BALB/c mice immunized by intraperitoneal injection of purified Pseudomonas aeruginosa lectin preparations are fully protected against a lethal dose of the live bacteria. Intraperitoneal inoculation of splenocytes or bone marrow cells obtained from actively immunized mice into naive syngeneic mice was shown to significantly increase their resistance to P. aeruginosa infection. A similar transfer of splenocytes or bone marrow cells from untreated control mice did not provide protection against a lethal Pseudomonas challenge. Administration of immunocompetent cells from immunized animals to naive mice by the i.v. route was less efficient than the i.p. inoculation, probably due to different homing of the cells. Only the i.p. injection concentrated the immune cells to the site of infection.

Animals↗

PubMedDate not supplied

Effects of human interleukin 3, macrophage and granulocyte-macrophage colony-stimulating factor on monocyte function following autologous bone marrow transplantation.

The effects of human interleukin 3 (IL-3), macrophage colony-stimulating factor (M-CSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) were studied on the functional activity of human peripheral blood monocytes from healthy individuals and from eight patients at 4, 8 and 12 weeks following autologous bone marrow transplantation (ABMT). Functions studied included superoxide production, phagocytosis of Candida albicans and reduction of 3-[4,5-dimethylthiazol-2-yl]-2.5-diphenyl tetrazolium bromide (MTT). IL-3 and GM-CSF significantly enhanced the oxidative metabolism of monocytes from healthy individuals, while the effect of M-CSF was moderate. A considerable variability between healthy individuals was found in both resting and cytokine-stimulated monocytes with regard to superoxide production. All three investigated CSFs, i.e. IL-3, M-CSF and GM-CSF did not affect phagocytosis of C. albicans by the cells or their metabolic activity (reduction of MTT). In ABMT patients no deficit in the functional activity of monocytes was found at any time after transplantation and all three CSFs investigated did not modulate the functional activity of the cells. These results suggest that monocytes do not have a major role in infectious complications post-ABMT.

Adolescent↗

PubMedDate not supplied

Failure of cyclosporine to induce graft-vs-host disease or graft-vs-leukemia after syngeneic bone marrow transplantation in mice.

Cyclosporine administration can result in graft-vs-host disease (GVHD) after syngeneic or autologous bone marrow transplantation (BMT). However, data on its anti-tumor effects are limited. We have tried to produce cyclosporine-induced GVHD or graft-vs-leukemia (GVL) against two Ia-bearing murine leukemias. BALB/c mice undergoing syngeneic BMT after total-body irradiation received 1 mg/kg or 10 mg/kg cyclosporine or dextrose intraperitoneally for 30 days post-transplant, followed by 5 x 10(3) BCL1 murine leukemia cells 1 or 3 weeks after stopping cyclosporine. Similarly, SJL/J mice undergoing syngeneic BMT received 10 mg/kg cyclosporine or dextrose intraperitoneally for 30 days post-transplant, and were inoculated with 5 x 10(3) or 10(5) murine acute myeloid leukemia (mAML) cells 3 weeks after stopping cyclosporine. No clinical or histological evidence of GVHD was seen, and leukemia-free and overall survival was similar with cyclosporine and dextrose. We conclude that under the experimental conditions used, it is not possible to induce syngeneic GVHD with cyclosporine in BALB/c or SJL/J mice. In the absence of GVHD, this approach is also not associated with any GVL effect against murine leukemias BCL1 and mAML.

Animals↗

PubMedDate not supplied

Non-myeloablative allogeneic stem cell transplantation focusing on immunotherapy of life-threatening malignant and non-malignant diseases.

Allogeneic bone marrow transplantation (BMT) represents an important therapeutic tool for treatment of otherwise incurable malignant and non-malignant diseases. Until recently, myeloablative regimens were considered mandatory for eradication of all undesirable host-derived hematopoietic elements. Our preclinical and ongoing clinical studies indicated that much more effective eradication of host immunohematopoietic system cells could be achieved by adoptive allogeneic cell therapy with donor lymphocyte infusion (DLI) following BMT. Thus, eradication of blood cancer cells, especially in patients with CML can be frequently accomplished despite complete resistance of such tumor cells to maximally tolerated doses of chemoradiotherapy. Our cumulative experience suggested that graft versus leukemia (GVL) effects might be a useful tool for eradication of otherwise resistant tumor cells of host origin. The latter working hypothesis suggested that effective BMT procedures may be accomplished without lethal conditioning of the host, using new well tolerated non-myeloablative regimen, thus possibly minimizing immediate and late side effects related to myeloablative procedures considered until recently mandatory for conditioning of BMT recipients. Recent clinical data that will be presented suggests that safe non-myeloablative stem cell transplantation (NST), with no major toxicity can replace the conventional BMT. Thus, NST may provide an option for cure for a large spectrum of clinical indications in children and elderly individuals without lower or upper age limit, while minimizing procedure-related toxicity and mortality.

