Drinker's own drinking, experience of alcohol-related harms, and concern for drinking predict drinker's attitudes towards non-drinkers.

Objective: While the stigma experienced by non-drinkers is well-documented, little is known about the factors that influence it. This study aims to test a sequential mediation model in which the amount of alcohol consumed by a drinker, predicts their experienced alcohol-related harm, which in turn predicts the concern they have for drinking and their negative attitudes towards non-drinkers.

Methods: A sample of 787 Australian drinkers (M = 38.

Development and Validation of the Cheers Attitudes towards Non-drinkers Scale (CANS).

Non-drinkers report experiencing stigma, which can act as a barrier to non-drinking. Two studies were undertaken to develop and test a new scale to measure attitudes towards non-drinkers. In Study 1, 29 items were presented to 426 Australian drinkers.

The 'sober eye': examining attitudes towards non-drinkers in Australia.

Background: The proportion of Australians who choose not to drink alcohol has increased in recent years; yet, non-drinkers report experiences of stigma and judgement from peers for this choice. This study aimed to explore the attitudes that exist towards non-drinkers and examine what drives this stigma.

Method: Thematic analysis of four focus groups was undertaken, comprising 37 drinking and non-drinking Australian adults.

Resident and monocyte-derived dendritic cells become dominant IL-12 producers under different conditions and signaling pathways.

IL-12 is such a pivotal cytokine that it has been called the third signal for T cell activation, TCR engagement being the first and costimulation being the second. It has been generally viewed that the resident CD8(+) dendritic cell (DC) subset is the predominant IL-12-producing cell type. In this study, we found, although this is so under steady state conditions, under inflammatory conditions monocyte-derived DC (mDC) became a major cell type producing IL-12.

Synergism between active listeriolysin O and dimethyldioctadecylammonium bromide to activate CD8(+) T cells.

Purified recombinant listeriolysin O (LLO) was assessed for its ability to induce T cell responses in mice. Intraperitoneal immunisation with LLO, as a fusion with glutathione-S-transferase (GST), induced the production of LLO-specific CD8(+) T cells, but not LLO-specific CD4(+) T cells. The generation of this response could be blocked by pre-treatment with cholesterol, indicating a requirement for LLO pore formation.

Bystander activation of CD8+ T lymphocytes during experimental mycobacterial infection.

Infection of C57BL/6 mice with Mycobacterium avium leads to the activation of both CD4+ and CD8+ gamma interferon (IFN-gamma)-producing T cells, although the CD8+ cells play no role in protection against infection. Using transfer of different lines of transgenic T cells with T-cell receptors (TCRs) which recognize irrelevant antigens, we show here that transferred CD8+ T cells from two of the three lines were activated to the same degree as the host cells, suggesting that the majority of the IFN-gamma-producing CD8+ T cells of the host represented bystander activation. The third line, specific for the male HY antigen, showed no activation.

A totally synthetic vaccine of generic structure that targets Toll-like receptor 2 on dendritic cells and promotes antibody or cytotoxic T cell responses.

A simple generic peptide-based vaccine structure that targets Toll-like receptor 2-expressing dendritic cells and causes their activation is described. The vaccines are totally synthetic, serve as their own adjuvant, and are composed of (i) a single helper T cell epitope, (ii) a target epitope that is either recognized by CD8+ T cells or B cells, and (iii) a Toll-like receptor 2-targeting lipid moiety, S-[2,3-bis(palmitoyloxy)propyl]cysteine, that is situated between the peptide epitopes to form a branched configuration. The different CD8+ T cell epitopes examined were from (i) influenza virus, (ii) the intracellular bacterium Listeria monocytogenes, and (iii) ovalbumin as a model tumor antigen.

Apoptosis of CD4+ and CD8+ T cells during experimental infection with Mycobacterium avium is controlled by Fas/FasL and Bcl-2-sensitive pathways, respectively.

Both CD4+ and CD8+ T cells from mice infected with Mycobacterium avium suffered a high rate of apoptosis, beginning with the onset of the immune response and culminating in the loss of T cells from the tissues and loss of IFN-gamma production. Fas expression increased over the course of infection on both T cell populations, as did their susceptibility to the induction of apoptosis in vitro by anti-Fas mAb. Nevertheless, although the rate of apoptosis among CD4+ T cells from infected mice was reduced to normal levels in lpr mice with a defective Fas, CD8+ T cells were unaffected, implying that Fas/FasL interaction was not important in these cells in vivo.

