Jane Calvert

Deregulated production of protective cytokines in response to Candida albicans infection in patients with chronic mucocutaneous candidiasis.

ABSTRACT Patients with chronic mucocutaneous candidiasis (CMC) are selectively unable to clear the yeast Candida, which results in persistent debilitating infections affecting the skin, nails, and mucous membranes. The underlying defect is unknown. Recent animal studies highlighted the importance of type 1 cytokines in protection against Candida, and previous work suggested that CMC patients may exhibit altered cytokine production in response to Candida. Based on these findings, in this study we investigated cytokine production in CMC patients by assessing a range of inflammatory, anti-inflammatory, type 1, and type 2 cytokines (interleukin-2 [IL-2], IL-4, IL-5, IL-6, IL-10, IL-12, gamma interferon [IFN-γ], tumor necrosis factor alpha [TNF-α]) in whole-blood cultures in response to five different fractions of Candida albicans (carbohydrate, purified mannan, and protein-rich fractions, etc.), as well as non-Candida antigens. Our results demonstrate that cytokine production is deregulated in a Candida-specific way for some cytokines (IL-2, IL-10), is deregulated more generally for other cytokines (IL-12, IL-6, IFN-γ), and is not markedly altered for still other cytokines (TNF-α, IL-4, IL-5). The most notable finding in CMC patients was the markedly impaired production of IL-12 in parallel with dramatically increased levels of IL-6 and IL-10 that occurred selectively in response to Candida. These results suggest that patients with CMC have impaired production of type 1-inducing cytokines (possibly a macrophage or dendritic cell defect?), which could result in an inability to mount protective cell-mediated responses and a failure to clear Candida. Continued tissue damage and inflammation may trigger production of high levels of inhibitory cytokines, such as the IL-10 production seen in our study, which would further reduce production of type 1-inducing cytokines in a positive feedback loop leading to persistent infection.

Cytokine Production Differs in Children and Adults

The susceptibility of normal, healthy children to infection has long been recognized, but the underlying mechanisms are poorly understood. As adequate cytokine production is crucial for optimal immune responses, we assessed antigen and mitogeninduced cytokine production in healthy children. Our results demonstrate that healthy children differ markedly compared with adults in their ability to produce cytokines (IL-2, interferon-γ, IL-4, and IL-6). Maximal stimulation with mitogen demonstrated impaired cytokine production with markedly lower levels of all four cytokines produced compared with adult levels. When stimulated with antigens, median levels of IL-2 and IL-4 remained lower than adult values, IL-6 production was increased as was interferon-γ, albeit not significantly. Although the study was carried out on peripheral blood mononuclear cells that represent a restricted compartment of the immune system, these data suggest that, in healthy children, cytokine production is decreased and/or altered and could result in a suboptimal immune response, which could be one of the factors underlying increased susceptibility to infection in children.

Intervening Early: Attendance and Performance Monitoring as a Trigger for First Year Support in the Biosciences.

Abstract A centralised system monitoring attendance and performance among first year students in Biomedical Sciences has been established at Newcastle University. Early signs of absence and poor performance trigger immediate intervention by academic staff, with the aim of providing support for students at risk of failure or withdrawal. Difficulties associated with monitoring attendance in large lecture classes are avoided by monitoring attendance only at ‘high stakes’ classes, namely practicals and seminars. Level of attendance at non-lecture classes was a predictor of academic achievement and the early intervention strategy was associated with improvements in attendance. Student perceptions of attendance monitoring were evaluated and found to be positive. Meeting with absent and underperforming students at the earliest possible opportunity has proved an effective way of promoting dialogue between staff and students who are experiencing difficulties.

Abnormal immunoglobulin G subclass production in response to keyhole limpet haemocyanin in atopic patients.

A proportion of patients with atopic dermatitis have elevated serum levels of IgG4. In order to investigate further this abnormality of IgG subclass production, atopic patients were immunized with the protein antigen keyhole limpet haemocyanin (KLH), and IgG subclass responses following primary and secondary immunization were analysed. In the primary response, titres of IgG 1, 2 and 3 antibodies were lower in the atopic patients than in the controls. In contrast, titres of IgG4 were much higher for the patient group. In both patients and controls, the kinetics of IgG4 antibody production following the initial immunization with KLH showed a slow rise reaching a peak at 30 weeks. This time course indicated that the high IgG4 response was unlikely to be due to previous exposure of the patients to a cross‐reacting antigen. A higher proportion of IgG4 was also seen in the atopic patients following secondary immunization; indeed, IgG4 was the major subclass in the secondary response in the patient group. In the controls, but not in the patients, titres of IgG4 anti‐KLH correlated with total serum levels of IgG4, and some of the highest IgG4 antibody responses were detected in atopic patients whose serum IgG4 concentration was in the normal range. The results suggest that raised serum levels of IgG4 in atopy may reflect abnormal isotype regulation in response to protein antigens.

Assessed Online Discussion Groups in Biology Education

Abstract Sophisticated software such as Virtual Learning Environments (VLEs) are rapidly being deployed by universities. Despite widespread use of such systems, experience shows that there is frequently poor pedagogic development, leading primarily to use of VLEs as electronic document repositories rather than as online learning systems in which the available suite of tools are used to their full potential. Online assessment is the major potential efficiency gain of such systems, but most staff do not scratch the surface of the full capabilities of the software. Based on our experience, we describe practical guidelines for a model of online assessment which promotes deep versus superficial learning, encourages higher level learning competencies and inclusivity.

