Impact of bone marrow transplantation on type I diabetes.

Type I diabetes is a systemic autoimmune disease. Evidence is accumulating that autoimmune diseases such as type I diabetes are linked to the bone marrow hematopoietic stem cell (HSC) itself rather than its derivatives. HSC chimerism achieved through bone marrow transplantation (BMT) may affect type I diabetes in two ways: first, to induce tolerance to pancreas and islet cell transplants; and second, to reverse the autoimmune process prior to the development of terminal complications. Transplantation of bone marrow from normal donors into patients with hematologic malignancy and coexistent type I diabetes has reversed the systemic diabetic autoimmune process. Donor HSCs can also be utilized for the induction of donor-specific tolerance to islet cell transplants. Islet or whole pancreas transplantation is the most physiologic approach to treating type I diabetes. Currently, this is limited by the requirement for high-dose chronic nonspecific immunosuppression to prevent rejection. Despite these agents, chronic rejection remains the primary cause for late graft loss. Donor-specific tolerance eliminates the requirement for immunosuppression and prevents the development of chronic rejection. Bone marrow transplantation does have limitations. In particular these limitations include the morbidity associated with lethal conditioning, graft-versus-host disease, and failure of engraftment. Currently the morbidity and mortality associated with lethal conditioning could not be justified for tolerance induction or interruption of the autoimmune state in type I diabetes. The goal of current research is to identify those factors in both recipient and donor that optimize engraftment to reverse the risk/benefit ratio associated with BMT. This article reviews the state of the art for HSC chimerism affecting diabetes.

Alpha beta TCR+ T cells play a nonredundant role in the rejection of heart allografts in mice.

BACKGROUND: Although the transplantation of solid organs and cellular grafts is a clinical routine, the morbidity and mortality associated with immunosuppression is significant. This could be avoided by the induction of donor-specific tolerance. To develop targeted antirejection strategies and regimens to induce donor-specific tolerance, cell populations in the recipient-mediating rejection of solid organ and cellular grafts must be defined. In this study we examined the role of alpha beta-TCR+ cells in the rejection of allogeneic heart grafts, by use of knockout (KO) mice deficient in the production of alpha beta-TCR+ T cells. METHODS: C57BL/6-TcrbtmlMom (alpha beta-KO) and C57BL6/J (B6) recipient mice were transplanted with B10.BR/SgSnJ (B10.BR) or BALB/c heart allografts. Animals also received bone marrow from normal B10.BR donors, followed by donor-specific or third-party heart transplants. RESULTS: Naive B6 control mice rejected B10.BR and BALB/c grafts within 16 days. In striking contrast, B10.BR and BALB/c heart allografts were indefinitely accepted in unmanipulated alpha beta-KO mice. The immune responsiveness was restored after bone marrow transplantation from normal donors. After bone marrow transplantation major histocompatibility-disparate BALB/c third-party heart grafts were rejected, whereas donor-specific grafts were still accepted. CONCLUSIONS: alpha beta-TCR+ T cells play a nonredundant role in the rejection of heart allografts in mice. Bone marrow chimerism is associated with donor-specific transplantation tolerance.

Clinical applications of mixed chimerism.

Bone marrow transplantation (BMT) is currently a procedure that is associated with high morbidity and mortality. Thus, the clinical application of this technique is limited to the treatment of life-threatening hematopoietic malignancies. The morbidity and mortality of BMT is mainly related to graft-versus-host disease (GVHD), failure of engraftment, and toxicity related to fully myeloablative conditioning. GVHD can be prevented by T-cell depletion. However, T-cell depletion increases the risk of failure of engraftment. With the identification of a facilitating cell population that enables engraftment of hematopoietic stem cells across major histocompatibility barriers, the dichotomy between GVHD and failure of engraftment has been resolved. If one could overcome the toxicity of conditioning with the development of partially ablative conditioning strategies, BMT could be used for the treatment of a variety of nonmalignant diseases, as well as in the induction of donor-specific transplantation tolerance. This review outlines the development and advantages of partially ablative conditioning strategies and illustrates possible applications of the technique. Forty years ago E.D. Thomas discussed the potential of BMT for treating immunodeficiencies and for the induction of transplantation tolerance. BMT can be viewed as a natural form of gene therapy to replace a defective cell or enzyme with a functional and normally regulated one.

Effect of FLT3 ligand and granulocyte colony-stimulating factor on expansion and mobilization of facilitating cells and hematopoietic stem cells in mice: kinetics and repopulating potential.

We have previously identified a cellular population in murine bone marrow that facilitates engraftment of highly purified hematopoietic stem cells (HSC) across major histocompatibility complex (MHC) barriers without causing graft-versus-host disease. Here we investigated the effect of flt3 ligand (FL) and granulocyte colony-stimulating factor (G-CSF) on the mobilization of facilitating cells (FC) and HSC into peripheral blood (PB). Mice were injected with FL alone (day 1 to 10), G-CSF alone (day 4 to 10), or both in combination. The number of FC (CD8(+)/alpha betaTCR-/gamma deltaTCR-) and HSC (lineage-/Sca-1(+)/c-kit+) was assessed daily by flow cytometry. Lethally irradiated allogeneic mice were reconstituted with PB mononuclear cells (PBMC). FL and G-CSF showed a highly significant synergy on the mobilization of FC and HSC. The peak efficiency for mobilization of FC (21-fold increase) and HSC (200-fold increase) was reached on day 10. Our data further suggest that the proliferation of FC and HSC induced by FL in addition to the mobilizing effect mediated by G-CSF might be responsible for the observed synergy of both growth factors. Finally, the engraftment potential of PBMC mobilized with FL and G-CSF or FL alone was superior to PBMC obtained from animals treated with G-CSF alone. Experiments comparing the engraftment potential of day 7 and day 10 mobilized PBMC indicate that day 10, during which both FC and HSC reached their maximum, might be the ideal time point for the collection of both populations.

