Lymphocyte Turnover Research Articles

P.403: Donor-recipient lymphocyte turnover kinetics is affected by immunosuppressive treatment in a model of intestinal transplantation acute cellular rejection.

P.403: Donor-recipient lymphocyte turnover kinetics is affected by immunosuppressive treatment in a model of intestinal transplantation acute cellular rejection.

Lymphopenia and high Ki-67 expression in peripheral blood CD4+ and CD8+ T cells associate with progressive sarcoidosis

BackgroundEarly identification of patients at risk for progressive sarcoidosis may improve intervention. High bronchoalveolar lavage fluid (BALF) lymphocytes and peripheral blood (PB) lymphopenia are associated with worse prognosis. The mechanisms...

The effects of repeated brain MRI on chromosomal damage

BackgroundMagnetic resonance imaging (MRI) is currently considered a safe imaging technique because, unlike computed tomography, MRI does not expose patients to ionising radiation. However, conflicting literature reports possible genotoxic effects of MRI. We herein examine the chromosomal effects of repeated MRI scans by performing a longitudinal follow-up of chromosomal integrity in volunteers.MethodsThis ethically approved study was performed on 13 healthy volunteers (mean age 33 years) exposed to up to 26 3-T MRI sessions. The characterisation of chromosome damage in peripheral blood lymphocytes was performed using the gold-standard biodosimetry technique augmented with telomere and centromere staining.ResultsCytogenetic analysis showed no detectable effect after a single MRI scan. However, repeated MRI sessions (from 10 to 20 scans) were associated with a small but significant increase in chromosomal breaks with the accumulation of cells with chromosomal terminal deletions with a coefficient of 9.5% (95% confidence interval 6.5–12.5%) per MRI (p < 0.001). Additional exposure did not result in any further increase. This plateauing of damage suggests lymphocyte turnover. Additionally, there was no significant induction of dicentric chromosomes, in contrast to what is observed following exposure to ionising radiation.ConclusionsOur study showed that MRI can affect chromosomal integrity. However, the amount of damage per cell might be so low that no chromosomal rearrangement by fusion of two deoxyribonucleic breaks is induced, unlike that seen after exposure to computed tomography. This study confirms that MRI is a safe imaging technique.

Resolving estimates of in vivo lymphocyte turnover from deuterated-glucose and heavy water in a murine model

Resolving estimates of in vivo lymphocyte turnover from deuterated-glucose and heavy water in a murine model

Promoter Targeting RNAs: Unexpected Contributors to the Control of HIV-1 Transcription

In spite of prolonged and intensive treatment with combined antiretroviral therapy (cART), which efficiently suppresses plasma viremia, the integrated provirus of HIV-1 persists in resting memory CD4+ T cells as latent infection. Treatment with cART does not substantially reduce the burden of latent infection. Once cART is ceased, HIV-1 replication recrudesces from these reservoirs in the overwhelming majority of patients. There is increasing evidence supporting a role for noncoding RNAs (ncRNA), including microRNAs (miRNAs), antisense (as)RNAs, and short interfering (si)RNA in the regulation of HIV-1 transcription. This appears to be mediated by interaction with the HIV-1 promoter region. Viral miRNAs have the potential to act as positive or negative regulators of HIV transcription. Moreover, inhibition of virally encoded long-asRNA can induce positive transcriptional regulation, while antisense strands of siRNA targeting the NF-κB region suppress viral transcription. An in-depth understanding of the interaction between ncRNAs and the HIV-1 U3 promoter region may lead to new approaches for the control of HIV reservoirs. This review focuses on promoter associated ncRNAs, with particular emphasis on their role in determining whether HIV-1 establishes active or latent infection.

Recipient Cell Turnover of Gut-Resident Lymphocytes in Intestinal Allografts - Association of Delayed Turnover With Non-Rejecting Allografts.

Recipient Cell Turnover of Gut-Resident Lymphocytes in Intestinal Allografts - Association of Delayed Turnover With Non-Rejecting Allografts.

