Greg J. Beilman

Persistent HIV-1 replication is associated with lower antiretroviral drug concentrations in lymphatic tissues

Significance We show that HIV continues to replicate in the lymphatic tissues of some individuals taking antiretroviral regimens considered fully suppressive, based on undetectable viral loads in peripheral blood, and that one mechanism for persistent replication in lymphatic tissues is the lower concentrations of the antiretroviral drugs in those tissues compared with peripheral blood. These findings are significant because they provide a rationale and framework for testing the efficacy of new agents and combinations of drugs that will fully suppress replication in lymphatic tissues. More suppressive regimens could improve immune reconstitution, as well as provide the effective regimens needed for functional cure and eradication of infection. Antiretroviral therapy can reduce HIV-1 to undetectable levels in peripheral blood, but the effectiveness of treatment in suppressing replication in lymphoid tissue reservoirs has not been determined. Here we show in lymph node samples obtained before and during 6 mo of treatment that the tissue concentrations of five of the most frequently used antiretroviral drugs are much lower than in peripheral blood. These lower concentrations correlated with continued virus replication measured by the slower decay or increases in the follicular dendritic cell network pool of virions and with detection of viral RNA in productively infected cells. The evidence of persistent replication associated with apparently suboptimal drug concentrations argues for development and evaluation of novel therapeutic strategies that will fully suppress viral replication in lymphatic tissues. These strategies could avert the long-term clinical consequences of chronic immune activation driven directly or indirectly by low-level viral replication to thereby improve immune reconstitution.

Noninvasive method for measuring local hemoglobin oxygen saturation in tissue using wide gap second derivative near-infrared spectroscopy.

A simple continuous wave near-infrared algorithm for estimating local hemoglobin oxygen saturation in tissue (%StO2) is described using single depth attenuation measurements at 680, 720, 760, and 800 nm. Second derivative spectroscopy was used to reduce light scattering effects, chromophores with constant absorption, baseline/instrumentation drift, and movement artifacts. Unlike previous second derivative methods which focused primarily on measuring deoxyhemoglobin concentration; a wide 40 nm wavelength gap used for calculating second derivative attenuation significantly improved sensitivity to oxyhemoglobin absorption. Scaled second derivative attenuation at 720 nm was correlated to in vitro hemoglobin oxygen saturation to generate a %StO2 calibration curve. The calibration curve was insensitive to total hemoglobin, optical path length, and optical scattering. Measurement error due to normal levels of carboxyhemoglobin, methemoglobin, and water absorption were less than 10 %StO2 units. Severe methemoglobinemia or edema combined with low blood volume could cause StO2 errors to exceed 10 StO2 units. Both a broadband and commercial four-wavelength spectrometer (InSpectra) measured %StO2. The InSpectra tissue spectrometer readily detected limb ischemia on 26 human volunteers for hand, forearm, and leg muscles. A strong linear correlation, r2>0.93, between StO2 and microvascular %SO2 was observed for isolated animal hind limb, kidney, and heart.

Simian Immunodeficiency virus-Induced Lymphatic Tissue Fibrosis Is Mediated by Transforming Growth Factor β1-positive Regulatory T Cells and Begins in Early Infection

In human immunodeficiency virus (HIV) infection, collagen deposition and fibrosis within the T cell zone disrupt the lymphatic tissue architecture, contributing to depletion of CD4(+) T cells and limiting immune reconstitution. We used relevant animal and in vitro models to investigate the kinetics and possible underlying mechanism(s) of this process. In the lymphatic tissue of simian immunodeficiency virus (SIV)-infected rhesus macaques, we observed parallel increases in immune activation, transforming growth factor (TGF) beta 1-positive regulatory T (T(reg)) cells, and collagen type I deposition by 7 days after inoculation, consistent with the hypothesis that early immune activation elicits a countering T(reg) cell response associated with TGF beta 1 expression and collagen deposition. In support of this hypothesis and the possible role of fibrosis in viral pathogenesis, we show (1) spatial colocalization and temporal concordance in levels of TGF beta 1(+) T(reg) cells and collagen deposition; (2) TGF beta 1(+) inducible T(reg) cell stimulation of primary lymphatic tissue fibroblasts to produce collagen type I in vitro; and (3) high levels of immune activation, TGF beta 1(+) T(reg) cells, and collagen deposition in pathogenic SIV infection of macaques, in contrast to apathogenic SIV infection in sooty mangabeys in which levels of immune activation, TGF beta 1(+) T(reg) cells, and collagen deposition were low. We thus conclude that the response of TGF beta 1(+) T(reg) cells to immune activation in early SIV/HIV infection is a double-edged sword: TGF beta 1(+) T(reg) cells normally have a positive effect by limiting immunopathological and autoreactive immune responses, but they also have a negative effect by dampening the antiviral immune response and, as we show here, causing deleterious effects on CD4(+) T cell homeostasis by inducing collagen deposition in lymphatic tissues.

