Intestine Transplantation Across a Positive Crossmatch With Preformed Donor-Specific Antibodies.

BACKGROUND: We describe our experience using a modified protocol for immunosuppression for intestine transplantation across a positive crossmatch. Patients who underwent transplantation in 2013 were evaluated over a 12-month period for rejection and infectious events with comparison to procedure-matched controls on our standard protocol of immunosuppression. PATIENTS AND METHODS: We used a modified protocol for intestine and multivisceral transplantation for patients with a positive flow crossmatch. In addition to our standard protocol, patients with positive crossmatch were given rituximab and intravenous immunoglobulin (IVIg) preoperatively. DSA was sent for clinical evaluation at monthly intervals. Patients were screened for rejection by endoscopic evaluation. RESULTS: Four patients underwent transplantation within a single year across a positive crossmatch. Two received isolated intestine transplants and 2 had multivisceral transplantation (MVT). During the 12-month follow-up, 1 patients had an episode of severe acute cellular rejection, which was managed with increased immunosuppression. None of the patients had episodes of cytomegalovirus infection. One patient developed major infection and 3 patients developed minor bacterial infections. Among procedure-matched controls with negative final crossmatch on standard management (no preoperative rituximab or IVIg), 2 developed mild acute cellular rejection and 2 developed minor infections. One developed cytomegalovirus viremia with invasion to the colonic mucosa. CONCLUSIONS: We report our protocol for immunosuppression for IT and MVT across a positive crossmatch. This allowed transplantation despite the presence of a positive crossmatch, with low rejection rates but potentially increased risk for major infections compared to the negative crossmatch controls on our standard protocol.

Carbohydrate supplementation affects blood granulocyte and monocyte trafficking but not function after 2.5 h or running.

This randomized, double-blind, placebo-controlled study was designed to determine the influence of carbohydrate supplementation on the granulocyte and monocyte response to 2.5 h of high-intensity running [76.7 +/- 0.4% of maximal oxygen consumption (VO2max)]. Thirty experienced marathon runners (VO2max 53.4 +/- 1.0 mL.kg-1.min-1, age 41.5 +/- 1.4 y) were randomly assigned to carbohydrate-supplement (n = 17) and placebo (n = 13) groups. Subjects rested for 10-15 min before a blood sample was taken at 0715, and then ingested 0.75 L carbohydrate beverage or placebo. At 0730 subjects began running at 75-80% of VO2max for 2.5 h, and drank 0.25 L carbohydrate or placebo fluid every 15 min. Immediately after the 2.5-h run (1000), another blood sample was taken, followed by 1.5-h, 3-h, and 6-h recovery samples. Carbohydrate supplementation had a significant effect compared with placebo on the pattern of change in plasma glucose and cortisol, and the blood concentration of neutrophils (F[14, 112] = 5.13, P = 0.001) and monocytes (F[14, 112] = 4.78, P = 0.001), but not on blood granulocyte and monocyte phagocytosis or oxidative burst activity after 2.5 h of intensive running.

The role of anti-pig antibody in pig-to-baboon cardiac xenotransplant rejection.

The role of naturally produced antibody in discordant xenograft rejection is still uncertain. Twelve orthotopic pig-to-baboon heart transplants (HTx) were performed. In 2 baboons, no antibody adsorption (AbA) was performed. In 5 baboons, AbA with a pig lung was performed during circulatory arrest. In 5 baboons, AbA and blood exsanguination at the beginning of cardiopulmonary bypass (CPB) were performed. Baboons were divided into 2 groups; group 1 (n = 4) died within 24 hr of HTx and group 2 (n = 8) survived more than 24 hr. Mean survival period was 9.8 +/- 3.0 hr in group 1 and 151 +/- 33 hr in group 2. Baboon anti-pig antibody (Ab) was measured before CPB, before circulatory arrest, during AbA, at the end of CPB, and daily after HTx. Anti-RBC Ab was measured by the titration method at temperatures of 4 degrees C and 37 degrees C (RAb-4 and RAb-37). Anti-endothelial cell Ab (EAb) and anti-white blood cell Ab (WAb) titers were measured with ELISA. RAb titration > or = 1/4 and EAB and WAb > or = 1/256 were determined to be seropositive (S(+)). S(+) rate of RAb-37 at the end of CPB (endCPB) in group 2 was significantly higher than that in group 1 (8/8 vs. 1/4; P < 0.05). The seronegative (S(-)) rates of RBC-4 and EAb (endCPB) in group 2 were higher than those in group 1 (7/8 vs. 1/4 and 6/8 vs. 1/4, respectively), but not significantly. There was no difference in S(-) rate of WAb (endCPB) between group 1 and group 2. More than 4-fold decrease in RAb-4 and RAb-37 by AbA with a pig lung was observed in 5 and 7 of 8 baboons, while EAb and WAb did not change by AbA. In all of group 2, RAb-4 reverted to S(+) within 3 days after HTx. One baboon had no rejection episode and died of infection 16 days after HTx (baboon 16); however, it also became S(+) for RAb-4 a day after HTx until death. In 4 of group 2, RAb-37 became S(+) 1 or 2 days before death by rejection. Baboon 16, however, became S(+) for RAb-37 7 days after HTx and S(-) again 9 days after HTx until death. EAb became S(+) in all of group 2, but 5 of them survived more than 5 days after seroconversion.(ABSTRACT TRUNCATED AT 400 WORDS)

