Mast Cell-Derived TNF Can Exacerbate Mortality during Severe Bacterial Infections in C57BL/6-Kit Mice
2009; Elsevier BV; Volume: 176; Issue: 2 Linguagem: Inglês
10.2353/ajpath.2010.090342
ISSN1525-2191
AutoresAdrian M. Piliponsky, Ching‐Cheng Chen, Michele A. Grimbaldeston, Stacy M. Burns-Guydish, Jonathan Hardy, Janet Kalesnikoff, Christopher H. Contag, Mindy Tsai, Stephen J. Galli,
Tópico(s)Food Allergy and Anaphylaxis Research
ResumoWe used mast cell-engrafted genetically mast cell-deficient C57BL/6-KitW-sh/W-sh mice to investigate the roles of mast cells and mast cell-derived tumor necrosis factor in two models of severe bacterial infection. In these mice, we confirmed findings derived from studies of mast cell-deficient WBB6F1-KitW/W-v mice indicating that mast cells can promote survival in cecal ligation and puncture (CLP) of moderate severity. However, we found that the beneficial role of mast cells in this setting can occur independently of mast cell-derived tumor necrosis factor. By contrast, using mast cell-engrafted C57BL/6-KitW-sh/W-sh mice, we found that mast cell-derived tumor necrosis factor can increase mortality during severe CLP and can also enhance bacterial growth and hasten death after intraperitoneal inoculation of Salmonella typhimurium. In WBB6F1-KitW-sh/W-sh mice, mast cells enhanced survival during moderately severe CLP but did not significantly change the survival observed in severe CLP. Our findings in three types of genetically mast cell-deficient mice thus support the hypothesis that, depending on the circumstances (including mouse strain background, the nature of the mutation resulting in a mast cell deficiency, and type and severity of infection), mast cells can have either no detectable effect or opposite effects on survival during bacterial infections, eg, promoting survival during moderately severe CLP associated with low mortality but, in C57BL/6-KitW-sh/W-sh mice, increasing mortality during severe CLP or infection with S. typhimurium. We used mast cell-engrafted genetically mast cell-deficient C57BL/6-KitW-sh/W-sh mice to investigate the roles of mast cells and mast cell-derived tumor necrosis factor in two models of severe bacterial infection. In these mice, we confirmed findings derived from studies of mast cell-deficient WBB6F1-KitW/W-v mice indicating that mast cells can promote survival in cecal ligation and puncture (CLP) of moderate severity. However, we found that the beneficial role of mast cells in this setting can occur independently of mast cell-derived tumor necrosis factor. By contrast, using mast cell-engrafted C57BL/6-KitW-sh/W-sh mice, we found that mast cell-derived tumor necrosis factor can increase mortality during severe CLP and can also enhance bacterial growth and hasten death after intraperitoneal inoculation of Salmonella typhimurium. In WBB6F1-KitW-sh/W-sh mice, mast cells enhanced survival during moderately severe CLP but did not significantly change the survival observed in severe CLP. Our findings in three types of genetically mast cell-deficient mice thus support the hypothesis that, depending on the circumstances (including mouse strain background, the nature of the mutation resulting in a mast cell deficiency, and type and severity of infection), mast cells can have either no detectable effect or opposite effects on survival during bacterial infections, eg, promoting survival during moderately severe CLP associated with low mortality but, in C57BL/6-KitW-sh/W-sh mice, increasing mortality during severe CLP or infection with S. typhimurium. The factors determining whether particular infections will be successfully controlled or progress to death are incompletely understood. Studies conducted using genetically mast cell-deficient (WB/ReJ × C57BL/6)F1-KitW/W-v mice (WBB6F1-KitW/W-v mice), the corresponding normal (WBB6F1-Kit+/+) mice, and mast cell-engrafted WBB6F1-KitW/W-v mice have indicated that mast cells can increase survival during various models of bacterial infection of moderate severity,1Echtenacher B Mannel DN Hultner L Critical protective role of mast cells in a model of acute septic peritonitis.Nature. 