Acute Disease↗

PubMedDate not supplied

Donor lymphocyte infusions to displace residual host hematopoietic cells after allogeneic bone marrow transplantation for beta-thalassemia major.

PURPOSE: Donor lymphocyte infusion (DLI) was used to reverse relapse after allogeneic bone marrow transplantation (BMT) in a patient with beta-thalassemia major. PATIENTS AND METHODS: The patient with unstable mixed chimerism after BMT was treated with graded increments of donor lymphocytes (10(5) T cells/kg to 5 x 10(7) T cells/kg) to displace residual hematopoietic host cells. RESULTS: DLI resulted in complete donor-derived reconstitution of the hematopoietic compartment. The patient developed mild graft-versus-host disease (GVHD) that could be controlled by steroid treatment. CONCLUSIONS: This case report shows that DLI can effectively eradicate host stem cells in mixed chimeras after BMT in nonmalignant hematopoietic diseases.

Bone Marrow Transplantation↗

PubMedDate not supplied

Fludarabine-based protocol for human umbilical cord blood transplantation in children with Fanconi anemia.

PURPOSE: A novel conditioning regimen of fludarabine monophosphate (FLM), anti-T-lymphocyte globulin (ATG), and low-dose cyclophosphamide with no irradiation for human umbilical cord blood transplantation (HUCBT) for the treatment of Fanconi anemia (FA) is described. PATIENT AND METHODS: A 12-year-old girl with FA received a human umbilical cord blood transplant from a fully matched sibling donor. After the HUCBT, the patient was given granulocyte colony stimulating factor in combination with erythropoietin. Pretransplant conditioning consisted of FLM (30 mg/m2/d) from day -10 to day -5, cyclophosphamide (10 mg/kg/d) on day -7 and -6, and rabbit ATG (ATG-Frasenius, 10 mg/kg/d) from day -4 to day -1. Cyclosporin A (3 mg/kg/d) was administered from day -1 as graft-versus-host disease prophylaxis. Cord blood from a sibling donor was used as a source of hematopoietic stem cells. RESULTS: Engraftment was normal and sustained. The regimen was well tolerated with very mild toxicity and no major transplant-related complications or >grade II graft-versus-host disease. Chimerism was 100% donor origin as determined by restriction fragment length polymorphism. CONCLUSIONS: It is possible to achieve sustained engraftment and only mild toxicity in FA after HUCBT with a conditioning regimen of FLM, ATG, and cyclophosphamide with no irradiation. These preliminary results with this novel conditioning protocol are encouraging and should be evaluated in a larger group of patients with FA undergoing HUCBT.

Child↗

PubMedDate not supplied

Successful treatment of diabetes in NOD mice with advanced disease by islet isografts following immunoregulation with Linomide (quinoline-3-carboxamide).

We have recently documented that oral Linomide (quinoline-3-carboxamide) prevents autoimmune insulitis, islet destruction, and diabetes in NOD mice treated at an early stage (5 wk of age) of the disease. In this report, we show that treatment of female NOD mice with advanced disease (age 23-24 wk) by syngeneic islet transplantation and oral Linomide administration results in prevention of graft insulitis and diabetes in the Linomide group up to 40 wk (diabetes at 40 wk: isograft recipients with Linomide n = 0 of 6; isograft recipients alone n = 5 of 6; p < 0.0001). The extent of protection from glucose intolerance by the combination of transplantation with Linomide was superior to that of Linomide alone [blood glucose (mean +/- SD) 60 min post-i.p. injection of 1 g/kg body weight glucose: Linomide plus isograft 6.7 +/- 1 mmol/L; Linomide alone 18.7 +/- 6.3 mmol/L; p < 0.0001]. Thus, Linomide should be considered a potential immunoregulatory modality in patients undergoing pancreatic islet or whole organ transplantation.

Adjuvants, Immunologic↗

PubMedDate not supplied

Cell therapy with preimmunized effector cells mismatched for minor histocompatible antigens in the treatment of a murine mammary carcinoma.

