Allospecific rejection of MHC class I-deficient bone marrow by CD8 T cells.

Avoidance of long-term immunosuppression is a desired goal in organ transplantation. Mixed chimerism offers a promising approach to tolerance induction, and we have aimed to develop low-toxicity, nonimmunodepleting approaches to achieve this outcome. In a mouse model achieving fully MHC-mismatched allogeneic bone marrow engraftment with minimal conditioning (3 Gy total body irradiation followed by anti-CD154 and T cell-depleted allogeneic bone marrow cells), CD4 T cells in the recipient are required to promote tolerance of preexisting alloreactive recipient CD8 T cells and thereby permit chimerism induction. We now demonstrate that mice devoid of CD4 T cells and NK cells reject MHC Class I-deficient and Class I/Class II-deficient marrow in a CD8 T cell-dependent manner. This rejection is specific for donor alloantigens, since recipient hematopoiesis is not affected by donor marrow rejection and MHC Class I-deficient bone marrow that is syngeneic to the recipient is not rejected. Recipient CD8 T cells are activated and develop cytotoxicity against MHC Class I-deficient donor cells in association with rejection. These data implicate a novel CD8 T cell-dependent bone marrow rejection pathway, wherein recipient CD8 T cells indirectly activated by donor alloantigens promote direct killing, in a T cell receptor-independent manner, of Class I-deficient donor cells.

Recipient dendritic cells, but not B cells, are required antigen-presenting cells for peripheral alloreactive CD8+ T-cell tolerance.

Induction of mixed allogeneic chimerism is a promising approach for achieving donor-specific tolerance, thereby obviating the need for life-long immunosuppression for solid organ allograft acceptance. In mice receiving a low dose (3Gy) of total body irradiation, allogeneic bone marrow transplantation combined with anti-CD154 tolerizes peripheral CD4 and CD8 T cells, allowing achievement of mixed chimerism with specific tolerance to donor. With this approach, peripheral CD8 T-cell tolerance requires recipient MHC class II, CD4 T cells, B cells and DCs. Recipient-type B cells from chimeras that were tolerant to donor still promoted CD8 T-cell tolerance, but their role could not be replaced by donor-type B cells. Using recipients whose B cells or DCs specifically lack MHC class I and/or class II or lack CD80 and CD86, we demonstrate that dendritic cells (DCs) must express CD80/86 and either MHC class I or class II to promote CD8 tolerance. In contrast, B cells, though required, did not need to express MHC class I or class II or CD80/86 to promote CD8 tolerance. Moreover, recipient IDO and IL-10 were not required. Thus, antigen presentation by recipient DCs and not by B cells is critical for peripheral alloreactive CD8 T cell tolerance.

Contrasting pulmonary blood flow profiles in children with atrial and ventricular septal defects.

Instantaneous pulmonary blood flow profiles and other measures of haemodynamic functioning were studied in children with atrial septal defects (ASD) and children with ventricular septal defects (VSD) and normal pulmonary pressures and resistance at the time of defect closure. Though most parameters measured were comparable in the patient populations, several significant differences were found in the shape of the flow profile.

Two-dimensional in vivo pressure/diameter relationships in the canine main pulmonary artery.

Ultrasonic measurement of blood flow within the main pulmonary artery (MPA) requires a precise knowledge of the mean blood velocity within this vessel and the cross-sectional area (CSA). Small conformational changes in the elliptical shape of the MPA have substantial effects on the calculation of CSA and, subsequently, flow. We examined the extent of these changes by measuring the pulsatile and mean elliptical dimensions of the MPA in nine anaesthetised, open-chested, mechanically ventilated mongrel dogs using two pairs of 10 MHz ultrasonic, piezoelectric crystals. These custom-made devices were sutured to the PA adventitia along the long and short cross-sectional axes 2 cm distal to the pulmonary valve. Axial dimensions were collected during normal, elevated (via noradrenaline and fluid additions) and reduced (via exsanguination) PA pressures. We confirmed the linear pressure/diameter response in 15/18 axial data sets (r greater than 0.80). Further, the linear axial responses of the long and short diameters were parallel (7/9, p less than 0.05) and have different zero-pressure intercepts (7/9, p less than 0.0001). A mathematical consequence of this parallelism is predictable, although non-constant, eccentricity. Finally, error analysis of multi-axial measurement techniques were shown to improve CSA accuracy by as much as 50% when compared with uni-axial determinations.

Velocity profile in the main pulmonary artery in a canine model.