Generalized immunological decline during long-term experimental infection with Mycobacterium avium.

Terminal loss of immune responsiveness in C57BL/10 mice intranasally infected with Mycobacterium avium was observed in both spleen and lung. It was nonspecific and related to the duration of infection, not the age of the mice. While there was loss of total T cells, the remaining cells were less efficient at gamma interferon production.

Identification of an I-Ad restricted peptide on the 65-kilodalton heat shock protein of Mycobacterium avium.

The 65 kilodalton heat shock protein (Hsp65) from mycobacterial species elicits immune responses and in some cases protective immunity. Here we have used a DNA sublibrary approach to identify antigenic fragments of Mycobacterium avium Hsp65 and a synthetic peptide approach to delineate CD4+ T cell determinants. A panel of Hsp65 reactive CD4+ T cell clones was established from lymph node cells obtained from BALB/c mice immunized with recombinant Hsp65.

T-cell activation, proliferation and apoptosis in primary Listeria monocytogenes infection.

Listeria monocytogenes infection of mice leads to a rapid expansion of activated T cells, followed by a decline in specific cells once the bacteria are eliminated. In order to define the relationship between T-cell proliferation and activation, and to investigate the role of apoptosis in limiting the expansion, the expression of activation markers, uptake of 5-bromo-2'-deoxyuridine (BrdU) in vivo and the incidence of apoptosis was investigated. Increased numbers of T cells expressing the activated phenotype CD25+, CD44hi and CD62Llo were detected 4 days after infection.

Oxidised mannan-listeriolysin O conjugates induce Th1/Th2 cytokine responses after intranasal immunisation.

Clearance of infectious organisms does not always require polarised Th1 or Th2 responses and it may be advantageous for both Th1 and Th2 responses to be elicited for effective protection against an invading pathogen. It was the aim of this study to investigate oxidised mannan as a possible Th1/Th2 adjuvant. Oxidised mannan was conjugated to two candidate antigens and delivered intranasally to mice.

Oxidised mannan as a novel adjuvant inducing mucosal IgA production.

Mannan, oxidatively coupled to recombinant protein antigens, has here been tested as a possible adjuvant for the production of antibody on the mucosa. Given intranasally, but not intraperitoneally, mannan markedly enhanced the production of IgA, IgG1 and IgG2a in the serum, and IgA locally in the lung and at remote mucosal sites, including tears, vaginal and salivary secretions. Oxidative coupling was critical to its action, since neither mannan simply mixed with protein nor mannan-protein conjugates which had been reduced by treatment with sodium borohydride, acted as adjuvants.

Interleukin-2 and loss of immunity in experimental Mycobacterium avium infection.

Experimental infection of mice with a virulent strain of Mycobacterium avium leads to a slowly progressive disease, which we have previously shown culminates in loss of gamma interferon (IFN-gamma) production by T lymphocytes and death of the animals approximately 40 weeks after infection. Here we investigated the changes in T-cell activation, the production of interleukin-2 (IL-2), and the response to IL-2 throughout M. avium infection as a possible explanation for this loss.

Molecular and immunological characterization of Mycobacterium avium 65 kDa heat shock protein (Hsp65).

The heat shock protein Hsp65 has been characterized previously in several mycobacterial species. This is the first report of the complete sequence of the coding region of the Mycobacterium avium homologue. The sequence was highly homologous to the Hsp65 of other mycobacterial species, as well as being related closely to the murine and human homologues.

Non-major histocompatibility complex control of antibody isotype and Th1 versus Th2 cytokines during experimental infection of mice with Mycobacterium avium.

Infection of different strains of mice with Mycobacterium avium has revealed genetic control of the immunoglobulin isotype induced and of the balance between Th1 and Th2 cytokines. Female BALB/c or C57BL/10 mice were infected intranasally with 10(5) M. avium organisms.

T lymphocytes from granulocyte colony-stimulating factor-/- mice produce large quantities of interferon-gamma in a chronic infection model.

Little is known about the role of granulocyte colony-stimulating factor (G-CSF) in the response to chronic bacterial infections. To address this we infected G-CSF knock out (G-CSF-/-) mice with Mycobacterium avium. Infection was not exacerbated in G-CSF-/- mice despite a deficiency in the total bone marrow cells, colony-forming haemopoietic cells, granulocytes and monocyte precursors in the bone marrow.