Immunotoxic effects of an industrial waste incineration site on groundwater in rainbow trout (Oncorhynchus mykiss)

The discharge of organic waste from the petrochemical industry into the Mercier lagoons caused major groundwater contamination. The objective of this study was to determine the immunotoxic potential of three groundwater wells at increasing distance from the incinerator dumping site (1.17, 2.74 and 5.40 km). Rainbow Trout were exposed to increasing concentrations of water from three groundwater wells for 14 days. Immunocompetence was characterized by phagocytosis, mitogen-stimulated proliferation of lymphocytes, cell cycle analysis and apoptosis. A significant increase in innate (phagocytosis) and specific immune response (B lymphocyte proliferation) was observed in trout exposed to water collected from the well at 2.74 km. However, phagocytosis activity was suppressed in groups at 1.17 and 5.40 km. The proportion of lymphocytes in S phase was significantly increased in groups at 2.74 and 5.40 km, while lymphocytes in G0/G1 phase were decreased in all three exposure groups. Additionally, dexamethasone (DEX)-induced apoptosis of lymphocytes was significantly reduced in the group at 2.74 km, which suggests decreased lymphocyte turnover. Furthermore, the ratio of DEX-induced apoptosis/apoptosis was lower in the groups at 2.74 and 5.40 km. In summary, our experiments have shown that exposure to the mixture of organic compounds present in Mercier groundwater modulates phagocytosis and cell proliferation, disrupts the cell cycle and reduces the ratio of DEX-induced apoptosis/apoptosis. It is concluded that groundwater collected in the vicinity of an incinerator containment field could impact immunocompetence in fish.

Hypertrophy of infected Peyer's patches arises from global, interferon-receptor, and CD69-independent shutdown of lymphocyte egress.

Lymphoid organ hypertrophy is a hallmark of localized infection. During the inflammatory response, massive changes in lymphocyte recirculation and turnover boost lymphoid organ cellularity. Intriguingly, the exact nature of these changes remains undefined to date. Here, we report that hypertrophy of Salmonella-infected Peyer's patches (PPs) ensues from a global “shutdown” of lymphocyte egress, which traps recirculating lymphocytes in PPs. Surprisingly, infection-induced lymphocyte sequestration did not require previously proposed mediators of lymphoid organ shutdown including type I interferon receptor and CD69. In contrast, following T-cell receptor-mediated priming, CD69 was essential to selectively block CD4+ effector T-cell egress. Our findings segregate two distinct lymphocyte sequestration mechanisms, which differentially rely on intrinsic modulation of lymphocyte egress capacity and inflammation-induced changes in the lymphoid organ environment.

Human T cell lifespan (P1284)

Abstract In humans, the lifespan and turnover of T cells have been debated for almost half a century. Until now, the analysis of T lymphocyte turnover have been primarily approached through short term labelling studies and nucleotide analogue incorporation. While much has been gained from these endeavours, there is still considerable debate about the half live of the different T lymphocyte subsets and it is apparent that neither approach alone can give the necessary information to reach a conclusive result. Taking advantage of the decreasing concentration of 14C in the atmosphere since the cold war, we have retrospectively assessed the birth date of the different T cell populations in the blood of healthy adult humans.

Divergent Kinetics of Proliferating T Cell Subsets in Simian Immunodeficiency Virus (SIV) Infection: SIV Eliminates the “First Responder” CD4 + T Cells in Primary Infection

Although increased lymphocyte turnover in chronic human immunodeficiency virus and simian immunodeficiency virus (SIV) infection has been reported in blood, there is little information on cell turnover in tissues, particularly in primary SIV infection. Here we examined the levels of proliferating T cell subsets in mucosal and peripheral lymphoid tissues of adult macaques throughout SIV infection. To specifically label cells in S-phase division, all animals were inoculated with bromodeoxyuridine 24 h prior to sampling. In healthy macaques, the highest levels of proliferating CD4(+) and CD8(+) T cells were in blood and, to a lesser extent, in spleen. Substantial percentages of proliferating cells were also found in intestinal tissues, including the jejunum, ileum, and colon, but very few proliferating cells were detected in lymph nodes (axillary and mesenteric). Moreover, essentially all proliferating T cells in uninfected animals coexpressed CD95 and many coexpressed CCR5 in the tissues examined. Confocal microscopy also demonstrated that proliferating cells were substantial viral target cells for SIV infection and viral replication. After acute SIV infection, percentages of proliferating CD4(+) and CD8(+) T cells were significantly higher in tissues of chronically infected macaques and macaques with AIDS than in those of the controls. Surprisingly, however, we found that proliferating CD4(+) T cells were selectively decreased in very early infection (8 to 10 days postinoculation [dpi]). In contrast, levels of proliferating CD8(+) T cells rapidly increased after SIV infection, peaked by 13 to 21 dpi, and thereafter remained significantly higher than those in the controls. Taken together, these findings suggest that SIV selectively infects and destroys dividing, nonspecific CD4(+) T cells in acute infection, resulting in homeostatic changes and perhaps continuing loss of replication capacity to respond to nonspecific and, later, SIV-specific antigens.