The velocity profile of the main pulmonary artery was determined in nine adult, open-chested, mechanically ventilated mongrel dogs using an intraluminal, needle-mounted, range-gated, pulsed Doppler technique. Mean phasic point velocities were determined at 2 mm intervals across the lumen of the vessel, 2 cm above the pulmonary valve, by recording the Doppler shift of an activated 20 MHz piezoelectric crystal, range-gated 3.5 mm in the direction of the pulmonary valve. Mean Reynolds numbers from the main pulmonary artery ranged from 275 to 1140. Radially normalised intraluminal distance versus mean phasic point velocity plots were constructed which demonstrated a curved profile in all 9 dogs. First order regression analysis demonstrated a poor fit (r: 0.05-0.68). Second order (r:0.61-0.97) and third order (r:0.72-0.99) regression analyses markedly improved the fit, confirming the non-linear nature of the velocity profile. Step-wise third order regression analysis to determine the importance of the entry sequence demonstrated that the most important term for determining the regression coefficient was the X2 term in six dogs. In addition, the velocity profile was noted to be shifted, with the highest velocities recorded between the centre of the vessel and the anterior wall in eight of nine dogs (location of highest velocity: +0.26 radius +/- 0.25 (mean +/- SD).

The velocity profile in the canine ascending aorta and its effects on the accuracy of pulsed Doppler determinations of mean blood velocity.

The velocity profile in the ascending aorta, which has been variously reported as parabolic, skewed or flat, was measured using an intraluminal probe equipped with a miniature piezoelectric crystal activated by a 20 MHz pulsed Doppler with a 1 mm3 sample volume size. Phasic velocities, determined via spectral analyses, were obtained at 2 mm intervals across the aortic lumen of anaesthetised open-chest dogs by sequentially moving the crystal between posterior and anterior walls, maintaining the ultrasound beam parallel to the flow axis. Range-gating allowed data acquisition 3.5 mm proximal to the crystal, thus eliminating probe induced turbulence. Time and spatial distributions of velocities over the cross-section were computed. Expected errors in vessel mean velocity estimates based on point measurements were calculated. Analysis of phasic waveforms showed increased posterior velocities during systole but third order regression analysis of normalised point mean velocities across the lumen showed the best fit was a statistically flat straight line. Centreline referenced analyses indicated the expected difference between point mean velocity and vessel mean velocity was maximal (19.3 +/- 10.1%) 4 mm toward the posterior wall and minimal (4.7 +/- 9.1%) 5 mm toward the anterior wall. Anterior posterior wall referenced analyses indicated the most reliable position for estimating vessel mean velocity from point mean velocity along this axis was 5 mm from the anterior wall.

Analysis of T cell receptor-gene rearrangement in T cells from the cerebrospinal fluid of patients with multiple sclerosis.

Characterization of T cells present in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) may contribute to an understanding of the immunopathologic role of these cells. To analyze the T cells in the CSF of MS patients, 30 cloned T cell lines from each of two MS patients were surveyed for their patterns of T cell receptor (TcR) beta-chain gene rearrangement. DNA from the (CSF-derived) T cell clones was digested with a number of restriction endonucleases and the gene rearrangement patterns were analyzed with a T cell receptor beta-chain probe. Southern blot analysis of the DNA of these T cell clones indicated that all had rearrangements of the TcR beta-chain genes, but none of the rearrangements were identical. These results suggest that, if a few clones of specific T cells are involved, they must form a tiny minority in comparison to the total number of T cells in the CSF of MS patients.

Maturation of pulmonary input impedance spectrum in infants and children with ventricular septal defect.

To determine whether pulmonary vascular disease can be detected in infants with ventricular septal defect (VSD) by the presence of an increase in the frequency of the impedance modulus minimum of the pulmonary input impedance spectrum, as has been implied for older children, spectra of 25 infants (2 years or younger) (group 1) were compared with spectra of 20 children (ages 2 to 7 years) (group 2). Groups were subdivided according to mean pulmonary artery (PA) pressure: those with moderate pressure levels (35 mm Hg or less, groups 1A and 2A) and those with high pressure levels (at least 40 mm Hg, groups 1B and 2B). Pulmonary vascular resistance, characteristic impedance and frequency of the modulus minimum were significantly lower in group 2A than in group 1A. The decrease in pulmonary vascular resistance and characteristic impedance with increasing age was consistent with body surface area increases; however, the shift in frequency of the modulus minimum could be more easily related to a decrease in the pulse wave velocity than to a shift in the primary reflection site. Pulmonary vascular resistance, characteristic impedance and the frequency of the first modulus minimum were comparable in groups 1B and 2B; however, none of the patients in group 1B had evidence of pulmonary damage, whereas 3 of 4 group 2B patients had microscopically apparent pulmonary vascular disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Intraluminal pulsed Doppler evaluation of the pulmonary artery velocity time curve in a canine model of acute pulmonary hypertension.

