Search PubMed⌕ Search

Biomedical subjects

R Aspinall

Publications and source records attributed to R Aspinall.

At least 19 recordsLinked to original sources
PubMed2002

Thymic activity in late-stage HIV-1 infected individuals receiving highly active antiretroviral therapy: potential effect of steroid therapy.

OBJECTIVE: Our objective was to monitor the effect of steroid therapy on the thymic output and function of late-stage HIV-1-infected patients undergoing highly active antiretroviral therapy (HAART). DESIGN: The indirect measurement of T cells that have recently emigrated from the thymus as a means of quantifying thymic output, and therefore thymic function, was achieved through use of the polymerase chain reaction-based signal joint T cell receptor rearrangement excision circles (sjTREC) assay. Proliferative capacity and interleukin (IL)-2 and IL-4 production by T cells after antigenic, mitogenic and IL-2 stimulation were also analysed. METHOD: Measurements were made of sjTREC levels in peripheral blood mononuclear cell DNA samples from five HIV-1 infected patients (one on steroid therapy prior to and at the time of sample extraction) receiving HAART. IL-2 and IL-4 production and proliferative capacity were also measured in three patients, including the patient receiving steroids. RESULT: The sjTREC assay gave an extremely weak result for the patient on steroids but, under the same assay conditions, provided clear, positive readings for the four patients not on steroids. Comparison of the patients' cytokine profiles revealed that IL-2 production was generally low or absent in all three patients tested but that IL-4 production was significantly higher in the patient given steroids. Functional potential as revealed by proliferation assays showed very low or absent cellular proliferation. CONCLUSION: The thymic contribution to the restoration of T lymphocyte numbers, particularly during the treatment of HIV-1 infection, may become compromised if thymic inhibitory factors such as steroids are used. Furthermore, the use of steroids may also favour the development of a T helper 2 response, which could prove particularly undesirable during HIV-1 infection.

Adult↗
PubMed2002

Use of a high-fidelity simulator to develop testing of the technical performance of novice anaesthetists.

BACKGROUND: We used the Delphi technique to gain a consensus from 26 consultant anaesthetists about technical tasks during general anaesthesia. We then developed a technical scoring system to assess anaesthetists undertaking general anaesthesia with rapid sequence induction. METHODS: We then followed the performance of six novice anaesthetists on five occasions during their first 3 months of training. At each, visit each novice 'anaesthetized' the Human Patient Simulator at Bristol Medical Simulator Centre. For comparison seven post-fellowship anaesthetists were scored on one occasion. RESULTS: Novice scores improved significantly over the 12-week period (P<0.01). A significant difference was also found between the final novice scores and the post-fellowship subjects (P<0.05). CONCLUSIONS: These findings suggest that simulation can be used to observe and quantify technical performance.

Adolescent↗
PubMed2001

Age-associated thymic atrophy is not associated with a deficiency in the CD44(+)CD25(-)CD3(-)CD4(-)CD8(-) thymocyte population.

Age-associated thymic atrophy has been proposed to be due to changes in both the thymic microenvironment and in the intrinsic properties of the early T cell progenitors, the CD44(+)CD25(-)CD3(-)CD4(-)CD8(-) cells. We have purified these cells from the thymus of both old and young mice and demonstrate no age-associated defect in their ability to differentiate into their progeny in vitro when used to reconstitute fetal thymic organ cultures. We also demonstrate that in the presence of anti-IL-7, CD44(+)CD25(-)CD3(-)CD4(-)CD8(-) cells from young mice show reduced thymocyte development in fetal thymic organ cultures compared with controls. Finally we have shown that old mice treated with IL-7 show improved thymopoiesis compared with control groups. The increased thymopoiesis seen in the old animals occurs in the sequential manner which would be anticipated for an agent working directly on the early stages, including the CD44(+)CD25(-)CD3(-)CD4(-)CD8(-) cells.

Aging↗
PubMed2001

Il-7 and not stem cell factor reverses both the increase in apoptosis and the decline in thymopoiesis seen in aged mice.