Assessment of three methods of evaluating soluble class I HLA molecules in cell culture supernatants and serum samples from the second international workshop on soluble human leukocyte antigens.

Three methodologies were compared in assessing sHLA specificities in cell culture supernatants and serum specimens from the Second International Workshop on sHLA: CDC inhibition, FC inhibition, and cellular ELISA inhibition. Initially, the CDC inhibition assay used polyclonal antisera in commercial HLA-phenotyping trays to confirm known specificities and screen for unknown specificities in 31 specimens. Although partly successful, critical limits were imposed by the variable antiserum titers. Thus, using pools of these same antisera and renal transplant recipient antisera, the FC inhibition assay was employed to determine the endpoint serum titers before confirming the known sHLA specificities. Of 25 specimens, four were not confirmed and five gave weak inhibitory reactions. The cellular ELISA inhibition assay, incorporating patient sera and mAbs toward three HLA, successfully confirmed all three known specificities in eight selected workshop specimens. Each methodology had advantages and disadvantages, but all three methods were successful in detecting and identifying sHLA class I specificities. Success, however, was dependent on the initial characterization (specificity and titer) and titration to end point (appropriate for each method's sensitivity) of each antibody preparation.

Effects of mode and carbohydrate on the granulocyte and monocyte response to intensive, prolonged exercise.

The influence of exercise mode and 6% carbohydrate (C) vs. placebo (P) beverage ingestion on granulocyte and monocyte phagocytosis and oxidative burst activity (GMPOB) after prolonged and intensive exertion was measured in 10 triathletes. The triathletes acted as their own controls and ran or cycled for 2.5 h at approximately 75% maximal O2 uptake, ingesting C or P (4 total sessions, random order, with beverages administered in double-blind fashion). During the 2. 5-h exercise bouts, C or P (4 ml/kg) was ingested every 15 min. Five blood samples were collected (15 min before exercise, immediately after exercise, and 1.5, 3, and 6 h after exercise). The pattern of change over time for GMPOB was significantly different between C and P conditions (P

Carbohydrate and the cytokine response to 2.5 h of running.

This randomized, double-blind, placebo-controlled study was designed to determine the influence of 6% carbohydrate (C) vs. placebo (P) beverage ingestion on cytokine responses (5 total samples over 9 h) to 2.5 h of high-intensity running (76.7 +/- 0.4% maximal O2 uptake) by 30 experienced marathon runners. For interleukin-6 (IL-6), a difference in the pattern of change between groups was found, highlighted by a greater increase in P vs. C immediately postrun (753 vs. 421%) and 1.5 h postrun (193 vs. 86%) [F(4,112) = 3.77, P = 0.006]. For interleukin-1-receptor antagonist (IL-1ra), a difference in the pattern of change between groups was found, highlighted by a greater increase in P vs. C 1.5 h postrun (231 vs. 72%) [F(2,50) = 6.38, P = 0.003]. No significant interaction effects were seen for bioactive IL-6 or IL-1 beta. The immediate postrun plasma glucose concentrations correlated negatively with those of plasma cortisol (r = -0.67, P < 0.001); postrun plasma cortisol (r = 0.70, P < 0.001) and IL-6 levels (r = 0.54, P = 0.003) correlated positively with levels of IL-1ra. Taken together, the data indicate that carbohydrate ingestion attenuates cytokine levels in the inflammatory cascade in response to heavy exertion.

Indomethacin does not alter natural killer cell response to 2.5 h of running.

