2001-emanuilidis-3919

On Fri, 06-Jul-2012   /   Intravenous Injection  
Emmanuilidis K, Weighardt H, Maier S, Gerauer K, Fleischmann T, Zheng XX, Hancock W, Holzmann B, Heidecke CD.
Critical Role of Kupffer Cell-Derived IL-10 for Host Defense in Septic Peritonitis.
J Immunol. 2001 Oct 1;167(7):3919–27.
[toggle_content title=”Abstract”]Intra-abdominal infection in patients following major visceral surgery is associated with high mortality. Using a macrophage depletion technique, we demonstrate that in murine septic peritonitis, Kupffer cells are a major source of systemic IL-10 levels. Kupffer cell-depleted mice were highly susceptible to the lethal effects of septic peritonitis and exhibited an increased bacterial load. Kupffer cell-depleted mice were protected by the administration of an IL-10-Fc fusion protein. Loss of Kupffer cell-derived IL-10 was associated with a weak increase in serum IL-12 levels, whereas TNF, IL-1, and IL-18 levels were not significantly elevated, suggesting that the loss of Kupffer cell-derived IL-10 did not result in a toxic cytokine release syndrome. Instead, loss of Kupffer cell-derived IL-10 was associated with a reduced splenocyte production of IFN- that is required for immune protection in murine septic peritonitis. Therefore, the results suggest that the protective function of IL-10 in septic peritonitis may not be restricted to the anti-inflammatory activities of IL-10. [/toggle_content] [toggle_content title=”Clodronate Liposome Parameters”] [custom_table]
Clodronate Concentration Total Lipid Concentration Lipid Composition Lipid Mole % Liposome Type Control Liposomes
5 mg/ml 23.4 mg/ml EPC/Chol 84/16 MLV none
[/custom_table] [/toggle_content] [toggle_content title=”Animals and Dosing”] [custom_table]
Animal Description Clodronate Dose Dosing Method/Site Target Phagocytes Local Dosing? Local Results
C57BL/6, female mice, 8-12 w 40 µl (diluted to 160 µl) i.v./not specified F4/80+ Kupffer cells no N/A
[/custom_table] [/toggle_content] [toggle_content title=”Notes”]
  1. Liposome prep method cited was a review, so standard clodronate liposome parameters assumed.
  2. Animals were dosed with clodronate liposomes 24 h prior to surgical procedure used to induce (peritonitis) sepsis.
  3. Some groups were splenectomized to determine the contribution of splenic macrophages.
[/toggle_content] [toggle_content title=”Results”]
  1. Peritoneal and aveolar macrophages were not reduced in number, but this was a 5X lower dose than often used in other papers (40 µl vs 200 µl).
  2. Authors report “complete” depletion of Kupffer cells along with splenic marginal macrophages and metallophilic macrophages, but not red pulp macrophages within 24 h by histology.
  3. Depletion lasted for at least 72 h.
  4. All other measurements on clodronate liposome-treated animals were either cytokine levels or IL-10 mRNA levels.
  5. The authors did not evaluate depletion in septic mice, therefore within the 12 h post-induction of sepsis (36 h post-clodronate liposome injection) before organs were harvested, would some migration of repopulating monocytes into tissues (including liver and spleen) occur?
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2009-zecher-7810

On Thu, 28-Jun-2012   /   Intravenous Injection  
Zecher D, van Rooijen N, Rothstein DM, Shlomchik WD, Lakkis FG.
An innate response to allogeneic nonself mediated by monocytes.
The Journal of Immunology. 2009;183(12):7810.
[toggle_content title=”Abstract”] The mammalian innate immune system has evolved diverse strategies to distinguish self from microbial nonself. How the innate immune system distinguishes self-tissues from those of other members of the same species (allogeneic nonself) is less clear. To address this question, we studied the cutaneous hypersensitivity response of lymphocyte-deficient RAG-/- mice to spleen cells transplanted from either allogeneic or syngeneic RAG-/- donors. We found that RAG-/- mice mount a specific response to allogeneic cells characterized by swelling and infiltration of the skin with host monocytes/macrophages and neutrophils. The response required prior priming with allogeneic splenocytes or skin grafts and exhibited features of memory as it could be elicited at least 4 wk after immunization. Neither depletion of host NK cells nor rechallenging immunized mice with F1 hybrid splenocytes inhibited the response, indicating that the response is not mediated by NK cells. Depletion of host monocytes/macrophages or neutrophils at the time of rechallenge significantly diminished the response and, importantly, the adoptive transfer of monocytes from alloimmunized RAG-/- mice conferred alloimmunity to naive RAG-/- hosts. Unlike NK- and T cell-dependent alloresponses, monocyte-mediated alloimmunity could be elicited only when donor and responder mice differed at non-MHC loci. These observations indicate that monocytes mount a response to allogeneic nonself, a function not previously attributed to them, and suggest the existence of mammalian innate allorecognition strategies distinct from detection of missing self-MHC molecules by NK cells. [/toggle_content] [toggle_content title=”Clodronate Liposome Parameters”] [custom_table]
Clodronate Concentration Total Lipid Concentration Lipid Composition Lipid Mole % Liposome Type Control Liposomes
10 mg/ml 1 46 mg/ml EPC/Chol 86/14 MLV PBS

1Clodronate and lipid concentrations assumed based on information in paper (dosed 2 mg in 0.2 ml).