1996; 381: 75-77Crossref PubMed Scopus (802) Google Scholar, 2Malaviya R Ikeda T Ross E Abraham SN Mast cell modulation of neutrophil influx and bacterial clearance at sites of infection through TNF-alpha.Nature. 1996; 381: 77-80Crossref PubMed Scopus (970) Google Scholar, 3Prodeus AP Zhou X Maurer M Galli SJ Carroll MC Impaired mast cell-dependent natural immunity in complement C3-deficient mice.Nature. 1997; 390: 172-175Crossref PubMed Scopus (234) Google Scholar, 4Maurer M Echtenacher B Hultner L Kollias G Mannel DN Langley KE Galli SJ The c-kit ligand, stem cell factor, can enhance innate immunity through effects on mast cells.J Exp Med. 1998; 188: 2343-2348Crossref PubMed Scopus (132) Google Scholar, 5Supajatura V Ushio H Nakao A Akira S Okumura K Ra C Ogawa H Differential responses of mast cell Toll-like receptors 2 and 4 in allergy and innate immunity.J Clin Invest. 2002; 109: 1351-1359Crossref PubMed Scopus (438) Google Scholar, 6Jippo T Morii E Ito A Kitamura Y Effect of anatomical distribution of mast cells on their defense function against bacterial infections: demonstration using partially mast cell-deficient tg/tg mice.J Exp Med. 2003; 197: 1417-1425Crossref PubMed Scopus (37) Google Scholar, 7McLachlan JB Hart JP Pizzo SV Shelburne CP Staats HF Gunn MD Abraham SN Mast cell-derived tumor necrosis factor induces hypertrophy of draining lymph nodes during infection.Nat Immunol. 2003; 4: 1199-1205Crossref PubMed Scopus (249) Google Scholar, 8Maurer M Wedemeyer J Metz M Piliponsky AM Weller K Chatterjea D Clouthier DE Yanagisawa MM Tsai M Galli SJ Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (255) Google Scholar, 9Xu X Zhang D Lyubynska N Wolters PJ Killeen NP Baluk P McDonald DM Hawgood S Caughey GH Mast cells protect mice from Mycoplasma pneumonia.Am J Respir Crit Care Med. 2006; 173: 219-225Crossref PubMed Scopus (72) Google Scholar, 10Orinska Z Maurer M Mirghomizadeh F Bulanova E Metz M Nashkevich N Schiemann F Schulmistrat J Budagian V Giron-Michel J Brandt E Paus R Bulfone-Paus S IL-15 constrains mast cell-dependent antibacterial defenses by suppressing chymase activities.Nat Med. 2007; 13: 927-934Crossref PubMed Scopus (92) Google Scholar, 11Siebenhaar F Syska W Weller K Magerl M Zuberbier T Metz M Maurer M Control of Pseudomonas aeruginosa skin infections in mice is mast cell-dependent.Am J Pathol. 2007; 170: 1910-1916Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar, 12Thakurdas SM Melicoff E Sansores-Garcia L Moreira DC Petrova Y Stevens RL Adachi R The mast cell-restricted tryptase mMCP-6 has a critical immunoprotective role in bacterial infections.J Biol Chem. 2007; 282: 20809-20815Crossref PubMed Scopus (151) Google Scholar, 13Piliponsky AM Chen CC Nishimura T Metz M Rios EJ Dobner PR Wada E Wada K Zacharias S Mohanasundaram UM Faix JD Abrink M Pejler G Pearl RG Tsai M Galli SJ Neurotensin increases mortality and mast cells reduce neurotensin levels in a mouse model of sepsis.Nat Med. 2008; 14: 392-398Crossref PubMed Scopus (97) Google Scholar defined herein as infections that result in relatively low mortality in normal mice. Although mast cells can contribute to host defense against bacteria by multiple direct and indirect mechanisms,1Echtenacher B Mannel DN Hultner L Critical protective role of mast cells in a model of acute septic peritonitis.Nature. 