iNKT Cells Suppress the CD8(+) T Cell Response to a Murine Burkitt's-Like B Cell Lymphoma

The T cell response to B cell lymphomas differs from the majority of solid tumors in that the malignant cells themselves are derived from B lymphocytes, key players in immune response. B cell lymphomas are therefore well situated to manipulate their surrounding microenvironment to enhance tumor growth and minimize anti-tumor T cell responses. We analyzed the effect of T cells on the growth of a transplantable B cell lymphoma and found that iNKT cells suppressed the anti-tumor CD8(+) T cell response. Lymphoma cells transplanted into syngeneic wild type (WT) mice or Jalpha18(-/-) mice that specifically lack iNKT cells grew initially at the same rate, but only the mice lacking iNKT cells were able to reject the lymphoma. This effect was due to the enhanced activity of tumor-specific CD8(+) T cells in the absence of iNKT cells, and could be partially reversed by reconstitution of iNKT cells in Jalpha 18(-/-) mice. Treatment of tumor-bearing WT mice with alpha -galactosyl ceramide, an activating ligand for iNKT cells, reduced the number of tumor-specific CD8(+) T cells. In contrast, lymphoma growth in CD1d1(-/-) mice that lack both iNKT and type II NKT cells was similar to that in WT mice, suggesting that type II NKT cells are required for full activation of the anti-tumor immune response. This study reveals a tumor-promoting role for iNKT cells and suggests their capacity to inhibit the CD8(+) T cell response to B cell lymphoma by opposing the effects of type II NKT cells.     

Cardiovascular GO Annotation Initiative Year 1 Report: Why Cardiovascular GO?

Gene Ontology (GO) vocabularies are an established standard for linking functional information to genes and gene products (www.geneontology.org/). A recent collaboration between University College London and the European Bioinformatics Institute is providing GO annotation to human cardiovascular-associated genes (http://www.ucl.ac.uk/medicine/cardiovascular-genetics/geneontology.html). This report outlines the aims of this collaboration and summarizes how the cardiovascular community can help improve the quality and quantity of GO annotations. This new initiative is funded by the British Heart Foundation and fully supported by the GO Consortium.     

Life depends upon two kinds of water

Background

Many well-documented biochemical processes lack a molecular mechanism. Examples are: how ATP hydrolysis and an enzyme contrive to perform work, such as active transport; how peptides are formed from amino acids and DNA from nucleotides; how proteases cleave peptide bonds, how bone mineralises; how enzymes distinguish between sodium and potassium; how chirality of biopolymers was established prebiotically.

Methodology/Principal Findings

It is shown that involvement of water in all these processes is mandatory, but the water must be of the simplified configuration in which there are only two strengths of water-water hydrogen bonds, and in which these two types of water coexist as microdomains throughout the liquid temperature range. Since they have different strengths of hydrogen bonds, the microdomains differ in all their physical and chemical properties. Solutes partition asymmetrically, generating osmotic pressure gradients which must be compensated for or abolished. Displacement of the equilibrium between high and low density waters incurs a thermodynamic cost which limits solubility, depresses ionisation of water, drives protein folding and prevents high density water from boiling at its intrinsic boiling point which appears to be below 0°C. Active processes in biochemistry take place in sequential partial reactions, most of which release small amounts of free energy as heat. This ensures that the system is never far from equilibrium so that efficiency is extremely high. Energy transduction is neither possible and nor necessary. Chirality was probably established in prebiotic clays which must have carried stable populations of high density and low density water domains. Bioactive enantiomorphs partition into low density water in which they polymerise spontaneously.

Conclusions/Significance

The simplified model of water has great explanatory power.     

Immunoglobulin genes expressed by B-lymphocytes infiltrating cervical carcinomas show evidence of antigen-driven selection

Lymphocyte infiltration is often present in cervical cancer lesions, possibly reflecting an ongoing (but ineffective) immune response to the tumour. B-lymphocytes are the predominant lymphocyte infiltrate in pre-malignant cervical lesions, where they are thought to comprise the host immune response to active human papillomavirus (HPV) infection. Although B cells are less frequently detected in cervical tumours, a high proportion of terminally differentiated plasma cells expressing tumour-specific immunoglobulin (Ig) remain. The antigen specificity and functional significance of the antibody response to cervical tumours is unknown. As part of a study to characterise the antibodies expressed by the tumour-infiltrating B cells (TIL-B) in cervical tumours using antibody phage display, we examined expressed Ig gene sequences to determine if there was molecular evidence of a selective response to antigenic changes in the transformed epithelial cells. We found that biased variable region gene usage by the B cells and the rate of somatic hypermutation in the rearranged Ig heavy chain variable regions (VH) both indicated antigenic selection of the B cells. We also found evidence of affinity maturation, as indicated by the detection of antibodies of the IgG1, IgG2 and IgA isotypes, and possible clonal selection of the Ig receptors. These data support the notion that B-lymphocytes and plasma cells infiltrating cervical carcinomas are the result of an antigen-induced response to HPV infection or transformation.     

