of LMIR5 were differentially regulated in mouse versus human cells.

Analysis of Mouse LMIR5/CLM-7 as An Activating Receptor: Differential Regulation of LMIR5/CLM-7 in Mouse Versus Human Cells

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4639-4639
Author(s):  
Yoshinori Yamanishi ◽  
Jiro Kitaura ◽  
Kumi Izawa ◽  
Toshihiko Oki ◽  
Toshiyuki Takai ◽  
...  
Keyword(s):  
Mast Cells ◽  
Cytokine Production ◽  
Fetal Liver ◽  
Human Cells ◽  
Cross Linking ◽  
Wild Type ◽  
Predominant Role ◽  
Innate Immunity Cells ◽  
Production Cell ◽  
Activating Receptor

Abstract We have analyzed leukocyte mono-Ig–like receptor 5 (LMIR5) as an activating receptor among paired LMIRs. Mouse LMIR5 (mLMIR5) is expressed in myeloid cells such as mast cells, granulocytes, macrophages, and dendritic cells. Cross-linking of transduced mLMIR5 in bone marrow–derived mast cells (BMMCs) caused activation events, including cytokine production, cell survival, degranulation, and adhesion to the extracellular matrix. mLMIR5 associated with DAP12 and to a lesser extent with DAP10, and mLMIR5-mediated functions of BMMCs were strongly inhibited by DAP12 deficiency. Importantly, cross-linking of endogenous mLMIR5 induced Syk-dependent activation of fetal liver–derived mast cells. Unlike mLMIR5, cross-linking of human LMIR5 (hLMIR5) induced cytokine production of BMMCs even in the absence of both DAP12 and DAP10, suggesting the existence of unidentified adaptors. Interestingly, hLMIR5 possessed a tyrosine residue (Y188) in the cytoplasmic region. Signaling via Y188 phosphorylation played a predominant role in hLMIR5-mediated cytokine production in DAP12-deficient, but not wild-type BMMCs. In addition, experiments using DAP10/DAP12 double-deficient BMMCs suggested the existence of Y188 phoshorylation-dependent and -independent signals from unidentified adaptors. Collectively, although both mouse and human LMIR5 play activatory roles in innate immunity cells, the functions of LMIR5 were differentially regulated in mouse versus human cells.

Apaf-1 and caspase-9 are required for cytokine withdrawal-induced apoptosis of mast cells but dispensable for their functional and clonogenic death

Blood ◽  
2006 ◽  
Vol 107 (5) ◽  
pp. 1872-1877 ◽  
Author(s):  
Vanessa S. Marsden ◽  
Thomas Kaufmann ◽  
Lorraine A. O'Reilly ◽  
Jerry M. Adams ◽  
Andreas Strasser
Keyword(s):  
Mast Cells ◽  
Fetal Liver ◽  
Wild Type ◽  
Caspase 9 ◽  
Receptor Stimulation ◽  
Apoptotic Pathway ◽  
Apoptotic Pathways ◽  
Induced Apoptosis ◽  
Cytokine Deprivation

Cytokines promote survival of mast cells by inhibiting apoptotic pathways regulated by the Bcl-2 protein family. We previously showed that lymphocyte apoptosis can proceed via a Bcl-2-inhibitable pathway independent of the canonical initiator caspase, caspase-9, and its adaptor, Apaf-1. Here we report that mast cells lacking caspase-9 or Apaf-1 are refractory to apoptosis after cytotoxic insults but still lose effector function and ability to proliferate. In response to cytokine deprivation or DNA damage, fetal liver-derived mast cells lacking Apaf-1 or caspase-9 failed to undergo apoptosis. Nevertheless, the cytokine-starved cells were not functionally alive, because, unlike those overexpressing Bcl-2, they could not degranulate on Fcϵ receptor stimulation or resume proliferation on re-addition of cytokine. Furthermore, mast cells lacking Apaf-1 or caspase-9 had no survival advantage over wild-type counterparts in vivo. These results indicate that the Apaf-1/caspase-9-independent apoptotic pathway observed in lymphocytes is ineffective in cytokine-deprived mast cells. However, although Apaf-1 and caspase-9 are essential for mast cell apoptosis, neither is required for the functional or clonogenic death of the cells, which may be due to mitochondrial dysfunction.

scholarly journals Leukocyte common antigen (CD45) is required for immunoglobulin E-mediated degranulation of mast cells.

