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Biomedical subjects

M A Epstein

Publications and source records attributed to M A Epstein.

At least 91 records · Page 5Linked to original sources
PubMed1982

Cytotoxic T cell recognition of Epstein-Barr virus-infected B cells. III. Establishment of HLA-restricted cytotoxic T cell lines using interleukin 2.

Epstein-Barr virus (EBV)-specific cytotoxic T cell precursors, present in the circulation of previously infected (seropositive) individuals, have been reactivated in vitro by challenging with autologous EBV-transformed cells, and the reactivated populations subsequently expanded as interleukin 2 (IL2)-dependent cell lines. These lines were dominated by T cells possessing the cytotoxic/suppressor cell surface phenotype and, when tested for effector function in chromium-release assays, demonstrated potent EBV-specific, HLA-A and -B antigen-restricted cytotoxicity even when derived from seropositive donors whose initial cytotoxic response to in vitro reactivation was relatively weak. With all the lines tested from 10 seropositive donors, strong killing of autologous EBV-transformed cells was observed in the absence of any significant lysis of autologous mitogen-stimulated lymphoblasts or of a panel of EBV genome-negative cell lines sensitive to natural killing. Furthermore, the availability of IL2-expanded effectors cell populations allowed their being tested upon a wide panel of allogeneic EBV-transformed targets such that the dominant HLA-restricted reactivities within these populations could be identified. Monoclonal antibody blocking experiments confirmed that lysis of the autologous EBV-transformed cell line by IL2-expanded effectors could be specifically inhibited (a) by pretreatment of the target cells with antibodies binding to the HLA/beta 2-microglobulin complex, and (b) by pretreatment of the effector cells with the cytotoxic/suppressor T cell-specific antibody Leu 2a.

Adult↗
PubMed1982

Epstein-Barr virus-specific cytotoxic T cell responses in rheumatoid arthritis patients.

The level of Epstein-Barr (EB) virus-specific cytotoxic T cell responsiveness was measured in 21 patients with active, progressive rheumatoid arthritis (RA). A significant number (8 out of 20) of EB virus sero-positive patients showed a markedly impaired responsiveness when compared with a control group of healthy individuals. Serological responses of the RA patients to EB virus antigens were not significantly different from the control group. The defect in EB virus-specific cellular immunity shown by these results is of interest in the light of previous evidence of an alteration in the virus-host balance in RA patients.

Adult↗
PubMed1982

A new ventilator for physiologic studies during high-frequency ventilation.

Research in the field of high frequency ventilation (HFV) has been hampered by the difficulty of collecting and analyzing expired gas, uncontaminated by gas of inspiratory composition, at ventilatory frequencies in the relevant range. A new mechanical ventilator is described, which has been designed to facilitate studies on the respiratory physiology of small animals during HFV. Ventilation is controlled by a single rotary valve, which alternately switches the airway to an approximately constant-flow gas source during inspiration, and to gas measurement and analysis apparatus during expiration, which takes place passively. Theoretical and experimental studies, using the techniques of mechanical and mathematical modelling, are used to measure the performance of the ventilator and to demonstrate that it allows the quantitative collection of expired gas during ventilation at rates up to at least 20 Hz. The advantages and limitations of this approach are discussed.

Animals↗
PubMed1982

Quantification of an Epstein-Barr virus-associated membrane antigen component.

A method is described for the preparation of a 125I-labelled membrane antigen (MA) component (gp340) from B95-8 cell membranes using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Good yields of antigenic material were obtained when renaturation of the [125I]gp340 was carried out by removal of SDS in the presence of urea and subsequent removal of the urea. The availability of purified, radiolabelled gp340 has provided the essential basis for the development of a radioimmunoassay which, for the first time, permits quantification of this antigen. The assay has been used to demonstrate that cell membrane MA is a better source of gp340 for large-scale work than is the Epstein-Barr virus envelope and to measure the increase in expression of gp340 following treatment of cells with 12-O-tetradecanoyl-phorbol-13-acetate (TPA).

Animals↗
PubMed1982

Purified Epstein-Barr virus Mr 340,000 glycoprotein induces potent virus-neutralizing antibodies when incorporated in liposomes.

The purified Mr 340,000 glycoprotein component of Epstein-Barr (EB) virus-induced membrane antigen complex incorporated into liposomes was shown to be a potent immunogen in mice. High-titer antisera were induced that (i) are specific for membrane antigen components without absorption, (ii) bind the antigens induced by three different EB virus isolates, and (iii) neutralize the ability of the virus to transform fetal cord blood lymphocytes in vitro. The development of this immunogenic form of purified antigen provides an important step towards a potential subunit vaccine against Epstein-Barr virus infection.

Antibodies, Viral↗
PubMed1982

Mechanisms of halothane adsorption by dry soda-lime.

