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F G Lakkis

Publications and source records attributed to F G Lakkis.

34 records · Page 2Linked to original sources
PubMed1997

Expression of recombinant rat interleukin-13 (IL-13) and generation of a neutralizing rat IL-13 antiserum.

Using baculoviral and bacterial systems, we expressed biologically active recombinant rat IL-13 and generated neutralizing rat IL-13 antiserum. Recombinant rat IL-13 produced by baculovirus-infected insect cells stimulated proliferation of TF-1 premyeloid cell line and induced expression of 15-lipoxygenase mRNA in human peripheral blood monocytes. Antiserum generated by immunizing a rabbit with recombinant bacterial rat IL-13 specifically inhibited TF-1 proliferation induced by baculoviral rat IL-13 but did not neutralize human IL-13 mitogenic activity. Western blotting with anti-rat IL-13 serum revealed a approximately 12 kD protein band in supernatants of insect cells infected with recombinant baculovirus carrying the rat IL-13 cDNA. The availability of recombinant rat IL-13 and rat IL-13 antibodies should facilitate studying the role of IL-13 in rat models of human inflammatory disorders.

Animals↗
PubMed1997

Blocking the CD28-B7 T cell costimulation pathway induces long term cardiac allograft acceptance in the absence of IL-4.

Blocking the CD28-B7 T lymphocyte costimulatory pathway with the recombinant protein CTLA4Ig induces long term allograft survival in rodents. It has been suggested that this results from selective activation of the Th2 immune pathway. To test this hypothesis, we compared vascularized cardiac allograft survival in wild-type (IL-4 +/+) and homozygous IL-4 gene-knockout (IL-4 -/-) mice. We report in this study that long term survival (>100 days) of fully allogeneic grafts can be induced readily in IL-4 -/- recipients treated with a short course of CTLA4Ig. We also demonstrate that IL-4 -/- mice are deficient in Th2-type cytokine expression following in vitro or in vivo allostimulation. These results suggest that IL-4 production and subsequent generation of a Th2-type immune response are not obligatory for CTLA4Ig-induced long term acceptance of vascularized allografts.

Abatacept↗
PubMed1997

Immunologic determinants of susceptibility to experimental glomerulonephritis: role of cellular immunity.

To identify the immunologic mechanisms that influence susceptibility to GN, we compared the severity of accelerated anti-glomerular basement membrane (GBM) nephritis between Lewis (LEW) and Brown Norway (BN) rats and analyzed differences in their immune responses to the nephritogenic immunoglobulin. Lewis (LEW) rats preimmunized with sheep IgG developed proliferative GN with marked proteinuria [peak protein excretion (mean +/- SEM) = 85.3 +/- 15.3 mg/24 hr; normal = 6.4 +/- 0.8 mg/24 hr] after receiving a subnephritogenic dose of sheep anti-rat GBM antiserum. Identically treated Brown Norway (BN) rats, on the other hand, had minimal renal pathology and minimal proteinuria (peak protein excretion = 22.6 +/- 3.1 mg/24 hr; normal = 13.0 +/- 0.6 mg/24 hr). Serum titers of rat anti-sheep IgG isotypes and intraglomerular binding of sheep IgG, rat IgG, and rat complement (C3) were comparable in both strains. In contrast, only LEW rats developed a strong cellular immune response to sheep IgG represented by intrarenal T lymphocyte (OX19+) and monocyte (ED1+) accumulation [LEW vs. BN (mean +/- SEM): OX19+ = 0.60 +/- 0.10 vs. 0.14 +/- 0.01 cells/glomerulus, control = 0.02 +/- 0.01; ED1+ = 4.0 +/- 0.4 vs. 1.0 +/- 0.2 cells/glom., control = 0.8 +/- 0.3] and a significant cutaneous delayed-type hypersensitivity (DTH) reaction [LEW versus BN (mean +/- SEM): delta ear thickness = 0.22 +/- 0.02 vs. 0.05 +/- 0.03 mm; control = 0.04 +/- 0.02 mm]. Upon rechallenge with sheep IgG in vitro, LEW splenocytes expressed a T helper 1 (Th1) cytokine pattern (IFN gamma and IL-2 mRNA, but little IL-4 mRNA) which is associated with delayed-type hypersensitivity reactions. BN splenocytes, on the other hand, expressed IL-4 in addition to IL-2 and IFN gamma mRNA that is consistent with an undifferentiated (Th0) cytokine profile. These studies suggest that humoral immunity to heterologous immunoglobulin planted in the kidney is not sufficient for full expression of accelerated anti-GBM nephritis, and that additional cellular immune mechanisms are required. We conclude that susceptibility to accelerated anti-GBM nephritis is strongly influenced by the host's propensity to mount a Th1-type response and DTH reaction to the disease-inciting immunoglobulin.

Animals↗
PubMed1996

Acute rejection of vascularized heart allografts in the absence of IFNgamma.

