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Biomedical subjects

J Gordon

Publications and source records attributed to J Gordon.

At least 91 records · Page 5Linked to original sources
PubMed1998

Prolonged phenotypic, functional, and molecular change in group I Burkitt lymphoma cells on short-term exposure to CD40 ligand.

Group I Burkitt lymphoma (BL) cell lines (L3055, Elijah, Louckes, BL2, and BL29) retaining the original biopsy phenotype were found to undergo prolonged phenotypic, functional, and molecular change on short-term exposure to soluble recombinant CD40L trimer. Sensitivity to, extent of, and duration of change appeared to reflect passage number in that the earliest passaged lines, L3055 and BL29, were generally the most susceptible. Culture of group I BL lines with CD40L resulted in significant growth arrest (without apoptosis) that, for L3055 cells, was sustained for 7 to 9 days after 72 hours of exposure. This was accompanied by the formation of large homotypic aggregates together with gross changes in individual cell morphology and a concomitant upregulation of CD54 (ICAM-1). Three of the five group I BL lines exhibited rapid downregulation of the hallmark CD77 surface antigen, which, for L3055 cells, was maintained for at least 12 days after 72 hours of incubation with CD40L. With the exception of BL2, all group I BL lines were induced to express CD40 homodimers on CD40-stimulation, whereas only monomers were detected in unstimulated cells. Experiments using CD40-transfected Rat-1 fibroblasts showed that the ability to signal for dimer formation required Thr234 of CD40. For L3055 and BL29 cells, an initial 72 hours of exposure to CD40L resulted in the maintenance of homodimers for up to 14 and 10 days, respectively. There was a close correlation between the induction and duration of CD40 homodimers and the appearance of Bcl-2. For L3055 cells, which are sensitive to apoptosis-induction on BCR-engagement, exposure to CD40L for 72 hours was found to provide considerable protection from anti-IgM, which was still significant to 20 days. The implications of such sustained effects on relatively short-term exposure of tumor B cells to CD40L are discussed.

Antibodies, Anti-Idiotypic↗
PubMed1998

Retinoblastoma-related protein pRb2/p130 and suppression of tumor growth in vivo.

BACKGROUND: The RB/p105 and p107 genes of the retinoblastoma family are tumor suppressor genes whose proteins are inactivated by interaction with T-antigen proteins encoded by polyomaviruses (e.g., simian virus 40 and human JC virus), which have been found to be highly tumorigenic in animals. A variety of indirect evidence suggests that another member of the retinoblastoma gene family, RB2/p130, is also a tumor suppressor gene. To investigate the putative tumor suppressor activity of RB2/p130 more directly, we utilized a tetracycline-regulated gene expression system to control expression of the encoded protein pRb2/p130 in JC virus-induced hamster brain tumor cells and to study the effects of pRb2/p130 on the growth of such tumor cells in nude mice. The ability of pRb2/p130 to interact with JC virus T antigen was also studied. METHODS: Northern blot hybridization analyses were performed on samples of total cellular RNA to measure RB2/p130 and beta-actin messenger RNA levels. Immunoprecipitation and western blot analyses were used to determine T-antigen and pRb2/p130 protein levels and to assess the phosphorylation status of these proteins. Tumor cells were injected subcutaneously into nude mice, and tumor growth, with or without induced expression of pRb2/p130, was monitored. RESULTS: Induction of pRb2/p130 expression brought about a 3.2-fold, or 69% (95% confidence interval = 64%-73%), reduction in final tumor mass in nude mice. We also demonstrated that JC virus T antigen binds hypophosphorylated pRb2/p130 and that stimulation of pRb2/p130 expression overcomes cellular transformation mediated by this antigen. CONCLUSION: Our findings support the hypothesis that RB2/p130 is a tumor suppressor gene.

Animals↗
PubMed1998

Release of clonal block in B cell chronic lymphocytic leukaemia by engagement of co-operative epitopes on CD40.