Genetic Labeling Reveals Altered Turnover and Stability of Innate Lymphocytes in Latent Mouse Cytomegalovirus Infection

Mouse CMV (MCMV) infection rapidly induces the proliferation of NK cells, which correlates with immunological protection. Whether NK cells primed during acute response against MCMV are maintained for the long term is not known. In this study, we used TcrdH2BeGFP mice in which maturing NK cells are genetically labeled with a pulse of very stable histone-2B-eGFP. In this system, we found that the reporter protein was diluted out upon NK cell division during acute MCMV infection. At the same time, mature NK cells in uninfected mice showed only very limited turnover in vivo. Three months after primary infection when MCMV latency was established, the majority of peripheral NK cells still displayed a higher record of proliferation than NK cells in mock-infected controls. This observation included both Ly49H(+) and Ly49H(-) NK cells. Conversely, naive NK cells did not show more proliferation after transfer into latently MCMV-infected mice than that after transfer into mock-infected control mice. This indicated that the observed alterations of the NK cell compartment in MCMV latency were "legacy" (i.e., resulting from prior events during the initial immune response). Together, these results suggest that antiviral immune responses induce sustained alterations of innate lymphocyte populations that extend far beyond the first days of acute infection.

191 Anti-FasL treatment preserves SIV-specific memory and slows progression to AIDS in rhesus macaques

Previously we reported an increased survival advantage in SIV-infected rhesus macaques after injection with a FasL-blocking monoclonal antibody (Salvato et al., Clin Dev Immunol. 2007: 93462). In the present study we analyzed the effect of such FasL blocking on immune parameters and disease progression in a cohort of SIV-infected rhesus macaques. Upon infection, various immune parameters in the blood developed highly significant differences between animals given anti-FasL RNOK203 antibody and those given an isotype control antibody. The anti-FasL treated animals had a clear preservation of central memory CD4 lymphocytes starting from acute infection through at least set point viremia. The total memory CD4 lymphocytes in these animals were also significantly higher throughout the 25-week study period. Anti-FasL treatment resulted in significantly higher percentages of memory CD8 lymphocytes in peripheral blood. Besides these changes in the memory lymphocyte compartment, the treated animals had a higher response to infection as evidenced by increased turnover of CD4 and CD8 lymphocytes expressing Ki67. Importantly, the antigen-specific cellular immune responses were significantly higher at 4, 6 and 17 weeks after infection in anti-FasL treated animals. Preservation of memory lymphocytes and cellular immunity are considered the main correlates of protection in SIV infection. Our study demonstrates that FasL is a major contributor towards depletion of anti-SIV immunity and that blocking this pathway is a beneficial strategy that should be considered for vaccine formulations.

Heightened T-cell proliferation without an elevation of CD4 + T cell spontaneous apoptosis in AIDS patients

T lymphocyte turnover has been studied extensively in HIV infection. The dynamic characteristics of various subsets of T cells in antiretroviral-naive, HIV-1-infected individuals, however, have not been well defined. Here, we performed a cross-sectional study using peripheral blood T cells from 39 antiretroviral-naive, chronically HIV-infected patients, as well as 16 healthy, HIV-negative controls. T-cell subset turnover rates were measured by Ki-67 antigen staining; levels of spontaneous apoptosis and activation in T-cell subsets were also determined by flow cytometry. Surprisingly, with disease progression, the level of T-cell spontaneous apoptosis did not increase significantly, despite a heightened rate of T-cell subset turnover and increased expression of the CD38 activation marker. These data refute the idea that increased T cell turnover is merely a homeostatic process in response to CD4 T cell loss during HIV disease progression, and suggest that future mechanistic studies may be needed for a comprehensive understanding of T-cell dynamics during HIV infection. Such understanding may help to develop new strategies for the immune modulation of clinical disease.

