DNA methyltransferase 3a limits the expression of interleukin-13 in T helper 2 cells and allergic airway inflammation

The inverse correlation between DNA methylation and lineage-specific gene expression during T helper cell development is well documented. However, the specific functions of the de novo methyltransferases Dnmt3a and Dnmt3b in cytokine gene regulation have not been defined. We demonstrate that the expression of Dnmt3a and Dnmt3b are induced to a greater extent in T helper 2 (Th2) cells than in T helper 1 cells during polarization. Using conditional mutant mice, we determined that Dnmt3a, but not Dnmt3b, regulated expression of T helper cell cytokine genes, with the Il13 gene most prominently affected. Dnmt3a deficiency was accompanied by decreases in DNA methylation and changes in the H3K27 acetylation/methylation status at the Il13 locus. Dnmt3a-dependent regulation of Il13 also occurred in vivo because Dnmt3a(fl/fl)Cd4cre mice exhibited increased lung inflammation in a murine asthma model, compared with littermate controls. Based on these observations, we conclude that Dnmt3a is required for controlling normal Il13 gene expression and functions as a rate-limiting factor to restrict T helper 2-mediated inflammation.     

Dichotomy between naïve and memory CD4+ T cell responses to Fas engagement

Engagement of Fas (APO-1, CD95), a member of the tumor necrosis factor receptor superfamily, can induce apoptotic cell death. However, Fas engagement also can costimulate lymphocyte proliferation. The physiologic regulation of these two outcomes is poorly understood. Here, we have used two systems, the first in vitro and the second in vivo, to demonstrate that na?ve and memory CD4(+) T cells display dichotomous responses to Fas ligation. Na?ve CD4(+) T cells (CD44(lo), CD45RB+, CD62L+) die as a consequence of Fas ligation in the presence of anti-CD3 antibody, whereas memory T cells (CD44(hi), CD45RB-, CD62L-), freshly isolated from the same starting population and subjected to the same stimulation conditions, are costimulated to proliferate by Fas ligation. In vitro, we demonstrate that CD28-mediated signals or T helper 1 and T helper 2 differentiation cytokines alter the response of na?ve T cells, but not of memory T cells, to Fas ligation. In vivo experiments in hen egg lysozyme (HEL) T cell receptor transgenic mice show that CD4(+) T cells from HEL-na?ve mice are killed by Fas ligation, but CD4(+) T cells from long-term HEL-exposed mice are costimulated by Fas ligation. Thus, the physiological outcome of Fas ligation in CD4(+) T cells is determined primarily by the antigenic history of the T cell.     

In TH2 cells the Il4 gene has a series of accessibility states associated with distinctive probabilities of IL-4 production

TH2 clones may produce very variable amounts of IL-4. Among six TH2 clones prepared from homozygous or heterozygous mice in which Gfp replaced the first exon of Il4, a range of patterns of CpG methylation in the Il4/Il13 locus was observed correlating with the degree of expression of IL-4 or green fluorescence protein. Patterns of histone acetylation also showed differences between "high" and "low" TH2 clones. These results indicate that in TH2 cells the Il4 locus may display variable patterns of chromatin accessibility associated with distinct degrees of IL-4 expression. This finding suggests a regulation of IL-4 expression keyed to the function of this cytokine in cell/cell interactions and in the regulation of threshold responses.     

IBD: of mice and men-shedding new light on IL-13 activity in IBD

a recent study in a mouse model of colitis has demonstrated that interleukin (Il)?13, through inhibition of the mixed type 1 and type 17 T?helper cell inflammatory response, has a protective effect. the decoyreceptor Il?13rα2 inhibits this protective effect, suggesting blockade of Il?13rα2 as a potential therapy for patients with IBD.     

Early intrahepatic antigen-specific retention of naïve CD8+ T cells is predominantly ICAM-1/LFA-1 dependent in mice

