Published online before print September 20, 2007
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Department of Surgery, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, Rhode Island, USA
1 Correspondence: Division of Surgical Research, Rhode Island Hospital, NAB 214, 593 Eddy Street, Providence, RI 02903, USA. E-mail: jdaley{at}lifespan.org
ABSTRACT
The anti-granulocyte receptor-1 (Gr-1) mAb, RB6-8C5, has been used extensively to deplete neutrophils in mice and to investigate the role of these cells in host defense. RB6-8C5 binds to Ly6G, which is present on neutrophils, and to Ly6C, which is expressed on neutrophils, dendritic cells, and subpopulations of lymphocytes and monocytes. It is thus likely that in vivo administration of RB6-8C5 may deplete not only neutrophils but also other Gr-l+ (Ly6C+) cells. This study describes the use of an Ly6G-specific mAb, 1A8, as an alternative means to deplete neutrophils. In vivo administration of RB6-8C5 reduced blood neutrophils and Gr-1+ monocytes, whereas administration of 1A8 reduced blood neutrophils but not Gr-1+ monocytes. Plasma TNF-
in endotoxemia was increased 20-fold by RB6-8C5 pretreatment and fourfold by 1A8 pretreatment. In a wound model, pretreatment with either antibody decreased wound neutrophils and macrophages. TNF- staining in brefeldin-treated wound leukocytes was increased by pretreatment with RB6-8C5, but not 1A8. Neutrophil depletion with 1A8 offers advantages over the use of RB6-8C5, as it preserves non-neutrophil Gr-1+ cells depleted by the anti-Gr-1 antibody. The loss of non-neutrophil Gr-1+ populations in RB6-8C5-treated animals is associated with increased TNF- responses, suggesting these cells may function to suppress TNF- production.
Key Words: monocytes • macrophages • Gr-1 • Ly6C • wound • endotoxemia • TNF-
INTRODUCTION
The depletion of cell lineage-specific immune effector cells has been used successfully to delineate their functions in immune responses. The development of genetically altered animals lacking one or more immune cell types has greatly expanded the understanding of innate and adaptive immunity. No genetic manipulation that eliminates neutrophils has been reported in viable animals. The lack of neutrophil "knockout" animals has mandated use of pharmacologic agents or antibody-mediated depletion strategies to investigate the role of neutrophils in animal models of disease.
Neutrophil depletion has been achieved with agents that suppress myelopoiesis, such as cyclophosphamide or vinblastine. However, these agents deplete not only neutrophils but also other leukocytes [1 ]. The mAb RB6-8C5 [2 ] has been used extensively to deplete neutrophils in murine disease models, including surgical injury [3 ], acute tubular necrosis [4 ], experimental encephalitis [5 ], pulmonary cryptococcosis [6 ], Legionella pneumonia [7 ], Toxoplasma gondii infection [8 ], carcinogenesis [9 ], amebiasis [10 ], and Chagas’ disease [11 ]. A search of the PubMed database using the terms "neutropenia" and "RB6-8C5" identified over 60 studies that used this antibody to determine neutrophil function in various disease models.
RB6-8C5 was described originally as binding a surface antigen, granulocyte receptor 1 (Gr-1), believed to be expressed only by mature granulocytes [2 ]. The Gr-1 antigen was identified subsequently as a member of the Ly6 gene family. Ly6G, a granulocyte surface marker, is the major antigen detected by RB6-8C5 [12 ]. However, RB6-8C5 also binds to Ly6C [12 ], which is expressed on neutrophils, dendritic cells (DCs), and subsets of monocytes, macrophages, and lymphocytes [13 14 15 16 17 ].
Recent studies have determined that Ly6C+ (Gr-1+) blood monocytes are precursors of inflammatory macrophages [18 19 20 ]. RB6-8C5 detects this subpopulation of blood monocytes, as well as monocytes from early peritoneal exudates, which maintain Gr-1 expression as they differentiate into macrophages [21 ].
