In-vivo-Effekte proliferationsbasierter T-Zellselektion bei allogener Knochenmarktransplantation in murinen Systemen

Für maligne Erkrankungen des hämatopoetischen Systems stellt die allogene Knochenmarktransplantation ein gut etabliertes Therapieverfahren dar. Der kurative Effekt einer solchen Behandlung liegt in der Erkennung und Zerstörung maligner Zellen durch im Transplantat enthaltene immunkompetente Zellen (Graft-versus-Leukämie (GvL)-Effekt). Hauptnebenwirkung der Knochenmarktransplantation stellt die Graft-versus-host Erkrankung (GvHD) dar, eine Immunreaktion der transplantierten Zellen gegen Empfängergewebe. Diese kann einen schweren bis tödlichen Verlauf nehmen. Im Rahmen dieser Arbeit wurde ein neues Verfahren geprüft, T-Zellen in vitro aufgrund von Proliferationsunterschieden gegenüber allogenen Stimuli zu trennen. Die Separation erfolgte mittels einer Farbstoffverdünnungsmethode („CFSE“) und anschließender FACS-Sortierung. Zytotoxizitätsmessungen zeigen, dass in vitro proliferierende Zellen („low“) im Gegensatz zur ruhenden Zellfraktion („high“) zytolytische Aktivität gegenüber Rezipientenzellen zeigen. Im Kontext allogener Knochen-marktransplantation im Mausmodell wurden die in vivo-Effekte der erhaltenen T-Zellpopulationen untersucht. Im Gegensatz zu alloreaktiven T-Zellen („low“) induzieren in vitro ruhende Zellen in zwei unterschiedlichen Transplantationsmodellen keine GvHD. Eine verbleibende Reaktivität der „high“-Zellen gegenüber unverwandten Antigenen konnte nach Reisolation demonstriert werden. Jedoch vermitteln die transplantierten „high“-Zellen – im MHC-identen Modellsystem – keine Tumorabstoßung (Verlust des GvL-Effektes). Die Gewinnung tumorreaktiver T-Zellen mittels dieses Separationsverfahrens und deren Transplantation scheinen jedoch erfolgversprechend, da durch Transfer dieser T-Zellen die Induktion einer GvHD vermieden werden kann, bei erwiesener Tumorreaktivität

Acyclovir therapy reduces the CD4+ T cell response against the immunodominant pp65 protein from cytomegalovirus in immune competent individuals

Cytomegalovirus (CMV) infects the majority of the global population and leads to the development of a strong virus-specific immune response. The CMV-specific CD4+ and CD8+ T cell immune response can comprise between 10 and 50% of the T cell pool within peripheral blood and there is concern that this may impair immunity to other pathogens. Elderly individuals with the highest magnitude of CMV-specific immune response have been demonstrated to be at increased risk of mortality and there is increasing interest in interventions that may serve to moderate this. Acyclovir is an anti-viral drug with activity against a range of herpes viruses and is used as long term treatment to suppress reactivation of herpes simplex virus. We studied the immune response to CMV in patients who were taking acyclovir to assess if therapy could be used to suppress the CMV-specific immune response. The T cell reactivity against the immunodominant late viral protein pp65 was reduced by 53% in people who were taking acyclovir. This effect was seen within one year of therapy and was observed primarily within the CD4+ response. Acyclovir treatment only modestly influenced the immune response to the IE-1 target protein. These data show that low dose acyclovir treatment has the potential to modulate components of the T cell response to CMV antigen proteins and indicate that anti-viral drugs should be further investigated as a means to reduce the magnitude of CMV-specific immune response and potentially improve overall immune function

The cellular localization of human cytomegalovirus glycoprotein expression greatly influences the frequency and functional phenotype of specific CD4+ T cell responses

CMV infection is a significant cause of morbidity and mortality in immunocompromised individuals, and the development of a vaccine is of high priority. Glycoprotein B (gB) is a leading vaccine candidate but the glycoprotein H (gH) pentameric complex is now recognized as the major target for neutralizing Abs. However, little is known about the T cell immune response against gH and glycoprotein L (gL) and this is likely to be an important attribute for vaccine immunogenicity. In this study, we examine and contrast the magnitude and phenotype of the T cell immune response against gB, gH, and gL within healthy donors. gB-specific CD4(+) T cells were found in 95% of donors, and 29 epitopes were defined with gB-specific response sizes ranging from 0.02 to 2.88% of the CD4(+) T cell pool. In contrast, only 20% of donors exhibited a T cell response against gH or gL. Additionally, gB-specific CD4(+) T cells exhibited a more cytotoxic phenotype, with high levels of granzyme B expression. Glycoproteins were effectively presented following delivery to APCs but only gB-derived epitopes were presented following endogenous synthesis. gB expression was observed exclusively within vesicular structures colocalizing with HLA-DM whereas gH was distributed evenly throughout the cytoplasm. Grafting of the C-terminal domain from gB onto gH could not transfer this pattern of presentation. These results reveal that gB is a uniquely immunogenic CMV glycoprotein and this is likely to reflect its unique pattern of endogenous Ag presentation. Consideration may be required toward mechanisms that boost cellular immunity to gH and gL within future subunit vaccines