We have previously shown that na?ve CD8+ T cells recognizing their cognate antigen within the liver are retained and undergo activation in situ, independent from lymphoid tissues. Intrahepatic primary T cell activation results in apoptosis and may play a crucial role in the ability of the liver to induce tolerance. Although adhesion molecules required for intrahepatic retention of T cells that have undergone previous extra-hepatic activation have been characterized, adhesive interactions involved in selective antigen-dependent intrahepatic retention of na?ve CD8+ T cells have not been investigated. By adoptively transferring radiolabeled T cell receptor (TCR)-transgenic CD8+ T cells into recipient animals ubiquitously expressing the relevant antigen, we show that 40% to 60 % of donor antigen-specific na?ve CD8+ T cells were retained in the liver within 1 hour after transfer, despite ubiquitous expression of the antigen. Intravital microscopy showed that most donor na?ve T cells slowed down and were irreversibly retained intrahepatically within the first few minutes after adoptive transfer, strongly suggesting that they were directly activated by liver cells in situ. This process was largely dependent on LFA-1 and ICAM-1, but was independent of blocking with antibodies against VCAM-1, alpha4 integrin, P-selectin, VAP-1, and beta1 integrin. ICAM-2 seemed to play only a minor role in this process. Interestingly, LFA-1 expressed by both donor T cells and liver cells was involved in retention of the antigen-reactive T cells. In conclusion, LFA-1-dependent intrahepatic T cell retention and activation are linked events that may play a crucial role in the establishment of liver-induced antigen-specific tolerance.     

Effect of improved zinc status on T helper cell activation and TH1/TH2 ratio in healthy elderly individuals

Mild zinc deficiency is a common condition in healthy elderly individuals leading to impaired cell-mediated immune response. Here we report the effect of improved zinc status on TH1/TH2 balance and on the activation status of T helper cells in 19 healthy elderly subjects aged 69.8 ± 5.1 years. Our investigations revealed a mild zinc deficiency which was adjusted by oral zinc supplementation for seven weeks. Improved serum zinc levels significantly reduced levels of activated T helper cells whereas changes in TH1/TH2 ratio (determined by CCR4 and CCR5 expression) were not observed. These findings suggest that elderly individuals may benefit from moderate zinc supplementation due to improved immune response leading to reduced incidences of autoimmune diseases and infections.Presented at the Zinc Age Conference, Madrid, Feburary 10–13, 2006.     

CD8+ T cells control the TH phenotype of MBP-reactive CD4+ T cells in EAE mice

Trimolecular interactions between the T cell antigen receptor and MHC/peptide complexes, together with costimulatory molecules and cytokines, control the initial activation of naive T cells and determine whether the helper precursor cell differentiates into either T helper (TH)1 or TH2 effector cells. We now present evidence that regulatory CD8(+) T cells provide another level of control of TH phenotype during further evolution of immune responses. These regulatory CD8(+) T cells are induced by antigen-triggered CD4(+) TH1 cells during T cell vaccination and, in vitro, distinguish mature TH1 from TH2 cells in a T cell antigen receptor Vbeta-specific and Qa-1-restricted manner. In vivo, protection from experimental autoimmune encephalomyelitis (EAE) induced by T cell vaccination depends on CD8(+) T cells, and myelin basic protein-reactive TH1 Vbeta8(+) clones, but not TH2 Vbeta8(+) clones, used as vaccine T cells, protect animals from subsequent induction of EAE. Moreover, in vivo depletion of CD8(+) T cells during the first episode of EAE results in skewing of the TH phenotype toward TH1 upon secondary myelin basic protein stimulation. These data provide evidence that CD8(+) T cells control autoimmune responses, in part, by regulating the TH phenotype of self-reactive CD4(+) T cells.     

Dissecting memory T cell responses to TB: Concerns using adoptive transfer into immunodeficient mice

Several studies have used adoptive transfer of purified T cell subsets into immunodeficient mice to determine the subset of T cells responsible for mediating protection against Mycobacterium tuberculosis. These studies suggested that CD62L(hi) memory CD4(+) T cells from BCG-vaccinated mice are key for protection against tuberculosis. Importantly, we observed that transfer of na?ve CD4(+) T cells into Rag1-/- recipients protected against a mycobacterial challenge as well as transfer of BCG-experienced CD4(+) T cells. We found that transfer of total CD4(+) T cells from na?ve mice or enriched CD62L(hi)CD4(+) T cells from BCG-vaccinated mice into Rag1-/- recipients induced severe colitis by 3 weeks post cell transfer, whereas transfer of CD62L(lo)CD4(+) T cells from BCG-vaccinated mice did not. Na?ve and CD62L(hi)CD4(+) T cells proliferated extensively upon transfer and developed an activated effector phenotype in the lung, even in the absence of infectious challenge. The induction of colitis and systemic cytokine response induced by the transfer and subsequent activation of CD4(+) T cells from na?ve mice or CD62L(hi)CD4(+) T cells from BCG-vaccinated mice, into immunodeficient recipients, may heighten their ability to protect against mycobacterial challenge. This raises doubts about the validity of this model to study CD4(+) T cell-mediated protection against tuberculosis.     