As RB6-8C5 binds to Gr-1+ monocytes/macrophages, systemic administration of the antibody could eliminate these cells, along with neutrophils. Data presented here demonstrate that RB6-8C5 depleted not only neutrophils but also Gr-l+ monocytes/macrophages. In addition, in vivo administration of RB6-8C5 has already been shown to deplete DCs [22
] and lymphocytes [23
], although there is some evidence that RB6-8C5 does not recognize Ly6C on lymphocytes [24
]. Thus, results of studies that use RB6-8C5 reflect not only the loss of neutrophils but also the loss of other Gr-1-expressing cells [25
]. This laboratory has used RB6-8C5 in experimental wounds and in endotoxemia. Pretreatment with this antibody decreased wound neutrophils and wound macrophages and was associated with increased TNF- responses. The present study re-examines the effects of in vivo administration of RB6-8C5 and proposes the Ly6G-specific mAb 1A8 [12
] as an improved strategy to deplete neutrophils in mice.
MATERIALS AND METHODS
Animals
B6D2F1 male mice (Taconic, Germantown, NY, USA), 8–12 weeks of age, were housed in the Central Research Facilities at Rhode Island Hospital (Providence, RI, USA) and fed mouse chow and water ad libitum. Athymic nude BALB/c male mice (Taconic), 6 weeks of age, were housed in a pathogen-free environment and fed mouse chow and water ad libitum. Mice were certified free of common pathogens by the suppliers and were monitored by Brown University/Rhode Island Hospital veterinary personnel. Animal protocols were approved by the Institutional Animal Care and Use Committee at Rhode Island Hospital.
Depletion of neutrophils
Anti-Gr-1 mAb (RB6-8C5 hybridoma originally produced by Robert L. Coffman, DNAX Research Institute, Palo Alto, CA, USA [2
]) was produced under serum-free conditions using a bioreactor (Cell Pharm Micro Mouse, UniSyn Technologies, Tustin, CA, USA). Neutrophils were depleted by a single i.p. injection of supernatant, containing 0.5 mg protein, 3 days prior to isolation of blood leukocytes or wounding. Anti-Ly6G mAb (1A8 hybridoma originally produced and generously provided by Thomas R. Malek, Miller School of Medicine, University of Miami, FL, USA [12
]) was produced in athymic nude BALB/c mice using the mouse ascites method [26
]. Antibody was purified by ammonium sulfate precipitation, dialyzed against PBS, and sterilized by 0.2 µ filtration. Mice were injected i.p. with ammonium sulfate-precipitated 1A8 ascites containing 1.0 mg protein 36 h prior to isolation of blood leukocytes or wounding. Control animals were not injected with antibody. Protein concentration was determined by bicinchoninic acid protein assay (Pierce Biotechnologies, Rockford, IL, USA). Neutrophil depletion was confirmed by differential leukocyte count in all experimental animals.
Endotoxemia model
Mice were injected i.p. with LPS (Escherichia coli serotype 055:B5, Sigma Chemical Co., St. Louis, MO, USA; 1 mg/kg). Ninety minutes after LPS injection, animals were killed by CO2 asphyxiation. Heparinized blood obtained by cardiac puncture was centrifuged immediately at 800 g for 10 min, and plasma was frozen at –70°C until analyzed for TNF-.
Polyvinyl alcohol (PVA) sponge wound model
Mice were anesthetized with pentobarbital, (50 mg/kg, i.p., Abbott Laboratories, North Chicago, IL, USA). Five PVA sponges (PVA Unlimited, Warsaw, IN, USA), measuring 1 x 1 x 0.6 cm, were inserted into individual, s.c. pockets through a midline dorsal incision under sterile conditions, and the skin closed with clips [27
]. Twenty-four hours later, mice were killed by CO2 asphyxiation, and the sponges were removed under sterile conditions. Culture of randomly selected PVA sponges at the time of removal confirmed sterility. Wound cells were isolated by rapid, repeated compression of sponges in a Stomacher (Tekmar, Cincinnati, OH, USA). When present, erythrocytes were removed by hypotonic lysis. Viable cells were identified by Trypan blue exclusion and counted using a hemocytometer; viability was greater than 95%. Differential cell counts were performed on cytocentrifuge preparations stained with Hema-3 (Biochemical Sciences, Swedesboro, NJ, USA).
