Activated Macrophage Conditioned Medium: Identification of the Soluble Factors Inducing Cytotoxicity and the L-Arginine Dependent Effector Mechanism

Amber, Ina J.; Hibbs, John B.; Parker, Charles J.; Johnson, Barbara B.; Taintor, Read R.; Vavrin, Zdenek

doi:10.1002/jlb.49.6.610

Cited by 54 publications

(31 citation statements)

References 44 publications

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“…ASE I and ASE II are encoded by two distinct genes and hydrolyze L-arginine into urea and L-ornithine, the latter being the main substrate for the production of polyamines (putrescine, spermidine, and spermine) that are required for cell cycle progression. L-Arginine is also metabolized in macrophages by iNOS to citrulline and nitric oxide, a highly reactive compound important in the cytotoxic mechanism of these cells (4,5). The importance of L-arginine on the immune response was suggested by the marked association of impaired T cell function in patients and rodents after liver transplantation or trauma (11,12,26,27), which rapidly recovered with the enteral or parenteral supplementation of L-arginine (14 -16, 28).…”

Section: Discussionmentioning

confidence: 99%

“…We recently showed that T cells cultured in low concentrations of L-arginine lose CD3 expression, are unable to proliferate, and have a decreased cytokine production (18,19). 4 However, it was unclear how L-arginine availability might be regulated during an immune response. We therefore asked which of the metabolic pathways that use L-arginine in macrophages could modulate the availability of L-arginine and whether this would alter the expression of CD3 chain and proliferation in normal T cells.…”

Section: Discussionmentioning

confidence: 99%

“…A similar effect has been seen in normal human T lymphocytes where the absence of arginine impairs the re-expression of chain by blocking its synthesis. 4 Inhibitors of ASE and iNOS and a hydrogen peroxide scavenger were then used to further determine the role of these enzymatic pathways in the regulation of extracellular L-arginine and the expression of CD3 . The addition of ASE inhibitors NOHA or Nor-NOHA, or the excess of L-arginine in the cultures, partially prevented the reduction of L-arginine in the tissue culture medium of IL-4 ϩ IL-13-stimulated PM (Table I).…”

Section: Ase I Depletes L-arginine and Induces Cd3 Down-regulationmentioning

confidence: 99%

“…Therefore, the decrease in L-arginine levels induced by IL-4 ϩ IL-13-stimulated PM could be the result of ASE I released into the tissue culture media. Interestingly, there was no 4 A. Zea. L-Arginine modulates CD3 expression and function in activated T cells.…”

Section: Ase I/cat-2b Increase L-arginine Uptake In Macrophagesmentioning

confidence: 99%

“…It is metabolized in macrophages, endothelial cells, hepatocytes, kidney cells, and certain tumors by three enzymatic pathways, the inducible nitric oxide synthase (iNOS), 3 arginase I (ASE I), and arginase II (ASE II) (3). In macrophages, L-arginine is metabolized by iNOS to produce citrulline and nitric oxide, which is one of the principal cytotoxic mechanisms in these cells (4,5). Alternatively, ASE I and ASE II metabolize L-arginine to L-ornithine and urea, the first being the precursor for the production of polyamines that are essential for cell proliferation and the second being an important mechanism for detoxification (3).…”

mentioning

confidence: 99%

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l-Arginine Consumption by Macrophages Modulates the Expression of CD3ζ Chain in T Lymphocytes

Rodríguez

Zea

DeSalvo

et al. 2003

The Journal of Immunology

423

355

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l-Arginine plays a central role in the normal function of several organs including the immune system. It is metabolized in macrophages by inducible nitric oxide synthase to produce nitric oxide, important in the cytotoxic mechanisms, and by arginase I (ASE I) and arginase II (ASE II) to synthesize l-ornithine and urea, the first being the precursor for the production of polyamines needed for cell proliferation. l-Arginine availability can modulate T cell function. Human T cells stimulated and cultured in the absence of l-arginine lose the expression of the TCR ζ-chain (CD3ζ) and have an impaired proliferation and a decreased cytokine production. The aim of this work was to test whether activated macrophages could modulate extracellular levels of l-arginine and alter T cell function, and to determine which metabolic pathway was responsible for this event. The results show that macrophages stimulated with IL-4 + IL-13 up-regulate ASE I and cationic amino acid transporter 2B, causing a rapid reduction of extracellular levels of l-arginine and inducing decreased expression of CD3ζ and diminished proliferation in normal T lymphocytes. Competitive inhibitors of ASE I or the addition of excess l-arginine lead to the re-expression of CD3ζ and recovery of T cell proliferation. In contrast, inducible nitric oxide synthase or ASE II failed to significantly reduce the extracellular levels of l-arginine and modulate CD3ζ expression. These results may provide new insights into the mechanisms leading to T cell dysfunction and the down-regulation of CD3ζ in cancer and chronic infectious diseases.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Ase I Depletes L-arginine and Induces Cd3 Down-regulationmentioning

confidence: 99%

Section: Ase I/cat-2b Increase L-arginine Uptake In Macrophagesmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

l-Arginine Consumption by Macrophages Modulates the Expression of CD3ζ Chain in T Lymphocytes