The clonal cells of patients with B-chronic lymphocytic leukaemia (B-CLL)--which essentially reside in a resting configuration--are characterised by a relative refractoriness to the normal signals for B cell growth and differentiation. Previously it has been shown that, using an in vitro culture system where CD40 is hyper-crosslinked by monoclonal antibody (mAb) held on CD32-transfected mouse L cells, the clonal block in B-CLL cells can be released with a resultant high rate of DNA synthesis ensuing. In the present study, we report that such release can be achieved purely with soluble reagents whereby co-operative epitopes on CD40 are targeted by the combined use of mAb and soluble recombinant CD40L. Substantial levels of DNA synthesis were induced under such conditions in 7/18 patients using CD40-targeted reagents alone and in 16/18 patients in the additional presence of interleukin 4. Possible extrapolation of these findings to novel therapeutic modalities could be envisaged.

Antibodies, Monoclonal↗
PubMed1998

Partner support as a mediator of intervention outcome.

Following up on previous work demonstrating that an intervention with at-risk mothers made a positive impact on the quality of mothers' partner support, responsiveness to the needs of the child, the child's expectation of being cared for, and child's response to a brief separation, the present paper examines whether quality of the mother's partner support mediated the impact of the intervention on these outcomes.

Adult↗
PubMed1998

Measuring the hospital experiences of junior doctors.

The development of an appraisal questionnaire which measures junior doctors' opinions about their hospital experiences is described. The first section of the questionnaire consists of seven reliable subscales which measure opinions about teaching and learning, registrar teaching, consultant teaching, staff support, workload, administration and overall experiences during a period of attachment or term. The second part of the survey contains 11 reliable questions about the hours spent on service and education during the term. The responses to this measure of 257 randomly selected Australian junior doctors are described. The questionnaire may be used to contrast the experiences of junior doctors in different types of terms, different hospitals or varying levels of training. The data generated from the instrument can provide useful information about hospitals, such as the work practices of junior staff and the effectiveness of educational programs.

Adult↗
PubMed1998

A system for maintaining the educational and training standards of junior doctors.

The development of junior doctors' competence is complex because the hospital environment in which doctors work places many demands on them. The need for quality education and training and personal development may be in direct conflict with the service commitments required from hospitals. This paper describes the methods by which the Postgraduate Medical. Council of New South Wales, Australia, addresses the needs of junior doctors in the state in order to improve the quality of their education. Key elements of the Council's function include the provision of hospital clinical supervisors who oversee junior doctor education and training, and central involvement in supplying the junior doctor workforce to all state hospitals who must meet defined accreditation standards. This paper also provides data on evaluation of those methods and some educational outcomes.

Education, Medical, Graduate↗
PubMed1998

Rapid synthesis of IFN-gamma by T cells in skin may play a pivotal role in the human skin immune system.

This study has shown that T lymphocytes with alphabeta TCR present in normal human skin possess unique characteristics which may allow them to fulfil a unique role in skin. It disclosed that unlike T cells from blood, they are devoid of proliferative capacity in response to any ligand including lectins and phorbol esters, and show a diminished Ca2+ flux in response to phytohemagglutinin (PHA). However, despite this apparent lack of reactivity, these cells respond to stimulation by synthesis of IFN-gamma. Furthermore, this response by T cells from the skin differs from that of T cells in blood in its tempo. Whereas IFN-gamma is first detectable in peripheral blood lymphocytes (PBL) 42 h following stimulation, T cells from skin produce IFN-gamma 2.5 h after activation, or 17 times faster. Whereas synthesis persists in PBL for several days, it only lasts in skin T cells for 90 min. This difference in the kinetics is a consequence of differences in the regulation of IFN-gamma synthesis in PBL and skin-derived T cells. Thus, unlike PBL, T cells in intact unimmunized skin or ex vivo express mRNA for IFN-gamma, transcribed in a constitutive manner. Activation of these cells turns on translation of IFN-gamma synthesis directly and at the same time turns off further transcription of the IFN-gamma gene: cessation of IFN-gamma synthesis coincides with the disappearance of IFN-gamma mRNA within 4 h of activation. These observations suggest that this pre-formed mRNA may be solely responsible for the early synthesis of IFN-gamma in skin, determining both the amount and the duration of its synthesis. We propose that these cells may represent the antigen-specific component of a first-line host defence system in human skin. The reported absence of delayed-type hypersensitivity reactivity in patients with a selective deficit of skin T cells supports this conclusion.

