Jodie H. Taylor

CD4+ T Cell Depletion during all Stages of HIV Disease Occurs Predominantly in the Gastrointestinal Tract

The mechanisms underlying CD4+ T cell depletion in human immunodeficiency virus (HIV) infection are not well understood. Comparative studies of lymphoid tissues, where the vast majority of T cells reside, and peripheral blood can potentially illuminate the pathogenesis of HIV-associated disease. Here, we studied the effect of HIV infection on the activation and depletion of defined subsets of CD4+ and CD8+ T cells in the blood, gastrointestinal (GI) tract, and lymph node (LN). We also measured HIV-specific T cell frequencies in LNs and blood, and LN collagen deposition to define architectural changes associated with chronic inflammation. The major findings to emerge are the following: the GI tract has the most substantial CD4+ T cell depletion at all stages of HIV disease; this depletion occurs preferentially within CCR5+ CD4+ T cells; HIV-associated immune activation results in abnormal accumulation of effector-type T cells within LNs; HIV-specific T cells in LNs do not account for all effector T cells; and T cell activation in LNs is associated with abnormal collagen deposition. Taken together, these findings define the nature and extent of CD4+ T cell depletion in lymphoid tissue and point to mechanisms of profound depletion of specific T cell subsets related to elimination of CCR5+ CD4+ T cell targets and disruption of T cell homeostasis that accompanies chronic immune activation.

Dynamic near-infrared spectroscopy measurements in patients with severe sepsis.

This study evaluated near-infrared spectroscopy (NIRS)-derived measurements in hemodynamically stable patients with severe sepsis, as compared with similar measurements in healthy age-matched volunteers. Prospective, preliminary, observational study in a surgical intensive care unit and clinical research center at a university health center. We enrolled 10 patients with severe sepsis and 9 healthy age-matched volunteers. For patients with severe sepsis, we obtained pulmonary artery catheter and laboratory values three times daily for 3 days and oxygen consumption values via metabolic cart once daily for 3 days. For healthy volunteers, we obtained all noninvasive measurements during a single session. We found lower values in patients with severe sepsis (versus healthy volunteers), in tissue oxygen saturation (StO2), in the StO2 recovery slope, in the tissue hemoglobin index, and in the total tissue hemoglobin increase on venous occlusion. Patients with severe sepsis had longer StO2 recovery times and lower NIRS-derived local oxygen consumption values versus healthy volunteers. In our preliminary study, NIRS provides a noninvasive continuous method to evaluate peripheral tissue oxygen metabolism in hemodynamically stable patients with severe sepsis. Further research is needed to demonstrate whether these values apply to broader populations of patients with systemic inflammatory response syndrome, sepsis, severe sepsis, and septic shock.

Lymphatic tissue fibrosis is associated with reduced numbers of naive CD4+ T cells in human immunodeficiency virus type 1 infection.

ABSTRACT The organized structure of lymphatic tissues (LTs) constitutes a microenvironment referred to as a niche that plays a critical role in immune system homeostasis by promoting cellular interactions and providing access to cytokines and growth factors on which cells are dependent for survival, proliferation, and differentiation. In chronic human immunodeficiency virus type 1 (HIV-1) infection, immune activation and inflammation result in collagen deposition and disruption of this LT niche. We have previously shown that these fibrotic changes correlate with a reduction in the size of the total population of CD4+ T cells. We now show that this reduction is most substantial within the naïve CD4+ T-cell population and is in proportion to the extent of LT collagen deposition in HIV-1 infection. Thus, the previously documented depletion of naïve CD4+ T cells in LTs in HIV-1 infection may be a consequence not only of a decreased supply of thymic emigrants or chronic immune activation but also of the decreased ability of those cells to survive in a scarred LT niche. We speculate that LT collagen deposition might therefore limit repopulation of naïve CD4+ T cells with highly active antiretroviral therapy, and thus, additional treatments directed to limiting or reversing inflammatory damage to the LT niche could potentially improve immune reconstitution.

Amount of lymphatic tissue fibrosis in HIV infection predicts magnitude of HAART-associated change in peripheral CD4 cell count.

The structure of lymphatic tissues is an important component of lymphatic tissue T-cell homeostasis. Collagen deposition in lymphatic tissues (common in HIV infection) disrupts the niche and limits the size of the resident CD4 cell population. In this report we show that a single measurement of lymphatic tissue collagen predicts the magnitude of recovery of the peripheral CD4 cell pool with HAART (P < 0.001). This suggests that collagen-targeted therapies might be of benefit.