Lymphoid Tissue Damage in HIV-1 Infection Depletes Naïve T Cells and Limits T Cell Reconstitution after Antiretroviral Therapy

Highly active antiretroviral therapy (HAART) can suppress HIV-1 replication and normalize the chronic immune activation associated with infection, but restoration of naïve CD4+ T cell populations is slow and usually incomplete for reasons that have yet to be determined. We tested the hypothesis that damage to the lymphoid tissue (LT) fibroblastic reticular cell (FRC) network contributes to naïve T cell loss in HIV-1 infection by restricting access to critical factors required for T cell survival. We show that collagen deposition and progressive loss of the FRC network in LTs prior to treatment restrict both access to and a major source of the survival factor interleukin-7 (IL-7). As a consequence, apoptosis within naïve T cell populations increases significantly, resulting in progressive depletion of both naïve CD4+ and CD8+ T cell populations. We further show that the extent of loss of the FRC network and collagen deposition predict the extent of restoration of the naïve T cell population after 6 month of HAART, and that restoration of FRC networks correlates with the stage of disease at which the therapy is initiated. Because restoration of the FRC network and reconstitution of naïve T cell populations are only optimal when therapy is initiated in the early/acute stage of infection, our findings strongly suggest that HAART should be initiated as soon as possible. Moreover, our findings also point to the potential use of adjunctive anti-fibrotic therapies to avert or moderate the pathological consequences of LT fibrosis, thereby improving immune reconstitution.

Early surgery for thoracolumbar spine injuries decreases complications.

BACKGROUND The proper timing for surgical fracture repair is controversial. Early repair of long bone and cervical fractures reduces complications and is safe. Few studies exist to compare time to surgery with outcomes in thoracolumbar (TL) spine injuries. METHODS Patients with TL spine injuries were identified from the trauma registry and divided into two cohorts on the basis of Injury Severity Score (ISS). Cohorts were compared for infectious, respiratory, and total complications in patients who had early (<72 hours from injury) versus late (>72 hours from injury) surgical repair. A retrospective chart review was performed on High ISS patients (> or =15) to identify differences in resuscitation needs and neurologic, respiratory, and infectious complications. RESULTS Early surgery, Low ISS patients were younger, received fewer anterior repairs, and had shorter hospitalizations. Early patients in the High ISS cohort had significantly fewer total complications and shorter hospital and intensive care unit lengths of stay. Resuscitative requirements were similar for both surgery groups. More late surgery patients required ventilator support for noninfectious reasons. There was no difference in admission or postoperative neurologic status or the incidence of head injury. CONCLUSION Early surgery in severely injured patients with thoracolumbar spine trauma was associated with fewer complications and shorter hospital and intensive care unit lengths of stay, required less ventilator support for noninfectious reasons, and did not increase neurologic deficits.