Circulating leukocytes
Heparinized blood was obtained by cardiac puncture. Leukocyte count was determined using a hemocytometer, after dilution and erythrocyte lysis with 0.01% crystal violet in 3% acetic acid. Differential cell counts were performed on blood smears stained with Hema-3. Blood leukocytes (buffy coat cells) were isolated by centrifugation of heparinized blood in Wintrobe tubes (800 g for 15 min at room temperature). Residual erythrocytes were removed by hypotonic lysis.
Cell staining and flow cytometry
Surface antigen staining
Nonspecific binding of antibody to FcRs was blocked with biotin-free FcR-blocking agent (Accurate Chemical, Westbury, NY, USA) or rat anti-mouse Fc
RIII/II (BD Biosciences, San Diego, CA, USA). Cells were incubated with predetermined optimal concentrations of fluorochrome-conjugated antibodies to cell surface antigens and/or isotype control antibodies and then washed in staining buffer [PBS without calcium and magnesium with 1% FBS (Hyclone, Logan, UT, USA) and 0.09% sodium azide]. Antibodies used for cell staining and flow cytometry are listed in Table 1
.
|
View this table: [in a new window] |
Table 1. Antibodies Used for Flow Cytometry
|
Intracellular TNF- staining
Cells were incubated (106 cells/ml) in RPMI (BD Biosciences) with 1% FBS and 100 U/ml penicillin-streptomycin for 6 h in the presence of Brefeldin A (1 µl/106 cells, Golgiplug, BD Biosciences). Cells were stained for intracellular antigen as described above, using anti-CD68, anti-TNF-, and/or the appropriate isotype control antibodies. CD68 was detected in permeable macrophages and to a lesser extent, in permeable neutrophils, thus providing excellent separation of wound neutrophil and wound macrophage populations [27
].
Cells were analyzed and sorted using a Becton-Dickinson FACSort and CellQuest software (BD Biosciences) or FCSPress software (Copyright 1995–2006, Ray Hicks).
Cytokines
Plasma TNF- concentration was measured by ELISA. The capture antibody was a hamster anti-mouse/rat TNF- mAb (BD Biosciences); the detection antibody was a polyclonal rabbit anti-mouse TNF- antibody (Pierce Biotechnologies).
Data analysis
All values are means ± SD, with the number (n) of subjects noted. Data were analyzed by appropriate parametric or nonparametric statistics as indicated in the figure legends and tables. Flow cytometry scatter plots are representative of replicated experiments.
RESULTS
mAb 1A8 binds to blood neutrophils but not blood monocytes
Blood leukocytes were stained for Gr-1 (RB6-8C5) and Ly6G (1A8), and cells in each gated region were sorted by FACS (Fig. 1
). Gr-1high Ly6G+ cells (R1) were neutrophils; Gr-1int Ly6Gneg cells (R2) were monocytes and a few eosinophils. Cells binding neither antibody (R3) were predominantly lymphocytes and some monocytes. As reported by others, 1A8 (anti-Ly6G) identified blood neutrophils but not blood monocytes or blood lymphocytes [12
]; RB6-8C5 (anti-Gr-1) identified blood neutrophils and a subpopulation of blood monocytes [18
, 19
]. Three-color staining demonstrated that Gr-1int Ly6Gneg cells (R2) expressed Ly6C and F4/80 antigen, confirming them as monocytes. Few Gr-1neg Ly6Gneg cells (R3) expressed Ly6C or F4/80. Addition of the Ly6C-specific mAb decreased the binding of RB6-8C5 to blood monocytes in a concentration-dependent manner, suggesting the two antibodies may compete for the Ly6C/Gr-1 antigen on these cells (not shown).
 View larger version (30K): [in a new window] |
Figure 1. Surface antigen expression of blood leukocytes. Blood leukocytes were stained for Gr-1 and Ly6G; cells in R1, R2, and R3 were sorted by FACS. Photomicrographs of cells from each gated region are shown. R1: Gr-1high Ly6G+ cells were neutrophils. R2: Gr-1int Ly6Gneg cells were monocytes and eosinophils. R3: Gr-1neg Ly6Gneg cells were predominantly lymphocytes with some monocytes. Histograms: Blood leukocytes were stained for Gr-1, Ly6G, and Ly6C or F4/80. Cells in R2 express Ly6C and F4/80. A small fraction of the cells in R3 expresses Ly6C and F4/80. Dotted line: Isotype control antibody. Solid line: Anti-Ly6C or anti-F4/80 mAb.