Using fresh soda-lime (15% water by weight) the soda lime/air partition coefficient of halothane was found to decrease as a function of vapour phase halothane concentration from 2.40 at 0.3% halothane by volume to 1.15 at 2.6%, but adsorption generally followed Henry's law. However, soda-lime dried to a constant weight and subsequently exposed to various concentrations of halothane adsorbed approximately 320 microlitre of vaporized liquid halothane per 100 g before a measurable concentration of halothane was detected in the vapour phase. Adding additional halothane then caused a linear increase in vapour concentration. We conclude that dry soda-lime can absorb large quantities of halothane by a mechanism which is similar to that of a molecular sieve. After these "high affinity" sites are satisfied, additional halothane is absorbed by a mechanism following Henry's law.

Adsorption↗
PubMed1982

Breath-to-breath variations in rate and depth of ventilation in sleeping infants.

Ventilatory measurements were made noninvasively over 2- to 3-h periods during sleep in each of nine normal infants at 1 mo of age. To assess the changes that occur in ventilation on a breath-to-breath basis, we 1) examined the variations of each of tidal volume (VT), respiratory cycle time (Ttot), expiratory time (TE), and inspiratory time (TI) and 2) studied their interrelationships. We found that the variations of VT, Ttot, and TE but not of TI were significantly greater in rapid-eye-movement (REM) than in quiet sleep. In addition, on a breath-to-breath basis, VT had a positive linear relationship and strong correlation with TI; however, the correlation between VT and TE was weak in both sleep states. VT/Ttot was found to be moderately and negatively correlated with Ttot in both REM and quiet sleep. VT was weakly correlated with Ttot in REM sleep and was, on the average, more correlated with Ttot in quiet sleep. We suggest that in infants 1) on a breath-to-breath basis, VT/Ttot is likely to drop if respiratory frequency is decreased and 2) VT is nonlinearly related to Ttot during sleep; this lack of linearity depends on the lack of constancy of VT/Ttot, which is in turn closely related to the variability of the "on-switching" of inspiratory activity.

Humans↗
PubMed1982

Interaction of Epstein-Barr virus with leukaemic B cells in vitro. I. Abortive infection and rare cell line establishment from chronic lymphocytic leukaemic cells.

Leukaemia B cell populations, each with an individual pattern of monoclonal surface immunoglobulin expression, were obtained from 23 patients with chronic lymphocytic leukaemia (CLL) and, following exposure to a potent dose of Epstein-Barr (EB) virus in vitro, were monitored for expression of the virus associated nuclear antigen EBNA, for activation of immunoglobulin synthesis and for virus-induced transformation to an established cell line. Although possessing the EB virus receptor, CLL cells were generally refractory (vis-à-vis normal adult B cells) to the full effects of the viral infection. All the leukaemic populations tested developed a small proportion of EBNA positive cells within a few days post-infection, but in most instances this disappeared with no subsequent evidence of viral activity. In certain cases, however, the EBNA staining became more intense, involving a larger fraction of the population and persisting for some weeks, but again this was not accompanied by virus-induced immunoglobulin synthesis or transformation. In contrast, the leukaemic cells from a single patient, tested on three separate occasions, regularly responded to EB virus infection with the rapid establishment of an EBNA positive B cell line in which the restricted pattern of surface and cytoplasmic immunoglobulin expression (gamma lambda) exactly matched that present on the original leukaemic cells.

Antigens, Nuclear↗
PubMed1981

Reactivation of Epstein-Barr virus-specific cytotoxic T cells by in vitro stimulation with the autologous lymphoblastoid cell line.

Unfractionated mononuclear (UM) cells and T cells freshly prepared from the blood of adult donors were co-cultivated in microtest plate wells with progressively lower numbers of cells from the autologous EB-virus-transformed B-cell line. The fresh cells present in co-cultures from EB virus antibody-negative (seronegative) donors regularly facilitated autologous cell line outgrowth, monitored after 4 weeks, whereas outgrowth was markedly inhibited in the corresponding co-cultures from seropositive donors. Larger-scale co-cultures, set up at a ratio of 80-100 fresh UM cells to one autologous virus-transformed B cell, were harvested after 8 to 12 days and the T-cell subpopulation was examined for cytotoxicity both by growth inhibition and by chromium release assays. Cytotoxic T cells were generated exclusively in seropositive donor co-cultures and were strongly active against the autologous virus-transformed cell line without affecting either autologous uninfected B cells or any of a range of EB virus genome-negative target cell lines chosen as sensitive indicators of non-specific cytotoxicity. Recognition of allogeneic EB-virus-transformed cells was restricted to those whose HLA-A and/or B and/or B and/or C antigen expression matched that of the effector cells themselves;; moreover target cell lysis was specifically inhibited in the presence of monoclonal antibodies binding to these HLA antigens. The results indicate that EB-virus-specific HLA-restricted memory T cells, present in the blood of previously-infected individuals, can be reactivated in vitro using the established autologous virus-transformed cell line as a stimulus. THe reactivated cytotoxic cells appear to recognize a virus-induced lymphocyte-detected membrane antigen, LYD-MA, analogous to that first invoked to explain the cytotoxic response to primary EB virus infection observed during infectious monoucleosis.