Reproducibility of anthropometric and body composition measurements: the HERITAGE Family Study.

OBJECTIVE: To determine the reproducibility of anthropometric and body composition measures using the HERITAGE Family Study protocol. DESIGN: Anthropometric and body composition measures were obtained on three separate days within a 3-wk period at each of the four HERITAGE Clinical Centers. SUBJECTS: Sixty men and women representative of the HERITAGE subject population, 15 from each of four Clinical Centers. MEASUREMENTS: Anthropometric measures included eight skinfolds, three girths and one length; and body composition measures included stature, mass, hydrostatic weight, residual volume, and body density, from which relative fat, fat mass and fat-free mass were estimated. RESULTS: Reproducibility as determined by technical error, coefficient of variation, and intraclass correlations was very high for the total sample. For example, intraclass correlations for the total sample generally ranged from 0.95-0.99 for the anthropometric measures, and from 0.97-1.00 for the body composition measures. The results across Clinical Centers were in close agreement with each other and with the pooled data. CONCLUSIONS: The reproducibility of anthropometric and body composition measures using the HERITAGE Family Study protocol is sufficiently high that it should be possible to detect small changes in any of these measures and to determine the genetic basis of these changes consequent to a 20 wk endurance training program.

Changes in stroke volume with beta-blockade before and after 10 days of exercise training in men and women.

We sought to determine whether 10 days of training would be a sufficient stimulus for cardiac adaptations that would allow a greater compensatory stroke volume during beta-blockade. We also sought to determine whether men and women had a similar cardiac reserve capacity for increasing stroke volume with beta-blockade during submaximal exercise. Eight men (age 29 +/- 2 yr, mean +/- SE) and eight women (25 +/- 2 yr) cycled at 65% of peak O2 consumption (unblocked) under placebo-control and beta-blockade (100 mg atenolol) conditions performed on separate days. These tests were repeated at the same power output after training (10 consecutive days, 1 h of cycling per day). Before training, beta-blockade significantly (P < 0.05) decreased heart rate (HR) and cardiac output and increased stroke volume in both men and women. After training, the increase in stroke volume and decrease in HR with beta-blockade was significantly less while cardiac output was reduced more. There were no gender differences in the effects of beta-blockade on HR, stroke volume, or cardiac output. These data indicate that, during exercise with beta-blockade, exercise training for 10 days does not enhance the compensatory increase in stroke volume and that men and women have a similar cardiac reserve capacity for increasing stroke volume.

Changes in stroke volume and maximal aerobic capacity with increased blood volume in men women.

This study determined whether the effects of acute plasma volume expansion (PVX) or 10 days of training on stroke volume during submaximal cycling and on treadmill maximal oxygen uptake (VO2max) were similar between men and women. Subjects performed a submaximal cycle test and a treadmill test to exhaustion under three conditions: control, PVX, and after training. Cycle peak VO2 (VO2peak) and blood volume were measured before and after training. Training consisted of daily 1-h bouts [30 min at 80% peak heart rate (HRpeak) and ten 2-min intervals at 95% HRpeak alternating with 1-min low-intensity pedaling] on a cycle ergometer for 10 consecutive days. Training increased cycle VO2peak in men [P < 0.05; 3.14 +/- 0.13 vs. 3.42 +/- 0.13 (SE) l/min] and women (2.11 +/- 0.10 vs. 2.37 +/- 0.12 l/min) and increased blood volume in men (67.6 +/- 3.0 vs. 72.3 +/- 3.1 ml/kg) and women (62.7 +/- 2.2 vs. 65.6 +/- 2.4 ml/kg). As a result of the greater blood volume with PVX and with training, stroke volume (ml) during submaximal cycling increased in men (control 110 +/- 4; PVX 123 +/- 4; trained 121 +/- 4) and women (control 87 +/- 5; PVX 95 +/- 6, trained 96 +/- 7). Treadmill VO2max (ml.kg-1.min-1) did not change with PVX despite a 6-7% reduction in hemoglobin concentration, whereas training resulted in an increase in VO2max in men (control 47.9 +/- 2.8; PVX 46.7 +/- 2.8; trained 49.9 +/- 2.6) and women (control 38.0 +/- 1.2; PVX 36.9 +/- 1.2; trained 39.2 +/- 1.2). The effects of PVX or training on stroke volume or VO2max did not differ between men and women. An additional finding was an increase in diastolic and mean blood pressures at 65% of cycle VO2peak with PVX and with training in women. Thus men and women hold similar cardiac reserve capacities for increasing stroke volume and, as a result, VO2max is maintained despite a reduction in hemoglobin concentration. However, gender differences in blood pressure regulation with increased blood volume might exist.