Functional reconstruction of the shoulder with a latissimus dorsi pedicle flap and a hemiarthroplasty: a case report.
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Adult↗
Biomedical subjects
T Owens
Publications and source records attributed to T Owens.
Th1 and Th2 help for B cells: differential capacity for induction of autonomous responsiveness to IL-2.
Sustained interaction with Th1 cells has been shown to induce IL-2 responsiveness by murine B cells. This is equivalently dependent on CD40, CD54/ICAM-1 and MHC II ligation, and co-cross-linking of CD54 and MHC II in the presence of IL-5 up-regulates a functional IL-2R on B cells. We now show that IL-5 (125 U/ml) synergizes with Th1 cells to induce B cell responses to IL-2, that are maintained following T-cell removal, e.g. autonomous. Th1 help in the absence of IL-5 resulted in weak or undetectable responses following T cell removal. The mechanism of IL-5 synergy involved persistence of IL-2R beta expression following T cell removal, as opposed to enhancement of IL-2R induction or function. The level of contact-induced IL-2R expression on B cells was not itself modified by IL-5. The effects of IL-5 did not overcome the requirement for T contact signals and treatment of B cells with soluble anti-Ig did not circumvent the need for IL-5 for autonomous IL-2 responses. Consistent with the above, interaction with an IL-5-producing Th2 clone induced strong autonomous B cell responses to IL-2. Qualitative differences of Th2 help over that of Th1 may thus be attributable to their differential ability to induce autonomous B cell responsiveness to cytokines. This may be representative of events in which maintenance of cell cycle is important, as is the case in germinal centers.
Animals↗
The immunology of multiple sclerosis and its animal model, experimental allergic encephalomyelitis.
Two questions were posed at the beginning of this article. Is EAE a good model for MS? And, is MS an autoimmune disease? The first question is easier to address than the second. EAE is the best available model for the inflammatory processes that occur in MS, and for the disease process. The latter depends somewhat on study of chronic relapsing EAE, rather than early or mono-episodic EAE, which, although of great immunological interest, is of less relevance to the established disease that presents as MS. The second question asks whether MS fulfills Koch's postulates as an autoimmune disease. MS has all the hallmarks of an inflammatory disease of the CNS. The question then is whether the inflammation is autoimmune. The evidence presented shows a considerable autoimmune component to MS inflammation, raising the subsidiary question of whether autoimmune reactivity induces MS. This remains unanswerable for the present, and it should be kept in mind that the same question also would be unanswerable by observation of EAE. The major postulate therefore remains unfulfilled. Diagnosis of MS as an autoimmune disease requires definitive identification of the autoantigen; otherwise, the possibility remains open that this is a disease resulting from the inadvertent activation and dysregulation of immune processes in the CNS that, themselves, are not directed at that organ.
Animals↗
CD54/intercellular adhesion molecule 1 and major histocompatibility complex II signaling induces B cells to express interleukin 2 receptors and complements help provided through CD40 ligation.
We have examined signaling roles for CD54 intercellular adhesion molecule 1 and major histocompatibility complex (MHC) II as contact ligands during T help for B cell activation. We used a T helper 1 (Th1)-dependent helper system that was previously shown to be contact as well as interleukin 2 (IL-2) dependent to demonstrate the relative roles of CD54, MHC II, and CD40 signaling in the events leading to the induction of B cell proliferation and responsiveness to IL-2. Paraformaldehyde-fixed activated Th1-induced expression of IL-2R alpha, IL-2R beta, and B7, and upregulated MHC II and CD54 on B cells. Anti-CD54 and MHC II mAbs as well as a CD8 alpha-CD40 ligand (L) soluble construct inhibited both the T-dependent induction of Ig secretion, and B cell phenotypic changes. We then compared the effects of activated Th1 cells with that of cross-linking these molecules. Cross-linking of CD54 and MHC II resulted in the upregulated expression of MHC II and of CD54 and B7, respectively, analogous to the effect of fixed activated Th1 cells. B7 expression was further enhanced by co-cross-linking CD54 and MHC II. Cross-linking of CD40 achieved comparable effects. Strikingly, cross-linking ligation of CD54 and MHC II in the presence of IL-5 induced expression of a functional IL-2R on small resting B cells. By contrast CD40 ligation, which induced B cell proliferation, did not induce IL-2 responsiveness. These data show that CD40 ligation is necessary but may not be sufficient for B cell differentiation and identify CD54 and MHC II as contact ligands that can complement CD40 signaling in the generation of T-dependent B cell responses to IL-2.