1994 ◽  
Vol 180 (2) ◽  
pp. 471-476 ◽  
Author(s):  
S A Berger ◽  
T W Mak ◽  
C J Paige
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Immunoglobulin E ◽  
Tyrosine Phosphatase ◽  
Cross Linking ◽  
Ionophore A23187 ◽  
Wild Type ◽  
Ige Receptor ◽  
High Affinity ◽  
Deficient Mice

We demonstrate using primary mast cell cultures derived from wild-type and CD45-deficient mice that mast cell triggering through the high-affinity immunoglobulin E (IgE) receptor requires the cell surface tyrosine phosphatase CD45. Unlike wild-type cells, cross-linking of surface-bound IgE in mast cells deficient in CD45 does not induce degranulation. Degranulation in these mutant cells does occur after treatment with the calcium ionophore A23187 indicating that the degranulation machinery is intact in these cells. We also demonstrate that the tyrosine phosphatase inhibitors orthoVanadate and perVanadate inhibit degranulation in wild-type mast cells, as does cross-linking of CD45 by anti-CD45 antibodies. Finally, we show that CD45-deficient mice are resistant to IgE-dependent systemic anaphylaxis. These results show that, like the T cell receptor and the antigen receptor on B cells, there is an absolute requirement for CD45 in signaling via the high affinity IgE receptor, expanding the number of receptors for which CD45 is an essential component.

Endogenous N-Terminal Truncated STAT5 Expressed from Alternative Start Codons Promotes SCF Signaling in Murine Primary Mast Cell Cultures.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 815-815
Author(s):  
Kevin D. Bunting ◽  
Christine Couldrey ◽  
Richard Moriggl ◽  
Yongzhi Cui ◽  
Harry Wright ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cell Cultures ◽  
Target Genes ◽  
Fetal Liver ◽  
Selective Advantage ◽  
Retroviral Vectors ◽  
Null Mouse ◽  
Wild Type

Abstract Signal transducer and activator of transcription (STAT5) has important functions in hematopoiesis. Our prior work with mice in which two coding exons of STAT5a and STAT5b have been deleted (STAT5ab−/−) has shown that mast cells derived in the presence of interleukin(IL)-3 and stem cell factor (SCF) have severe defects in survival and proliferation (Shelburne et al. Blood102:1290; 2002). However, the mechanism for STAT5 activation by SCF is unclear. In erythroid cells, STAT5 is not tyrosine phosphorylated by SCF alone. We have set out to determine whether some SCF-induced functions of STAT5 could be rescued in c-Kit+Fcγ+ primary bone marrow (BM) or fetal liver (FL) derived mast cell cultures from STAT5ab−/− mice. STAT5ab−/− mouse mast cells had a normal chemotactic response to SCF compared with wild-type mast cells, however we found they highly expressed N-terminal truncated STAT5 isoforms (STAT5ΔN) from position 102 and 136. In contrast, expression of STAT5ΔN in wild-type mast cells and some STAT5ab−/− tissues such as spleen and brain was barely detectable. While naturally occuring C-terminal STAT5 variants bind DNA but do not transactivate target genes, N-terminal variants of STAT5 have defective tetramerization. Generally, the N-terminus of STATs is also believed essential for function. Therefore, we have used mast cells to test whether N-terminal STAT5 mutants (STAT5ΔN1–136 or STAT5W37A) could functionally restore previously reported defects in vitro. Both of the STAT5 mutants were found to be dimerization-competent but tetramerization-deficient. MSCV-based retroviral vectors expressing these STAT5 mutants upstream of IRES-GFP were stably transduced into STAT5ab−/− mast cells. A strong selective advantage for GFP+ cells was observed for all vectors containing STAT5wt, STAT5ΔN, or STAT5W37A but not for IRES-GFP control. Inversely, in wild-type mast cells, only the STAT5ΔN mutant conferred a selective advantage, suggesting that the heterodimer (ΔN/wt) between STAT5ΔN and STAT5wt was most active. In apoptosis experiments, STAT5ΔN protected following cytokine withdrawal. We found the homodimer (ΔN/ΔN) to be intermediate in terms of growth reconstitution potential than the heterodimer. However, retroviral overexpression of the homodimer above that of the endogenous STAT5ΔN alone was sufficient to correct growth defects. To further explore the role of the endogenous STAT5ΔN, we obtained FL-derived mast cells from a new STAT5null/null mouse where the entire STAT5ab locus was deleted using the Cre-LoxP system (Cui et al. MCB, in press). These mast cells lacked STAT5 and STAT5ΔN. Unlike the STAT5ab−/− mast cells, the STAT5null/null mast cells showed a 9-fold reduction in SCF-induced chemotaxis relative to wild-type controls. Importantly, the chemotactic defects were completely rescued by gene transfer of wild-type STAT5a (P=0.018) or STAT5ΔN (P=0.02) relative to IRGFP control. Therefore, 1) endogenous STAT5ΔN is active but to a different degree depending on the specific function and cell type 2) co-expression along with full-length STAT5 confers maximal SCF-mediated responsiveness in mast cells. This work uncovers a potential mechanism by which STAT5-mediated SCF responses are regulated via formation of functional heterodimers.

