Homocysteine (Hcy) is the immediate precursor of the amino acid, methionine. In humans, blood concentrations of Hcy may become elevated as a result of deficiency in folate, vitamin B6 or vitamin B12 and has recently been identified as a putative risk factor for a number of age-associated disease states including arteriosclerosis, myocardial infarction, arterial occlusive disease, Alzheimer's disease and neural tube defects. Plasma Hcy levels have also been shown to be affected by a number of biological, environmental or pathological factors including aging, inflammation, renal insufficiency, and smoking. A specific role for Hcy or any of its metabolites, such as S-adenosyl Hcy (Ado-Hcy) or Hcy thiolactone, in these conditions has not yet been firmly established. Particularly absent is a description of the effects of elevated Hcy levels on immune function. Several studies have examined the effects of Hcy on monocyte, neutrophil and B cell function, inflammation and chemokine production; however, little is known about the Hcy effects on T lymphocytes. Given the increased levels of Hcy in elderly subjects and various inflammatory disease states, we initially sought to determine whether Hcy exhibits any direct cytotoxic or pro-apoptotic effects on human T lymphocytes. Our studies revealed that treatment of resting human T cells with Hcy (10mM to 1000 mM) resulted in a dose-dependent increase in apoptotic cell death. D, L Hcy was more potent than Hcy thiolactone in this respect while Ado-Hcy was found to be significantly less active or inactive in many cases. To examine the mechanism(s) and pathways by which Hcy mediates T cell death, we utilized a number of inhibitors of established apoptotic mediators and pathways to determine their role in our observed effects. We found that the pro-apoptotic effects of Hcy were abrogated with the addition of serum, pan-caspase inhibitors, and poly-ADP-ribose polymerase (PARP) inhibitors to the cell cultures. These results suggest that Hcy, like other apoptotic stressors, leads to the activation of the caspase cascade and eventually to the cleavage of the key cellular proteins, like PARP, eventually leading to the typical morphological changes observed in cells undergoing apoptosis. We have also found that Hcy-mediated apoptosis is inhibited by the phosphatase inhibitor, sodium orthovanadate, the intracellular calcium chelator, BAPTA-AM, and the protein synthesis inhibitor, cycloheximide. Moreover, we have also found that Hcy appears to potentiate cellular death induced by a number of other established apoptotic signals including activation-induced cell death (AICD), heat shock, and Fas ligand- and HIV-mediated T cell death. In addition to the pro-apoptotic effects of Hcy, we also examined the ability of Hcy to influence T cell activation and cytokine secretion. Stimulation of mononuclear cells or isolated T cells with immobilized anti-CD3 mAb in the presence of Hcy or thiolactone but not Ado-Hcy resulted in a significant increase in several type 1 cytokines including IL-2, IFN-gamma, TNF-alpha and IL-10 but not the type 2 cytokines, IL-4 or IL-5. More detailed examination of the Hcy effects on T cell activation revealed that this type 1 cytokine production profile is mediated, in part, through the production of IL-18 and possibly IL-12. The precise mechanism involved in the generation of these cytokines is currently under investigation but we believe the Hcy effect is being mediated, in part, by specific stress-associated signals post Hcy treatment. Data from our laboratory using microarray technology has revealed significant increase in the gene expression of a variety of molecules involved in cell cycle, apoptosis, inflammation, metabolism, stress and cell growth in Hcy-treated lymphocytes. The significance of these gene expression changes and their relationship to cellular and oxidative stress and cytokine production is currently under investigation. Overall, Hcy appears to exert a number of differential effects on immune cells, which may alter immune function in the circulation and tissue microenvironment with age and disease pathology. A greater understanding of the potential modulatory effects of Hcy and its metabolites on immune function may result in the development of potential therapeutic strategies to control and optimize immune responses with age and in various age-associated disease states.
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| Dawson, Harry; Collins, Gary; Pyle, Robert et al. (2004) The immunoregulatory effects of homocysteine and its intermediates on T-lymphocyte function. Mech Ageing Dev 125:107-10 |
| Kalayjian, Robert C; Landay, Alan; Pollard, Richard B et al. (2003) Age-related immune dysfunction in health and in human immunodeficiency virus (HIV) disease: association of age and HIV infection with naive CD8+ cell depletion, reduced expression of CD28 on CD8+ cells, and reduced thymic volumes. J Infect Dis 187:1924-33 |