Viral reservoirs, residual viremia, and the potential of highly active antiretroviral therapy to eradicate HIV infection

Although highly active antiretroviral therapy (HAART) can reduce HIV-1 viremia to levels that are below the limit of detection of clinical assays, the virus persists in reservoirs, and trace levels of free virions can be found in the plasma. Whether this residual viremia represents ongoing cycles of replication continuing despite HAART or simply the release of virus from stable reservoirs has been controversial. Here we summarize the evidence that HAART can stop ongoing cycles of replication. The evidence comes from a detailed analysis of the residual viremia, which shows it to be archival and nonevolving in character. In addition, new pharmacodynamic measures incorporating a previously ignored slope parameter have provided the first real indication of how well HAART actually suppresses viral replication in vivo. Together, these results argue that the ultimate theoretical potential of HAART to control viral replication has already been reached. Progress toward eradication of the infection will require novel approaches to target the stable reservoirs that persist even when viral replication is completely halted.

Dynamics of T- and B-Lymphocyte Turnover in a Natural Host of Simian Immunodeficiency Virus

Increased lymphocyte turnover is a hallmark of pathogenic lentiviral infection. To investigate perturbations in lymphocyte dynamics in natural hosts with nonpathogenic simian immunodeficiency virus (SIV) infection, the nucleoside analog bromodeoxyuridine (BrdU) was administered to six naturally SIV-infected and five SIV-negative sooty mangabeys. As a measure of lymphocyte turnover, we estimated the mean death rate by fitting a mathematical model to the fraction of BrdU-labeled cells during a 2-week labeling and a median 10-week delabeling period. Despite significantly lower total T- and B-lymphocyte counts in SIV-infected sooty mangabeys than in SIV-negative mangabeys, the turnover rate of B lymphocytes and CD4(+) and CD8(+) T lymphocytes was not increased in the SIV-infected animals. A small, rapidly proliferating CD45RA(+) memory subset and a large, slower-proliferating CD45RA(-) central memory subset of CD4(+) T lymphocytes identified in the peripheral blood of sooty mangabeys also did not show evidence of increased turnover in the context of SIV infection. Independently of SIV infection, the turnover of CD4(+) T lymphocytes in sooty mangabeys was significantly higher (P < 0.01) than that of CD8(+) T lymphocytes, a finding hitherto not reported in rhesus macaques or humans. The absence of aberrant T-lymphocyte turnover along with an inherently high rate of CD4(+) T-lymphocyte turnover may help to preserve the pool of central memory CD4(+) T lymphocytes in viremic SIV-infected sooty mangabeys and protect against progression to AIDS.

Intestinal Lymphocyte Subsets and Turnover Are Affected by Chronic Alcohol Consumption

We recently demonstrated that simian immunodeficiency virus (SIV) viral loads were significantly higher in the plasma of rhesus macaques consuming alcohol compared with controls following intrarectal SIV infection. To understand the possible reasons behind increased viral replication, here we assessed the effects of chronic alcohol consumption on distribution and cycling of various lymphocyte subsets in the intestine. Macaques were administered alcohol (n = 11) or sucrose (n = 12), and percentages of memory and naive and activated lymphocyte subsets were compared in the blood, lymph nodes, and intestines. Although minimal differences were detected in blood or lymph nodes, there were significantly higher percentages of central memory (CD95+CD28+) CD4+ lymphocytes in the intestines from alcohol-receiving animals before infection compared with controls. In addition, higher percentages of naive (CD45RA+CD95-) as well as CXCR4+CD4 cells were detected in intestines of alcohol-treated macaques. Moreover, alcohol consumption resulted in significantly lower percentages of effector memory (CD95+CD28-) CD8 lymphocytes as well as activated Ki67+CD8 cells in the intestines. A subset (7 receiving alcohol and 8 receiving sucrose) were then intrarectally inoculated with SIV(mac251). Viral RNA was compared in different tissues using real-time PCR and in situ hybridization. Higher levels of SIV replication were observed in tissues from alcohol-consuming macaques compared with controls. Central memory CD4 lymphocytes were significantly depleted in intestines and mesenteric lymph nodes from all alcohol animals at 8 weeks postinfection. Thus, changes in the mucosal immune compartment (intestines) in response to alcohol are likely the major reasons behind higher replication of SIV observed in these animals.