Defective monocyte enzymatic function and an inhibitory immune phenotype in HIV-exposed uninfected African infants in the era of antiretroviral therapy

BACKGROUND: HIV-Exposed Uninfected (HEU) infants are a rapidly expanding population in sub-Saharan Africa, highly susceptible to encapsulated bacterial disease in the first year of life. The mechanism of this increased risk is still poorly understood. We investigated if HIV-exposure dysregulates HEU immunity, vaccine-antibody production and human herpes virus (HHV) amplify this effect. METHODS: 34 HIV-infected and 44 HIV-uninfected pregnant women were recruited into the birth cohort, followed up to 6 weeks of age; and 43 HIV-infected and 61 HIV-uninfected mother-infant pairs into a longitudinal infant cohort, at either: 5-7 to 14-15; or 14-15 to 18-23 weeks of age. We compared monocyte function, innate and adaptive immune cell phenotype, and vaccine-induced antibody responses between HEU and HU infants. RESULTS: We demonstrate altered monocyte phagosomal function and B cell subset homeostasis, and lower vaccine-induced anti-Haemophilus influenzae type b (Hib) and anti-Tetanus Toxoid (TT) IgG titers in HEU compared to HU infants. HHV infection was similar between HEU and HU infants. CONCLUSION: In the era of antiretroviral therapy (ART)-mediated viral suppression, HIV-exposure may dysregulate monocyte and B cell function, during the vulnerable period of immune maturation. This may contribute to the high rates of invasive bacterial disease and pneumonia in HEU infants

Asymptomatic primary infection with Epstein-Barr virus: observations on young adult cases

Epstein-Barr virus (EBV) is typically acquired asymptomatically in childhood. By contrast, infection later in life often leads to infectious mononucleosis (IM), a febrile illness characterised by anti-EBV IgM antibody-positivity, high loads of circulating latently-infected B cells, and a marked lymphocytosis caused by hyper-expansion of EBV-specific CD8+ T cells plus milder expansion of CD56dim NKG2A+ KIR– NK cells. How the two situations compare is unclear due to the paucity of studies on clinically-silent infection. Here we describe five prospectively-studied asymptomatic infections identified in a sero-epidemiological survey of University entrants. In each case the key blood sample had high cell-associated viral loads without marked IM-like CD8 lymphocytosis or NK cell disturbance. Two of the highest viral load cases showed a coincident expansion of activated EBV-specific CD8+ T cells but overall CD8+ T cell numbers were either unaffected or only mildly increased. Two slightly lower load cases, which serology suggests may have been caught earlier in the course of infection, also showed no T or NK cell expansion at the time. Interestingly, in another higher load case where T and NK cell responses were undetectable in the primary infection bleed, EBV-specific T cell responses did not appear until several months later, by which time virus loads in the blood had already fallen. Thus some asymptomatic primary infections have very high circulating viral loads and a cell-mediated immune response that is qualitatively similar to IM but of lower magnitude. However, others may be quite different and ultimately could reveal novel mechanisms of host control

Defective Monocyte Enzymatic Function and an Inhibitory Immune Phenotype in Human Immunodeficiency Virus-Exposed Uninfected African Infants in the Era of Antiretroviral Therapy

Abstract Background Human immunodeficiency virus-exposed uninfected (HEU) infants are a rapidly expanding population in sub-Saharan Africa and are highly susceptible to encapsulated bacterial disease in the first year of life. The mechanism of this increased risk is still poorly understood. We investigated whether human immunodeficiency virus (HIV)-exposure dysregulates HEU immunity, vaccine-antibody production, and human herpes virus amplify this effect. Methods Thirty-four HIV-infected and 44 HIV-uninfected pregnant women were recruited into the birth cohort and observed up to 6 weeks of age; and then a subsequent 43 HIV-infected and 61 HIV-uninfected mother-infant pairs were recruited into a longitudinal infant cohort at either: 5–7 to 14–15; or 14–15 to 18–23 weeks of age. We compared monocyte function, innate and adaptive immune cell phenotype, and vaccine-induced antibody responses between HEU and HIV-unexposed uninfected (HU) infants. Results We demonstrate (1) altered monocyte phagosomal function and B-cell subset homeostasis and (2) lower vaccine-induced anti-Haemophilus influenzae type b (Hib) and anti-tetanus toxoid immunoglobulin G titers in HEU compared with HU infants. Human herpes virus infection was similar between HEU and HU infants. Conclusions In the era of antiretroviral therapy-mediated viral suppression, HIV exposure may dysregulate monocyte and B-cell function, during the vulnerable period of immune maturation. This may contribute to the high rates of invasive bacterial disease and pneumonia in HEU infants. </jats:sec

Report from the second cytomegalovirus and immunosenescence workshop.