Cell therapy with allogeneic donor cells mismatched for minor histocompatible (MiHC) antigens was applied to a murine mammary carcinoma (4T1) model to test the feasibility of graft versus tumor (GVT) effect against metastatic epithelial tumor cells. BALB/c mice bearing a 4T1 tumor of BALB/c origin were given syngeneic or MiHC-mismatched splenocytes. GVT effects were determined in secondary recipients of adoptively transferred lung cells derived from primary hosts who had previously been inoculated intravenously with 4T1 cells, and injected with one of the following: 1) naive BALB/c splenocytes, 2) naive DBA/2 splenocytes, 3) 4T1-immune DBA/2 splenocytes, or 4) DBA/2 splenocytes immunized with host-derived BABL/c spleen cells. Naive DBA/2 splenocytes inhibited tumor growth only slightly and only slightly prolonged the survival of secondary recipients, in comparison with fully matched tumor/host BALB/c spleen cells. An efficient GVT reaction was demonstrated in vitro and in vivo with MiHC-mismatched DBA/2 splenocytes from mice presensitized by multiple injections of irradiated tumor or BALB/c-derived spleen cells. All 30 mice adoptively inoculated with lung cells from primary hosts that had previously been treated with these presensitized effector cells were tumor free for >250 days. Secondary recipients inoculated with lung cells from mice given naive BALB/c or DBA/2 spleen cells died of metastatic tumors within 33 to 46 days. These results suggest that preimmunized donor cells represent an effective tool against metastatic disease; hence, the next goal should be to control graft-versus-host disease while exploiting the GVT potential.

Animals↗

PubMedDate not supplied

T-cell depletion in allogeneic bone marrow transplantation: progress and problems.

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Animals↗

PubMedDate not supplied

Combination of magnetic and chemocytotoxic cancer cell depletion for autologous bone marrow transplantation.

An artificial mixture of breast cancer cell line cells (T-47D) and normal human marrow cells was used to investigate optimal approaches for autologous BMT. The experiments were designed to test the applicability of chemocytotoxic agents, a dye-mediated photolytic agent and SBA for in vitro purging in autologous BMT of patients with advanced malignancies. Treatment with high concentration of etoposide (VP-16) (10 to 80 micrograms/ml) resulted in a maximal depletion of 1.5 log, whereas more efficient tumor cell eradication (2.5 log) was achieved by 30-min incubation with 100 micrograms/ml4HC. Photosensitization by exposure for 90 min to daylight in the presence of MC-540 could remove only 1 log of T-47D cells. The chemocytotoxic treatment with 4HC was chosen to follow initial tumor cell separation by SBA bound to polystyrene magnetic beads that had previously been shown to bind to several cancer cell types while sparing marrow progenitor cells. Artificial mixtures containing 10 to 14% T-47D cells in fresh normal BM cells were subjected to SBA-magnetic beads, and the SBA-negative fraction was further treated with 4HC. The combined two-step procedure resulted in a consistent tumor cell depletion of greater than 4 logs. The purging procedure appears acceptable for clinical marrow purging prior to cyropreservation and autotransplantation in patients with documented BM involvement of neoplastic cells sensitive to 4HC with positive binding to SBA such as breast cancer.

Bone Marrow Transplantation↗

PubMedDate not supplied

New approaches for the prevention of rejection and graft-vs.-host disease in clinical bone marrow transplantation.

The two major barriers to successful allogeneic bone marrow transplantation (BMT) in animals and man are graft-vs.-host disease (GVHD) and the risk of graft rejection. GVHD is the result of alloreactivity of mature donor T-lymphocytes present in the graft-vs.-host tissues and can be completely prevented by pregraft depletion of T-lymphocytes. Graft rejection results from residual host immunocompetent lymphocytes that survive heavy chemoradiotherapy prior to allogeneic BMT. Host resistance to allograft cannot be eradicated even by conventional conditioning with high-dose cyclophosphamide (120 mg/kg) and lethal whole body irradiation (1,200 rad). In the present report we have utilized two new techniques to overcome GVHD and graft rejection following allogeneic BMT. GVHD can be prevented by a new monoclonal rat antihuman lymphocyte antibody, CAMPATH-1, which binds human complement, enabling donor serum to serve as the source of complement. Prevention of rejection of T-lymphocyte-depleted marrow allografts can be achieved by the application of total lymphoid irradiation (TLI) in addition to conventional chemoradiotherapy, prior to allogeneic BMT. TLI causes potent immunosuppression with minimal side effects. A combination of TLI for overcoming host resistance to allograft, and CAMPATH-1 for overcoming GVHD, leads to a relatively smooth posttransplant outcome with no evidence of GVHD and with no need for posttransplant immunosuppression.

Antilymphocyte Serum↗