Hepatitis C Virus Phylogenetic Clustering Is Associated with the Social-Injecting Network in a Cohort of People Who Inject Drugs

It is hypothesized that social networks facilitate transmission of the hepatitis C virus (HCV). We tested for association between HCV phylogeny and reported injecting relationships using longitudinal data from a social network design study. People who inject drugs were recruited from street drug markets in Melbourne, Australia. Interviews and blood tests took place three monthly (during 2005–2008), with participants asked to nominate up to five injecting partners at each interview. The HCV core region of individual isolates was then sequenced and phylogenetic trees were constructed. Genetic clusters were identified using bootstrapping (cut-off: 70%). An adjusted Jaccard similarity coefficient was used to measure the association between the reported injecting relationships and relationships defined by clustering in the phylogenetic analysis (statistical significance assessed using the quadratic assignment procedure). 402 participants consented to participate; 244 HCV infections were observed in 238 individuals. 26 genetic clusters were identified, with 2–7 infections per cluster. Newly acquired infection (AOR = 2.03, 95% CI: 1.04–3.96, p = 0.037, and HCV genotype 3 (vs. genotype 1, AOR = 2.72, 95% CI: 1.48–4.99) were independent predictors of being in a cluster. 54% of participants whose infections were part of a cluster in the phylogenetic analysis reported injecting with at least one other participant in that cluster during the study. Overall, 16% of participants who were infected at study entry and 40% of participants with newly acquired infections had molecular evidence of related infections with at least one injecting partner. Likely transmission clusters identified in phylogenetic analysis correlated with reported injecting relationships (adjusted Jaccard coefficient: 0.300; p<0.001). This is the first study to show that HCV phylogeny is associated with the injecting network, highlighting the importance of the injecting network in HCV transmission.     

Intracellular pH of frog sartorius muscle

A weak base, morpholine, has been labelled with ³H and tested for its suitability as an indicator for intracellular pH, by distribution in the tissue water of frog sartorius muscle in the species Hyla litoria. Its pK'a at 20°C in a solution of the same of ionic strength as frog Ringer was found to be 8.45 ± 0.02, which is in the range of maximal sensitivity. Morpholine equilibrated with the tissue in 17 h; it was shown that it was not bound to intracellular constituents, that it was not metabolised nor toxic in the concentrations used; it was therefore judged suitable as a pH indicator. Intracellular pH was then measured by distribution of morpholine (6.985 ± 0.08), nicotine (6.915 ± 0.03) and the weak acid 5,5′-dimethyl-2,4-oxazolidinedione (7.10 ± 0.05) and with pH-sensitive microelectrodes (5.9, the equilibrium value). It was shown that the four significantly different values could not be reconciled in terms of experimental error, heterogeneity of intracellular pH, liquid junction potential differences, or binding of indicator molecules inside the fibre. They could, however, be reconciled if the fibre water had different structure and solvent properties from the extracellular water and ions were distributed across the membrane as between two liquid phases containing different solvents. Then the H⁺ would be in equilibrium, as shown by the microelectrode measurement, but intracellular pH would be indeterminable and probably greater than 6.     

Dimerization of CtIP may stabilize in vivo interactions with the Retinoblastoma-pocket domain

CtIP is a tumor suppressor that interacts with Retinoblastoma protein (Rb) to regulate the G1/S-phase transition of the cell cycle. Despite its large size (897 residues) CtIP has few known structured regions. Rather it contains several linear motifs that interact with known binding partners, including an LXCXE motif that binds the pocket domain of Rb-family proteins. This LXCXE motif lies at the C-terminus of the only known structured domain, an N-terminal coiled-coil dimerization domain (DD; residues 45-160). Yeast two-hybrid (Y2H) and GST-pulldown analyses showed that CtIP requires the LXCXE motif to bind the Rb-pocket. Although isothermal titration calorimetry data indicates that the LXCXE motif is the sole determinant of binding affinity for the Rb-pocket domain (K(A) approximately 10(6)M(-1)), Y2H data indicates that the DD is required to stabilize the interaction in vivo. Thus dimerization may increase the apparent stability of the proteins and/or the lifetime of the complexes.     

New Zealanders working non-standard hours also have greater exposure to other workplace hazards

Exposure to workplace hazards, such as dust, solvents, and fumes, has the potential to adversely affect the health of people. However, the effects of workplace hazards on health may differ when exposure occurs at different times in the circadian cycle, and among people who work longer hours or who do not obtain adequate sleep. The aim of the present study was to document exposures to workplace hazards across a national sample of New Zealanders, comparing people who work a standard 08:00 ?17:00 h Monday-to-Friday working week (Std hours) and those who do not (N-Std hours). New Zealanders (n = 10 000) aged 20–64 yrs were randomly selected from the Electoral Roll to take part in a nationwide survey of workplace exposures. Telephone interviews were conducted between 2004 and 2006, using a six-part questionnaire addressing demographics, detailed information on the current or most recent job (including exposures to a range of workplace hazards), sleep, sleepiness, and health status. N-Std hours were categorised on the basis of: being required to start work prior to 07:00 h or finish work after 21:00 h and/or; having a regular on-call commitment (at least once per week) and/or; working rotating shifts and/or; working night shift(s) in the last month. The response rate was 37% (n = 3003), with 22.2% of participants (n = 656) categorised as working N-Std hours. Industry sectors with the highest numbers of participants working N-Std hours were manufacturing, health and community services, and agriculture, fishing, and forestry. Response rate was 37% (n = 3003) with 22.2% (n = 656) categorised as working N-Std hours. Participants working N-Std hours were more likely to be exposed to all identified hazards, including multiple hazards (OR = 2.45, 95% CI = 2.01–3.0) compared to those working Std hours. Participants working N-Std hours were also more likely to report ‘never/rarely’ getting enough sleep (OR = 1.38, 95% CI = 1.15–1.65), ‘never/rarely’ waking refreshed (OR = 1.23, 95% CI = 1.04–1.47), and excessive sleepiness (OR = 1.77, 95% CI = 1.29–2.42). New Zealanders working N-Std hours are more likely to be exposed to hazards in the workplace, to be exposed to multiple hazards, and to report inadequate sleep and excessive sleepiness than their colleagues working a standard 08:00?17:00 h Monday-to-Friday working week. More research is needed on the effects of exposure to hazardous substances outside the usual waking day, on the effects of exposure to multiple hazards, and on the combination of hazard exposure and sleep restriction as a result of shift work.