Hilary MacQueen

Immunofluorescent localisation of tumour necrosis factor‐α receptors on the popliteal lymph node and the surrounding adipose tissue following a simulated immune challenge

We used immunohistochemical techniques to demonstrate the distribution of receptors for the cytokine tumour necrosis factor‐α on the popliteal lymph node and the adipose tissue surrounding it for 5 d following a simulated immune challenge to one hind leg in the rat. We found different patterns of expression of receptors on adipocytes surrounding a lymph node to a distance of about 1 mm, and on those more remote from the node. Sites recognised by an antibody to type I tumour necrosis factor receptors appeared on the challenged node and the adipocytes surrounding it within 30 min of an injection of bacterial lipopolysaccharide, but appeared on adipocytes surrounding the unchallenged popliteal node only 24 h later. Adipocytes distant from the node, both within the same depot and in the contralateral depot, showed no response. Sites recognised by an antibody to type II tumour necrosis factor receptors were present at all times on lymph nodes and the adipocytes close to them, but appeared on more distant adipocytes only 24 h after immune challenge, in both challenged and unchallenged legs. These data support the proposal, based on in vitro studies, that the adipose tissue surrounding major lymph nodes is specialised to respond to cytokines derived from lymphoid cells, and participates in the immune responses of the adjacent node.

Vascularisation in adipose depots surrounding immune-stimulated lymph nodes

We report a change in the vascularisation of the adipose depots surrounding the popliteal lymph node that has, and the contralateral node that has not, been exposed to a simulated immune challenge. The percentage of the depot that consists of vessels, as measured by image analysis, decreases over a period of 2 d after immune stimulus, then increases in a biphasic manner over the next 2–3 wk. By 1 mo after the stimulus, the vascularisation has returned to baseline values. The adipose tissue surrounding both the stimulated and the unstimulated lymph nodes shows a similar pattern, but the unstimulated depot lags by 3–6 d in reaching its maximum vascularisation. These data support the hypothesis that perinodal adipose tissue is involved in peripheral immune responses.

Dietary fatty acids influence the appearance of tumour necrosis factor-α receptors on adipocytes following an immune challenge

Rats were fed from weaning on chow supplemented with suet or sunflower oil (10 % (w/w) each). The appearance of receptors for tumour necrosis factor-alpha on perinodal adipocytes from the popliteal depot following a subcutaneous injection of bacterial lipopolysaccharide was examined. In rats fed on sunflower oil-supplemented chow receptors appeared at a time similar to that described in rats fed unsupplemented chow, but in rats fed on chow supplemented with suet receptor appearance was significantly delayed. The popliteal adipocytes were found to contain different proportions of fatty acids as assessed by GLC. These preliminary results suggest that the fatty acid component of the diet can, by influencing the triacylglycerol-fatty acids within adipocytes, directly alter the time course of an early inflammatory immune response.

Quantitative Analysis of Cytokine Receptors Using Single-Photon Counting

Fluorescent molecules are widely used as labels or probes in biological research [1,2]. In particular, they can be chemically attached to antibodies which, by binding to their specific antigens, can identify the location of particular molecules of biological interest. We have used this technique to identify cell surface receptors to a cytokine, tumor necrosis factor-α (TNFα), that are produced as a result of an immune challenge [3]. This approach works well for qualitative studies, but hitherto it has been difficult to quantify the technique so that the numbers per cell of the molecules of interest can be assessed, mainly because of the sensitivity required. One of us has previously used the technique of single-photon counting to quantify fluorescent molecules [4] and showed it to be highly sensitive and reproducible. We report here the application of this technique to the quantitation of fluoresceinated bound antibodies in small biological samples. We have measured the numbers TNFα receptors appearing on the surface of adipocytes (fat cells) surrounding a rat lymph node at various times after an immune challenge to that node. The change in receptor number confirms and refines our earlier qualitative results and lends support to our hypothesis that adipocytes are locally specialized to modulate the mammalian immune response [5].

