NOD.Cd1-/- mice have increased numbers of CD4+CD25+Foxp3+ T regulatory cells in the periphery

Recent studies indicate Natural Killer T (NKT) lymphocytes and Regulatory T (Treg) cells interact to regulate immune responses. NOD mice bearing a targeted deletion of the NKT cell restriction molecule, CD1d, lack NKT cells. To determine whether absence of NKT cells has an effect on Treg numbers, NOD/Lt and NOD.Cd1-/- mice were examined for differences in their Treg population. Flow cytometric analysis of 10-week old female mice of both strains revealed that NOD.Cd1-/- mice have a higher number of CD4+CD25+Foxp3+ T cells in both the spleen and liver. Further studies on differences in regulatory T cells between these strains at different ages and the effect of adoptive transfer of NKT cells on Treg cells would give us a better understanding about interactions of these two populations

An Essential Role for Tumor Necrosis Factor in Natural Killer Cell–mediated Tumor Rejection in the Peritoneum

Natural killer (NK) cells are thought to provide the first line of defence against tumors, particularly major histocompatibility complex (MHC) class I− variants. We have confirmed in C57BL/6 (B6) mice lacking perforin that peritoneal growth of MHC class I− RMA-S tumor cells in unprimed mice is controlled by perforin-dependent cytotoxicity mediated by CD3− NK1.1+ cells. Furthermore, we demonstrate that B6 mice lacking tumor necrosis factor (TNF) are also significantly defective in their rejection of RMA-S, despite the fact that RMA-S is insensitive to TNF in vitro and that spleen NK cells from B6 and TNF-deficient mice are equally lytic towards RMA-S. NK cell recruitment into the peritoneum was abrogated in TNF-deficient mice challenged with RMA-S or RM-1, a B6 MHC class I− prostate carcinoma, compared with B6 or perforin-deficient mice. The reduced NK cell migration to the peritoneum of TNF-deficient mice correlated with the defective NK cell response to tumor in these mice. By contrast, a lack of TNF did not affect peptide-specific cytotoxic T lymphocyte–mediated rejection of tumor from the peritoneum of preimmunized mice. Overall, these data show that NK cells delivering perforin are the major effectors of class I− tumor rejection in the peritoneum, and that TNF is specifically critical for their recruitment to the peritoneum

Double deficiency of toll-like receptors 2 and 4 alters long-term neurological sequelae in mice cured of pneumococcal meningitis

Toll-like receptor (TLR) 2 and 4 signalling pathways are central to the body's defence against invading pathogens during pneumococcal meningitis. Whereas several studies support their importance in innate immunity, thereby preventing host mortality, any role in protecting neurological function during meningeal infection is ill-understood. Here we investigated both the acute immunological reaction and the long-term neurobehavioural consequences of experimental pneumococcal meningitis in mice lacking both TLR2 and TLR4. The absence of these TLRs significantly impaired survival in mice inoculated intracerebroventricularly with Streptococcus pneumoniae. During the acute phase of infection, TLR2/4-deficient mice had lower cerebrospinal fluid concentrations of interleukin-1 beta, and higher interferon-gamma, than their wild-type counterparts. After antibiotic cure, TLR2/4 double deficiency was associated with aggravation of behavioural impairment in mice, as shown by diurnal hypolocomotion throughout the adaptation phases in the Intellicage of TLR-deficient mice compared to their wild-type counterparts. While TLR2/4 double deficiency did not affect the cognitive ability of mice in a patrolling task, it aggravated the impairment of cognitive flexibility. We conclude that TLR2 and TLR4 are central to regulating the host inflammatory response in pneumococcal meningitis, which may mediate diverse compensatory mechanisms that protect the host not only against mortality but also long-term neurological complications

Ferromagnetic phase transition for the spanning-forest model (q \to 0 limit of the Potts model) in three or more dimensions

We present Monte Carlo simulations of the spanning-forest model (q \to 0 limit of the ferromagnetic Potts model) in spatial dimensions d=3,4,5. We show that, in contrast to the two-dimensional case, the model has a "ferromagnetic" second-order phase transition at a finite positive value w_c. We present numerical estimates of w_c and of the thermal and magnetic critical exponents. We conjecture that the upper critical dimension is 6.Comment: LaTex2e, 4 pages; includes 6 Postscript figures; Version 2 has expanded title as published in PR

Critical speeding-up in a local dynamics for the random-cluster model

We study the dynamic critical behavior of the local bond-update (Sweeny) dynamics for the Fortuin-Kasteleyn random-cluster model in dimensions d=2,3, by Monte Carlo simulation. We show that, for a suitable range of q values, the global observable S_2 exhibits "critical speeding-up": it decorrelates well on time scales much less than one sweep, so that the integrated autocorrelation time tends to zero as the critical point is approached. We also show that the dynamic critical exponent z_{exp} is very close (possibly equal) to the rigorous lower bound \alpha/\nu, and quite possibly smaller than the corresponding exponent for the Chayes-Machta-Swendsen-Wang cluster dynamics.Comment: LaTex2e/revtex4, 4 pages, includes 5 figure