Early hypothermia in severely injured trauma patients is a significant risk factor for multiple organ dysfunction syndrome but not mortality

Objective:To evaluate the relationship of early hypothermia to multiple organ failure and mortality in a prospectively-collected database of severely injured trauma patients. Methods:This prospective observational study was performed at 7 level I trauma centers over a 16-month period. Severely injured trauma patients with signs of hypoperfusion (eg, base deficit, hypotension) and need for blood transfusion during their early hospital course were followed for 24 hours with near infrared spectroscopy-derived tissue oxygen saturation (StO2) and other variables for 28 days to evaluate outcomes including multiple organ dysfunction syndrome (MODS) and death. Early hypothermia was defined as the presence of a temperature ≥35°C anytime within the first 6 hours of hospitalization. Comparisons between groups were made using the Wilcoxon Two-Sample test for continuous variables and either the Fisher exact or &khgr;2 test for categorical variables. Multivariate logistic regression was utilized to understand the effect of hypothermia on outcome (MODS and mortality). Results:Hypothermia was very common in this cohort of patients, present in 43% of patients enrolled (155/359). Hypothermic patients were 3 times more likely than normothermic patients to develop MODS (21% vs. 9%, P = 0.003). Hypothermic patients did not have an increased incidence of mortality (16% vs. 12%, P= 0.2826). Base deficit in hypothermic patients did not discriminate between patients who did or did not develop MODS (9.8 + 4.6 mEq/L vs. 9.4 + 4.4 mEq/L), but had good discrimination for mortality in both hypothermic and normothermic patients. Significant predictors of MODS using multivariate analysis included minimum StO2 (P= 0.0014) and hypothermia (P = 0.0371). Predictors for mortality using multivariate analysis included minimum StO2 (P= 0.0021) and base deficit (P= 0.0454), but not hypothermia (P= 0.5289). Hypothermia remained a significant risk factor for MODS when systolic blood pressure, volume of fluid, and volume of blood infused were included in the multivariate model. Conclusion:Hypothermia is common in severely injured trauma patients (nearly half of patients in this series) and is a significant risk factor for MODS but not mortality. The predictive value of base deficit for development of MODS is blunted in the presence of hypothermia. A low StO2 value predicts MODS and mortality in trauma patients and is a durable measure in both normothermic and hypothermic patient groups.

Moderately elevated serum troponin concentrations are associated with increased morbidity and mortality rates in surgical intensive care unit patients.

ObjectiveTo determine the significance of moderate elevations of serum troponin I in a surgical intensive care unit patient population in terms of its impact on indexes of outcome including mortality, morbidity, and hospital and intensive care unit length of stay. DesignRetrospective chart review and analysis of clinical data. SettingA surgical intensive care unit at a tertiary care hospital. PatientsFrom the 27-month surgical intensive care unit database of admissions, 869 patients with serum troponin I determinations during their admission were identified. Patients who had cardiac surgery were excluded. InterventionsNone. Measurements and ResultsPatients were divided into four groups based on their maximum serum troponin I concentrations. Hospital mortality, incidence of myocardial infarction, and hospital and intensive care unit length of stays were compared. Patients with moderate elevations of serum troponin I (0.4–2.0 &mgr;g/L) had a significantly higher mortality rate (chi-square = 32.57, p < .0001) and longer length of intensive care unit and hospital stays (p < .0005) when compared with patients without similar elevations. Within the range of moderately elevated troponin concentrations, higher titers were associated with increasing mortality risk, longer hospital and intensive care unit stays, and a higher incidence of myocardial infarction. The use of a &bgr;-blocker and aspirin was associated with better survival for patients with maximum serum troponin concentrations ≥2 &mgr;g/L. ConclusionModerate elevations of serum troponin I, which are below the threshold required to diagnose overt myocardial infarction, may reflect ongoing myocardial injury in the critically ill and are associated with a higher mortality rate and longer hospital and intensive care unit length of stays. The use of &bgr;-blockers and aspirin is associated with better outcomes for this subset of patients.