1996; 381: 75-77Crossref PubMed Scopus (802) Google Scholar, 2Malaviya R Ikeda T Ross E Abraham SN Mast cell modulation of neutrophil influx and bacterial clearance at sites of infection through TNF-alpha.Nature. 1996; 381: 77-80Crossref PubMed Scopus (970) Google Scholar, 3Prodeus AP Zhou X Maurer M Galli SJ Carroll MC Impaired mast cell-dependent natural immunity in complement C3-deficient mice.Nature. 1997; 390: 172-175Crossref PubMed Scopus (234) Google Scholar, 4Maurer M Echtenacher B Hultner L Kollias G Mannel DN Langley KE Galli SJ The c-kit ligand, stem cell factor, can enhance innate immunity through effects on mast cells.J Exp Med. 1998; 188: 2343-2348Crossref PubMed Scopus (132) Google Scholar, 5Supajatura V Ushio H Nakao A Akira S Okumura K Ra C Ogawa H Differential responses of mast cell Toll-like receptors 2 and 4 in allergy and innate immunity.J Clin Invest. 2002; 109: 1351-1359Crossref PubMed Scopus (438) Google Scholar, 7McLachlan JB Hart JP Pizzo SV Shelburne CP Staats HF Gunn MD Abraham SN Mast cell-derived tumor necrosis factor induces hypertrophy of draining lymph nodes during infection.Nat Immunol. 2003; 4: 1199-1205Crossref PubMed Scopus (249) Google Scholar, 8Maurer M Wedemeyer J Metz M Piliponsky AM Weller K Chatterjea D Clouthier DE Yanagisawa MM Tsai M Galli SJ Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (255) Google Scholar, 9Xu X Zhang D Lyubynska N Wolters PJ Killeen NP Baluk P McDonald DM Hawgood S Caughey GH Mast cells protect mice from Mycoplasma pneumonia.Am J Respir Crit Care Med. 2006; 173: 219-225Crossref PubMed Scopus (72) Google Scholar, 10Orinska Z Maurer M Mirghomizadeh F Bulanova E Metz M Nashkevich N Schiemann F Schulmistrat J Budagian V Giron-Michel J Brandt E Paus R Bulfone-Paus S IL-15 constrains mast cell-dependent antibacterial defenses by suppressing chymase activities.Nat Med. 2007; 13: 927-934Crossref PubMed Scopus (92) Google Scholar, 11Siebenhaar F Syska W Weller K Magerl M Zuberbier T Metz M Maurer M Control of Pseudomonas aeruginosa skin infections in mice is mast cell-dependent.Am J Pathol. 2007; 170: 1910-1916Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar, 12Thakurdas SM Melicoff E Sansores-Garcia L Moreira DC Petrova Y Stevens RL Adachi R The mast cell-restricted tryptase mMCP-6 has a critical immunoprotective role in bacterial infections.J Biol Chem. 2007; 282: 20809-20815Crossref PubMed Scopus (151) Google Scholar, 13Piliponsky AM Chen CC Nishimura T Metz M Rios EJ Dobner PR Wada E Wada K Zacharias S Mohanasundaram UM Faix JD Abrink M Pejler G Pearl RG Tsai M Galli SJ Neurotensin increases mortality and mast cells reduce neurotensin levels in a mouse model of sepsis.Nat Med. 2008; 14: 392-398Crossref PubMed Scopus (97) Google Scholar several groups have focused on the potential role of mast cell-derived tumor necrosis factor (TNF) in such settings.1Echtenacher B Mannel DN Hultner L Critical protective role of mast cells in a model of acute septic peritonitis.Nature. 1996; 381: 75-77Crossref PubMed Scopus (802) Google Scholar, 2Malaviya R Ikeda T Ross E Abraham SN Mast cell modulation of neutrophil influx and bacterial clearance at sites of infection through TNF-alpha.Nature. 1996; 381: 77-80Crossref PubMed Scopus (970) Google Scholar, 3Prodeus AP Zhou X Maurer M Galli SJ Carroll MC Impaired mast cell-dependent natural immunity in complement C3-deficient mice.Nature. 1997; 390: 172-175Crossref PubMed Scopus (234) Google Scholar, 4Maurer M Echtenacher B Hultner L Kollias G Mannel DN Langley KE Galli SJ The c-kit ligand, stem cell factor, can enhance innate immunity through effects on mast cells.J Exp Med. 1998; 188: 2343-2348Crossref PubMed Scopus (132) Google Scholar, 5Supajatura V Ushio H Nakao A Akira S Okumura K Ra C Ogawa H Differential responses of mast cell Toll-like receptors 2 and 4 in allergy and innate immunity.J Clin Invest. 