Adult↗
PubMed1981

Cytotoxic T cell recognition of Epstein-Barr virus-infected B cells. I. Specificity and HLA restriction of effector cells reactivated in vitro.

The experiments show that the phenomenon of regression, seen exclusively in Epstein-Barr (EB) virus-infected cultures of mononuclear cells from EB virus antibody-positive donors, is mediated by cytotoxic T cells reactivated in vitro and specifically recognizing an EB virus-induced lymphocyte-detected membrane antigen LYDMA. Thus, effector T cells from regressing cultures kill autologous EB virus-transformed cells but not autologous pokeweed mitogen-stimulates lymphoblasts nor any of a range of EB virus genome-negative human hemopoietic cell lines (K562, HSB2, BJAB, EB4) particularly sensitive to nonspecific natural killer-like activities. Moreover, these reactivated effector cells exhibit classical HLA restriction of target cell recognition; in a survey of 14 effector cell donors, preferential lysis of the autologous virus-transformed line was a consistent feature, while the relative degree of lysis of allogeneic lines was in general directly related to the number of HLA-A and B antigens shared between effector and target cells. The pattern of reactivity shown by effector T cell preparations from any one donor was strikingly reproducible, and the results from a number of donors revealed differences between particular HLA-A and B antigens with respect to the level of EB virus-specific killing which was associated with sharing through these determinants.

B-Lymphocytes↗
PubMed1981

Cytotoxic T cell recognition of Epstein-Barr virus-infected B cells. II. Blocking studies with monoclonal antibodies to HLA determinants.

Monoclonal antibodies specifically binding common determinants on all HLA-A, B anc C antigen molecules blocked the lysis of EB virus-transformed target cells by EB virus-specific cytotoxic T cells reactivated in vitro. Blocking was mediated through binding of the antibodies to the target rather than to the effector cells and was maximal (75 to 85% inhibition of lysis) at saturating antibody concentrations. A similar blocking effect was also shown by a monoclonal antibody binding to beta 2-microglobulin, a molecule physically associated with HLA-A, B and C antigens on the target cell surface, but not by a monoclonal antibody binding to the non-HLA-associated leukocyte-common antigen. Saturating concentrations of monoclonal antibodies specific for common determinants on all HLA-DRw antigen molecules either had no effect at all upon EB virus-specific T cell cytotoxicity or caused a slight, but nonspecific, inhibition. The results demonstrate unequivocally that HLA-A, B and C antigens on the target cell surface are indeed the polymorphic elements which impose genetic restriction upon EB virus-specific cytotoxic T cell function.

Antibodies, Monoclonal↗
PubMed1981

Antigenic differences between the membrane antigen polypeptides determined by different EB virus isolates.

The antigenic specificities of the membrane antigen (MA) complex of three Epstein-Barr (EB) virus-producing cell lines have been compared by complete absorption of an anti-MA antiserum with P3HR-1 cells, followed by testing for residual anti-MA antibody activity against B95-8 and QIMR-WIL cells. Indirect membrane immunofluorescence showed that the absorbed serum, which had lost the capacity to bind P3HR-1 cells, nevertheless gave bright staining on a small proportion of B95-8 cells. SDS-polyacrylamide gel electrophoretic analysis of 125I-labelled MA polypeptides showed that the absorbed serum was able to isolate MA polypeptides from both B95-7 and QIMR-WIL cells, but not from P3HR-2 cells. The results demonstrate that there is substantial sharing of antigenic specificities between the MA determined by three isolates of EB virus, and some antigenic divergence.

Animals↗
PubMed1981

Flow to lung compartments with different time constants: effect of choice of model.

The thesis that an accelerating flow generator provides more even ventilation than a constant flow generator originated from studies using mechanical lung models. We simulated such studies by use of mathematical models. We found that the conclusions of these previous studies were dependent on the particular characteristics of their mechanical models. Studies employing more commonly used models indicate that constant flow generators, in fact, tend to provide more uniform ventilation, especially with long inspiratory times.

Humans↗
PubMed1980

HLA-restricted T-cell recognition of Epstein-Barr virus-infected B cells.

In mice the cytotoxic T-cell response to several types of virus is influenced by genes within the major histocompatibility complex; in particular, genetic control is exercised at the effector cell level through a requirement that virus-specific cytotoxic T cells recognise viral antigens in association with H-2K and H=2D region gene products on the surface of infected cells. In man the restriction which the analogous HLA-A, -B and -C-region gene products might place on virus-specific T-cell function is still in dispute. The earliest and most controversial evidence concerns the Epstein-Barr virus (EBV), a B lymphotropic agent which causes infectious mononucleosis (IM) and which induces an unusually vigorous T-cell response; cytotoxic T cells from IM patients' blood were shown to be EBV-specific yet, in contrast to mouse systems, apparently free of any obvious HLA restriction. Since then T-cell recognition of EBV-infected B cells has assumed particular significance as a model system for the study of cytotoxic T-cell function in man. This report describes the results of a new approach clearly indicating that HLA-A and -B region products do indeed have a role in this system.

Antibodies, Viral↗