The velocity pattern of the blood flow in the pulmonary artery was investigated in an animal model of acute pulmonary hypertension. Nine anesthetized, open-chest dogs were embolized with polystyrene microspheres, and the velocity pattern of the blood flow in the pulmonary artery was studied with use of an invasive pulsed Doppler technique. Phasic intraluminal velocity was recorded with use of a miniature piezoelectric crystal activated by 20-MHz Doppler pulses and mounted on the tip of a needle probe introduced into the pulmonary artery. The recorded Doppler quadrature signals were processed by spectral analysis. Significant increases occurred in mean, systolic, and diastolic pulmonary arterial pressures (p less than 0.0002), in pulmonary vascular resistance (p less than 0.005), and in negative velocity time (duration in milliseconds that the mean velocity was directed toward the pulmonic valve) (p less than 0.002). Significant decreases occurred in right ventricular ejection time (p less than 0.006) and in positive velocity time (duration in milliseconds that the mean velocity was directed away from the pulmonic valve) (p less than 0.005). A significant shortening in the time to peak velocity (acceleration time) was found (p less than 0.005). Second-order regression analyses demonstrated an inverse correlation between the ratio of positive velocity time to negative velocity time and the mean pulmonary artery pressure in all animals (r = 0.71). These findings should be compared with the velocity patterns of the blood flow in the pulmonary artery obtained under pulmonary hypertensive conditions due to various causes to facilitate interpretation and understanding of clinical investigations.

Constant postoperative monitoring of cardiac output after correction of congenital heart defects.

A new method has been developed that permits constant postoperative monitoring of mean and phasic cardiac output in patients after correction of congenital heart defects. A miniature ultrasound probe is attached to the adventitia of the ascending aorta at the conclusion of the operative procedure. This is connected to the monitoring equipment by means of polyurethane-covered wires that exit the chest wall through a small stab wound. The probe can easily be removed by gentle traction when the patient's condition is stable. The technique was developed, validated, and refined in extensive animal studies, and this report describes the first series of 20 consecutive human implants, performed between August 1984 and September 1985, in which the absolute cardiac output determination obtained with the ultrasound probe at the time of its application was correlated with cardiac output as measured with a standard electromagnetic flow probe. Fourteen male and six female patients (mean age 5.5 years) were studied. Operations performed included eight atrial septal defect repairs, four procedures for tetralogy of Fallot, three ventricular septal defect repairs, three stenotic valve corrections, and two Senning operations. One operative death occurred, but no complications were related to probe application or removal. The average cardiac output in the 20 patients as measured with the ultrasound probe was 2.2 +/- 1.1 L/min (range 0.67 to 5.27 L/min). This is nearly identical to the results noted with the electromagnetic flow probe, where the mean cardiac output was 2.3 +/- 1.2 L/min (range 0.7 to 6 L/min). Regression analysis revealed a high linear correlation (r = 0.9) between the two techniques. A monitor can display the cardiac output trend with 1 minute updates, which greatly enhance management of intravenous drug therapy and volume administration. In conclusion, this new extraluminal removable probe allows virtually continuous monitoring of the postoperative cardiac output after correction of congenital heart defects and should become a standard technique in the postoperative care of these patients.

Effects of pneumonectomy on pulmonary input impedance.

The long-term effects of pneumonectomy on the pulmonary circulation quantifiable through pulmonary input impedance analysis were studied. Excision of the left lung was performed in purebred beagle dogs aged 6 to 10 weeks (n = 6 group I) or 1 year (n = 8 group II). Unoperated beagles served as controls (n = 8 group III). When the dogs were 5 years of age, pulmonary pressure and flow were measured and the impedance spectra calculated. Characteristic impedance (Zo) (indicative of changes in proximal vessel physical properties) and pulmonary vascular resistance (PVR) (indicative of the distal response) were estimated. In group III the cardiac output (CO) was 1.7 +/- 0.4 L/min, mean pressure 16 +/- 5 mm Hg, PVR 605 +/- 448 dyne-sec/cm, and Zo 204 +/- 76 dyne-sec/cm. Group I results exhibited bimodal distributions that were not statistically different from results of groups II or III; four dogs had spectra comparable to those of group III, while two dogs had developed moderate hypertension and high PVR and Zo. Group II results were more normally distributed, and comparison with group III indicated statistically significant differences (P less than 0.05) in CO (1.1 +/- 2 L/min), PVR (1396 +/- 573 dynes-sec/cm), and Zo (543 +/- 273 dynes-sec/cm). Doubling of PVR and Zo in group II indicated that proximal vessel compliance and peripheral perfusion radius had not increased following pneumonectomy in adult beagles. Group I results indicate that marked facilitory adaptation can occur when pneumonectomy is performed in puppies; however, the adaptation may not be based on true lung growth and, therefore, may not be sustained indefinitely.