Thymic atrophy is an age-associated decline in commitment to the T cell lineage considered to be associated with defective TCR beta-chain rearrangement. Both IL-7 and stem cell factor (SCF) have dominant roles at this stage of triple negative (TN) thymocyte development. Because there is no age-associated decrease in the number of CD44(+)CD25(-)CD3(-)CD4(-)CD8(-) cells, this study investigated whether alterations in apoptosis within the TN pathway accounted for diminishing thymocyte numbers with age. Here we show significant age-associated increases in apoptotic TN thymocytes, specifically within CD44(+)CD25(+) and CD44(-)CD25(+) subpopulations, known to be the location of TCR beta-chain rearrangement. IL-7 added to TN cultures established from old mice significantly both reduces apoptosis and increases the percentage of live cells within CD44(+)CD25(+) and CD44(-)CD25(+) subpopulations after 24 h, with prosurvival effects remaining after 5 days. SCF failed to demonstrate prosurvival effects in old or young cultures, and IL-7 and SCF together did not improve upon IL-7 alone. IL-7R expression did not decline with age, ruling out the possibility that the age-associated increase in apoptosis was attributed to reduced IL-7R expression. Compared with PBS, treatment of old mice with IL-7 produced significant increases in live TN cells. By comparison, treatment with SCF failed to increase live TN numbers, and IL-7 and SCF together failed to significantly improve thymopoiesis above that shown by IL-7 alone. Thus, treatment with IL-7 alone can reverse the age-associated defect in TN thymocyte development revealed by in vitro studies to be located at the stages of TCR beta-chain rearrangement.

Aging↗
PubMed2001

Effect of CD4(+) and CD8(+) cell depletion on wound healing.

BACKGROUND: Depression of the immune system can result in poor or delayed wound healing. METHODS: Thymectomized rats were depleted of CD4(+) and CD8(+) lymphocytes by intraperitoneal injection of Medical Research Council Oxford (MRC OX)38 antibodies and MRC OX8. Significant depletion was demonstrated throughout the wound healing process by immunofluorescence studies of peripheral blood. Following depletion the rats underwent laparotomy incisions which were allowed to heal for 10 weeks. Differences in healing were demonstrated by analysing the wounds biomechanically by tensiometry to obtain values of ultimate strength, resilience, toughness, maximum extension and elastic constant. RESULTS: Wounds of animals depleted of CD4+ lymphocytes showed a significant decrease in ultimate strength, resilience and toughness. Wounds of animals depleted of CD8(+) lymphocytes showed a significant increase in ultimate strength, resilience and toughness. CONCLUSION: Wounds healed in the absence of T lymphocytes. However, the subsets have an opposing regulatory role, with CD4(+) lymphocytes upregulating and CD8(+) lymphocytes downregulating wound healing. Presented to the Surgical Research Society in Nottingham, UK, 11 July 1997 and published in abstract form as Br J Surg 1997; 84: 1618

Animals↗
PubMed2001

Both age and gender affect thymic output: more recent thymic migrants in females than males as they age.

The thymus undergoes age-associated involution, with studies showing thymic size decreasing from birth at a rate of approximately 3% per year until middle age, and at a rate of 1% per year thereafter. The aim of this study was to determine the effect of thymic atrophy on T-lymphocyte production by the thymus, and to clarify the ongoing uncertainty regarding gender differences in thymic function. We quantified recent thymic emigrants (RTEs) in blood through the measurement of signal joint T-cell receptor rearrangement excision circles (sjTRECs), and showed that the decline in the number of RTEs in the blood with increasing age is gender-linked. Peripheral blood from females contained significantly higher levels of sjTRECs per CD3+ T cell than blood from males (P = 0.002), despite there being no significant gender difference in the absolute number of CD3+ T cells in the populations analysed (P > 0.10). Our findings suggest better thymic function in females compared with males, providing females with a higher number of recent thymic emigrants for longer periods of life. Such a finding provides a plausible explanation for the immunological gender differences observed in previous studies and possibly, for the general longer life expectancy in females compared with males.

Adult↗
PubMed2001

Gender-related differences in the rates of age associated thymic atrophy.

Age associated thymic atrophy has been shown to be linked to problems with rearrangement of the beta chain of the T cell receptor (TCR) in male mice during the early phases of the intrathymic T cell developmental pathway. In this study, thymic atrophy in female mice was found to occur at a different rate than in male mice. At 9 months of age there was a significantly greater number of cells in the thymus of female mice compared with male mice, with the major difference found in the CD4+CD8+ populations. The thymii of female mice at 9 months of age contained double the number of these cells compared with male mice. Analysis of the CD4+CD8+ cells at 9 months of age demonstrated increased numbers of cells expressing higher levels of CD3 in females compared with males indicating that in females more of these cells were producing successful alphabetaTCR pairings. In F5 transgenic mice comparison of the CD4+CD8+ population revealed no significant difference in their absolute numbers at 9 months of age. These results indicate that the gender differences at this time point were due to fewer permitted divisions prior to the expression of a selectable TCR alpha chain within the CD4+CD8+ populations in male compared with female mice. This gender difference was not due to the action of testosterone and unlikely to be due to differences in the level of oestrogen. The potential mechanisms of this difference may be related to a regulatory feedback of peripheral T cells on the developing thymocyte populations. Such age related changes in the numbers of cells within distinct thymic subpopulations leads to the possibility that the potential repertoire in females is greater than in males later in life.