2002; 109: 1351-1359Crossref PubMed Scopus (438) Google Scholar, 7McLachlan JB Hart JP Pizzo SV Shelburne CP Staats HF Gunn MD Abraham SN Mast cell-derived tumor necrosis factor induces hypertrophy of draining lymph nodes during infection.Nat Immunol. 2003; 4: 1199-1205Crossref PubMed Scopus (249) Google Scholar Results obtained in work using TNF-deficient mice4Maurer M Echtenacher B Hultner L Kollias G Mannel DN Langley KE Galli SJ The c-kit ligand, stem cell factor, can enhance innate immunity through effects on mast cells.J Exp Med. 1998; 188: 2343-2348Crossref PubMed Scopus (132) Google Scholar or mice in which the actions of TNF are blocked by neutralizing antibodies1Echtenacher B Mannel DN Hultner L Critical protective role of mast cells in a model of acute septic peritonitis.Nature. 1996; 381: 75-77Crossref PubMed Scopus (802) Google Scholar, 2Malaviya R Ikeda T Ross E Abraham SN Mast cell modulation of neutrophil influx and bacterial clearance at sites of infection through TNF-alpha.Nature. 1996; 381: 77-80Crossref PubMed Scopus (970) Google Scholar have clearly demonstrated that TNF can have protective functions during some bacterial infections, and that such TNF-dependent effects may include the enhancement of neutrophil recruitment and/or function and the promotion of bacterial clearance. However, such genetic or neutralizing antibody-based approaches eliminate or reduce the function of TNF derived from all cellular sources, not just mast cell-derived TNF. Thus, there has been no direct evidence yet published that shows that mast cells represent a critical source of TNF in such settings. Moreover, WBB6F1-KitW/W-v mice have a modest deficiency in neutrophils,14Chervenick PA Boggs DR Decreased neutrophils and megakaryocytes in anemic mice of genotype W/W.J Cell Physiol. 1969; 73: 25-30Crossref PubMed Scopus (63) Google Scholar, 15Zhou JS Xing W Friend DS Austen KF Katz HR Mast cell deficiency in Kit(W-sh) mice does not impair antibody-mediated arthritis.J Exp Med. 2007; 204: 2797-2802Crossref PubMed Scopus (148) Google Scholar, 16Nigrovic PA Gray DH Jones T Hallgren J Kuo FC Chaletzky B Gurish M Mathis D Benoist C Lee DM Genetic inversion in mast cell-deficient (W(sh)) mice interrupts corin and manifests as hematopoietic and cardiac aberrancy.Am J Pathol. 2008; 173: 1693-1701Abstract Full Text Full Text PDF PubMed Scopus (169) Google Scholar and this abnormality, as well as their virtual lack of mast cells, might contribute to the increased pathology observed in these mice during certain models of bacterial infection. Therefore, in the present study, we used genetically mast cell-deficient C57BL/6-KitW-sh/W-sh mice (produced as in ref.17Grimbaldeston MA Chen CC Piliponsky AM Tsai M Tam SY Galli SJ Mast cell-deficient W-sash c-kit mutant Kit W-sh/W-sh mice as a model for investigating mast cell biology in vivo.Am J Pathol. 2005; 167: 835-848Abstract Full Text Full Text PDF PubMed Scopus (462) Google Scholar) engrafted with either wild-type mast cells or mast cells unable to produce TNF to re-examine the roles of mast cells, and mast cell-derived TNF, in two different models of bacterial infection. Adult C57BL/6-KitW-sh/W-sh mice are profoundly mast cell-deficient,17Grimbaldeston MA Chen CC Piliponsky AM Tsai M Tam SY Galli SJ Mast cell-deficient W-sash c-kit mutant Kit W-sh/W-sh mice as a model for investigating mast cell biology in vivo.Am J Pathol. 2005; 167: 835-848Abstract Full Text Full Text PDF PubMed Scopus (462) Google Scholar, 18Tono T Tsujimura T Koshimizu U Kasugai T Adachi S Isozaki K Nishikawa S Morimoto M Nishimune Y Nomura S c-kit Gene was not transcribed in cultured mast cells of mast cell-deficient Wsh/Wsh mice that have a normal number of erythrocytes and a normal c-kit coding region.Blood. 