The Second International Workshop on CMV & Immunosenescence was held in Cambridge, UK, 2-4th December, 2010. The presentations covered four separate sessions: cytomegalovirus and T cell phenotypes; T cell memory frequency, inflation and immunosenescence; cytomegalovirus in aging, mortality and disease states; and the immunobiology of cytomegalovirus-specific T cells and effects of the virus on vaccination. This commentary summarizes the major findings of these presentations and references subsequently published work from the presenter laboratory where appropriate and draws together major themes that were subsequently discussed along with new areas of interest that were highlighted by this discussion.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

The exponent of Hölder calmness for polynomial systems

Diese Arbeit befasst sich mit Untersuchung der Hölder Calmness, eines Stabilitätskonzeptes das man als Verallgemeinerung des Begriffs der Calmness erhält. Ausgehend von Charakterisierungen dieser Eigenschaft für Niveaumengen von Funktionen, werden, unter der Voraussetzung der Hölder Calmness, Prozeduren zur Bestimmung von Elementen dieser Mengen analysiert. Ebenso werden hinreichende Bedingungen für Hölder Calmness studiert. Da Hölder Calmness (nichtleerer) Lösungsmengen endlicher Ungleichungssysteme mittels (lokaler) Fehlerabschätzungen beschrieben werden kann, werden auch Erweiterungen der lokalen zu globalen Ergebnissen diskutiert. Als Anwendung betrachten wir speziell den Fall von Niveaumengen von Polynomen bzw. allgemeine Lösungsmengen polynomialer Gleichungen und Ungleichungen. Eine konkrete Frage, die wir beantworten wollen, ist die nach dem Zusammenhang zwischen dem größten Grad der beteiligten Polynome sowie dem Typ, d.h. dem auftretenden Exponenten, der Hölder Calmness des entsprechenden Systems.This thesis is concerned with an analysis of Hölder calmness, a stability property derived from the concept of calmness. On the basis of its characterization for (sub)level sets, we will cogitate about procedures to determine points in such sets under a Hölder calmness assumption. Also sufficient conditions for Hölder calmness of (sub)level sets and of inequality systems will be given and examined. Further, since Hölder calmness of (nonempty) solution sets of finite inequality systems may be described in terms of (local) error bounds, we will as well amplify the local propositions to global ones. As an application we investigate the case of (sub)level sets of polynomials and of general solution sets of polynomial equations and inequalities. A concrete question we want to answer here is, in which way the maximal degree of the involved polynomials is connected to the exponent of Hölder calmness or of the error bound for the system in question

Antiviral therapy can reverse the development of immune senescence in elderly mice with latent cytomegalovirus infection

Cytomegalovirus (CMV) infection leads to the development of adaptive and humoral immune responses that are among the largest for any pathogen, and intriguingly, the magnitude of the immune response increases with age, a phenomenon termed “memory inflation.” Elevated CMV-specific immunity has been correlated with an increased mortality rate in elderly individuals and with impaired vaccination responses. The latent phase of CMV infection is characterized by intermittent episodes of subclinical viral reactivation and the production of immunogenic transcripts that may maintain memory inflation of virus-specific cytotoxic lymphocytes. However, the relative importance of CMV reactivation in the development of memory inflation is uncertain, as is the potential for antiviral treatment to reverse this effect. Here, we administered valaciclovir for up to 12 months in mice with established murine CMV (MCMV) infection. Treatment reduced the magnitude of the MCMV-specific CD8(+) T-lymphocyte response by 80%, and the residual MCMV tetramer-specific lymphocytes exhibited a less differentiated phenotype. In addition, latent MCMV infection suppressed the proportion of naïve CD8(+) T cells by 60% compared to antiviral-treated mice or MCMV-negative animals. Furthermore, treatment led to a reduction in influenza A viral loads following a challenge in elderly MCMV-infected animals and also reduced the differentiation of influenza virus-specific cytotoxic lymphocytes. These observations demonstrate that MCMV-specific memory inflation is maintained by viral replication and that therapeutic intervention could lead to improved immune function