Near-Infrared Spectroscopy in Patients with Severe Sepsis: Correlation with Invasive Hemodynamic Measurements

BACKGROUND Clinicians have begun using near-infrared spectroscopy (NIRS) to monitor tissue perfusion in hemorrhagic shock, as the technique allows continuous noninvasive monitoring of tissue hemoglobin oxygen saturation (StO(2)) and the tissue hemoglobin index (THI). We hypothesized that StO(2) measurements in patients with severe sepsis would be associated with the severity of their illness and would correlate with invasive hemodynamic measurements. METHODS We measured mean arterial pressure (MAP), serum lactate concentration, blood hemoglobin concentration, StO(2), and THI in nine healthy volunteers and ten patients with septic shock in a surgical intensive care unit (ICU). Enrolled patients had a pulmonary artery catheter, and had family able to give informed consent. The average Acute Physiology and Chronic Health Evaluation (APACHE) II score at enrollment for the patients was 19 +/- 5 (standard deviation) points. Volunteers and patients were similar with respect to age and sex. To collect NIRS data, we used the InSpectra Tissue Spectrometer, Model 325 (Hutchinson Technology, Inc., Hutchinson, MN). For three consecutive days, we obtained invasive hemodynamic measurements three times daily, simultaneously with NIRS measurements, and metabolic cart measurements once daily. RESULTS Patients with severe sepsis had significantly lower thenar muscle StO(2) values (p = 0.031) than healthy volunteers. Near-infrared spectroscopy-derived mixed venous oxygen saturation (NIRSvO(2)) and StO(2) measured from the thenar eminence in patients with severe sepsis correlated with SvO(2) from the pulmonary artery catheter (p < 0.05). In this group of patients, StO(2) did not correlate significantly with lactate concentration, base deficit, or APACHE II score. CONCLUSIONS Near-infrared spectroscopic measurements of StO(2) correlated with invasive hemodynamic measurements in patients with severe sepsis but did not correlate with severity of illness. These findings suggest that NIRStO(2) may be a clinically useful measurement in monitoring patients with severe sepsis. Further study of this device in early resuscitation of patients with sepsis is necessary.

Ringer's ethyl pyruvate in hemorrhagic shock and resuscitation does not improve early hemodynamics or tissue energetics

Reactive oxygen species (ROS) have been implicated in the pathogenesis of hemorrhagic shock. Ethyl pyruvate, a derivative of pyruvate and a proposed oxygen radical scavenger, is attractive as a possible resuscitation fluid. We investigated whether resuscitation with lactated Ringers (LR) containing ethyl pyruvate (REP) had any hemodynamic or tissue energetic benefits compared with LR alone for hemorrhagic shock. Hemorrhagic shock was induced in splenectomized pigs via inferior vena cava cannula. After 90 min of shock, animals were resuscitated in a stepwise fashion with LR or REP (30 mg/kg/dose, given as 1.5 mg/mL in LR) at 20 cc/kg/step for four steps. Data collected during this experiment included physiologic and hemodynamic parameters, near-infrared reflectance spectroscopy measurements of tissue hemoglobin oxygen (StO2) of the stomach, liver, and hind limb, and nuclear magnetic resonance phosphorus spectra of the liver and hind limb at each time point. In both resuscitative groups, heart rate, and lactate and pyruvate values increased during shock and began to drop toward baseline values during resuscitation. Mean arterial pressure, oxygen delivery, and oxygen consumption decreased during shock and increased toward baseline levels during the resuscitative process. There were no significant changes in physiologic parameters between the LR- and REP-resuscitated animals. There was a significantly lower stomach StO2 and hind limb cellular cytoplasmic pH during later resuscitative endpoints in REP-resuscitated animals. The clinical significance of these findings are unclear. There is no short-term hemodynamic or tissue energetic advantage to using REP as a resuscitation fluid when compared with LR. Long-term outcome studies are needed to further evaluate any potential benefits of use of REP in hemorrhagic shock.

Tissue energetics as measured by nuclear magnetic resonance spectroscopy during hemorrhagic shock

The defect in energy production in an organism during shock states may be related to the impairment of mitochondrial respiration early in shock. The aim of this study was to investigate the timing and degree of cellular energetic changes during hemorrhagic shock in real time. Instrumented, splenectomized swine were randomized to undergo hemorrhagic shock, induced by a 35% blood volume bleed, for 90 min with (n = 10) or without (n = 9) subsequent resuscitation. Resuscitated animals received shed blood in two increments followed by two normal saline boluses (20 mL/kg/bolus). Throughout experimentation, tissue phosphoenergetics of liver and skeletal muscle were monitored using 31P nuclear magnetic resonance (NMR) spectroscopy via NMR coils on the liver and hindlimb. Near-infrared spectroscopy probes were used to measure liver, stomach, and skeletal muscle oxyhemoglobin saturation (StO2). Hemorrhagic shock induced an increase in phosphomonoesters in skeletal muscle (baseline: 7.09%, 90 min: 9.94% (P < 0.05); expressed as percent total phosphorus). This increase resolved in animals receiving resuscitation (n = 10) but remained elevated in those in unresuscitated shock (n = 9). Inorganic phosphate levels increased and &bgr;ATP levels decreased significantly in the liver of animals in shock as compared with baseline. StO2 in skeletal muscle, stomach, and liver correlated with whole organism oxygen delivery (r2 = 0.356, 0.368, and 0.432, respectively). We conclude that hemorrhagic shock induces early elevation of phosphomonoesters in skeletal muscle, which correlates with the severity of shock. This implies an early transition to anaerobic glycolysis during hemorrhagic shock, which may be indicative of early mitochondrial dysfunction.