Calcium↗
PubMed1998

The use of metoprolol and glycopyrrolate to prevent hypotensive/bradycardic events during shoulder arthroscopy in the sitting position under interscalene block.

UNLABELLED: Sudden profound hypotensive and/or bradycardic events (HBE) have been reported in >20% of patients undergoing shoulder arthroscopy in the sitting position under interscalene block anesthesia. Retrospective studies suggest that the administration of beta-blockers is safe and may decrease the incidence of these episodes. We performed a randomized, prospective study to evaluate prophylaxis of these events. One hundred fifty patients were randomized to one of three groups (placebo; prophylactic metoprolol to achieve a heart rate <60 bpm or a maximal dose of 10 mg; or prophylactic glycopyrrolate to achieve a heart rate >100 bpm or a maximal dose of 6 microg/kg) immediately after the administration of the interscalene block. Blood pressure control was achieved with IV enalaprilat as needed. The incidence of HBE was 28% in the placebo group versus 5% in the metoprolol group (P = 0.004). The rate of 22% in the glycopyrrolate group was not significantly different from placebo. Preoperative heart rate and arterial blood pressure, intraoperative sedation score, IV fluids, and enalaprilat use were similar in those patients who had a HBE compared with those who did not. Many aspects of this clinical setting are similar to tilttable testing for patients with recurrent vasovagal syncope, in which beta-adrenergic blockade with metoprolol has also been shown to be effective. We conclude that the Bezold-Jarisch reflex is the most likely mechanism for these events. IMPLICATIONS: Episodes of acute hypotension and bradycardia occur during shoulder arthroscopy in the sitting position under interscalene block. In this study, we demonstrate that metoprolol, but not glycopyrrolate, markedly decreases the incidence of these episodes when given prophylactically immediately after the administration of the block.

Adrenergic beta-Antagonists↗
PubMed1998

Tumor pathogenesis of human neurotropic JC virus in the CNS.

In the last several years, studies have reported the detection of sequences similar to the polyomavirus, Simian Virus 40 (SV40) in human tumors including choroid plexus papillomas and ependymomas. Taken together with well-established evidence that SV40, as well as the human polyomaviruses JC virus (JCV) and BK virus (BKV), are oncogenic in several animal models, new interest has resurfaced regarding a possible role for these viruses in human tumors. In particular, a strong case can be made for re-evaluating the oncogenic potential of JCV in a neurotropic polyomavirus which is the causative agent of the fatal human demyelinating disease, Progressive Multifocal Leukoencephalopathy (PML). In this review, we discuss the transforming capability of JCV in vitro, JCV's ability to induce neural origin tumors in non-human primates as well as in rodents, and several case reports which suggest a possible role for JCV in tumor pathogenesis in the central nervous system.

Animals↗
PubMed1998

The human polyomavirus, JCV, and neurological diseases (review).

JC virus, a human neurotropic polyomavirus, is the established etiologic agent of the fatal demyelinating disease, progressive multifocal leukoencephalopathy, which usually affects individuals with defects in cell-mediated immunity. Cytolytic destruction of oligodendrocytes, the myelin-producing cells of the central nervous system is attributed as the mechanism by which JCV induces demyelination. PML was at one time a rare complication however it is now a much more common disease affecting patients of all ages due to the prevalence of AIDS. Of interest, in vitro evidence points to a cooperative interaction between JCV and HIV-1, via the HIV-1 regulatory protein, Tat. In addition, JCV has been demonstrated to induce tumors of neural origin in several animal models, including rodents and non-human primates, and several clinical reports have suggested a possible association between JCV and glial-origin tumors in humans. The viral regulatory protein, T-antigen, which has been shown to play a key role in orchestrating the events of the viral lytic cycle, is also capable of altering cellular functions, by nature of its direct interaction with cellular regulatory proteins and by its effect on cellular transcription. In this review, we discuss clinical aspects of PML, the ability of JCV to induce tumors in animal models, and the ability of JCV T-antigen to alter cellular function in vitro.