It is generally assumed that IFNgamma plays a central role in acute allograft rejection. To test this hypothesis, we transplanted fully allogeneic (MHC class I and II incompatible) C3H/HeJ (H2k) murine hearts to IFNgamma-/- (IFNgamma gene-knockout) and IFNgamma+/+ BALB/c (H2d) mice. The phenotype of IFNgamma-/- mice was confirmed by demonstrating absent IFNgamma protein production by Con A stimulated IFNgamma-/- splenocytes. Both IFNgamma-/- and IFNgamma+/+ strains rejected transplanted hearts acutely: graft survival (mean +/- SD) was 5.2+/-0.4 and 6.0+/-0.0 days, respectively. Histologic examination revealed similar patterns of acute cellular rejection in both mouse groups. IFNgamma mRNA was present in hearts rejected by IFNgamma+/+ mice but was absent in those rejected by IFNgamma-/- mice. IL-2, IL-4, IL-10, and TNFalpha mRNA expression, on the other hand, was similar in grafts rejected by either strain. We also observed that hapten-induced delayed-type hypersensitivity (DTH) response was significantly reduced but not absent in IFNgamma-/- mice. Our results demonstrate that IFNgamma is not required for acute cellular rejection of fully allogeneic murine hearts. We propose that non-DTH mechanisms of allograft destruction could be enhanced in the absence of IFNgamma and thus lead to robust acute rejection.

Acute Disease↗
PubMed1996

Anti-inflammatory lymphokine mRNA expression in antibody-induced glomerulonephritis.

T-helper subset 2 (Th2) lymphocytes produce interleukin 4 (IL-4) and IL-10, which exert anti-inflammatory actions on monocytes and macrophages. Th1 lymphocytes, on the other hand, secrete interferon-gamma (IFN gamma) which promotes tissue inflammation. The functional dichotomy between TH1 and Th2 lymphocyte subsets suggests that these cells play a regulatory role in inflammatory disease. The participation of Th subpopulations and their lymphokine products in experimental glomerulonephritis (GN) has not been previously evaluated. In this study, we examined renal expression of Th1 and Th2-type lymphokines in the first 48 hours of passive anti-glomerular basement membrane (anti-GBM) GN in the rate. Using the reverse transcriptase-polymerase chain reaction (RT-PCR) method, apparent increase in expression of both TH1-type (IL-2 and IFN gamma) and Th2-type (IL-4 and IL-10) lymphokine mRNA was observed in glomerular-enriched renal tissue obtained from nephritic rats. Induction of monocyte-derived IL-1 alpha and IL-1 receptor antagonist (IL-1RA) mRNA expression was also detected shorted after initiation of GN. Evidence for influx of mononuclear cells including T lymphocytes into the kidney was noted during the same time period as cytokine mRNA expression. Utilizing a monoclonal anti-rat IL-4 antibody, we also detected interleukin 4-producing cells in the renal cortex 24 hours following induction of GN. these experiments demonstrate for the first time anti-inflammatory lymphokine (IL-4 and IL-10) mRNA expression and IL-4 protein production in the kidney during antibody-mediated GN. WE hypothesize that Th lymphocyte subsets modulate glomerular inflammation by producing lymphokines with opposing actions.

Animals↗
PubMed1996

Atheroembolic renal disease.

Atheroembolic renal disease is an important and often underdiagnosed cause of renal insufficiency in the elderly. Renal damage results from embolization of cholesterol crystals from atherosclerotic plaques in large vessels such as the abdominal aorta to small arteries of the kidney. The typical patient is a white man older than 60 years who has an insidious increase in serum creatinine levels after an arteriographic procedure or vascular surgery. Renal outcome ranges from partial recovery of kidney function to end stage renal disease requiring replacement therapy. Tissue injury from cholesterol crystal embolization is not restricted to the kidneys but also involves the skin, muscles, abdominal organs, and central nervous system resulting in significant morbidity and mortality. Accurate diagnosis is often made by performing a skin, muscle, or renal biopsy. Although effective therapy is lacking, heightened awareness of atheroembolic renal disease is necessary for prompt diagnosis and institution of supportive care.

Embolism, Cholesterol↗
PubMed1995

The role of cytokines in inflammatory glomerular injury.

T lymphocyte and macrophage-derived cytokines play an important role in the pathogenesis of inflammatory glomerular injury by exerting both effector and regulatory actions. In this article, evidence implicating these cytokines in the pathogenesis of experimental and human glomerulonephritis will be reviewed. The possibility that cytokines derived from different T lymphocyte subpopulations modulate glomerular inflammation by exerting opposite actions on macrophages will be discussed.

Animals↗
PubMed1994

Lipoxygenase products in normal and diseased glomeruli.