1992; 80: 1448-1453PubMed Google Scholar, 19Wolters PJ Mallen-St Clair J Lewis CC Villalta SA Baluk P Erle DJ Caughey GH Tissue-selective mast cell reconstitution and differential lung gene expression in mast cell-deficient Kit(W-sh)/Kit(W-sh) sash mice.Clin Exp Allergy. 2005; 535: 82-88Crossref Scopus (129) Google Scholar as are WBB6F1-KitW/W-v mice,20Kitamura Y Go S Hatanaka K Decrease of mast cells in W/Wv mice and their increase by bone marrow transplantation.Blood. 1978; 52: 447-452Crossref PubMed Google Scholar, 21Galli SJ Kitamura Y Genetically mast-cell-deficient W/Wv and Sl/Sld mice. Their value for the analysis of the roles of mast cells in biologic responses in vivo.Am J Pathol. 1987; 127: 191-198PubMed Google Scholar but, unlike WBB6F1-KitW/W-v mice, C57BL/6-KitW-sh/W-sh mice have been reported to have increased levels of blood and bone marrow neutrophils.15Zhou JS Xing W Friend DS Austen KF Katz HR Mast cell deficiency in Kit(W-sh) mice does not impair antibody-mediated arthritis.J Exp Med. 2007; 204: 2797-2802Crossref PubMed Scopus (148) Google Scholar, 16Nigrovic PA Gray DH Jones T Hallgren J Kuo FC Chaletzky B Gurish M Mathis D Benoist C Lee DM Genetic inversion in mast cell-deficient (W(sh)) mice interrupts corin and manifests as hematopoietic and cardiac aberrancy.Am J Pathol. 2008; 173: 1693-1701Abstract Full Text Full Text PDF PubMed Scopus (169) Google Scholar We used both WBB6F1-KitW/W-v mice and C57BL/6-KitW-sh/W-sh mice to investigate the role of mast cells in moderately severe versus severe cecal ligation and puncture (CLP), with moderately severe versus severe CLP defined herein as models of CLP in which no more than 50% (in the moderately severe CLP model) vs. more than 50% (in the severe CLP model) of normal mice succumb during the first 4 days after surgery to induce acute bacterial peritonitis. Our observations in C57BL/6-KitW-sh/W-sh mice confirm work in WBB6F1-KitW/W-v mice1Echtenacher B Mannel DN Hultner L Critical protective role of mast cells in a model of acute septic peritonitis.Nature. 1996; 381: 75-77Crossref PubMed Scopus (802) Google Scholar, 3Prodeus AP Zhou X Maurer M Galli SJ Carroll MC Impaired mast cell-dependent natural immunity in complement C3-deficient mice.Nature. 1997; 390: 172-175Crossref PubMed Scopus (234) Google Scholar, 4Maurer M Echtenacher B Hultner L Kollias G Mannel DN Langley KE Galli SJ The c-kit ligand, stem cell factor, can enhance innate immunity through effects on mast cells.J Exp Med. 1998; 188: 2343-2348Crossref PubMed Scopus (132) Google Scholar, 5Supajatura V Ushio H Nakao A Akira S Okumura K Ra C Ogawa H Differential responses of mast cell Toll-like receptors 2 and 4 in allergy and innate immunity.J Clin Invest. 2002; 109: 1351-1359Crossref PubMed Scopus (438) Google Scholar, 6Jippo T Morii E Ito A Kitamura Y Effect of anatomical distribution of mast cells on their defense function against bacterial infections: demonstration using partially mast cell-deficient tg/tg mice.J Exp Med. 2003; 197: 1417-1425Crossref PubMed Scopus (37) Google Scholar, 8Maurer M Wedemeyer J Metz M Piliponsky AM Weller K Chatterjea D Clouthier DE Yanagisawa MM Tsai M Galli SJ Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (255) Google Scholar in providing evidence that mast cells can enhance host resistance and survival during moderately severe CLP, a well known model of mouse bacterial peritonitis, but provide evidence that mast cells can enhance survival in this model independently of their ability to produce TNF. By contrast, mast cell-derived TNF increased mortality during severe CLP in C57BL/6-KitW-sh/W-sh mice. We found that mast cell-derived TNF also can hasten mortality in C57BL/6-KitW-sh/W-sh