The acute effects of pneumonectomy on pulmonary vascular impedance in the dog.

Pulmonary vascular impedance is a measure of the pulsatile characteristic of pressure and flow that occurs in the proximal pulmonary arteries. Pulmonary vascular resistance (PVR) is most influenced by the distal circulation of the lung. This study was performed to evaluate the changes that occurred in pulmonary vascular impedance, as well as in other hemodynamic variables, following pneumonectomy by a closed-chest method in 10 anesthetized dogs. The following observations were made (numbers compare mean values for the 10 dogs before and after pneumonectomy): (1) PVR increased from 447 to 761 dyne sec cm-5 (p = .02); (2) the oscillatory work of the right ventricle increased from 1.23 to 1.76 J/min (p = .006); (3) the mean pulmonary artery pressure increased from 14 to 18.8 mm Hg (p = .0001); and (4) cardiac output and heart rate remained unchanged. Surprisingly, the estimated characteristic impedance (the impedance to oscillatory flow in the proximal bed) did not change significantly (279 to 296 dyne sec cm-5). This observation cannot be explained by the usual lumped compartmental models classically used to characterize the pulmonary vascular bed.

Clinical and experimental evaluation of left ventriculoiliac shunt bypass during repair of lesions of the descending thoracic aorta.

Temporary ventriculoiliac bypass with a tridodecylmethylammonium chloride-coated shunt has been used routinely at the University of North Carolina for the past seven years for repair of lesions of the descending thoracic aorta. Although the technic appears to be safe and reliable, the hemodynamic effects of prolonged nonvalved apical diversion on left ventricular function are not defined. To evaluate left ventricular performance during ventriculoiliac shunt bypass, the procedure was investigated in adult sheep. Systolic flow through the shunt was pulsatile and accounted for approximately 35% of the total cardiac output. Reversed flow was minimal. No significant change occurred in cardiac output, left ventricular end-diastolic pressure, or left atrial pressure. Perfusion of the abdominal viscera through the shunt was sufficient to prevent intestinal and renal ischemia. Our results indicate that the shunt provides left ventricular decompression without evidence of deterioration in left ventricular performance for up to three hours of apical bypass and aortic occlusion. It is included that bypass with a left ventriculoiliac shunt provides safe and effective diversion during repair of lesions of the descending thoracic aorta and offers and excellent alternative to methods involving greater technical hazard or requiring systemic anticoagulation.

Cardiovascular effects of positive end-expiratory pressure (PEEP) after pneumonectomy in dogs.

Little is known regarding the hemodynamic effect of positive end-expiratory pressure (PEEP) following pneumonectomy. To investigate this, 9 mongrel dogs underwent PEEP before and after lung resection. With the chest closed, the dog anesthetized, and partial pressure of carbon dioxide constant, PEEP was added in increments of 2 mm Hg until the animal's condition became hemodynamically unstable. At each level of PEEP, aortic, pulmonary, left atrial, and central venous pressures were monitored while aortic flow (cardiac output) was determined with an electromagnetic probe and airway pressure was measured with a Millar catheter in the respiratory tubing. Pneumonectomy was then performed, PEEP was again sequentially added, and the same measurements were recorded. Both before and after pneumonectomy, a strong positive linear correlation exists between the level of PEEP and pulmonary vascular resistance (PVR) (r greater than 0.74; p less than 0.05). Also, there is a high negative linear correlation between the level of PEEP and cardiac output (r greater than -0.76; p less than 0.05). At 0 mm Hg of PEEP, the PVR is higher after pneumonectomy than before (p less than 0.02). The incremental elevation in PVR persists after pneumonectomy at each level of PEEP, and in 5 of the 9 dogs the slope of the linear regression line relating PVR to PEEP was steeper following resection (p less than 0.05), thereby demonstrating an exaggerated effect of PEEP on PVR. In addition, all animals had a lower cardiac output at each comparable level of PEEP following pneumonectomy (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Elective pulmonary lobectomy: factors associated with morbidity and operative mortality.