Animals↗
Co-stimulation by anti-immunoglobulin is required for B cell activation by CD40Llow T cells.
During cognate B:T interactions, B cells encounter antigen (Ag) through surface immuno-globulin (sIg) and present antigenic peptides to T helper (Th) cells. However, most in vitro systems used to study contact events involved in the delivery of T help for B cells circumvent the requirement for T cell Ag specificity by using anti-CD3/T cell receptor (TcR) monoclonal antibodies (mAb) to activate T cells. To study the role of sIg engagement in the responsiveness of B cells to T help, we pre-treated small resting B cells with soluble anti-kappa mAb prior to contact with an activated Th1 clone. By reducing the concentration of anti-TcR mAb we obtained low levels of CD40 ligand (CD40Llow) on Th cells, comparable to those expressed by lymph node T cells activated in vitro (ex vivo T cells). In contrast to untreated B cells, which did not respond to CD40Llow Th, anti-Ig-treated B cells responded strongly. Low buoyant density B cells also responded to CD40Llow Th cells. There was no B cell response to resting Th cells. mAb against CD54/intercellular adhesion molecule-1 or major histocompatibility complex (MHC) class II completely inhibited B cell responses to CD40Llow Th1 cells, equivalent to the effects of blocking CD40 interactions. This contrasts with mAb blocking responses to CD40Lhigh Th, where CD40 effects predominate. Our data show that sIg engagement is necessary for the induction of B cell response to CD40Llow Th cells. Anti-CD3-activated ex vivo T cells that were also CD40Llow did not provide help to small resting B cells, but did induce responses from sIg-stimulated B cells. Thus, our data support a requirement for sIg signaling in physiological B cell activation, and further confirm previous work showing CD40 ligation to be necessary but not sufficient for delivery of T help to B cells.
Animals↗
Cytokine production by cells in cerebrospinal fluid during experimental allergic encephalomyelitis in SJL/J mice.
Cytokine production by T cells in the cerebrospinal fluid (CSF) and central nervous system (CNS) of SJL/J mice during myelin basic protein (MBP)-induced experimental allergic encephalomyelitis (EAE) was examined. Reverse transcriptase/polymerase chain reaction (RT/PCR) was used to measure interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) mRNA levels from perfused CNS tissue (brain and spinal cord) and from cells isolated from CSF. Animals were grouped according to EAE severity, ranging from asymptomatic (adjuvant only) to severe disease (paralysis or severe paresis). Cytokine signals, normalized to actin, were almost undetectable in control tissues, and only slightly elevated in whole CNS tissue from animals with mild EAE. Both cytokine messages were strongly upregulated in CNS tissues derived from severely affected animals, consistent with previous observations correlating disease progression with infiltration by memory/effector CD4+ T cells, the major source of these cytokines. This cytokine upregulation was specific to the CNS, since other organs from the same animals did not express significant levels of IL-2 and IFN-gamma. CSF was obtained from the cisterna magna of unperfused mice and verified as such by absence of red blood cells (RBCs) and by immunoglobulin concentration orders of magnitude lower than in serum. Cytokine message was measured in RNA isolated from cells in CSF. Levels of IL-2 and IFN-gamma mRNA in CSF cells were significantly elevated in mild EAE and strongly upregulated in severe disease, correlating with those in total CNS tissue. These results confirm the CSF as representative of the immune status of the CNS and indicate a role for IL-2 and IFN-gamma in inflammatory CNS disease.
Animals↗
Inflammatory cytokines in the brain: does the CNS shape immune responses?
Immune responses in the central nervous system (CNS) have traditionally been regarded as representing the intrusion of an unruly, ill-behaved mob of leukocytes into the well-ordered and organized domain of thought and reason. However, results accumulated over the past few years suggest that, far from being an immunologically privileged organ, T lymphocytes may be regular and frequent visitors to the CNS, for purposes of immune surveillance. Here, Trevor Owens and colleagues propose that the brain itself can regulate or shape immune responses therein. Furthermore, given that the immune cells may be subverted to autoimmunity, they suggest that the study of inflammatory autoimmune disease in the brain may shed light on the ability of the local environment to regulate immune responses.