mice subjected to a different model of severe bacterial infection: that induced by the intraperitoneal inoculation of Salmonella typhimurium. c-kit mutant genetically mast cell-deficient (WB/ReJ-KitW/+ × C57BL/6J-KitW-v/+) F1-KitW/W-v (WBB6F1-KitW/W-v or KitW/W-v) mice and the congenic normal WBB6F1+/+ (WBB6F1- Kit+/+) mice, and C57BL/6J (B6J) mice, were purchased from Jackson Laboratories (Bar Harbor, ME). The Wsh or "sash" mutation was named after the broad white belt or sash observed on the coat of a single female offspring of a cross between two inbred strains (C3H/HeH × 101/H).22Lyon MF Glenister PH A new allele sash (Wsh) at the W-locus and a spontaneous recessive lethal in mice.Genet Res. 1982; 39: 315-322Crossref PubMed Scopus (72) Google ScholarWsh homozygotes were characterized as black-eyed white mice (some of them with patches of pigment on the ear pinnae and/or base of the tail) that were viable, non-anemic and fertile,22Lyon MF Glenister PH A new allele sash (Wsh) at the W-locus and a spontaneous recessive lethal in mice.Genet Res. 1982; 39: 315-322Crossref PubMed Scopus (72) Google Scholar but which had a marked reduction in mast cells in multiple anatomical sites.23Stevens J Loutit JF Mast cells in spotted mutant mice (W Ph mi).Proc R Soc Lond B Biol Sci. 1982; 215: 405-409Crossref PubMed Scopus (74) Google Scholar Genetically mast cell-deficient C57BL/6-KitW-sh/+ mice (B6-KitW-sh/+ mice)24Duttlinger R Manova K Chu TY Gyssler C Zelenetz AD Bachvarova RF Besmer P W-sash affects positive and negative elements controlling c-kit expression: ectopic c-kit expression at sites of kit-ligand expression affects melanogenesis.Development. 1993; 118: 705-717PubMed Google Scholar were the generous gift of Peter Besmer (Molecular Biology Program, Memorial Sloan-Kettering Cancer Center and Cornell University Graduate School of Medical Sciences, New York, NY); while these mice were reported to be on the C57BL/6 background, it is not clear from the publication whether, and if so how many times, these mice had been backcrossed to mice from the Jackson Laboratories' C57BL/6J population. We used such B6-KitW-sh/+ mice to produce B6-KitW-sh/W-sh mice for some of our initial CLP experiments; for these experiments, we used the B6-Kit+/+ littermates as wild-type controls. In addition, we backcrossed B6-KitW-sh/+ mice with C57BL/6J (B6J) mice for 11 generations to produce B6J-KitW-sh/W-sh mice; for CLP experiments using these mice, we used B6J-Kit+/+ mice purchased from Jackson Laboratories as wild-type controls. Like B6-KitW-sh/W-sh mice,17Grimbaldeston MA Chen CC Piliponsky AM Tsai M Tam SY Galli SJ Mast cell-deficient W-sash c-kit mutant Kit W-sh/W-sh mice as a model for investigating mast cell biology in vivo.Am J Pathol. 2005; 167: 835-848Abstract Full Text Full Text PDF PubMed Scopus (462) Google Scholar, 19Wolters PJ Mallen-St Clair J Lewis CC Villalta SA Baluk P Erle DJ Caughey GH Tissue-selective mast cell reconstitution and differential lung gene expression in mast cell-deficient Kit(W-sh)/Kit(W-sh) sash mice.Clin Exp Allergy. 2005; 535: 82-88Crossref Scopus (129) Google Scholar we found that adult (∼12-week-old) B6J- KitW-sh/W-sh mice lacked any detectable mast cells in the peritoneal cavity or mesentery. Based on reverse transcription-PCR analysis and sequencing of the resultant PCR products, we confirmed that the Wsh inversion 3′ breakpoint in our B6J-KitW-sh/W-sh mice and that in the B6-KitW-sh/W-sh mice that were characterized by Nigrovic et al16Nigrovic PA Gray DH Jones T Hallgren J Kuo FC Chaletzky B Gurish M Mathis D Benoist C Lee DM Genetic inversion in mast cell-deficient (W(sh)) mice interrupts corin and manifests as hematopoietic and cardiac aberrancy.Am J Pathol. 