Critical role of CD4 T cells in maintaining lymphoid tissue structure for immune cell homeostasis and reconstitution

Loss of the fibroblastic reticular cell (FRC) network in lymphoid tissues during HIV-1 infection has been shown to impair the survival of naive T cells and limit immune reconstitution after antiretroviral therapy. What causes this FRC loss is unknown. Because FRC loss correlates with loss of both naive CD4 and CD8 T-cell subsets and decreased lymphotoxin-β, a key factor for maintenance of FRC network, we hypothesized that loss of naive T cells is responsible for loss of the FRC network. To test this hypothesis, we assessed the consequences of antibody-mediated depletion of CD4 and CD8 T cells in rhesus macaques and sooty mangabeys. We found that only CD4 T-cell depletion resulted in FRC loss in both species and that this loss was caused by decreased lymphotoxin-β mainly produced by the CD4 T cells. We further found the same dependence of the FRC network on CD4 T cells in HIV-1-infected patients before and after antiretroviral therapy and in other immunodeficiency conditions, such as CD4 depletion in cancer patients induced by chemotherapy and irradiation. CD4 T cells thus play a central role in the maintenance of lymphoid tissue structure necessary for their own homeostasis and reconstitution.

Glycemic control and prevention of perioperative infection.

Purpose of reviewHyperglycemia is common during perioperative management of surgical and critically ill patients. There is extensive ongoing study of detrimental effects associated with hyperglycemia, with several remaining unanswered questions. This review discusses recent literature on tight glucose control with insulin therapy and its effects in prevention and management of infection. Recent findingsHyperglycemia affects multiple pathways of the immune system, resulting in decreased phagocytic and chemotactic functions in neutrophils and monocytes, as well as increased rates of apoptosis of the former and decreased ability of the latter to present antigen. Intensive insulin therapy has been shown to counteract many of these deleterious effects. Clinically, the benefits of tight glucose control have been evaluated in different patient populations with conclusions that remain varied. Hypoglycemia as a complication of tight glucose control continues to be an issue and has led to discontinuation of two large-scale studies. The clinical relevance of hypoglycemic events remains unclear. SummaryHyperglycemia impairs the cellular immune system, stimulates inflammatory cytokines, and affects the microcirculation, thus increasing risk for infection and preventing normal wound healing. Additional investigation is needed to define appropriate patient populations and to develop effective treatment strategies for preventing perioperative morbidity.

Tissue hemoglobin index: a non-invasive optical measure of total tissue hemoglobin.

IntroductionThe tissue hemoglobin index (THI) is a hemoglobin signal strength metric provided on the InSpectra™ StO2 Tissue Oxygenation Monitor, Model 650. There is growing interest regarding the physiologic meaning of THI and whether a clinically useful correlation between THI and blood hemoglobin concentration exists. A series of in vitro and in vivo experiments was performed to evaluate whether THI has potential utility beyond its primary purpose of helping InSpectra™ device users optimally position a StO2 sensor over muscle tissue.MethodsThe THI and tissue hemoglobin oxygen saturation (StO2) were measured using the InSpectra™ StO2 Tissue Oxygenation Monitor, Model 650, with a 15 mm optical sensor. A THI normal reference range was established in the thenar eminence (hand) for 434 nonhospitalized human volunteers. In 30 subjects, the thenar THI was also evaluated during 5-minute arterial and venous blood flow occlusions, and with blood volume exsanguination in the hand induced with an Esmarch bandage. In addition, correlation of the THI to blood total hemoglobin concentration (Hbt) was studied in five pigs whose Hbt was isovolumetrically diluted from 13 to 4 g/dl systemically and 0.5 g/dl locally in the hind limb. The sensitivity and specificity of the THI to measure tissue hemoglobin concentration (THC) were characterized in vitro using isolated blood tissue phantoms.ResultsIn human thenar tissue, the average THI was 14.1 ± 1.6 (mean ± standard deviation). The THI extrapolated to 100% blood volume exsanguination was 3.7 ± 2.0 units presumably from myoglobin. On average, the THI increased 1.5 ± 1.0 units with venous occlusion and decreased 4.0 ± 2.0 units with arterial occlusion. In porcine hind limbs, the THI weakly correlated with Hbt (r2 = 0.26) while ΔTHI during venous occlusion had a stronger correlation (r2 = 0.62). In vitro tests indicated that THI strongly correlated (r2 > 0.99) to phantom THC and was insensitive to StO2 changes.ConclusionsSteady-state THI values do not reliably indicate Hbt. The THI is a reproducible quantitative index for THC, and THI trends can discriminate between arterial or venous blood flow occlusions. The THI magnitude permits the estimation of myoglobins contribution to StO2.