Animals↗
Cystic periventricular leukomalacia and type of cerebral palsy in preterm infants.
A case series design was used to identify cases of cystic periventricular leukomalacia (N = 31) identified by neurosonography at one regional tertiary intensive care nursery. Patients were preterm infants born at < or = 32 weeks of gestation who had cysts involving predominantly the middle-posterior or posterior periventricular regions. Neurodevelopmental evaluations were made for 26 (96%) of 27 survivors. All infants assessed had cerebral palsy (i.e., 54% quadriplegia, 42% diplegia, and 4% hemiplegia). Most cognitive delays and all sensory impairments occurred in children with quadriplegia. Periventricular cysts were most extensive on parasagittal, anteroposterior views. The parasagittal, anteroposterior extent of periventricular cysts was most accurate in predicting the type and severity of motor and cognitive disabilities. Quadriplegia was associated with larger and more extensive cysts.
Cerebral Palsy↗
Selective enrichment of Th1 CD45RBlow CD4+ T cells in autoimmune infiltrates in experimental allergic encephalomyelitis.
The cytokine effector status of CD4+ T cells from lymph nodes (LN) and the central nervous system (CNS) of SJL/J mice immunized with autoantigen in adjuvant for the induction of experimental allergic encephalomyelitis (EAE) was compared. CD4+ T cells were FACS sorted based on the levels of expression of the activation marker CD45RB. Low levels of expression of this surface marker are induced by antigen recognition and are associated with 'effector' T cell function. Reverse transcriptase polymerase chain reaction (PCR) was used to analyze the expression of different T cell cytokine genes in the sorted populations. CD45RBlow cells constituted a minority of CD4+ cells in the LN and expressed elevated levels of IL-2, IFN-gamma, and IL-4 mRNA, whereas the CD45RBlow CD4+ population did not express detectable message for these cytokines under linear PCR conditions. By contrast to the LN, CD4+ cells from the CNS were predominantly CD45RBlow and expressed readily detectable levels of IL-2 and IFN-gamma mRNA, but almost no IL-4 transcription could be detected. IL-4 mRNA levels in CNS were 100- to 250-fold lower than in LN. Also, IL-4 message could not be detected in the CNS 1 week after remission. A cytokine-specific immunocytochemical single cell staining technique was used to enumerate cytokine-producing cells in LN cell populations and in CNS infiltrates. Between 1 and 5% cells in isolated LN cells produced detectable IL-2 and IFN-gamma. By contrast, the frequency of cytokine-producing cells stained in perivascular infiltrates in frozen sections from the brains of animals with active EAE was 10-fold higher.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Myocardial metabolism during increased work states in the porcine left ventricle in vivo.
It is not known whether myocardial energy requirements can be increased to the degree that they exceed myocardial O2 availability in the absence of abnormalities of coronary blood flow or coronary reserve. To determine whether this form of "demand ischemia" occurs, 10 swine were subjected to pressure overload induced by aortic constriction, inotropic and chronotropic stimulation by dobutamine, and the combination of these interventions. In an additional 9 animals, intravenous adenosine was administered during the combination of constriction and dobutamine to determine whether further increases in coronary flow could be achieved and if they would attenuate the metabolic changes. Left ventricular anterior wall transmural blood flow was measured by radioactive microspheres. Energy phosphates were assessed by 31P magnetic resonance spectroscopy using the Fourier series window technique to increase the proportion of signal derived from the subendocardium. Myocardial lactate release was quantified independent of net lactate uptake using an isotopic tracer technique. The three interventions produced 39% to 195% increases in myocardial O2 uptake from control measurements. The phosphocreatine to ATP ratio (PCr/ATP), uncorrected for partial saturation, fell significantly, from 1.39 +/- 0.10 at control conditions to 1.25 +/- 0.10 with dobutamine alone and 1.15 +/- 0.08 with dobutamine plus constriction (P < .05 for both). Myocardial lactate release rose from 0.21 +/- 0.03 mumol.g-1.min-1 at control conditions to 0.45 +/- 0.05 and 0.59 +/- 0.10 mumol.g-1.min-1, respectively (P < .05 for both), with these two interventions. Although transmurally averaged left ventricular blood flow rose from 0.97 +/- 0.09 mL.g-1.min-1 at control conditions to 3.25 +/- 0.47 mL.g-1.min-1 (P < .001) and subendocardial blood flow increased from 1.02 +/- 0.09 to 2.92 +/- 0.45 mL.g-1.min-1 (P < .001) at the highest of the three increased work states, the subendocardial to subepicardial flow ratio declined progressively from 1.13 +/- 0.08 to 0.87 +/- 0.04 (P < .05). With a further increase in aortic constriction, myocardial O2 uptake and subepicardial blood flow rose, whereas subendocardial blood flow did not change, and there was a further decline in PCr/ATP and a rise in lactate release. Although adenosine increased the average myocardial blood flow during high work state from 3.79 +/- 0.91 to 6.29 +/- 1.08 mL.g-1.min-1 (P < .001), the further rise in subendocardial flow from 3.08 +/- 0.62 to 3.78 +/- 0.68 mL.g-1.min-1 was not significant, nor were the accompanying changes in PCr/ATP or lactate metabolism.(ABSTRACT TRUNCATED AT 400 WORDS)
Adenosine↗
[MBP P89-101 induced experimental allergic encephalomyelitis in mice and establishment of CD4+ T cell line specific to MBP P89-101].