2008; 173: 1693-1701Abstract Full Text Full Text PDF PubMed Scopus (169) Google Scholar are identical (data not shown). To generate WB/ReJ-KitW-sh/+ mice, B6J-KitW-sh/+ mice were crossed with WB/ReJ-Kit+/+ mice for 11 generations. Some genetically mast cell-deficient B6J-KitW-sh/+ mice were then crossed with WB/ReJ-KitW-sh/+ mice to produce WBB6F1- KitW-sh/W-sh mice (WB/ReJ-KitW-sh/+ × B6-KitW-sh/+) F1-KitW-sh/W-sh and the normal wild-type littermates (ie, WBB6F1-Kit+/+ mice). TNF-deficient mice (Tnf−/−) on the C57BL/6J background25Korner H Cook M Riminton DS Lemckert FA Hoek RM Ledermann B Kontgen F Fazekas de St Groth B Sedgwick JD Distinct roles for lymphotoxin-alpha and tumor necrosis factor in organogenesis and spatial organization of lymphoid tissue.Eur J Immunol. 1997; 27: 2600-2609Crossref PubMed Scopus (268) Google Scholar were bred and maintained at the Stanford University Research Animal Facility. Unless specified otherwise, all experiments were performed using 12-week-old male mice. All animal care and experimentation was conducted in accord with current National Institutes of Health guidelines and with the approval of the Stanford University Institutional Animal Care and Use Committee. Some WBB6F1-KitW/W-v, B6-KitW-sh/W-sh, and WBB6F1- KitW-sh/W-sh mice (4 to 6 weeks old) were repaired of their mast cell deficiency selectively and locally by the i.p. injection of growth factor-dependent, congenic mouse bone marrow-derived cultured mast cells (BMCMCs). Briefly, femoral bone marrow cells from WBB6F1-Kit+/+ or B6-Kit+/+ mice were maintained in vitro for ∼4 weeks in interleukin-3 (Peprotech, Rocky Hill, NJ)-containing medium until mast cells represented >95% of the total cells according to staining with May-Grünwald-Giemsa. 2.0 × 106 BMCMCs, in 200 μl of PBS, were injected i.p. (via a 26G needle) and the mice were used for experiments, together with strain-, gender-, and age-matched mast cell-deficient mice, 4 to 6 weeks after adoptive transfer of BMCMCs. Other B6-KitW-sh/W-sh mice received, 4 to 6 weeks before CLP, 2.0 × 106 BMCMCs generated from B6-Tnf−/− mice. The numbers of peritoneal mast cells were similar in C57BL/6-KitW-sh/W-sh mice that had been engrafted with either wild-type or TNF-deficient mast cells (3.8 + 0.8% or 3.6 + 0.7% of total cells) or in WBB6F1-KitW-sh/W-sh mice that had been engrafted with WBB6F1 wild-type mast cells (6.1 + 0.5% of total cells), as was the distribution of mast cells in the mesentery (ie, mesenteric windows) of these three groups of mice (6.2 + 0.96, 6.4 + 0.64, or 6.1 + 0.55 mast cells/mm2, respectively). The wild-type virulent S. typhimurium strain, SL1344, was obtained from Bruce Stocker (deceased) of Stanford University and labeled with the Lux operon to create bioluminescent SL1344lux (SMB500) as reported.26Burns-Guydish SM Olomu IN Zhao H Wong RJ Stevenson DK Contag CH Monitoring age-related susceptibility of young mice to oral Salmonella enterica serovar Typhimurium infection using an in vivo murine model.Pediatr Res. 2005; 58: 153-158Crossref PubMed Scopus (51) Google Scholar Bacteria were grown as previously described27Chatterjea D Burns-Guydish SM Sciuto TE Dvorak A Contag CH Galli SJ Adoptive transfer of mast cells does not enhance the impaired survival of Kit(W)/Kit(W-v) mice in a model of low dose intraperitoneal infection with bioluminescent Salmonella typhimurium.Immunol Lett. 2005; 99: 122-129Crossref PubMed Scopus (14) Google Scholar and mice were infected by i.p. injection of 25 colony forming units (CFUs) of the labeled (ie, bioluminescent) bacteria. Mice were imaged as previously described.27Chatterjea D Burns-Guydish SM Sciuto TE Dvorak A Contag CH Galli SJ Adoptive transfer of mast cells does not enhance the impaired survival of Kit(W)/Kit(W-v) mice in a model