Cytokine production in human intestinal mast cells triggered by Fc∈ receptor cross-linking is dependent on both protein kinase C and MAP kinase

2001 ◽  
Vol 120 (5) ◽  
pp. A497
Author(s):  
Axel Lorentz ◽  
Ilka Klopp ◽  
Thomas Gebhardt ◽  
Michael P. Manns ◽  
Stephan C. Bischoff
Keyword(s):  
Protein Kinase C ◽  
Mast Cells ◽  
Protein Kinase ◽  
Map Kinase ◽  
Cytokine Production ◽  
Fc Receptor ◽  
Cross Linking ◽  
Kinase C

scholarly journals Impaired degranulation but enhanced cytokine production after FcεRI stimulation of diacylglycerol kinase ζ–deficient mast cells

2006 ◽  
Vol 203 (6) ◽  
pp. 1471-1480 ◽  
Author(s):  
Benjamin A. Olenchock ◽  
Rishu Guo ◽  
Michael A. Silverman ◽  
Jennifer N. Wu ◽  
Jeffery H. Carpenter ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cytokine Production ◽  
Cell Function ◽  
Second Messengers ◽  
Diacylglycerol Kinase ◽  
Negative Regulator ◽  
Calcium Flux ◽  
Cross Linking

Calcium and diacylglycerol are critical second messengers that together effect mast cell degranulation after allergen cross-linking of immunoglobulin (Ig)E-bound FcεRI. Diacylglycerol kinase (DGK)ζ is a negative regulator of diacylglycerol-dependent signaling that acts by converting diacylglycerol to phosphatidic acid. We reported previously that DGKζ−/− mice have enhanced in vivo T cell function. Here, we demonstrate that these mice have diminished in vivo mast cell function, as revealed by impaired local anaphylactic responses. Concordantly, DGKζ−/− bone marrow–derived mast cells (BMMCs) demonstrate impaired degranulation after FcεRI cross-linking, associated with diminished phospholipase Cγ activity, calcium flux, and protein kinase C–βII membrane recruitment. In contrast, Ras-Erk signals and interleukin-6 production are enhanced, both during IgE sensitization and after antigen cross-linking of FcεRI. Our data demonstrate dissociation between cytokine production and degranulation in mast cells and reveal the importance of DGK activity during IgE sensitization for proper attenuation of FcεRI signals.