Rodríguez

Zea

DeSalvo

et al. 2003

The Journal of Immunology

423

355

View full text Add to dashboard Cite

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Nitric oxide production by tumor targets in response to TNF: paradoxical correlation with susceptibility to TNF-mediated cytotoxicity without direct involvement in the cytotoxic mechanism

Fast

Lynch

Leu

1992

Journal of Leukocyte Biology

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Tumor necrosis factor (TNF) is selectively cytotoxic for some tumor cells in vivo and in vitro. We determined whether TNF-mediated cytotoxicity for TNF-sensitive tumor targets was related to TNF-stimulated production of NO by the tumor cell itself. We found that a cell line that was sensitive to TNF-mediated cytotoxicity produced NO in response to TNF as measured by the accumulation of nitrite in the supernatants of TNF-stimulated cells. Production of NO in response to TNF was inhibited by the competitive substrate inhibitor, NG-monomethyl-L-arginine (NMMA). The kinetics of NO production in response to TNF indicated that most of the NO was produced during the first 24 h and peaked after 48 h of culture and that TNF-stimulated NO production was dose dependent. TNF-resistant cell lines produced less NO than a TNF-sensitive cell line, and the amount of nitrite produced correlated with the relative sensitivity of each cell line to TNF-mediated cytotoxicity. In addition, recombinant interferon-gamma augmented the amount of NO produced in response to TNF by both sensitive and resistant cells and correspondingly enhanced the susceptibility of resistant cells to TNF cytotoxicity. Both sensitive and resistant cells were sensitive to NO, however, in that NO generated exogenously by culture in the presence of sodium nitroprusside was cytotoxic for both sensitive and resistant cells in a dose-dependent manner. We were unable, however, to demonstrate directly a role for NO in TNF-mediated cytotoxicity as NMMA- and arginine-free media provided little protection from TNF-mediated cytotoxicity. We tentatively conclude that the ability of adherent murine tumor cells to produce nitric oxide in response to TNF correlates directly with their level of sensitivity to TNF-mediated cytotoxicity, although NO thus produced appears not to be directly involved in the cytotoxic mechanism.

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Inhibition of tumor cell mitochondrial respiration by macrophage cytotoxic mediators distinct from interferon-γ

Tucker

Auzenne

Sivaramakrishnan

1993

Journal of Leukocyte Biology

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Macrophage-mediated inhibition of mitochondrial respiration in EMT-6 murine mammary adenocarcinoma cells can be mimicked in vitro by treatment of the cells with interferon-gamma (IFN-gamma) in combination with tumor necrosis factor, interleukin-1, or lipopolysaccharide. Conditioned supernatants obtained from activated macrophages appear to contain interferon-gamma, suggesting that inhibition of mitochondrial respiration in tumor cells was caused by synergy of IFN-gamma with other cytokines. To further characterize monokines that cause inhibition of mitochondrial respiration in tumor cells, EA13.5 macrophage-like cells were isolated and selected for inhibition of mitochondrial respiration in EMT-6 tumor cells. After stimulation with IFN-gamma and lipopolysaccharide, the EA13.5 cells released into conditioned supernatants a cytotoxic mediator that induced nitric oxide synthesis and caused lesions in the electron transport chain of EMT-6 cells similar to the lesions caused by activated peritoneal macrophages. Enzyme-linked immunosorbent assay demonstrated that the conditioned supernatants produced by EA13.5 macrophage cells did not contain IFN-gamma. Treatment of the EA13.5 cell-conditioned supernatants with neutralizing antibody against IFN-gamma did not abrogate the inhibition of mitochondrial respiration in EMT-6 cells caused by these conditioned supernatants. This study demonstrated that unidentified macrophage cytotoxic mediators distinct from IFN-gamma are involved in the induction of nitric oxide synthesis and inhibition of mitochondrial respiration in tumor cells.

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Activated Macrophage Conditioned Medium: Identification of the Soluble Factors Inducing Cytotoxicity and the L-Arginine Dependent Effector Mechanism

Cited by 54 publications

References 44 publications

l-Arginine Consumption by Macrophages Modulates the Expression of CD3ζ Chain in T Lymphocytes

l-Arginine Consumption by Macrophages Modulates the Expression of CD3ζ Chain in T Lymphocytes

Nitric oxide production by tumor targets in response to TNF: paradoxical correlation with susceptibility to TNF-mediated cytotoxicity without direct involvement in the cytotoxic mechanism

Inhibition of tumor cell mitochondrial respiration by macrophage cytotoxic mediators distinct from interferon-γ

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