of low dose intraperitoneal infection with bioluminescent Salmonella typhimurium.Immunol Lett. 2005; 99: 122-129Crossref PubMed Scopus (14) Google Scholar Briefly, mice were placed in an in vivo Imaging System (IVIS, a Xenogen product from Caliper LifeSciences, Alameda, CA). A grayscale reference image was taken under low light illumination followed by a 5 minutes image, in complete dark, of the bioluminescent signal originating from the labeled bacteria within the animals. The data were expressed as photons emitted per mouse, or region of interest, per second. A pseudocolor representation of the photon collection (red signifying most intense and blue signifying least intense) was superimposed over the grayscale reference image. Acquisition and analysis of data were performed using the LivingImage (a Xenogen product from Caliper LifeSciences) software, which runs as an overlay on the IgorPro image analysis package (Wavemetrics, Lake Oswego, OR). Mice were imaged before and at selected time points after infection. CLP was performed as described.1Echtenacher B Mannel DN Hultner L Critical protective role of mast cells in a model of acute septic peritonitis.Nature. 1996; 381: 75-77Crossref PubMed Scopus (802) Google Scholar Briefly, mice were deeply anesthetized by i.m. injection of 100 mg/kg ketamine and 20 mg/kg xylazine. The cecum was exposed by a 1- to 2-cm midline incision on the anterior abdomen and subjected to ligation of its distal portion and a single needle puncture of the ligated segment. Protocols for eliciting severe or moderately severe CLP were developed so that >50% or 20% to 50% of the wild-type mice died within 4 days after CLP. We used survival as the criterion to classify the severity of CLP, identifying procedures of performing "moderately severe" or "severe" CLP, which resulted in the death of no more than 50% or >50% of wild-type mice, respectively. However, we found, as expected, that the hypothermia induced in wild-type mice by CLP was more pronounced in mice subjected to severe CLP than in mice subjected to moderately severe CLP (a reduction in body temperature of 4.6 + 0.86°C vs. 1.6 + 0.48°C [mean + SD] for severe vs. moderately severe CLP, P < 0.05, n = 5/group) at 18 to 24 hours after surgery. For severe CLP, we ligated the distal half or two thirds of the cecum and made a single puncture with a 22G needle; for moderately severe CLP, we ligated the distal half of the cecum and made a single puncture with a 22G needle. The cecum was then replaced into the abdomen, 1 ml of sterile saline (pyrogen-free 0.9% NaCl) was administrated into the peritoneal cavity, and the incision was closed using 9-mm steel wound clips. Mice were observed for mortality at least four times daily. Mice that were clearly moribund were euthanized by CO2 inhalation. Absolute numbers of neutrophils and platelets in the blood were counted using the Abbott Cell-Dyn 3500 automated hematology analyzer. Neutrophils (Gr-1high/F4/80− cells) in the bone marrow, spleen and peritoneal fluid, macrophages (Gr-1−/F4/80+ cells) in the peritoneal fluid, and basophils (FcεRI+/c-Kit−/DX5+ cells) in the blood were analyzed by flow cytometry. Briefly, red cells were lysed with AKC lysis buffer for 5 minutes. Total cell numbers were counted using a hemocytometer. Cells were blocked with unconjugated anti-CD16/CD32 on ice for 5 minutes and then stained with a combination of fluorescein isothiocyanate-labeled anti-Gr-1 (RB6-8C5, 2.5 μg/ml) and allophycocyanin-labeled anti-F4/80 (BM8, 4 μg/ml of each) antibodies (eBioscience San Diego, CA) on ice for 15 minutes for the detection of neutrophils and macrophages. To identify basophils (as c-Kit−CD49b+FcεRI α chain+ cells28Mukai K Matsu
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