Haemopoiesis in mice genetically lacking granulocyte-macrophage colony stimulating factor during chronic infection with Mycobacterium avium.

In order to test the role of granulocyte-macrophage colony stimulating factor (GM-CSF) in haemopoiesis during chronic infection, mice with a targeted disruption of the gene for GM-CSF were infected intraperitoneally with the facultative intracellular pathogen, Mycobacterium avium. The bacteria spread to lungs, liver and spleen and persisted for more than 10 weeks at levels between 105 and 106 CFU. Bacterial numbers did not differ significantly between infected GM-CSF-/- and wild-type mice, making this an excellent model in which to study the effects of GM-CSF deficiency on haemopoietic cells without complications of interpretation relating to differences in bacterial load.

Use of recombinant viruses to deliver cytokines influencing the course of experimental bacterial infection.

The feasibility of using viral constructs expressing cytokine genes to influence the course of bacterial infection was tested in mice. The mice were first infected with vaccinia or fowlpox viruses expressing the cytokine of interest, then challenged with the facultative intracellular bacterial pathogen Listeria monocytogenes. The course of infection was assessed by subsequent bacterial counts.

Anergy, IFN-gamma production, and apoptosis in terminal infection of mice with Mycobacterium avium.

We have followed the course of experimental infection of mice with Mycobacterium avium over an extended period, assessing bacterial numbers and T cell responsiveness. When mice were infected intranasally, bacteria spread to the spleen and liver, but remained in highest numbers in the lungs. Both CD4+ and CD8+ T cells, assayed at any time from 6-28 wk after infection, produced IFN-gamma.

Functional deficiencies of peritoneal cells from gene-targeted mice lacking G-CSF or GM-CSF.

Gene-targeted mice lacking the hemopoietic growth factors, granulocyte colony-stimulating factor (G-CSF) or granulocyte-macrophage (GM)-CSF, show increased susceptibility to infection with the facultative intracellular bacterium, Listeria monocytogenes. The resident peritoneal cell populations from G-CSF(-/-) and GM-CSF(-/-) mice showed reduced production of the bactericidal molecule nitric oxide. Macrophage-mediated tumoricidal activity and phagocytosis of Listeria were reduced in G-CSF(-/-), but not in GM-CSF(-/-), mice.

Control of IL-12 and IFN-gamma production in response to live or dead bacteria by TNF and other factors.

When mice were infected i.v. with either Listeria monocytogenes or Brucella abortus, bioactive IL-12 was briefly detected in serum and supernatants of spleen homogenates immediately ex vivo.

Essential roles for granulocyte-macrophage colony-stimulating factor (GM-CSF) and G-CSF in the sustained hematopoietic response of Listeria monocytogenes-infected mice.

The in vivo roles of granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte (G)-CSF were studied in factor-deficient gene-targeted knockout mice infected with the facultative intracellular bacterium Listeria monocytogenes. Previous results showed that G-CSF-/- mice had an underlying selective deficiency in granulopoiesis, but GM-CSF-/- mice had little disturbance in resting hematopoiesis. Nevertheless, in this study it is revealed that 3 days after intraperitoneal infection with 2 x 10(5) Listeria, GM-CSF-/- mice harbored 50-fold more organisms in their spleen and liver than similarly infected wild-type mice.

The Rel subunit of NF-kappaB-like transcription factors is a positive and negative regulator of macrophage gene expression: distinct roles for Rel in different macrophage populations.

The role of Rel in the monocyte/macrophage lineage was examined in mice with an inactivated c-rel gene. Although the frequency of monocytic cells was normal in Rel-/- mice, we show that Rel serves distinct roles in regulating gene expression and immune effector function in different mature macrophage populations. Stimulated Rel-/- resident peritoneal macrophages produced higher than normal levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF) and interleukin-6 (IL-6), but tumour necrosis factor-alpha (TNF-alpha) production was not induced.

Either B7-1 or B7-2 is required for Listeria monocytogenes-specific production of gamma interferon and interleukin-2.

Listeria infection results in the induction of gamma interferon (IFN-gamma)-producing T lymphocytes. Blocking of the costimulatory molecule B7 in vivo led to a marked decrease in antigen-specific production of IFN-gamma and interleukin-2 by lymphocytes. Blocking of both B7-1 (CD80) and B7-2 (CD86) was required in order to inhibit cytokine production, indicating that either molecule could act alone.