Aging↗
PubMed2000

A simple method for the measurement of sjTREC levels in blood.

We have developed a relatively rapid, safe and simple method for the quantification of sjTREC levels in samples of peripheral blood. The assay uses an image analysis package to measure the brightness of PCR product bands on an image of the standard agarose gel. Comparison of the brightness of the band with that obtained from a standard curve provides a read-out of the amount of sjTRECs in the sample. We have compared the sjTREC levels we obtained with this method with those obtained from real time analysis PCR using a Lightcycler and found that they are comparable.

Adult↗
PubMed2000

Thymic atrophy in the mouse is a soluble problem of the thymic environment.

Age related deterioration in the function of the immune system has been recognised in many species. The clinical presentations of such immune dysfunction are an age-related increased susceptibility to certain infections, and an increased incidence of autoimmune disease and certain cancers. Laboratory investigations reveal a reduced ability of the cells from older individuals, compared with younger individuals, to perform in functional in vitro assays. These manifestations are thought to be causally linked to an age associated involution of the thymus, which precedes the onset of immune dysfunction. Hypotheses to account for the age-related changes in the thymus include: (i) an age related decline in the supply of T cell progenitors from the bone marrow (ii) an intrinsic defect in the marrow progenitors, or (iii) problems with rearrangement of the TCR beta chain because of a defect in the environment provided by the thymus. We have analysed these possible options in normal mice and also in mice carrying a transgenic T cell receptor. The results from these studies reveal no age related decline either in the number of function of T cell progenitors in the thymus, but changes in the thymic environment in terms of the cytokines produced. We have shown that specific cytokine replacement therapy leads to an increase in thymopoiesis in old animals.

Aging↗
PubMed2000

Thymic involution in aging.

The size of the naive T-cell pool is governed by output from the thymus and not by replication. This pool contributes cells to the activated/memory T-cell pool whose size can be increased through cell multiplication; both pools together constitute the peripheral T-cell pool. Aging is associated with involution of the thymus leading to a reduction in its contribution to the naive T-cell pool; however, despite this diminished thymic output, there is no significant decline in the total number of T cells in the peripheral T-cell pool. There are, however, considerable shifts in the ratios of both pools of cells, with an increase in the number of activated/memory T cells and the accumulation in older individuals of cells that fail to respond to stimuli as efficiently as T cells from younger individuals. Aging is also associated with a greater susceptibility to some infections and some cancers. An understanding of the causal mechanism of thymic involution could lead to the design of a rational therapy to reverse the loss of thymic tissue, renew thymic function, increase thymic output, and potentially improve immune function in aged individuals.

Aged↗
PubMed2000

Longevity and the immune response.

The domino hypothesis of the onset of age associated immune insufficiency suggests that it is the consequence of a cascade of events beginning with involution of the thymus. Involution is associated with a reduced thymic output leading to fewer naïve T cells contributing to the peripheral T-cell pool. Homeostatic mechanisms, which maintain the number of T cells in the peripheral pool within precise limits, induce the proliferation and prolong the survival of resident T cells to fill the niches left vacant by the absent naïve T cells. In this hypothesis, falling thymic output would be matched by resident T-cell proliferation and with age these proliferating cells will reach their replicative limit. Their prolonged survival will lead to the accumulation of cells unable to replicate, producing a decline in immune function and a susceptibility to infection, or certain cancers. Comparison of gender differences in life-span and rates of death in each age group due to infectious or parasitic disease suggests that the immune system in females works more efficiently and effectively for longer than the immune system in males. This leads to the suggestion that involution of the thymus, and hence thymic output, occurs more rapidly in males than in females.

Animals↗
PubMed2000

Immunosenescence: potential causes and strategies for reversal.

Age-related deterioration in immune function has been recognized in many species. In humans the clinical manifestation of such immune dysfunction is age-related increases in the susceptibility to certain infections and in the incidence of some autoimmune disease and certain cancers. Laboratory investigations reveal age-related changes in the peripheral T cell pool, in the predominant phenotype, cytokine production profiles, signalling function and in replicative ability following stimulus with antigen, mitogens or anti-CD3 antibody. These changes in the properties of peripheral T cells are thought to be causally linked to an age-associated involution in the thymus. Our analysis reveals that thymic involution is due to a change in the thymic microenvironment linked to a reduction in the level of available interleukin 7. Treatment with interleukin 7 leads to a reversal of thymic atrophy with increased thymopoiesis. This provides the potential to reverse the immune dysfunction seen in the peripheral T cell pool by replacing old cells with new output generated in the thymus. Problems to overcome in order for such an experimental therapy to be successful require careful analysis in order to provide an optimal strategy to ensure that new T cell emigrants from the thymus have a broad range of specificities and are able to enter the peripheral T cell pool.