T-cell repopulation and thymic volume in HIV-1-infected adult patients after highly active antiretroviral therapy

The origin of T cells after highly active antiretroviral therapy (HAART) in patients infected with human immunodeficiency virus 1 (HIV-1) is now under discussion. The possibility of renewed lymphopoiesis in aged thymuses is still controversial. In this work we combine the analysis of na?ve T cells, T-cell receptor excision circles (TRECs), and computed tomography scanning of thymic tissue to further assess whether the thymus is involved in immune reconstitution. Fifteen antiretroviral-na?ve HIV-1-infected patients were evaluated during 48 weeks of HAART. At baseline, significant correlation was present among age and both thymic volume and TRECs, and between na?ve T cells and TRECs. After starting HAART, there was a significant increase at week 12 in na?ve CD4(+) and CD8(+) T cells, TRECs, and thymic volume. The initial net increases in na?ve T cells and TREC counts were significantly correlated. Changes in thymic volume and TRECs were also indirectly related; splitting the population into 2 groups of high and low baseline TREC levels, only the group with low TREC levels had significant increases in both TRECs and thymic volume. Thus, the increase in thymic volume might be functional, in response to depleted TREC levels. Taken together, our data strongly suggest a thymic role in immune reconstitution, at least in patients with depleted baseline TREC levels. (Blood. 2002;99:3702-3706)     

T cells from elderly persons respond to neoantigenic stimulation with an unimpaired IL-2 production and an enhanced differentiation into effector cells

We analysed the capacity of T cells from young and elderly persons to produce IL-2 and IFN-gamma after in vitro stimulation with two neoantigens, namely inactivated rabies virus and recombinant Etr protein of tick-borne encephalitis virus (TBEV). Soluble antigens should per definition primarly stimulate CD4(+) na?ve T cells. Cytokine production was analysed by ELISPOT technology. T cells from elderly and young donors produced similar amounts of IL-2 after priming with both neoantigens. In contrast, IFN-gamma production was induced earlier and at lower antigenic concentrations in T cells from elderly persons than from young controls indicating an enhanced capacity of primed T cells to differentiate into effector cells. In both age groups the response pattern to neoantigenic stimulation was the same whether unfractionated blood mononuclear cells or purified CD4(+)CD45RA(+) T cells with autologous DC as APC were used. The magnitude of the response was, however slightly lower in isolated cells. Autologous DC still induced an MLR in purified CD4(+)CD45RA(+) cells, which was more pronounced in the young than in the elderly group. Our results demonstrate that the ability of CD4(+) T cells from elderly persons to respond to neoantigenic stimulation is intact and that their capacity to differentiate into effector cells is even enhanced. This is in good agreement with earlier reports on alterations in the homing receptor pattern of na?ve T cells in old age. Rapid generation of effector cells from na?ve cells may at least partly counterbalance the decreasing size of the na?ve T cell pool in elderly persons.     

Immunologic recovery after hematopoietic cell transplantation with nonmyeloablative conditioning

OBJECTIVE: We studied immune reconstitution in 51 recipients of HLA-identical hematopoietic cellular transplant (HCT) after nonmyeloablative conditioning compared to a reference group of 67 recipients after myeloablative conditioning. METHODS: Nonmyeloablative conditioning consisted of 2 Gy total-body irradiation+/-fludarabine and postgrafting cyclosporine and mycophenolate mofetil. All patients received G-CSF-mobilized peripheral blood mononuclear cells. Patients were followed with serial assessments of lymphocyte subset counts, antibody levels, virus-induced lymphoproliferation, and limiting-dilution assays for cytomegalovirus (CMV) T helper (T(H)) cells. Rates of infections over the first year after transplant were calculated. RESULTS: During the first 180 days, absolute lymphocyte subset counts were similar (except higher total and memory B cell counts on day 80 in nonmyeloablative patients). At 1 year, however, total and na?ve CD4 counts, and na?ve CD8 counts, were higher in myeloablative patients. The levels of antibodies were similar at all time points and after vaccinations. The function of CD4 cells assessed by virus-induced lymphoproliferation was similar. However, the absolute counts of CMV T(H) cells were higher at days 30 and 90 (p=0.002 and p=0.0003, respectively) after nonmyeloablative conditioning. The rates of definite infections were lower for nonmyeloablative patients during the first 90 days, but were higher later. The higher number of CMV-specific T cells days 30 and 90 after nonmyeloablative HCT coincided with a lower rate of CMV infections during that time. CONCLUSION: The immunity of nonmyeloablative HCT recipients appears better than the immunity of conventional HCT recipients early, but not late, after HCT.