|
 View larger version (26K): [in a new window] |
Figure 2. Surface antigen expression of wound leukocytes. Wound cells were stained for Gr-1 and F4/80; cells in R1 and R2 were sorted by FACS. Photomicrographs of cells from each gated region are shown. R1: Gr-1high F4/80neg cells were neutrophils. R2: Gr-1int F4/80+ cells were macrophages and a few eosinophils. Histograms: Wound cells were stained with Gr-1, F4/80, and Ly6C or Ly6G. Neutrophils (R1, Gr-1high F4/80neg cells) express Ly6C and Ly6G; macrophages (R2, Gr-1int F4/80+ cells) express Ly6C but not Ly6G. Dotted line: Isotype control antibody. Solid line: Anti-Ly6C or anti-Ly6G mAb.
|
|
View this table: [in a new window] |
Table 2. Effect of 1A8 or RB6-8C5 on Blood Leukocyte Counts (x106/ml)
|
|
View this table: [in a new window] |
Table 3. Effect of 1A8 or RB6-8C5 on Blood Monocyte Subpopulations
|
in endotoxemia concentration, measured 90 min after endotoxin injection, was increased 20-fold in RB6-8C5-treated mice and fourfold in 1A8-treated mice compared with controls (Fig. 3
).
 View larger version (14K): [in a new window] |
Figure 3. Effect of RB6-8C5 or 1A8 on plasma TNF- in endotoxemia. Mice were pretreated or not with RB6-8C5 or 1A8 mAb. Ninety minutes after LPS injection (1 mg/kg, i.p.), animals were killed, and blood was obtained by cardiac puncture for determination of plasma TNF-. Each group differs from all other groups (P < 0.05) by ANOVA and Newman-Keuls after logarithmic transformation of the data. n = 4 for each group.
|
 View larger version (19K): [in a new window] |
Figure 4. Effect of RB6-8C5 or 1A8 on wound cellularity. Mice were pretreated or not with RB6-8C5 or 1A8 mAb. PVA sponges were inserted s.c. under sterile conditions. Sponges were removed at 24 h, and total and differential cell counts were obtained. Experimental groups were compared by Kruskall-Wallis test with post hoc Mann-Whitney U-test. n  16 in each group.
|
staining in wound leukocytes [27
]. Results shown in Figure 5
demonstrate a 30% increase in TNF- staining in wound macrophages from RB6-8C5-pretreated mice compared with controls. TNF- staining in wound neutrophils from RB6-8C5-treated mice was also increased. Pretreatment with 1A8 did not increase intracellular TNF- staining in wound macrophages or wound neutrophils.
 View larger version (22K): [in a new window] |
Figure 5. Effect of RB6-8C5 or 1A8 on TNF- production in wound leukocytes. Wound cells were isolated from control or RB6-8C5-treated or 1A8-treated animals, incubated with Brefeldin A, and stained for CD68 and TNF-. Wound neutrophils are CD68neg/low cells; wound macrophages are CD68high cells. Median channel fluorescence is indicated in each quadrant.
|
Neutrophil depletion has been used to elucidate the role of neutrophils in a variety of immune responses. Many studies used the anti-Gr-1 mAb RB6-8C5 as a neutrophil-depleting agent. The expression of Gr-1 on non-neutrophils has raised concern regarding the use of RB6-8C5 to induce neutropenia, as the results of studies that used this antibody may be confounded by the unintended depletion of other Gr-l-expressing cells. The present work demonstrates that in vivo administration of RB6-8C5 depleted Gr-1-expressing blood monocytes. These results, along with previous reports demonstrating reduction of blood lymphocytes [23 ] and splenic DCs [22 ] in RB6-8C5-treated animals, indicate that a more specific means to deplete neutrophils is needed.
This work describes the use of an Ly6G-specific mAb, 1A8, to deplete neutrophils in mice. This antibody identified blood neutrophils and wound neutrophils but not monocyte/macrophage populations. Administration of 1A8 decreased blood neutrophils as predicted (Table 2) but preserved Gr-1-expressing blood monocytes (Table 3) .