Aging↗
PubMed2000

Erythrocyte transfusion causes immunosuppression after total hip replacement.

A prospective study of 34 patients undergoing total hip replacement was done to determine whether homologous erythrocyte transfusion causes postoperative immunosuppression. In the transfused patient group (14 patients), there was a reduction in CD3+ and CD3+4+ cell numbers at Day 2, returning to preoperative levels by Day 7. In contrast, in the untransfused patient group (20 patients), there was no significant depression in these lymphocyte subgroups at Day 2 and an increase in total lymphocyte, CD3+, CD3+4+, and CD3+4-8- cell numbers at Day 7. In both patient groups there was postoperative leukocytosis, granulocytosis, and monocytosis at Days 2 and 7, with no significant change in postoperative B (CD19+) cell numbers, natural killer cells, or the minor T cell populations of CD3+4+8+ and CD3+25+. The lymphocyte functional test of Candida recall was significantly impaired at Day 7 in the transfused patient group, where Candida recall is a memory T cell response to an antigen extracted from the yeast Candida Albicans. These findings suggest that homologous erythrocyte transfusion after hip replacement surgery causes cell mediated immune suppression. The main clinical implication of the current study is that perioperative homologous erythrocyte transfusion may place patients at greater risk of infectious complications, including infection of the prosthesis.

Arthroplasty, Replacement, Hip↗
PubMed1999

Production of human allotype-specific anti-CD45 monoclonal antibodies.

The aim of this work was to raise allotype specific monoclonal antibodies to human CD45, with the long-term objective of producing a reagent which could be used to prolong graft survival in renal transplantation through removal of passenger leukocytes from the graft. At present there are no anti-CD45 monoclonal antibodies able to distinguish between host and donor leukocytes. An in vitro immunisation technique has been developed through which donated human leukocytes are sensitised to CD45 prior to fusion with a myeloma cell line. IgM was produced by all the anti-CD45-positive clones. Flow cytometric analysis using these antibodies showed their ability to differentiate between blood from individual donors, indicating the existence of allotypic forms of human CD45, in conformity with the findings in rats and pigs. Therefore, a reagent which could be used in renal transplantation is a technical possibility.

Antibodies, Monoclonal↗
PubMed1999

Does the immune system of a mouse age faster than the immune system of a human?

One of the characteristics of all somatic cells is a finite life span. Cells may proliferate until they reach a point after which, although they are metabolically active, they can no longer produce daughter cells. This observation is central to the clonal exhaustion hypothesis, a mechanism cited to explain age-associated immune dysfunction. In this hypothesis, repeated division of lymphocytes leads to a replicative limit, after which they enter the senescent phase but are not lost from the pool of T cells. Advancing age would then be associated with an increase in the number of T cells that are unable to proliferate to a stimulus which induces a proliferative response in T cells from younger individuals. This hypothesis seems both logical and reasonable and is supported by data from both humans and mice with the demonstration of an age-related accumulation of senescent T cells in both species. However, there is an apparent paradox. The paradox arises because the onset of immunosenescence appears to be more closely linked to the life span of the animal rather than the life span of the lymphocyte.

Aging↗
PubMed1998

Age-related changes in the absolute number of CD95 positive cells in T cell subsets in the blood.

Comparison of the absolute number of cells in distinct T cells subsets expressing CD95 (Fas) was carried out in two populations of healthy female volunteers. In one population, the average age was 30 +/- 5 years, and in the second population the average age was 73 +/- 13 years. No significant difference was noted in the total number of lymphocytes, CD3+, CD4+, or CD8+ cells per microL of blood between the two age groups, but major differences were noted in the number of cells expressing CD95. A significant reduction was seen in the number of cells per microL of blood in both the CD4+ CD45RA+ CD95+ and CD8+ CD45RA+ CD95+ populations in the older group compared with the younger group. Within the memory pool significantly fewer CD8+ CD45RO+ CD95+ cells were found in the older population compared with the younger group. No such difference were found in the number of CD4+ CD45RO+ CD95+ cells between groups. Such a significant decline in the number of CD95+ cells, whose expression is known to be linked with activation, may be implicated as a mechanism by which cells that have reached a stage of replicative senescence remain in the peripheral T cell pool. Anti-CD3-mediated activation of cells from both groups revealed much lower proliferative responses from the older group, supporting the idea that there is an age-associated increase in the number of cells that have reached their replicative limit. These cells may not be lost from the peripheral pool because they fail to express CD95.

Adult↗