Thesis (M.S.P.H.)--University of Michigan, 1940.

Thesis (M.S.P.H.)--University of Michigan, 1940.

No description available.

Thesis (doctor of medicine)--Rijksuniversiteit te Groningen, 1979.

Epstein-Barr virus (EBV) induced infectious mononucleosis (IM) is characterized by the activation and expansion of T lymphocytes and the induction of cytotoxic responses able to mediate the lysis of EBV-uninfected, allogeneic MHC mismatched and EBV-infected autologous target cells. Freshly isolated peripheral blood mononuclear cells (PBMC) were used to examine the nature of these cellular immune responses. Activated lymphocytes, as identified by HLA-DR expression, associated with EBV induced IM were shown to be a heterogeneous population containing significantly elevated cytotoxic/suppressor (CD8+) T cells, helper/inducer (CD4+) T cells and natural killer (NK, CD16+) cells. CD8+ T cells were the primary activated population, representing 24% of the total lymphocyte population and 60-70% of the CD8+ T cell population. The activated CD4+ T cells and natural killer (NK) cells accounted for 7% and 4% of the total lymphocyte population, respectively. Analysis of serum soluble interleukin 2 receptors (IL-2R) and CD8 molecules demonstrated significantly (p<0.001) elevated levels in the sera of IM patients compared with normal controls. These elevated levels of serum IL-2R am CD8 molecules correlated, (r=0. 67 and r=0.82, respectively) with increased percentages of CD8/HLA-DR positive T cells (i.e., activated CD8 T cells). Increased levels of soluble cell surface molecules peaked during the acute phase and normalized as the patients progressed toward convalescence. Individual patients demonstrated strong correlations between the percentage of CD8/HLA-DR positive cells and soluble CD8 levels. The functional significance of the serum IL-2R and CD8 molecules is presently unknown. However, the strong correlative data between serum CD8, and to a lesser extent IL-2R, and CD8 T cell activation suggests that serum CD8 levels may provide a sensitive measure of CD8 T cell activation in systemic infections. The ability of freshly isolated acute IM PBMC to lyse allogeneic, EBV-infected lymphoblastoid cell lines (LCL), demonstrated the ability of acute IM effector cells to lyse MHC mismatched target cells. Effector cells from acute IM patients lysed allogeneic DM-LCL and AF-LCL target cells by 34% (n=7) and 23% (n=6), respectively, compared with 4% (n=5) and 0% (n=5), respectively, for normal controls. MAb-dependent complement depletion of CD3+ or CD8+ cells with anti-CD3 and anti-CD8 mAb decreased the non-MHC restricted cytolysis of LCL by 96% and 89%, respectively. In contrast, complement depletion with NK-cell specific mAbs Leu 11b and NKH-1, resulted in only a slight decrease (<35%) in the lysis of these LCL (46%). Depletion with anti-HLA-DR also significantly (p<0.001) decreased the lysis of LCL. Depletions with anti-CD4 demonstrated no decrease in LCL-lysis. MAbs OKT3 and OKT8 inhibited the non-MHC restricted cytolysis by 87% and 82%, respectively. We interpret these results as evidence that, 1) lysis of allogeneic cells is mediated primarily by CD3+, CD8+, HLA-DR+, cytotoxic T lymphocytes (CTL); and 2) these acute IM cytotoxic T cells utilize the T cell receptor and the CD8 antigen as an accessory molecule. An active role for target cell MHC class I molecules in the recognition and subsequent lysis of target cells is supported by a number of observations: 1) the MHC class I reactive mAbs W6/32 and BBM.1 significantly (p<0.05) inhibited the lysis of 63463-LCL by 65% and 57%, respectively; 2) acute IM effector T cells did not lyse the MHC class I negative Daudi cell line; 3) allogeneic MHC class I matched LCL mediated strong competitive inhibition (72% at 10:1 competitor to target cell ratio) vs 29% competitive inhibition for an allogeneic MHC class I mismatched LCL; and 4) NK-cell depleted effector cells from one patient mediated preferential lysis of the K562 cell line expressing MHC class I. HLA-A2 molecules. We interpret these results as evidence that target cell MHC class I molecules (or associated determinants) are the target antigen(s) for the allogeneic MHC cytotoxic response. The role of EBV in this acute allogeneic response was examined using target cell lines devoid of EBV genome. Acute IM CTL mediated lysis of the allogeneic HSB-2 T cell line (45%), and allogeneic HTLV-I transformed T cell lines (16%). The lysis of the HSB-2 T cell line was inhibited by anti-OKT3 (58% inhibition), W6/32 (53%) and BBM.1 (42%). Similarily, lysis of HTLV-I T cell lines was inhibited by W6/32 (69% inhibition), BBM.1 (69%) and OKT3 (38%). These data demonstrate that EBV antigenic expression is not required for allogeneic recognition and subsequent lysis of these allogeneic target cells. Effector cells from acute IM patients (n=5) were able to lyse their autologous EBV-infected LCL (mean lysis=21%), but were unable to lyse the EBV-uninfected autologous HTLV-I T cell line. These same effectors, however, were able to mediate lysis of both allogeneic B cell lines (21% lysis) and allogeneic T cell lines (8% lysis). These data are consistent with the observations by Strang et al. (1987a), who recently cloned virus specific/MHC-restricted CTL cloned from acute IM PBMC. These virus specific/MHC-restricted T cells presumably mediate the lysis of the autologous EBV-transformed B cell lines but not the autologous EBV-uninfected T cell lines. Whether the CTL which lyse the autologous EBV-transformed LCL are also responsible for the observed allogeneic reactivity was examined with cold target competition using autologous and allogeneic LCL. Lysis of autologous LCL was inhibited only by autologous competitor cells (64% inhibition compared with 24% for allogeneic LCL). Likewise, lysis of the allogeneic LCL was inhibited only by the allogeneic competitor cells (85% inhibition compared with 30% for autologous LCL). These data demonstrated no competition between allogeneic and autologous LCL and therefore support the concept that lysis of autologous LCL and allogeneic target cells is mediated by distinct effector populations. These data help us to understand the unusual immune response observed during acute IM. The strong allogeneic cytotoxic response is thought to represent polyclonal CD8 T cell activities induced by EBV-infected and transformed B cells which circulate in vivo. In addition, a population of CD8 CTL exist which mediate the lysis of autologous EBV-transformed B cells. These CTL likely represent virus-specific/MHC-restricted CTL and presumably play a major role in the control of EBV infections. The role, if any, of the markedly expanded alloreactive CTL population in the elimination of EBV infected and transformed B cells remains to be clarified.