AIDS-Related Opportunistic Infections↗
PubMed1998

Oncogenic potential of human neurotropic papovavirus, JCV, in CNS.

The human polyomavirus, JCV, is the causative agent of Progressive Multifocal Leukoencephalopathy (PML), a fatal human demyelinating disease. PML results from the cytolytic destruction of oligodendrocytes, the myelin-producing cells of the nervous system. JCV has also been shown to be tumorigenic in several animal models. Transgenic mice expressing the JCV early protein, T-antigen, develop poorly differentiated neural crest origin tumours. Intracerebral inoculation of JCV into newborn hamsters induces medulloblastomas, astrocytomas, and primitive neuroectodermal tumours. Further, inoculation of the virus into the brains of non-human primates, owl and squirrel monkeys, results in astrocytomas and glioblastoma multiforme. Several case reports have associated JCV with human CNS tumours in patients with concomitant PML, and one such report has detected JCV in a glial tumour in the absence of PML. The induction of neural origin tumours by JCV has been studied in transgenic mice harbouring the early genome of the virus. Alterations in the level and function of tumour suppressor proteins p53 and Rb, as well as associated cell cycle regulators, have been detected in tumour tissue from JCV T-antigen transgenic mice. Possible mechanisms by which JCV may exert its oncogenic potential by alteration of cellular growth control pathways in both humans and experimental animals are discussed.

Animals↗
PubMed1997

Mechanisms of antigen receptor-dependent apoptosis of human B lymphoma cells probed with a panel of 27 monoclonal antibodies.

The present study has used a panel of 23 monoclonal antibodies to IgM and 4 to IgD in order to probe parameters influencing sIg-dependent apoptosis in an IgM/IgD-expressing Burkitt lymphoma line. No direct correlation was observed between the capacity of the different anti-mu to drive cells into apoptosis and either their domain specificity or their affinity for sIgM. There was, however, a direct correlation between the functional outcome and the ability of the monoclonal antibodies to elicit a rise in intracellular Ca2+. For apoptosis to occur, the Ca2+ response had to attain a threshold value of approximately 100 nM. A direct role for Ca2+ in the delivery of the apoptotic signal was demonstrated using thapsigargin to raise intracellular Ca2+ levels. Antigen receptor ligation was linked to Ca2+ increases by tyrosine kinases as revealed by direct analysis of protein tyrosine phosphorylation and the effects of selective protein tyrosine kinase-inhibiting tyrphostins. These findings reveal a central role for the antigen receptor-generated Ca2+ signal in driving apoptosis in human B lymphoma cells and stresses the need to use a panel of reagents when probing function with presumed ligand-mimetic monoclonal antibodies.

Antibodies, Monoclonal↗
PubMed1997

Role of cell cycle regulators in tumor formation in transgenic mice expressing the human neurotropic virus, JCV, early protein.

Transgenic mice harboring the early genome from the human neurotropic JC virus, JCV, develop massive abdominal tumors of neural crest origin during 6-8 months after birth and succumb to death a few weeks later. The viral early protein, T-antigen, which possesses the ability to transform cells of neural origin, is highly expressed in the tumor cells. Immunoblot analysis of protein extract from tumor tissue shows high level expression of the tumor suppressor protein, p53, in complex with T-antigen. Expression of p21, a downstream target for p53, which controls cell cycle progression by regulating the activity of cyclins and their associated kinases during the G1 phase, is extremely low in the tumor cells. Whereas the level of expression and activity of cyclin D1 and its associated kinase, cdk6, was modest in tumor cells, both cyclin A and E, and their kinase partners, cdk2 and cdk4, were highly expressed and exhibited significant kinase activity. The retinoblastoma gene product, pRb, which upon phosphorylation by cyclins:cdk induces rapid cell proliferation, was found in the phosphorylated state in tumor cell extracts, and was detected in association with JCV T-antigen. The transcription factor, E2F-1, which dissociates from the pRb-E2F-1 complex and stimulates S phase-specific genes upon phosphorylation of pRb and/or complexation of pRb with the viral transforming protein, was highly expressed in tumor cells. Accordingly, high level expression of the E2F-1-responsive gene, proliferating cell nuclear antigen (PCNA), was detected in the tumor cells. These observations suggest a potential regulating pathway that, upon expression of JCV T-antigen, induces formation and progression of tumors of neural origin in a whole animal system.