Figure 4 is a schematic summary of current evidence implicating products of the 15-lipoxygenase pathway of arachidonic acid metabolism, principally 15-S-HETE and LXA4, as endogenous antagonists for the proinflammatory actions of leukotrienes. Here, we have presented evidence for the pathophysiologic relevance of leukotrienes in glomerular immune injury and the emerging data on the multifaceted counterinflammatory actions of 15-lipoxygenase products as they relate specifically to the renal glomerulus. Clearly, these concepts are of a broader nature and would be expected to pertain to inflammatory reactions in general, be they in the glomerulus, the renal interstitium, or in extrarenal sites. The extent to which these early observations can be exploited to design strategies for the control of self-destructive inflammatory reactions in the kidney and elsewhere will be determined by future studies. Imaginative design of molecular tools for the manipulation of these enzyme systems in vivo, however, represents a potentially fruitful area of research towards the attainment of a highly worthwhile goal: the cure of glomerulonephritis.

Animals↗
PubMed1994

Induction of 15-lipoxygenase by interleukin-13 in human blood monocytes.

15-Lipoxygenase (15-LO) catalyzes hydroperoxidation of fatty acids, a reaction of potential relevance to inflammation, membrane remodeling, and atherosclerosis. In human leukocytes, 15-lipoxygenation of arachidonic acid produces 15-(S)-hydroxyeicosatetraenoic acid and lipoxin A4, which suppress white cell chemotaxis, adherence, and activation, and antagonize proinflammatory leukotrienes. Interleukin (IL)-13, produced by T-helper subset 2 (TH-2) lymphocytes, specifically and potently induced 15-LO gene expression and enzyme activity in human monocytes. Among other TH-2 lymphokines, this induction of 15-LO is shared by IL-4 but not by IL-10. Interferon-gamma, a product of TH-1 lymphocytes, blocked IL-13-mediated induction of 15-LO. The induction of the anti-inflammatory 15-LO pathway by IL-13 reveals a new facet of IL-13 biology that supports its role as a cytokine with potential to down-regulate inflammatory pathways. The contrasting effects of interferon-gamma and IL-13 on 15-LO induction demonstrate mechanisms by which T-lymphocyte subsets may modulate macrophage/monocyte function in inflammation or atherosclerosis.

Arachidonate 15-Lipoxygenase↗
PubMed1994

Co-regulated expression of glomerular 12/15-lipoxygenase and interleukin-4 mRNAs in rat nephrotoxic nephritis.

Arachidonate 12- and 15-lipoxygenase (LO) products are generated in experimental glomerulonephritis. 15-S-HETE (a 15-LO product) and lipoxins (interaction products between 5-LO and either 12-LO or 15-LO) counteract the proinflammatory actions of leukotrienes. IL-4 has been shown to up-regulate 15-LO gene expression in human leukocytes. Based on homology with human 15-LO, we cloned a 0.76 kbp fragment of a rat LO cDNA from leukocytes stimulated by interleukin-4 (IL-4). The deduced amino acid sequence shows 71.0% and 60.1% homology to human 15-LO and 12-LO, respectively, and 100% homology to a recently cloned "leukocyte type" rat 12-lipoxygenase enzyme, which possesses significant 15-lipoxygenase activity (heretofore referred to as "12/15-LO"). A deletion mutant was utilized to generate internal standard cRNA in quantitative PCR assays. Glomerular 12/15-LO mRNA increased significantly over controls 24 and 48 hours after NTS injection, then decreased at 72 hours. RNA from NTS glomeruli contained higher levels of 12/15-LO mRNA than that from unstimulated peripheral leukocytes, suggesting that 12/15-LO transcription is up-regulated locally in native and/or infiltrating glomerular cells. Glomerular IL-4 mRNA increased markedly 16 hours post-NTS, and was then reduced, suggesting a potential role for T cell-derived IL-4 in directing the expression of 12/15-LO during glomerulonephritis. This represents the first demonstration of tandem regulated in vivo gene expression for a lymphokine (IL-4) and a lipoxygenase, both of which promote counter-inflammatory influences in immune complex-mediated injury.

Amino Acid Sequence↗
PubMed1993

Cloning of rat interleukin-13 (IL-13) cDNA and analysis of IL-13 gene expression in experimental glomerulonephritis.

Using a reverse transcriptase-polymerase chain reaction method, a cDNA for the complete coding region and part of the 5'-untranslated region of rat IL-13 was cloned from rat renal cortex RNA following the induction of anti-glomerular basement membrane antibody-induced glomerulonephritis. The coding region of IL-13 cDNA displays 74% and 87% sequence identity with the coding regions of human and mouse IL-13 cDNA, respectively. The deduced amino acid sequence of rat IL-13 reveals 63% and 79% homology with the human and mouse proteins, respectively. Using the rat IL-13 cDNA as a molecular probe, increased production of IL-13 mRNA in renal and splenic tissue is demonstrated in the first 48 hours of antibody-induced glomerulonephritis in the rat. The data suggest a role for IL-13 in the inflammatory response in vivo.

Amino Acid Sequence↗