The increasing popularity of traditional Chinese medicine (TCM) therapies as health care options warrants thorough examination of the efficacy and safety evidence around these therapies. This thesis explores the intersection of TCM and fatigue using two rigorous methodologies: systematic reviews (SRs) and a randomized controlled clinical trial (RCT). In order to inform the development of an RCT of acupuncture for infectious mononucleosis (mono), a common condition with no known cure, characterized by profound fatigue, we examined the literature on TCM treatment of mono as well as another fatigue condition, idiopathic chronic fatigue (ICF). Additionally, we investigated the literature on the safety of pediatric acupuncture. SRs of TCM efficacy in mono and ICF determined that although clinical trials exist, methodological flaws compromised their validity. In particular, studies published as RCTs were found to lack proper randomization. Inclusion of these studies in the SRs would have been inappropriate and demonstrates the importance of verifying RCT methods. We also present the results of the first known SR of pediatric acupuncture safety. This review was comprehensive, including a large number of databases and publications in any language. Synthesis of the results from those studies that included a denominator produced mild adverse event estimates of 16.3/100 (95% CI 11.221.5) per patient, for RCTs, and 6.3/100 (95% CI 4.97.7) per patient, for cohort studies, with a combined estimate of 7.8/100 (95% CI 6.49.2) per patient. We developed and conducted an RCT of acupuncture for mono, although limitations to recruitment resulted in the enrolment of only three participants. The primary result of the small sample size was to restrict the determination of treatment effect, however, successful implementation of other elements is informative to further research in this area. In addition, we determined the local 5-year Monospot positive incidence rate to be 1.11/1000 (95% CI 0.953.2) for all ages and 5.46/1000 (95% CI 0.8910.0) for the 15-25 year old group. This dissertation examined the evidence around TCM and fatigue and provides recommendations that are aimed at increasing the value of research and the safety and efficacy of practice in this area.