As mentioned in Results, a number of studies have reported a proinflammatory phenotype in animals made neutropenic through a variety of cell-depleting strategies, including the systemic administration of RB6-8C5 [27
28
29
30
31
]. A salient component of this proinflammatory phenotype is an increase in TNF- responses, in either endotoxemia or tissue injury, which has been attributed to the elimination of neutrophils. To determine whether these observations stemmed from the elimination of neutrophils, or alternatively, were a consequence of the elimination of Gr-1+ non-neutrophil populations, experiments were performed in RB6-8C5- and 1A8-treated animals. RB6-8C5- and 1A8-pretreated animals had increased plasma TNF- in endotoxemia compared with controls; however, as shown in Figure 3
, plasma TNF- in endotoxemia was substantially less in 1A8-treated animals than in RB6-8C5-treated animals. The depletion of neutrophils and non-neutrophil Gr-1+ cells (by RB6-8C5) had a greater impact on the development of a proinflammatory phenotype than did the depletion of neutrophils alone.
In wounds, pretreatment with either antibody reduced the number of neutrophils and macrophages at the site of injury. The decrease in the number of wound macrophages in 1A8-pretreated animals, despite a normal number of Gr-1+ blood monocytes, suggests that the accumulation of macrophages in wounds depends not only on a normal complement of Gr-1+ blood monocytes but also on normal numbers of neutrophils in the wound. A decrease in wound neutrophils may alter macrophage migration to the wound, perhaps as a result of altered chemotactic signals in the neutrophil-depleted wound milieu.
Although 1A8 pretreatment decreased macrophage accumulation in the wound, it did not impact on TNF- production by wound macrophages, as determined by intracellular staining for this cytokine. This is in contrast to the increased TNF- production by wound cells in RB6-8C5-treated animals, which lack not only neutrophils but also Gr-1+ monocytes.
Results suggest that non-neutrophil Gr-1+ cells may suppress TNF- production locally and systemically. It is interesting that early wound macrophages express Gr-1 and CD11b (unpublished observation) and contain arginase I, all of which are characteristic of the recently described myeloid suppressor cells, which cause T cell dysfunction and which are detected in tumors, viral infections, and graft-versus-host disease [32
]. The possibility that macrophages exert an immunomodulatory function in wounds, as myeloid suppressor cells do in tumors, deserves further investigation.
Results presented here demonstrate that 1A8 mAb is a preferable alternative to RB6-8C5 mAb for neutrophil-depletion studies, as 1A8 selectively depletes neutrophils and preserves Gr-1-expressing blood monocytes. The use of 1A8 or other Ly6G-specific mAb for neutrophil depletion should clarify the role of neutrophils versus other Gr-1+ cells in host defense.
ACKNOWLEDGEMENTS
This work was supported by National Institutes of Health (NIH) grants GM-79227 (to J. M. D.), GM-42859 (to J. E. A.), and GM-66194 (to J. S. R.), the Carter Family Charitable Trust (Armand D. Versaci Research Scholar in Surgical Sciences Award to A. A. T.), and funds allocated to the Department of Surgery by Rhode Island Hospital, a Lifespan partner. M. D. C. was supported by NIH grant T32GM-65085. The authors thank Dr. Thomas Malek for the gift of the 1A8 hybridoma cells; Dr. Stephen Gregory for his helpful discussion; William L. Henry Jr., Balduino Mastrofrancesco, and Sara Spangenberger for technical assistance; and Patricia Young for assistance with preparation of the manuscript.
Received April 25, 2007; revised August 9, 2007; accepted August 29, 2007.