A comparison of monoclonal antibody immobilization of platelet antigen (MAIPA) and immunobead methods for detection of GPIIb/IIIa antiplatelet antibodies in immune thrombocytopenic purpura

In this study we have compared two assays for the detection of autoantibodies GpIIb/IIIa, platelet bound and in serum, in immune thrombocytopenic purpura (ITP). Both assays were found to have a similar sensitivity, but the monoclonal antibody immobilization of platelet antigen (MAIPA) assay was more reproducible than the immunobead assay. The MAIPA and immunobead assay demonstrated an 81% concordance of results for serum antibody detection and a 78% concordance for platelet‐associated antibody detection, with an 8–12% incidence of false positive or negative results.

The CD5+ B cell: a B cell lineage with a central role in autoimmune disease?

It is apparent that B cells are heterogeneous with respect to, for example, the antigens they express on their surface, and the stimuli to which they can respond. It is still unclear to what extent these differences relate to the stage of differentiation (eg. virgin B cells differing from activated B cells or memory cells), or whether distinct developmental lineages might exist. It has been proposed by some authors that, in the mouse, B cells expressing the ly-1 antigen constitute a separate lineage. In man also, a minor population of B cells expresses detectable levels of the CD5 antigen, but far less information is available about these cells. Interest in the CD5+ and ly-1+ B cell subpopulations has been further stimulated by the suggestion that these cells might play a special role in autoimmune disease. Although, in mouse, ly-1+ B cells differ in several respects from ly-1- B cells, the main evidence that they form a separate lineage derives from experiments in which ly-1+ B cells could not be reconstituted with adult bone marrow. It should be borne in mind that the situation is quite different in humans where, following bone marrow transplantation, CD5+ B cells are rapidly restored. Moreover, in the irradiated mice, at least in some of the experiments ly-1+ B cells were in fact reconstituted by adult bone marrow. Furthermore, at least in humans, expression of CD5 can sometimes be induced. There is, as yet, no good evidence that human CD5+ B cells form a distinct lineage, and it is possible that CD5 expression depends upon microenvironmental influences acting on the B cell during its differentiation. Several interesting properties have been attributed to ly-1+ B cells, including the ability to provide help to other B cells, and the secretion of autocrine factors. However there is also evidence that these features are not exclusive to B cells expressing ly-1. It has also been suggested that ly-1+ B cells might be long-lived. It is not yet known whether some of the properties of ly-1+ B cells might be a direct result of their expressing this antigen; this may become more clear when the function of CD5 is elucidated. The suggestion that the repertoire of ly-1+ B cells might be biased towards the expression of certain V genes is very interesting. Many of the hybridomas from neonatal mice produce antibodies which are multi-specific, and therefore well suited to form a first line of defence against potential pathogens.(ABSTRACT TRUNCATED AT 400 WORDS)

Characterization of the Impaired Antipneumococcal Polysaccharide Antibody Production in Immunosuppressed Pediatric Patients Following Cardiac Transplantation

We previously have demonstrated impaired pneumococcal polysaccharide IgG antibody responses in children immunosuppressed following cardiac transplantation in early childhood. We have further characterized the antibody defect. To further investigate the production of antibody, anti-pneumococcal polysaccharide (PPS) specific IgM, IgG, IgG subclasses, and IgA were measured in postvaccination sera by enzyme-linked immunosorbent assay. Two groups were studied: posttransplant children who made pneumococcal antibody in vivo following natural exposure or PPS immunization (R) and those with an impaired response (NR).There was no difference in IgM or IgA levels between R and NR. IgG and IgG2 levels were higher in R than NR (P = 0.002), even after adsorption of nonspecific common cell wall antigen antibody. Differences in anti-pneumococcal antibody levels suggest that immunoglobulin isotype switching from IgM to IgG and particularly IgG2 is impaired in patients immunosuppressed at a young age. These findings confirm data regarding the effect of immunosuppressive agents derived from animal models in humans.

Reinforcing the links between teaching and research: evaluation of a scheme to employ undergraduate students as laboratory assistants

Abstract A scheme was introduced to offer opportunities to second-year bioscience students to undertake part-time paid work in research laboratories. The aim was to provide students with a greater appreciation of bioscience research, to reinforce their laboratory skills and to encourage them to consider a research-based career. Students worked for 8 hours per week during term time and were paid at a minimal wage. Hours were negotiated with the supervisor to fit with the student’s timetable commitments. The scheme has run for five years employing 74 undergraduates and we have undertaken an evaluation of its impact on students and staff. Our findings indicate that the scheme has been well-received by students, with the overwhelming majority reporting that they enjoyed the experience, were well-supervised, and would recommend this scheme to friends. Students also reported a positive effect on their studies and that the experience had encouraged them to consider a career in research. Reports from supervisors were also highly positive. It is difficult to assess the impact of the scheme on students’ career choices. However, where destinations are known, it appears that a relatively high proportion of students have opted for higher degrees.

Levels of CD5+ B Cells are Not Increased in Probands or Relatives in a Family Study of Primary Sjögren's Syndrome

Levels of CD5+ B lymphocytes were assayed in a large family study of Primary Sjögrens syndrome. There was no significant difference in CD5 expression by index cases or their relatives when compared to controls. No association between CD5 expression, serological abnormalities or HLA haplotype was found and, furthermore, no evidence of linkage with HLA was observed. There was, however, variation in the expression of CD5+ B cells between the families. Levels in spouses were lower and reached statistical significance. The role for genetic and environmental factors influencing CD5 expression is discussed. Any genetic influence does not appear to involve the HLA region or genes in linkage disequilibrium.