No description available.

Epstein-Barr virus (EBV) is a ubiquitous herpesvirus that establishes a life-long latent infection of B cells. It is usually asymptomatic in healthy individuals; however, individuals with compromised immunity often develop EBV-induced lymphoma. EBV also encodes potential oncogenes that can contribute to tumorigenesis. Therefore, vaccine and immunotherapeutic strategies targeting EBV are desirable. Recent studies have shown that infusion of EBV-specific CD8+T cells can elicit remission of lymphomas arising after administration of immunosuppressive drugs during transplantation, suggesting an important role for T cells in the prevention of EBV-induced malignancy. A better understanding of the cellular immune components involved in the control of EBV will aid in the development of methods to prevent infection and/or treat EBV-associated disease. While EBV infection is usually acquired asymptomatically during childhood, primary infection of adolescents and young adults can result in an illness termed acute infectious mononucleosis (AIM). Because of the characteristic symptoms of the illness, individuals with AIM can be readily identified and diagnosed with acute EBV infection. Thus, primary CD4+ and CD8+ T cell responses against the virus can be evaluated. It has been previously found that there is a marked expansion of lytic EBV protein-specific CD8+ T cells early during AIM, with delayed detection of lower frequencies of latent EBV protein-specific CD8+ T cells. The magnitude and specificity of CD4+T cell responses during AIM has been less well characterized. This thesis dissertation presents data from both functional assays and direct staining experiments documenting the timing, magnitude, and antigen-specificity of CD4+ T cells over the course of primary EBV infection. Lytic and latent protein-specific CD4+ T cells were readily detected by intracellular IFN-γ production at presentation with AIM and declined rapidly thereafter. Blood EBV load was also quantitated and found to decrease over time following AIM. By contrast, CD8+T cell IFN-y responses remained high for several weeks following presentation with AIM. Direct staining of lytic epitope-specific CD4+ T cells during AIM revealed high frequencies of virus-specific cells with low proliferative and IFN-γ-producing potential. Blood EBV load in these patients was persistently high through 6 wk following AIM. These data suggest a relationship between high EBV load during acute infection and impaired EBV-specific CD4+ T cell responses, which are compatible with impaired CD4+ T cell responses reported during high viremia associated with other viral infections. This may represent a mechanism by which persistent viruses, such as EBV, are able to establish a life-long infection in their hosts.

Epstein-Barr Virus (EBV) is a ubiquitous human virus linked to several diseases, including cancers. CD8 T cells are important for controlling EBV replication. Generation and maintenance of virus-specific CD8 T cells is dependent on specific interaction between MHC-peptide complexes on the infected cell and the CD8 T cell receptor (TCR). Several lines of evidence suggest that the TCR repertoire is an essential component of the CD8 T-cell immune response. The current work focuses on delineating the features of the TCR repertoire that drive the selection of EBV-specific CD8 T cells into the memory phase. We used bulk and single-cell TCRαβ sequencing to analyze the TCR repertoire of human CD8 T cells specific for two immunodominant HLA-A02:01-restricted EBV-derived epitopes: BRLF1109-117 (YVLDHLIVV) and BMLF1280-288 (GLCTLVAML) during the acute and memory phases of primary EBV infection in humans. We showed that persistent EBV-specific clonotypes accounted for only 9% of unique clonotypes but were highly expanded in acute EBV infection and more commonly expressed identifiable features than non-persistent clonotypes. The other 91% of highly diverse unique clonotypes disappeared and were replaced in convalescence by equally diverse “de-novo” clonotypes. We provide evidence suggesting that recognition of BRLF1109-117may be driven by the TCRα. We identified a highly dominant and degenerate BRLF1109-117-specific TCRα sequence, AV8.1-CAVKDTDKLIF-AJ34, that was shared by all donors studied and identified conserved residues within this sequence that were important for antigen recognition. These findings are relevant to current efforts to develop or optimize the efficacy of T cell based therapies or vaccines.