Periodic review of clinical results is essential to ensure that high-quality patient care is maintained. To that end, we reviewed the morbidity and operative mortality in a consecutive series of 369 pulmonary lobectomies performed between January 1, 1970, and December 31, 1983. There were 251 male and 118 female patients with a mean age of 50.6 years. The thirty-day operative mortality was 2.2% (8/369), with 6 of these deaths related primarily to respiratory insufficiency. Two hundred twenty-four postoperative management problems occurred in 151 patients and included arrhythmia, air leak, pneumothorax, respiratory difficulties, postoperative bleeding, pleural effusion, wound infection, myocardial infarction, pulmonary embolus, empyema, bronchial stump leak, and lobar gangrene. Multiple factors were related to the occurrence of postoperative morbidity and mortality using both chi-square analysis to examine each individual item and discriminant analysis to evaluate their interaction. Chi-square tabulation showed no difference in the occurrence of major postoperative complications (p greater than 0.05) related to the side of operation, an abnormal preoperative electrocardiogram, a forced vital capacity of 2.8 liters or less, a one-second forced expiratory volume (FEV1) of less than 1.7 liters, an oxygen tension of less than 60 mm Hg, or the seniority of the surgeon (resident versus attending). An increased number of complications (p less than 0.05) was found in male patients, in patients operated on for carcinoma, and in patients older than 60 years. Stepwise discriminant analysis included FEV1 as a significant predictor of postoperative complications.(ABSTRACT TRUNCATED AT 250 WORDS)

Fluid mechanics of the pulmonary circulation.

Assessment of the fluid mechanics of the pulmonary circulation has been hindered by the relative inaccessibility of the pulmonary vascular bed. Current understanding is based largely on analyses of mathematical models which have been used to simulate hemodynamic characteristics within an architectural representation of that bed. These representations have ranged from simple, lumped parameter structures to anatomically realistic structures based on measurements of casts of the irregular branching network of pulmonary arteries and veins. Nonlinear models have been studied to determine the effects of vessel taper and within beat geometrical variations as cross-sectional shapes change from circular to elliptical and small vessels undergo recruitment, distention, and collapse as a result of the shifting balance in alveolar, pleural, and capillary pressures. This article reviews current concepts of pulmonary circulatory fluid mechanics focusing on mathematical models and relevant hemodynamic studies.

Software analysis of 20 MHz pulsed Doppler quadrature data.

A computer software procedure for processing quadrature data obtained from a 20 MHz pulsed Doppler system is described and the quality of results obtained is illustrated. The system is implemented on a standard laboratory minicomputer system. Software flexibility expands use of this high resolution device for measuring blood velocity to larger vessels than previously demonstrated. Techniques for dealing with high frequency aliasing, time delays, digital filtering and sampling intervals are included.

Characteristics of blood flow velocity in the hypertensive canine pulmonary artery.

Pulmonary artery blood flow velocity was measured in 15 dogs by a recently developed direct intraluminal pulsed Doppler technique. Changes in velocity characteristics under conditions of experimentally induced hypoxic pulmonary hypertension were observed. Experimental conditions (fractional inspired oxygen concentration = 0.10) produced significant increases in mean pulmonary artery pressure and pulmonary vascular resistance. Overall and maximal negative velocity increased with pulmonary hypertension. Negative velocity occurred predominantly in the posterior half of the pulmonary artery during both control and experimental conditions. With pulmonary hypertension, diastolic negative velocity increased only in the posterior half of the pulmonary artery and systolic negative velocity decreased only in the anterior half. More basic knowledge of pulmonary artery blood flow characteristics may facilitate an informed approach to noninvasive detection of pulmonary hypertension. Direct measurements by this recently developed intraluminal technique will be useful in studying various conditions with altered pulmonary blood flow.

HVEM/LIGHT/BTLA/CD160 cosignaling pathways as targets for immune regulation.

Immunosuppression is currently the treatment of choice to attenuate the chronic deterioration of tissue function as a result of the effector mechanisms of the immunological response in transplant rejection and autoimmune diseases. However, global immunosuppression greatly increases the risk of acquiring life-threatening infections and is associated with organ toxicity when used long-term. Thus, alternative approaches that inhibit only the unwanted immune responses and preserve general immunity are highly desirable. The receptor/ligand pairs involved in the cross-talk between DC and T cells have been the focus of intense and exciting research during the last decade. The HVEM/LIGHT/BTLA/CD160 costimulatory/coinhibitory pathway has emerged as a potential target for the development of immune therapeutic interventions. Herein, we will summarize and discuss how blockade of the costimulatory HVEM/LIGHT interaction or agonist signaling through the inhibitory BTLA and CD160 receptors could contribute to the control of deleterious immune responses.