[/custom_table] [/toggle_content] [toggle_content title=”Animals and Dosing”] [custom_table]
Animal Description Clodronate Dose Dosing Method/Site Target Phagocytes Adjunct Dosing? Adjunct Dosing Route
C57BL/6, C57BL/10 mice 2 , 8-20 w 250 µl intravenous/tail vein CD115+ F4/80int monocytes; F4/80+
Gr-1int splenic monocytes/MΦ; CD115+ F4/80+ bone marrow MΦ		 no NA

2 Base or background strain. Variants, mutants or otherwise genetically altered strains were also used in this study.

[/custom_table] [/toggle_content] [toggle_content title=”Notes”]
  1. Clodronate and lipid concentrations assumed based on referenced paper, although references provide little information as to final clodronate concentrations.
  2. Reference for clodronate liposome preparation – van Rooijen N, Sanders A. Liposome mediated depletion of macrophages: mechanism of action, preparation of liposomes and applications. Journal of immunological methods. 1994;174(1-2):83–93.
[/toggle_content][toggle_content title=”Results”]
  1. % Control Cells Remaining Post-Clodronate Liposome Treatment [custom_table]
    Tissue 16 h 24 h 48 h
    Bone Marrow 40 75 130
    Blood Monocytes 5 10 110
    Spleen 1 1 1

    Numbers extrapolated from supplemental figure 1 in paper.[/custom_table]

  2. MΦ/monocytes or neutrophil depletion reduces the allogenic non-self response (pinnae swelling or DTH) by 50%.
  3. PBS control liposome administration does not affect the response.
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1998-leenen-2166

On Tue, 26-Jun-2012   /   Intravenous Injection  
Leenen PJM, Radosevic K, Voerman JSA, Salomon B, van Rooijen N, Klatzmann D, van Ewijk W.
Heterogeneity of Mouse Spleen Dendritic Cells: In Vivo Phagocytic Activity, Expression of Macrophage Markers, and Subpopulation Turnover.
The Journal of Immunology. 1998;160(5):2166–73.
[toggle_content title=”Abstract”] In the normal mouse spleen, two distinct populations of dendritic cells (DC) are present that differ in microanatomical location. The major population of marginal DC is found in the “marginal zone bridging channels” and extends into the red pulp. The interdigitating cells (IDC) are localized in the T cell areas in the white pulp. The aim of the present study was to characterize these two splenic DC populations with regard to their phenotype, in vivo phagocytic function, and turnover. Both marginal DC and IDC are CD11c+ and CD13+, but only IDC are NLDC-145+ and CD8α+. Notably, both populations, when freshly isolated, express the macrophage markers F4/80, BM8, and Mac-1. To study the phagocytic capacity of these cells, we employed the macrophage “suicide” technique by injecting liposomes loaded with clodronate i.v. Marginal DC, but not IDC, were eliminated by this treatment. Phagocytosis of DiI-labeled liposomes by DC confirmed this finding. The two DC populations differed significantly with regard to their turnover rates, as studied in a transgenic mouse model of conditional depletion of DC populations with high turnover. In these mice, marginal DC were completely eliminated, but the IDC population remained virtually intact. From these data we conclude that the marginal DC population has a high turnover, in contrast to the IDC population. Taken together, the present results indicate that marginal DC and IDC represent two essentially distinct populations of DC in the mouse spleen. They differ not only in location, but also in phenotype, phagocytic ability, and turnover. [/toggle_content] [toggle_content title=”Clodronate Liposome Parameters”] [custom_table]
Clodronate Concentration Total Lipid Concentration Lipid Composition Lipid Mole % Liposome Type Control Liposomes
10 mg/ml 1 46 mg/ml EPC/Chol 86/14 MLV none

1Clodronate and lipid concentrations assumed based on information in paper (dosed 2 mg in 0.2 ml).

[/custom_table] [/toggle_content] [toggle_content title=”Animals and Dosing”] [custom_table]
Animal Description Clodronate Dose Dosing Method/Site Target Phagocytes Adjunct Dosing? Adjunct Dosing Route
C57BL/6, C57BL/10 mice 2, 8-20 w 200 µl intravenous CD11c+, CD13+, F4/80+, etc. splenic DC / IDC no NA

2Base or background strain. Variants, mutants or otherwise genetically altered strains were also used in this study.

[/custom_table] [/toggle_content] [toggle_content title=”Notes”]
  1. Clodronate and lipid concentrations assumed based on referenced paper, although references provide little information as to final clodronate concentrations.
  2. Reference for clodronate liposome preparation – van Rooijen N, Sanders A. Liposome mediated depletion of macrophages: mechanism of action, preparation of liposomes and applications. Journal of immunological methods. 1994;174(1-2):83–93.
[/toggle_content][toggle_content title=”Results”]
  1. Splenic marginal, but not interdigitated, dendritic cells were completely depleted at 48 h post-liposomal clodronate treatment as judged by histological analysis.
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