The effect of 2.5 h of treadmill running at 75.6 +/- 0.9% maximal O2 uptake (VO2max) on natural killer (NK) cell cytotoxic activity (NKCA) was investigated in 22 experienced marathon runners (VO2max 57.9 +/- 1.1 ml.kg-1.min-1, age 38.7 +/- 1.5 yr). Blood samples were taken before (0715) and immediately after exercise (1000), with three more samples taken during 6 h of recovery (1130, 1300, and 1600). Ten sedentary controls (VO2max 34.7 +/- 1.0 ml.kg-1.min-1, age 45.3 +/- 2.3 yr) sat in the laboratory during testing and had their blood sampled at the same time points. The pattern of change in NKCA over time was significantly different between groups [F(4,27) = 6.53; P = 0.001], with the runner's NKCA dropping 51-61% below preexercise levels throughout 6 h of recovery. Preincubation of blood mononuclear cells in vitro with indomethacin had no effect on the difference in pattern of change in NKCA between groups [F(4,17) = 8.59; P = 0.001] and did not attenuate the postexercise reduction in the runners. When NKCA was adjusted on a per-NK cell basis, group differences and the postexercise decline in NKCA were eliminated [F(4,80) = 0.65; P = 0.63]. Serum cortisol and plasma epinephrine in the runners were elevated relative to control subjects during recovery from exercise, but no significant correlation with changes in NK cells or NKCA was found. These data indicate that NKCA is decreased significantly during recovery from 2.5 h of running due to a numerical redistribution of NK cells.

Influence of obesity on immune function.

OBJECTIVE: To compare immune function in obese and nonobese subjects. DESIGN: Obese and nonobese subjects were compared cross-sectionally. To test for the influence of other factors on immunity, aerobic fitness, psychological well-being, and serum levels of glucose, triglycerides, and cholesterol were measured and included in multiple regression models to determine their comparative effects. SUBJECTS/SETTING: Community-based subjects included 116 obese women (age = 44.3 +/- 9.7 years, body mass index = 33.2 +/- 6.5) and 41 nonobese women (age = 42.2 +/- 10.9 years, body mass index = 21.2 +/- 1.9). STATISTICAL ANALYSES PERFORMED: Independent t tests, Pearson product moment correlations, and stepwise multiple regression procedures. RESULTS: Obesity was linked to elevated leukocyte and lymphocyte subset counts (except for natural killer and cytotoxic/suppressor T cells), suppressed mitogen-induced lymphocyte proliferation (an index of T- and B-cell function), higher monocyte and granulocyte phagocytosis and oxidative burst activity, and normal activity of natural killer cells. APPLICATIONS/CONCLUSIONS: These data support the contention that obesity is associated with alterations in immune function. Further research is needed to link immunosuppression with the previously reported elevated risk of infection among the obese.

Influence of mode and carbohydrate on the cytokine response to heavy exertion.

OBJECTIVE AND METHODS: This randomized, double-blind, placebo-controlled study was designed to determine the influence of exercise mode and 6% carbohydrate (C) versus placebo (P) beverage ingestion, on blood cell counts, plasma glucose, hormone, and inflammatory cytokine responses (five total samples over 9 h) to 2.5 h of high-intensity running and cycling (approximately 75% VO2max) by 10 triathletes who acted as their own controls. Statistical significance was set at P < or = 0.05. RESULTS: C relative to P ingestion (but not exercise mode) was associated with higher plasma levels of glucose and insulin, lower plasma cortisol and growth hormone, and diminished perturbation in blood immune cell counts. The pattern of change over time for interleukin (IL)-6 was significantly different between C and P conditions (P = 0.021) and between running and cycling modes (P < 0.001), with the lowest postexercise values seen in the C-cycling sessions (10.7 +/- 1.8 pg x mL(-1)) and the highest in the P-running sessions (51.6 +/- 14.2 pg x mL(-1)). The pattern of change over time between C and P conditions (but not modes) was significantly different for IL-1 receptor antagonist (P = 0.003), with values once again lowest for the C-cycling sessions (1.5 h postexercise, 301 +/- 114 pg x mL(-1)) and highest for the P-running sessions (1171 +/- 439 pg x mL(-1)). CONCLUSION: These data indicate that carbohydrate versus placebo ingestion (4 mL x kg(-1) carbohydrate or placebo every 15 min of the 2.5-h exercise bout) is associated with higher plasma glucose levels, an attenuated cortisol response, and a diminished pro- and anti-inflammatory cytokine response.

Immune response to exercise training and/or energy restriction in obese women.