REFERENCES
Eur. J. Immunol. 18,1819-1826[Medline] and neutrophils in endotoxin-induced liver injury Am. J. Physiol. 265,G1011-G1015[Medline] production in neutropenia Am. J. Physiol. Regul. Integr. Comp. Physiol. 288,R409-R412This article has been cited by other articles:
 |
|
 |
|
  A. M. Piliponsky, C.-C. Chen, M. A. Grimbaldeston, S. M. Burns-Guydish, J. Hardy, J. Kalesnikoff, C. H. Contag, M. Tsai, and S. J. Galli Mast Cell-Derived TNF Can Exacerbate Mortality during Severe Bacterial Infections in C57BL/6-KitW-sh/W-sh Mice Am. J. Pathol., February 1, 2010; 176(2): 926 - 938. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Ingersoll, R. Spanbroek, C. Lottaz, E. L. Gautier, M. Frankenberger, R. Hoffmann, R. Lang, M. Haniffa, M. Collin, F. Tacke, et al. Comparison of gene expression profiles between human and mouse monocyte subsets Blood, January 21, 2010; 115(3): e10 - e19. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Daley, S. K. Brancato, A. A. Thomay, J. S. Reichner, and J. E. Albina The phenotype of murine wound macrophages J. Leukoc. Biol., January 1, 2010; 87(1): 59 - 67. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. T. Silva When two is better than one: macrophages and neutrophils work in concert in innate immunity as complementary and cooperative partners of a myeloid phagocyte system J. Leukoc. Biol., January 1, 2010; 87(1): 93 - 106. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. D. Meeks, A. N. Sieve, J. K. Kolls, N. Ghilardi, and R. E. Berg IL-23 Is Required for Protection against Systemic Infection with Listeria monocytogenes J. Immunol., December 15, 2009; 183(12): 8026 - 8034. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. D. Tate, Y.-M. Deng, J. E. Jones, G. P. Anderson, A. G. Brooks, and P. C. Reading Neutrophils Ameliorate Lung Injury and the Development of Severe Disease during Influenza Infection J. Immunol., December 1, 2009; 183(11): 7441 - 7450. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E Ryschich, V Kerkadze, O Deduchovas, O Salnikova, A Parseliunas, A Marten, W Hartwig, M Sperandio, and J Schmidt Intracapillary leucocyte accumulation as a novel antihaemorrhagic mechanism in acute pancreatitis in mice Gut, November 1, 2009; 58(11): 1508 - 1516. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Kumar, D. A. Allen, J. E. Kieswich, N. S. A. Patel, S. Harwood, E. Mazzon, S. Cuzzocrea, M. J. Raftery, C. Thiemermann, and M. M. Yaqoob Dexamethasone Ameliorates Renal Ischemia-Reperfusion Injury J. Am. Soc. Nephrol., November 1, 2009; 20(11): 2412 - 2425. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. C. Chin, B. Fournier, E. J. Peatman, T. A. Reaves, W. Y. Lee, and C. A. Parkos CD47 and TLR-2 Cross-Talk Regulates Neutrophil Transmigration J. Immunol., November 1, 2009; 183(9): 5957 - 5963. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Fahlen-Yrlid, T. Gustafsson, J. Westlund, A. Holmberg, A. Strombeck, M. Blomquist, G. G. MacPherson, J. Holmgren, and U. Yrlid CD11chigh Dendritic Cells Are Essential for Activation of CD4+ T Cells and Generation of Specific Antibodies following Mucosal Immunization J. Immunol., October 15, 2009; 183(8): 5032 - 5041. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Ho-Tin-Noe, C. Carbo, M. Demers, S. M. Cifuni, T. Goerge, and D. D. Wagner Innate Immune Cells Induce Hemorrhage in Tumors during Thrombocytopenia Am. J. Pathol., October 1, 2009; 175(4): 1699 - 1708. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. E. Woodman, A. E. Cooley, R. Avdiushko, A. Bowman, M. Botto, R. M. Wooten, N. van Rooijen, D. A. Cohen, and B. Stevenson Roles for phagocytic cells and complement in controlling relapsing fever infection J. Leukoc. Biol., September 1, 2009; 86(3): 727 - 736. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. C. Kirby, L. Beattie, A. Maroof, N. van Rooijen, and P. M. Kaye SIGNR1-Negative Red Pulp Macrophages Protect against Acute Streptococcal Sepsis after Leishmania donovani-Induced Loss of Marginal Zone Macrophages Am. J. Pathol., September 1, 2009; 175(3): 1107 - 1115. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Bodhankar, M. D. Woolard, X. Sun, and J. W. Simecka NK Cells Interfere with the Generation of Resistance against Mycoplasma Respiratory Infection following Nasal-Pulmonary Immunization J. Immunol., August 15, 2009; 183(4): 2622 - 2631. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. E. Thaxton, R. Romero, and S. Sharma TLR9 Activation Coupled to IL-10 Deficiency Induces Adverse Pregnancy Outcomes J. Immunol., July 15, 2009; 183(2): 1144 - 1154. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Yin and T. A. Ferguson Identification of an IFN-{gamma}-Producing Neutrophil Early in the Response to Listeria monocytogenes J. Immunol., June 1, 2009; 182(11): 7069 - 7073. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. A. Thomay, J. M. Daley, E. Sabo, P. J. Worth, L. J. Shelton, M. W. Harty, J. S. Reichner, and J. E. Albina Disruption of Interleukin-1 Signaling Improves the Quality of Wound Healing Am. J. Pathol., June 1, 2009; 174(6): 2129 - 2136. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. R. Wilson, K. P. O'Dea, D. Zhang, A. D. Shearman, N. van Rooijen, and M. Takata Role of Lung-marginated Monocytes in an In Vivo Mouse Model of Ventilator-induced Lung Injury Am. J. Respir. Crit. Care Med., May 15, 2009; 179(10): 914 - 922. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. C. Timmons, A.-M. Fairhurst, and M. S. Mahendroo Temporal Changes in Myeloid Cells in the Cervix during Pregnancy and Parturition J. Immunol., March 1, 2009; 182(5): 2700 - 2707. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. P. Stirling, S. Liu, P. Kubes, and V. W. Yong Depletion of Ly6G/Gr-1 Leukocytes after Spinal Cord Injury in Mice Alters Wound Healing and Worsens Neurological Outcome J. Neurosci., January 21, 2009; 29(3): 753 - 764. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. P. O'Dea, M. R. Wilson, J. O. Dokpesi, K. Wakabayashi, L. Tatton, N. van Rooijen, and M. Takata Mobilization and Margination of Bone Marrow Gr-1high Monocytes during Subclinical Endotoxemia Predisposes the Lungs toward Acute Injury J. Immunol., January 15, 2009; 182(2): 1155 - 1166. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Leendertse, R. J. L. Willems, I. A. J. Giebelen, J. J. T. H. Roelofs, M. J. M. Bonten, and T. van der Poll Neutrophils Are Essential for Rapid Clearance of Enterococcus faecium in Mice Infect. Immun., January 1, 2009; 77(1): 485 - 491. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. Eyles, M. J. Hickey, M. U. Norman, B. A. Croker, A. W. Roberts, S. F. Drake, W. G. James, D. Metcalf, I. K. Campbell, and I. P. Wicks A key role for G-CSF-induced neutrophil production and trafficking during inflammatory arthritis Blood, December 15, 2008; 112(13): 5193 - 5201. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. A. Zola, E. S. Lysenko, and J. N. Weiser Mucosal Clearance of Capsule-Expressing Bacteria Requires Both TLR and Nucleotide-Binding Oligomerization Domain 1 Signaling J. Immunol., December 1, 2008; 181(11): 7909 - 7916. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Dumont, P. Bouchard, and J. Frenette Neutrophil-induced skeletal muscle damage: a calculated and controlled response following hindlimb unloading and reloading Am J Physiol Regulatory Integrative Comp Physiol, December 1, 2008; 295(6): R1831 - R1838. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Sawanobori, S. Ueha, M. Kurachi, T. Shimaoka, J. E. Talmadge, J. Abe, Y. Shono, M. Kitabatake, K. Kakimi, N. Mukaida, et al. Chemokine-mediated rapid turnover of myeloid-derived suppressor cells in tumor-bearing mice Blood, June 15, 2008; 111(12): 5457 - 5466. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. D. Woolard, L. L. Hensley, T. H. Kawula, and J. A. Frelinger Respiratory Francisella tularensis Live Vaccine Strain Infection Induces Th17 Cells and Prostaglandin E2, Which Inhibits Generation of Gamma Interferon-Positive T Cells Infect. Immun., June 1, 2008; 76(6): 2651 - 2659. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. C. Lockhart and S. V. Brooks Neutrophil accumulation following passive stretches contributes to adaptations that reduce contraction-induced skeletal muscle injury in mice J Appl Physiol, April 1, 2008; 104(4): 1109 - 1115. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