Marked telomere fluctuation of leukocytes during graft-versus-host disease in allogeneic stem cell transplantation

Immune dysfunction after allogeneic stem cell transplantation (SCT) is closely associated with cell turnover of lymphocytes and homeostasis of hematopoietic stem cells. Telomeres, repetitive sequences (TTAGGG)n on the end of linear chromosomes, reflect the mitotic history of stem cells. Using telomere fluorescence in situ hybridization (FISH) and flow cytometry (flow-FISH), we measured telomere length in lymphocytes and neutrophils at various intervals to analyze the relationship between telomere length change and clinical features in 5 patients who underwent allogeneic bone marrow transplantation. During the first year after allogeneic stem cell transplantation, a marked fluctuation of telomere length in peripheral blood leukocytes was observed in all recipients, and in 3 patients there was a reduction of telomere length during chronic graft-versus-host disease (GvHD) or during post-transplant lymphoproliferative disorder. The reduction of telomere length during GvHD was evident in lymphocytes and neutrophils, but telomere length in neutrophils tended to recover earlier than that observed in lymphocytes. The rapid reduction of telomere length in leukocytes during GvHD was too extensive to be explained by the end-replication problem, suggesting the presence of a telomerically unstable hematopoietic condition after transplant in vivo.

Cellular immune response to HTLV-1

There is strong evidence at the individual level and the population level that an efficient cytotoxic T lymphocyte (CTL) response to HTLV-1 limits the proviral load and the risk of associated inflammatory diseases such as HAM/TSP. This evidence comes from host population genetics, viral genetics, DNA expression microarrays and assays of lymphocyte function. However, until now there has been no satisfactory and rigorous means to define or to measure the efficiency of an antiviral CTL response. Recently, methods have been developed to quantify lymphocyte turnover rates in vivo and the efficiency of anti-HTLV-1 CTLs ex vivo. Data from these new techniques appear to substantiate the conclusion that variation between individual hosts in the rate at which a single CTL kills HTLV-1-infected lymphocytes is an important determinant, perhaps the decisive determinant, of the proviral load and the risk of HAM/TSP. With these experimental data, it is becoming possible to refine, parameterize and test mathematical models of the immune control of HTLV-1, which are a necessary part of an understanding of this complex dynamic system.

Lymphocyte homeostasis and the antitumor immune response

Lymphocyte homeostasis is regulated by proliferation of antigen-responsive T-cells in the peripheral circulation and their apoptosis. Patients with cancer have altered lymphocyte homeostasis. Spontaneous apoptosis of circulating CD8+ antigen-responding effector T-cells contributes to rapid lymphocyte turnover and depressed absolute numbers of T-cell subsets observed in patients with cancer. A rapid transit of naive CD8+ T-cells to the expanded memory pool and enhanced apoptosis of antitumor effector T-cells in the peripheral circulation of patients with cancer are partly responsible for this rapid lymphocyte turnover. Future strategies for restoration of normal lymphocyte homeostasis in cancer will involve therapies with survival cytokines and factors selected for extending survival of antitumor effector cells and establishing long-term immunologic memory.

The role of mutation accumulation in HIV progression

The onset of AIDS is characterized by the collapse of the immune system after a prolonged asymptomatic period. The mechanistic basis of this disease progression has remained obscure, hindering the development of effective therapies. Here I present a mechanism that underlies the deterioration of the immune system during HIV infection. The elevated turnover of lymphocytes throughout the asymptomatic period is postulated to result in the accumulation of deleterious mutations, which impairs immunological function, replicative ability and viability of lymphocytes. This mutational meltdown is proposed to occur throughout the hierarchy of lymphocyte progenitors, resulting in the deterioration of lymphocyte regeneration and an ensuing rise in viral loads. A mathematical model is used to illustrate this mechanism of progressive immunological deterioration. Mutation accumulation may explain not only the decline in CD4+T cells, but also the functional deterioration of CD4+T cells, CD8+T cells and B cells, and the exhaustion of lymphocyte regeneration.