Dynamic critical behavior of the Chayes-Machta-Swendsen-Wang algorithm

We study the dynamic critical behavior of the Chayes-Machta dynamics for the Fortuin-Kasteleyn random-cluster model, which generalizes the Swendsen-Wang dynamics for the q-state Potts model to noninteger q, in two and three spatial dimensions, by Monte Carlo simulation. We show that the Li-Sokal bound z \ge \alpha/\nu is close to but probably not sharp in d=2, and is far from sharp in d=3, for all q. The conjecture z \ge \beta/\nu is false (for some values of q) in both d=2 and d=3.Comment: Revtex4, 4 pages including 4 figure

CD1d-restricted NKT cells: an interstrain comparison

CD1d-restricted Va14-Ja281 invariant abTCR1 (NKT) cells are well defined in the C57BL/6 mouse strain, but they remain poorly characterized in non-NK1.1-expressing strains. Surrogate markers for NKT cells such as abTCR1CD42CD82 and DX51CD31 have been used in many studies, although their effectiveness in defining this lineage remains to be verified. Here, we compare NKT cells among C57BL/6, NK1.1-congenic BALB/c, and NK1.1-congenic nonobese diabetic mice. NKT cells were identified and compared using a range of approaches: NK1.1 expression, surrogate phenotypes used in previous studies, labeling with CD1d/a-galactosylceramide tetramers, and cytokine production. Our results demonstrate that NKT cells and their CD4/CD8-defined subsets are present in all three strains, and confirm that nonobese diabetic mice have a numerical and functional deficiency in these cells. We also highlight the hazards of using surrogate phenotypes, none of which accurately identify NKT cells,and one in particular (DX51CD31) actually excludes these cells. Finally, our results support the concept that NK1.1 expression may not be an ideal marker for CD1d-restricted NKT cells, many of which are NK1.1-negative, especially within the CD41 subset and particularly in NK1.1-congenic BALB/c mice

Modulation of TCR signalling components occurs prior to positive selection and lineage commitment in iNKT cells

iNKT cells play a critical role in controlling the strength and character of adaptive and innate immune responses. Their unique functional characteristics are induced by a transcriptional program initiated by positive selection mediated by CD1d expressed by CD4+CD8+ (double positive, DP) thymocytes. Here, using a novel Vα14 TCR transgenic strain bearing greatly expanded numbers of CD24hiCD44loNKT cells, we examined transcriptional events in four immature thymic iNKT cell subsets. A transcriptional regulatory network approach identified transcriptional changes in proximal components of the TCR signalling cascade in DP NKT cells. Subsequently, positive and negative selection, and lineage commitment, occurred at the transition from DP NKT to CD4 NKT. Thus, this study introduces previously unrecognised steps in early NKT cell development and separates, the events associated with modulation of the T cell signalling cascade prior to changes associated with positive selection and lineage commitment

Causes of Multiple Sclerosis: a functional genomics approach

Multiple Sclerosis (MS) is the most common disabling neurological disease affecting young adults in Western Society. To date, 55 strongly associated single nucleotide polymorphisms have been discovered. We now need to identify causal genes. While T-cells as targets for therapeutic intervention have rarely proven useful, there is strong clinical and in-vitro data identifying NK cell deficiencies in patients, and key roles for monocytes in myelin and axon destruction and autoantigen presentation. RNA extracted from magnetic bead sorted monocytes and NK cells, of healthy controls (HC) and untreated patients with relapsing remitting MS (RRMS), was labelled and hybridised to Affymetrix Human Gene 1.0 ST arrays. Expression values were standardized across chips using RMA and quantile normalization as implemented in GenePattern. Genes were ranked by expression difference significance by Mann Whitney U test and ANOVA. To date, we have analysed monocytes of 30 patients and 39 HC, and NK cells from 25 patients and 32 HC. Expression differences of those genes adjacent to MS associated risk SNPs lying between 110kb upstream and 40kb downstream of a candidate gene were considered. We have identified three genes worthy of further analysis on this basis: RGS1, HHEX and THEMIS. To test the relevance of these candidates to central nervous system (CNS) autoimmunity, we aim to mimic phenotypes associated with these expression quantitative trait loci (eQTL) in in-vitro cultures of purified NK cells and monocytes, and in-vivo in a mouse model of MS - experimental autoimmune encephalomyelitis (EAE)