Teaching Biology at a Distance: Pleasures, Pitfalls, and Possibilities

Abstract The Open University (OU) has a long and distinguished history of teaching biology at a distance by a supported open learning model. This article examines some of the challenges in delivering biology via distance teaching, explores some of the lessons learned, and discusses the opportunities and hazards of new teaching technologies that the OU has helped pioneer.

Changes in lymphokine receptor expression and fatty acid composition of phospholipids and triacylglycerols in rat adipocytes associated with lymph nodes following a transient immune challenge

Single‐photon counting fluorimetry was used to record the time course of the expression of interleukin‐10 receptors labelled with fluorescent antibodies on the surface of adipocytes over 24 h, following an immune challenge to the rat popliteal lymph node. Homologous perinodal and remote‐from‐node samples from the stimulated and unstimulated popliteal depots were compared in rats fed on plain chow and chow supplemented with 10% w/w suet, fish or vegetable oils. Receptor expression was maximal 6 h after stimulation, and returned to baseline after 24 h, and was similar in the stimulated and unstimulated depots. Fewer receptors were elicited in tissues from rats fed lipid‐supplemented diets compared with the control diet, with fewest of all following the fish oil diet. These data suggest that interleukin‐10 is involved in local interactions between perinodal adipocytes and lymph node lymphoid cells. Both triacylglycerols and phospholipids contained more polyunsaturates and fewer saturates in perinodal adipose tissue than in samples from sites not associated with lymphoid tissue. These data are consistent with paracrine interactions between perinodal adipocytes and activated lymphoid cells.

Investigating interactions between epicardial adipose tissue and cardiac myocytes: what can we learn from different approaches?

Heart disease is a major cause of morbidity and mortality throughout the world. Some cardiovascular conditions can be modulated by lifestyle factors such as increased exercise or a healthier diet, but many require surgical or pharmacological interventions for their management. More targeted and less invasive therapies would be beneficial. Recently, it has become apparent that epicardial adipose tissue plays an important role in normal and pathological cardiac function, and it is now the focus of considerable research. Epicardial adipose tissue can be studied by imaging of various kinds, and these approaches have yielded much useful information. However, at a molecular level, it is more difficult to study as it is relatively scarce in animal models and, for practical and ethical reasons, not always available in sufficient quantities from patients. What is needed is a robust model system in which the interactions between epicardial adipocytes and cardiac myocytes can be studied, and physiologically relevant manipulations performed. There are drawbacks to conventional culture methods, not least the difficulty of culturing both cardiac myocytes and adipocytes, each of which has special requirements. We discuss the benefits of a three‐dimensional co‐culture model in which in vivo interactions can be replicated.

Predictive Value of Biomarkers for Normal and Pathological Ageing in a Rat Model

Sub-optimal diets are widely known to be associated with disease. This study investigated two questions: first, whether a Western affluent diet affects long-term health in the rat, and second, whether, and to what extent, early changes in blood biomarkers can predict specific pathologies. A rat model was used in which animals were fed from weaning either a control diet or a high fat, low protein, energy dense Western affluent diet. Rats were harvested at either 12 or 18 months, and at these time points blood samples were taken and various clinical biomarkers measured using a hospital analyser or by ELISA. Predictive biomarkers should show small changes at early time points and larger changes at later time-points, and be associ- ated with disease(s). Tissues were examined for overt pathology, and data were mined to establish links between pathologies and specific blood markers that might be used predictively. Results showed that the Western affluent diet is associated with ill-health, with a relative risk of developing disease 4.5 times higher than for the control diet. Furthermore we conclude that triacylglycerol, HDL cholesterol, the ratio of total cholesterol to HDL cholesterol, creatinine and alanine aminotransferase were good predictors of disease as across the cohorts they showed altered levels before the clinical development of patholo- gies, in agreement with our hypothesis.