Phosphomonoesters predict early mortality in porcine hemorrhagic shock.

BACKGROUND Hemodynamic, laboratory, and tissue energetics were measured in a porcine model of hemorrhagic shock to evaluate variables as predictors of early mortality from shock. We hypothesized that elevated phosphomonoesters would predict early mortality in hemorrhagic shock. METHODS Pigs (n = 36) were subjected to 35% hemorrhage for 90 minutes in a 1.5-T nuclear magnetic resonance (NMR) magnet. Measurements included base deficit (BD); lactate; oxygen consumption/delivery; near-infrared spectroscopy of liver, stomach, and skeletal muscle tissue oxyhemoglobin saturation; and NMR spectroscopic measurements of high-energy phosphates of liver and skeletal muscle. Variables were compared between nonsurvivors and survivors to resuscitation after 90-minute measurements. RESULTS Ninety-minute mortality was 25%. Muscle phosphomonoesters (PMEs) and oxygen consumption differed significantly between survivors and nonsurvivors at baseline. Regression analysis identified baseline muscle PME levels, baseline BD, and 30-minute BD as early predictors of mortality before resuscitation (r2 = 0.304). CONCLUSION Baseline elevation in muscle PME levels predicts mortality in an animal model of severe hemorrhagic shock.

Mononuclear cell function and energetics in early traumatic injury.

BACKGROUND The reason for increased infectious complications after traumatic injury is complex and incompletely understood. We propose a relationship between the energetic state of circulating immune cells and immune cell function in traumatic injury. To examine this relationship, cellular adenosine triphosphate (ATP) concentration and cellular functions were measured in peripheral blood mononuclear cells from trauma patients and normal subjects. MATERIALS AND METHODS Mononuclear cells were isolated within 24 h of injury from trauma patients without (n = 12) or with (n = 10) hypotension (groups 1 and 2, respectively), and a group of normal control subjects (group 3, n = 13). Mononuclear cells were assayed for ATP levels using bioluminescence. Phagocytosis was quantified via flow cytometry after ingestion of fluorescent microspheres and phagocytic index (PI) was calculated (average number of particles ingested per monocyte). Protein synthesis was quantified using incorporation of (35)S-labeled methionine into cultured cells. Comparisons between groups were performed using one-way analysis of variance (ANOVA) with adjustment for multiple comparisons. RESULTS All but one trauma patient suffered blunt injury. There were significantly more transfusions in group 2 (hypotensive) patients (p = 0.0005). Nosocomial infections, length of stay (LOS), and mortality did not differ between groups 1 and 2. Concentrations of ATP and PI in both groups of traum a patients did not significantly differ from controls. In hypotensive trauma patients, increased duration of hypotension was associated with increased mononuclear cell ATP levels (r(2) = 0.227). A negative correlation between PI and ATP levels in trauma patients was discovered. Incorporation of (35)S was significantly greater in normotensive trauma patients than controls. CONCLUSION Mononuclear cell ATP levels and measured functions are preserved in early traumatic injury.

Microwave Processing for Sample Preparation to Evaluate Mitochondrial Ultrastructural Damage in Hemorrhagic Shock

This is a report of the adaptation of microwave processing in the preparation of liver biopsies for transmission electron microscopy (TEM) to examine ultrastructural damage of mitochondria in the setting of metabolic stress. Hemorrhagic shock was induced in pigs via 35% total blood volume bleed and a 90-min period of shock followed by resuscitation. Hepatic biopsies were collected before shock and after resuscitation. Following collection, biopsies were processed for TEM by a rapid method involving microwave irradiation (Giberson, 2001). Samples pre- and postshock of each of two animals were viewed and scored using the mitochondrial ultrastructure scoring system (Crouser et al., 2002), a system used to quantify the severity of ultrastructural damage during shock. Results showed evidence of increased ultrastructural damage in the postshock samples, which scored 4.00 and 3.42, versus their preshock controls, which scored 1.18 and 1.27. The results of this analysis were similar to those obtained in another model of shock (Crouser et al., 2002). However, the amount of time used to process the samples was significantly shortened with methods involving microwave irradiation.