SJL/J mice were immunized with peptide 89-101 of myelin basic protein in FCA and developed EAE around 13th day after immunization. Lymph-nodes were collected at the time of onset and lymphocytes were prepared and cultured with MBP P89-101 in vitro for 3 weeks. X-ray irradiated (3000R) syngeneic spleen cells were added as antigen presenting cells. Antigen stimulated blast cells were isolated with Ficoll-Hypaque. MBP P89-101 specificity of cells were measured by 3H-TdR incorporation. MBP P89-101 activated cells were stained with PE-CD4 and analyzed by FACS. The results showed that all of the blast cells were of CD4 positive phenotype, specific to MBP P89-101.
Animals↗
The acquisition of cytokine responsiveness by murine B cells: a role for antigen and IL-5 in the induction of IL-2 receptors.
The mechanism whereby small resting (high buoyant density) murine B cells are induced to express interleukin-2 receptors (IL-2R) and to respond to IL-2 was addressed by staining with anti-IL-2R alpha and -IL-2R beta monoclonal antibodies (mAb), and using receptor-specific cDNA probes. Resting B cells expressed undetectable levels of both IL-2R alpha and beta chains on their surface and did not respond to IL-2, even at supra-physiological concentrations. Sepharose-coupled, but not streptavidin-cross-linked, plastic-adsorbed or soluble, anti-mu up-regulated the expression of IL-2R alpha and beta chains and mRNA to levels comparable to those seen in activated T cells. Anti-mu-stimulated B cells responded to IL-2 by incorporation of [3H]thymidine and high rate immunoglobulin (Ig) secretion. Both IL-5 (at optimal concentration) and suboptimal lipopolysaccharide (LPS; 20 ng/ml) induced surface expression of IL-2R alpha. The level of expression induced by IL-5 was equivalent to that on anti-Ig-activated B cells. Neither stimulus induced detectable expression of IL-2R beta, and neither induced B cells to respond to IL-2. IL-2R alpha expression was strongly enhanced, and low levels of IL-2R beta staining and mRNA were induced by the combination of LPS plus IL-5. LPS+IL-5-treated B cells responded to IL-2 by Ig secretion. This indicates that B cells regulate their responsiveness to IL-2 similarly to T cells, via the combined level of expression of IL-2R beta and IL-2R alpha. The synergy between IL-5 and LPS for B-cell responses shows a requirement for complementary stimuli such as would be provided by cytokines, and either cellular interaction or antigen recognition in regulation of B-cell responsiveness to IL-2.
Animals↗
The attraction of adhesion molecules.
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Cell Adhesion Molecules↗
Review of day case surgery.
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Ambulatory Surgical Procedures↗
Loss rather than downregulation of CD4+ T cells as a mechanism for remission from experimental allergic encephalomyelitis.