Animals↗
PubMed1997

Terminal differentiation of human germinal center B cells in vitro.

In order to define an in vitro culture system allowing growth of single human germinal center B cells (GC-B), we have studied the proliferation and differentiation of human tonsillar GC-B, and subsets thereof, when cultured together with murine EL-4 thymoma cells in the "EL-4 system." The cells were analyzed and compared to resting tonsillar B cells with respect to phenotypic changes, proliferation, Ig secretion, intracellular Ig levels, and growth abilities under limiting dilution conditions. It was found that GC-B differentiated terminally to Ig-secreting cells with the phenotypic features of plasma cells in a similar manner to tonsillar resting B cells. The GC-B proliferated for 4-5 days, followed by a loss of GC-B phenotype and an increase in intracellular immunoglobulin levels. Over a 10-day culture period a larger proportion of the Ig produced by GC-B was IgG and IgA, as compared to resting B cells, indicating that these cells switched isotype more easily or had already switched in the germinal center prior to the culture period. Analysis of frequencies of Ig-producing cells revealed that 1/3.8 of GC-B and less than 1/10 of the centroblast B cell subpopulation (CB-B) differentiated toward Ig-producing cells when cultured in the EL-4 system whereas 1/1.25 and 1/1.5 of peripheral blood B cells (PBL-B) and resting tonsillar B cells did so, respectively. Taken together, these findings show that tonsillar GC-B differentiate in a similar manner to resting B cells when cultured in the EL-4 system, and we conclude that these conditions allow manipulation of GC-B in single cell cultures in vitro.

Animals↗
PubMed1997

Maintenance of human germinal center B cells in vitro.

The ability to maintain germinal center (GC) B cells in culture should facilitate studies on the molecular and cellular events which accompany affinity maturation and the generation of memory in T-dependent responses. We have investigated the ability of cytokines to maintain human tonsillar GC B cells (IgD-/CD39-/CD38+/CD77+) in the "CD40 culture system". In the absence of added cytokines, CD40 monoclonal antibody held on CD32-transfected L cells effectively sustained DNA synthesis in GC B cells for a maximum 3 to 4 days. Of the following cytokines (interleukin-1 beta [IL-1 beta], IL-2, IL-3, IL-4, IL-6, IL-7, IL-10, and stem cell factor), only IL-2 and IL-4 provided a significant enhancement to DNA synthesis in the CD40 culture system; this was modest and short-term. Following a study on the cooperative activity between pairs of cytokines, triple combinations were identified that could maintain high levels of GC B-cell stimulation for at least 10 days. IL-10 was a common component of these synergistic cytokine cocktails, which were IL-10 + IL-4 + IL-7; IL-10 + IL-3 + IL-7; IL-10 + IL-1 beta + IL-2; IL-10 + IL-1 beta + IL-3, and IL-10 + IL-3 + IL-6. Culture of GC B cells with these cytokine combinations resulted in a net increase in viable cell numbers of 50% to 100% whereas total cell numbers increased up to fourfold. Cells recovered from these cultures retained a GC B-cell phenotype with a significant proportion being CD38+/CD44-, features characteristic of centroblasts. Studies with metabolically inactive CD32-L cells supported a role for stromal cell-derived soluble factors in maintaining GC B cells in vitro.

Animals↗