PURPOSE: The effect of exercise training (five 45-min walking sessions/wk at 60-75% maximum heart rate) and/or moderate energy restriction (4.19-5.44 MJ or 1,200-1,300 kcal x d(-1)) on innate and adaptive immunity (including mitogen-stimulated lymphocyte proliferation (MSLP), natural killer cell activity (NKCA), and monocyte and granulocyte phagocytosis and oxidative burst (MGPOB) was studied in obese women (N = 91, age 45.6 +/- 1.1 yr, body mass index 33.1 +/- 0.6 kg x m(-2)) randomized to one of four groups: control (C), exercise (E), diet (D), exercise, and diet (ED). METHODS: Aerobic power, body composition, and immune function were measured in all subjects before and after a 12-wk diet intervention period, with data analyzed using a 4 x 2 repeated measures design. All subjects self-reported symptoms of sickness in health logs using a precoded checklist. Statistical significance was set at P < or = 0.05. RESULTS: Data from this study indicate that although exercise training was unrelated to any significant changes in resting immune function, the number of days with symptoms of upper respiratory tract infection (URTI) was reduced relative to subjects in the nonexercise groups (5.6 +/- 0.9 and 9.4 +/- 1.1 sickness days, respectively, P < 0.05). Energy restriction and weight loss (7.9 +/- 0.7 kg) was associated with a significant decrease in MSLP, but no change in NKCA, MGPOB, or URTI. CONCLUSION: The data are consistent the viewpoint that weight loss, even at a moderate rate, is associated with a decrease in mitogen-stimulated lymphocyte proliferation without a change in various measures of innate immunity of the blood compartment.

Immune function in marathon runners versus sedentary controls.

Marathon runners (N = 22) who had completed at least seven marathons (X +/- SEM = 23.6 +/- 5.7) and had been training for marathon race events for at least 4 yr (12.3 +/- 1.3) were compared with sedentary controls (N = 18). Although the two groups were of similar age (38.7 +/- 1.5 and 43.9 +/- 2.2 yr, respectively) and height, the marathon runners were significantly leaner and possessed a VO2max 60% higher than that of the controls. Neutrophil counts tended to be lower in the group of marathoners, while other leukocyte and lymphocyte subsets were similar to controls. Mitogen-induced lymphocyte proliferation did not differ between groups. Natural killer cell cytotoxic activity (NKCA) was significantly higher in the marathoners versus controls (373 +/- 38 vs 237 +/- 41 total lytic units, respectively, a 57% difference, P = 0.02). For all subjects combined (N = 40) and within the group of marathon runners (N = 22), percent body fat was negatively correlated with NKCA (r = -0.48, P = 0.002; r = -0.49, P = 0.019, respectively), and age was negatively correlated with Con A-induced lymphocyte proliferation (r = -0.41, P = 0.009; r = -0.53, P = 0.011, respectively). These data indicate that NKCA but not mitogen-induced lymphocyte proliferation is higher in marathon runners relative to sedentary controls.

Carbohydrate affects natural killer cell redistribution but not activity after running.

This randomized, double-blind, placebo-controlled study was designed to determine the influence of carbohydrate supplementation on the natural killer cell response to 2.5 h of high-intensity running (76.7 +/- 0.4% VO2max). Thirty experienced marathon runners (VO2max 53.4 +/- 1.0 mL x kg[-1] x min[-1], age 41.5 +/- 1.4 yr) were randomized into carbohydrate supplement (N = 17) and placebo (N = 13) groups. Subjects rested for 10-15 min before a blood sample at 0715, and then ingested 0.75 L of carbohydrate beverage (Gatorade) or placebo. At 0730, subjects began running at 75-80% VO2max for 2.5 h and drank 0.25 L of carbohydrate or placebo fluid every 15 min. Immediately after the 2.5 h run (1000), another blood sample was taken, followed by 1.5 h, 3 h, and 6-h recovery samples. Carbohydrate supplementation versus placebo had a significant effect on the pattern of change in glucose, cortisol, and the blood concentration of natural killer cells ([F (4,25) = 3.79, P = 0.015], but not natural killer cell activity following 2.5 h of intensive running.

Influence of carbohydrate on cytokine and phagocytic responses to 2 h of rowing.

PURPOSE: This study examined the influence of carbohydrate (C) versus placebo (P) beverage ingestion on the phagocytic and cytokine responses to normal rowing training by 15 elite female rowers. METHODS: Athletes received C or P before, during and after, two, 2-h bouts of rowing performed on consecutive days. Blood was collected before and 5-10 min and 1.5 h after rowing. Metabolic measures indicated that training was performed at moderate intensities, with some high-intensity intervals interspersed throughout the sessions. RESULTS: Concentrations of blood neutrophils and monocytes, phagocytic activity, and plasma IL-1ra were significantly lower postexercise after C versus P ingestion. No differences were observed for oxidative burst activity, IL-6, IL-8, or TNFalpha. Glucose was significantly higher after 2 h of rowing with C ingestion; however, cortisol, growth hormone, epinephrine, norepinephrine, and CRP were not affected by carbohydrate. CONCLUSIONS: These data indicate that carbohydrate compared with placebo ingestion attenuated the moderate rise in blood neutrophils, monocytes, phagocytosis, and plasma IL-1ra concentrations that followed 2-h bouts of training in elite female rowers. No changes in blood hormone concentrations were found.