CCR5 deficiency decreases susceptibility to experimental cerebral malaria

Blood ◽  
2003 ◽  
Vol 101 (11) ◽  
pp. 4253-4259 ◽  
Author(s):  
Elodie Belnoue ◽  
Michèle Kayibanda ◽  
Jean-Christophe Deschemin ◽  
Mireille Viguier ◽  
Matthias Mack ◽  
...  
Keyword(s):  
T Cells ◽  
Cerebral Malaria ◽  
Cd8 T Cells ◽  
Cytokine Production ◽  
Wild Type ◽  
Experimental Cerebral Malaria ◽  
Pathologic Features ◽  
Th1 Cytokine ◽  
The Brain ◽  
Deficient Mice

Abstract Infection of susceptible mouse strains with Plasmodium berghei ANKA (PbA) is a valuable experimental model of cerebral malaria (CM). Two major pathologic features of CM are the intravascular sequestration of infected erythrocytes and leukocytes inside brain microvessels. We have recently shown that only the CD8+ T-cell subset of these brain-sequestered leukocytes is critical for progression to CM. Chemokine receptor–5 (CCR5) is an important regulator of leukocyte trafficking in the brain in response to fungal and viral infection. Therefore, we investigated whether CCR5 plays a role in the pathogenesis of experimental CM. Approximately 70% to 85% of wild-type and CCR5+/- mice infected with PbA developed CM, whereas only about 20% of PbA-infected CCR5-deficient mice exhibited the characteristic neurologic signs of CM. The brains of wild-type mice with CM showed significant increases in CCR5+ leukocytes, particularly CCR5+ CD8+ T cells, as well as increases in T-helper 1 (Th1) cytokine production. The few PbA-infected CCR5-deficient mice that developed CM exhibited a similar increase in CD8+ T cells. Significant leukocyte accumulation in the brain and Th1 cytokine production did not occur in PbA-infected CCR5-deficient mice that did not develop CM. Moreover, experiments using bone marrow (BM)–chimeric mice showed that a reduced but significant proportion of deficient mice grafted with CCR5+ BM develop CM, indicating that CCR5 expression on a radiation-resistant brain cell population is necessary for CM to occur. Taken together, these results suggest that CCR5 is an important factor in the development of experimental CM.

scholarly journals Prominent Indomethacin-Induced Enteropathy in Fcgriib Defi-cient lupus Mice: An Impact of Macrophage Responses and Immune Deposition in Gut

2021 ◽  
Vol 22 (3) ◽  
pp. 1377
Author(s):  
Thansita Bhunyakarnjanarat ◽  
Kanyarat Udompornpitak ◽  
Wilasinee Saisorn ◽  
Bhumdhanin Chantraprapawat ◽  
Peerapat Visitchanakun ◽  
...  
Keyword(s):  
Cytokine Production ◽  
High Dose ◽  
Flux Analysis ◽  
Wild Type ◽  
Fc Gamma Receptor ◽  
Serum Cytokines ◽  
Extracellular Flux ◽  
Gamma Receptor ◽  
Gut Leakage ◽  
Gastrointestinal Permeability

A high dose of NSAIDs, a common analgesic, might induce lupus activity through several NSAIDs adverse effects including gastrointestinal permeability defect (gut leakage) and endotoxemia. Indomethacin (25 mg/day) was orally administered for 7 days in 24-wk-old Fc gamma receptor IIb deficient (FcgRIIb-/-) mice, an asymptomatic lupus model (increased anti-dsDNA without lupus nephritis), and age-matched wild-type (WT) mice. Severity of indomethacin-induced enteropathy in FcgRIIb-/- mice was higher than WT mice as demonstrated by survival analysis, intestinal injury (histology, immune-deposition, and intestinal cytokines), gut leakage (FITC-dextran assay and endotoxemia), serum cytokines, and lupus characteristics (anti-dsDNA, renal injury, and proteinuria). Prominent responses of FcgRIIb-/- macrophages toward lipopolysaccharide (LPS) compared to WT cells due to the expression of only activating-FcgRs without inhibitory-FcgRIIb were demonstrated. Extracellular flux analysis indicated the greater mitochondria activity (increased respiratory capacity and respiratory reserve) in FcgRIIb-/- macrophages with a concordant decrease in glycolysis activity when compared to WT cells. In conclusion, gut leakage-induced endotoxemia is more severe in indomethacin-administered FcgRIIb-/- mice than WT, possibly due to the enhanced indomethacin toxicity from lupus-induced intestinal immune-deposition. Due to a lack of inhibitory-FcgRIIb expression, mitochondrial function, and cytokine production of FcgRIIb-/- macrophages were more prominent than WT cells. Hence, lupus disease-activation from NSAIDs-enteropathy-induced gut leakage is possible.

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