SJL/J mice recover from clinical signs of experimental allergic encephalomyelitis (EAE) 2 to 3 days following the onset of the initial attack. The immunoregulatory events that induce clinical recovery are not well understood. In this paper we have compared the activation state of the T cells infiltrating the central nervous system (CNS) in symptomatic and remitted mice. We isolated mononuclear cells from the CNS at various time points during the course of EAE and used flow cytometry to describe the kinetics of CNS infiltration by CD45+, CD2+, CD3+, TCR alpha beta+, CD4+ cells. There was a 30-fold reduction in the number of CNS CD4+ T cells in remitted mice 10 days following the initial attack. More than 60% of CNS CD4+ cells were of a CD44high, CD45RBlow memory/effector phenotype both in active EAE, peak EAE and in remission, in contrast to lymph nodes where this phenotype never constituted more than 17%. The proportion of CD8+ T cells was not increased in remitted mice, and we detected no TCR gamma delta+ cells within the CNS. Our findings demonstrate an overt loss of CD4+ T cells from the CNS and the maintenance of an activated state by T cells within the CNS and during remission from EAE. This argues against downregulation of T cell function as a mechanism for remission.
Animals↗
Enhanced response to antigen within lymph nodes of SJL/J mice that were protected against experimental allergic encephalomyelitis by T cell vaccination.
The effects of T cell vaccination on peripheral immune responsiveness are not yet fully understood. We have induced resistance to rat spinal cord homogenate (RSCH)-induced experimental allergic encephalomyelitis (EAE) in SJL/J mice by vaccination with four T cell lines (RZ8, RZ15, RZ16, and A51) which were reactive to myelin basic protein (MBP) but not to proteolipid protein (PLP). The effect was relatively neuroantigen-specific since vaccination with ovalbumin (OVA)-reactive and alloantigen-specific cells did not prevent EAE induction. Alloantigen-reactive cells reduced the rate of relapse. The number of central nervous system (CNS) infiltrates and mean clinical EAE scores were significantly reduced. This is the first report demonstrating T cell vaccination in the SJL/J mouse, a strain in which PLP is the predominant encephalitogen in RSCH. The vaccinating cells were of the memory/effector (CD44high, CD45RBlow) surface phenotype. We examined the effect of T cell vaccination on lymph node T cell proliferative responses to MBP, encephalitogenic peptides of PLP and MBP, OVA and anti-CD3. With the exception of polyclonal cytokine responses to anti-CD3, which remained unchanged, vaccination led to a 5-10-fold augmentation in all, including background, responses. By comparison with lymph node cell (LNC) responses from naive mice and mice primed with OVA, it appeared that T cell vaccination restored cellular activation levels which had been depleted in peripheral lymphoid tissues of unvaccinated animals with EAE.
Animals↗
Infusion delays through multilumen catheters using standard syringe pumps.
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Catheterization, Peripheral↗
Direct demonstration of the infiltration of murine central nervous system by Pgp-1/CD44high CD45RB(low) CD4+ T cells that induce experimental allergic encephalomyelitis.
In experimental allergic encephalomyelitis (EAE), autoimmune T cells infiltrate the central nervous system (CNS) and initiate demyelinating pathology. We have used flow cytometry to directly analyse the migration to the CNS of MBP-reactive CD4+ T cells labelled with a lipophilic fluorescent dye (PKH2), in SJL/J mice with passively transferred EAE. Labelled cells constituted about 45% of the CNS CD4+ population at the time of EAE onset. Almost all (greater than 90%) of the PKH2-labelled CD4+ T cells from EAE CNS were blasts and were alpha/beta T cell receptor (TCR)+, CD44(Pgp-1)high, and the majority were CD45RB(low). By contrast, most PKH2-labelled CD4+ T cells in lymph nodes, although CD44high, were CD45RBhigh cells. The cells that were transferred to induce EAE were essentially similar to antigen-primed lymph node cell populations, containing less than 15% CD44high cells, and most of them were CD45RBhigh. The CD44high CD45RB(low) phenotype is characteristic of memory/effector T cells that have been activated by antigen recognition. The difference in CD45RB expression between CNS and LN could therefore reflect differential exposure and/or response to antigen. Consistent with this, PKH2-labelled CD4+ cells isolated from the CNS were responsive to MBP in vitro, whereas PKH2+ CD4+ cells from lymph nodes showed almost undetectable responses. In control experiments in which ovalbumin (OVA)-reactive T cells were transferred, a small number of fluorescent-labelled CD4+ T cells were also detected in CNS, but there were very few blasts, and these remained CD45RBhigh. These results argue for induction of the memory/effector phenotype of CD4+ T cells, and their selective retention in the CNS, as a consequence of antigen recognition.
Animals↗