Background and objectives: Busulfan is the most commonly used myeloablative alkylating agent, but... more Background and objectives: Busulfan is the most commonly used myeloablative alkylating agent, but is considered a poor anti-lymphocyte agent. Since engraftment of allogeneic stem cells depends not only on adequate immunosuppression but also on successful hematopoietic competition, and considering the fact that residual lymphocytes of host origin may play a beneficial role in preventing graft-versus-host disease (GVHD), we used low doses of oral busulfan as a single agent for conditioning prior to stem cell transplantation (SCT) in recipients of transplants from a variety of donors. Design and methods: Fifteen heavily pretreated high-risk patients (age 25-66, median 42 years) with hematologic malignancies were conditioned with busulfan alone, 4mg/kg/day for 2, 3, or 4 consecutive days. No additional pre- or post-transplant immunosuppressive agents were used in order to exploit the capacity of donor lymphocytes to induce graft-versus-malignancy (GVM) effects. Results: Conditioning was well tolerated, trilineage engraftment was documented in all patients and none exhibited immune-mediated rejection. Time to recovery of absolute neutrophil count >0.5x10(9)/L and 1.0x10(9)/L was 12 - 38 (median 15) days and 12 - 41 (median 15) days, respectively. The time to platelet recovery >or=20 and >or=50x10(9)/L ranged from 0 to 26 (median 11) days, and from 0 to 83 (median 14) days, respectively. Acute GVHD (<or=grade I) occurred in 13/15 patients. Three patients benefited from long-term survival. Interpretation and conclusions: We suggest that using busulfan alone for the preparation of patients for SCT may be sufficient for engraftment, in very high-risk heavily pre-treated patients.
N-acetylcysteine (NAC) is a known antioxidant and induces modulation of glutathione cellular cont... more N-acetylcysteine (NAC) is a known antioxidant and induces modulation of glutathione cellular content effects. It has been suggested that in the context of stem cell transplantation (SCT), NAC can prevent and treat graft-vs.-host disease, veno-occlusive disease and idiopathic pneumonia syndrome. We investigated the possible effect of NAC on graft-vs.-leukemia effect (GVL) and lymphokine activated cells (LAK) activity in murine models. After 10 days of NAC treatment, the cytotoxic activity of the LAK cells did not significantly differ from LAK activity generated from spleen cells obtained from untreated controls. However, NAC mildly suppressed GVL (appearance of leukemia in 8/36 animals treated with NAC as compared to 0/20 in the SCT control group, p=0.023). In spite of this mild suppression of GVL, no negative effect on achievement of donor chimerism was seen. We conclude that NAC usage in SCT may be relatively safe with regard to the GVL effect, yet further clinical studies are warranted.
Persistent influenza virus replication during antiviral therapy in patients undergoing hematopoie... more Persistent influenza virus replication during antiviral therapy in patients undergoing hematopoietic stem cell transplantation (HSCT) could promote the emergence of antiviral drug resistance. To follow the viral genotypic and drug susceptibility changes in a patient who developed progressive influenza A/H3N2 pneumonia despite oseltamivir therapy after haploidentical HSCT. Direct genotypic analysis of the neuraminidase (NA) and hemagglutinin (HA) genes in successive bronchoalveolar lavage specimens was employed in combination with hemagglutination and NA enzymatic activity assays of the corresponding viral isolates. The emergence of NA oseltamivir-resistance mutation R292K was detected by 12 days of oseltamivir treatment with 44,286-fold increase in oseltamivir IC50. Resurgence of wild type viral population was identified by 7 days after cessation of oseltamivir. Sequential HA mutations R228S and A138S were identified and associated with a shift in the HA receptor binding pattern reflected by loss of the ability to agglutinate chicken erythrocytes. These rapid evolutionary changes warrant close virologic monitoring of immunocompromised patients treated for influenza infection, and raise concern about the efficacy of mono-drug therapy for influenza-associated disease in HSCT recipients.
Critical Reviews in Oncology Hematology, Oct 1, 2007
The development of reduced intensity or non-myeloablative conditioning (NST) in preparation for a... more The development of reduced intensity or non-myeloablative conditioning (NST) in preparation for allogeneic stem cell transplantation (SCT) revolutionized the field and led to reconsideration of the dogma of upper age limit that was set up by the transplant centers as an eligibility parameter. Analysis of the literature data showed that NST regimens are associated with decreased transplant related mortality, and graft-versus-host disease, in comparison with standard myeloablative conditioning, in patients above the age of 50-55 years, or in younger patients with significant comorbidities. However we have to mention, that our considerations are based on the retrospective analysis of the literature data, and that well controlled prospective randomized studies are needed in order to definitely assess the role of NST. Comorbidity indices might be proved as the most important parameters for the choice of the most proper regimen for each patient in need and should be included in future trials.
Recent studies suggest that myocardial administration of stem cells improves perfusion and functi... more Recent studies suggest that myocardial administration of stem cells improves perfusion and function of ischemic myocardium. The present study evaluated the safety and efficacy of simple intracoronary administration of mononuclear autologous bone marrow (BM) cells in patients with ischemic cardiomyopathy without revascularization option. We enrolled 6 consecutive patients with ischemic cardiomyopathy, who were in New York Heart Association classes III to IV despite optimal medical treatment without revascularization options and who, on dobutamine stress echocardiograph (DSE), were found to have left ventricular ejection fraction &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 35% with significant hibernation and ischemia in at least 2 myocardial segments. BM cell suspension was collected, and on the next day, during coronary angiography, mild ischemia was induced by a short balloon inflation in each coronary conduit with a TIMI flow of &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt; or = 2 followed by slow infusion of up to 50 mL of BM cells suspension to each conduit. At baseline and 4 months&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; follow-up, patients underwent clinical evaluation, Holter monitoring, and DSE. BM infusion was successful in all patients. One patient developed postprocedure hypotension and troponin increase. At 4 months&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; follow-up New York Heart Association class improved from 3.5 +/- 0.5 to 2.3 +/- 1.0, P = .04, and resting ejection fraction improved from 25% +/- 7% to 28% +/- 8%, P = .055. We observed improvement in resting wall motion score only in the segments with hibernation in baseline DSE (2.3 +/- 0.5 to 2.0 +/- 0.6, P = .03) and improvement in high-dose dobutamine wall motion score, only in segments showing significant ischemia at baseline DSE (2.5 +/- 0.5 to 2.0 +/- 0.6, P = .001). There were no clinical arrhythmias or increased arrhythmia burden by Holter monitoring. In patients with severe symptomatic ischemic cardiomyopathy, mild induction of ischemia followed by intracoronary infusion of unmanipulated autologous BM is feasible and safe and may improve hibernation and ischemia.
Although allogenenic stem cell transplantation may provide a cure for a growing number of patient... more Although allogenenic stem cell transplantation may provide a cure for a growing number of patients with hematologic malignancies and several metastatic solid tumors, several problems remain to be solved. In routine medical practice transplant can be offered for patients with a matched donor available whereas the large majority of patients in need have no matched donor available. Although alloreactive lymphocytes may eliminate residual malignant cells, such an effect is accompanied by acute and chronic GVHD which may be hazardous even in recipients with perfectly matched allografts, and prohibitive in recipients treated with haploidentically mismatched allografts. On the other hand immunotherapy with intentionally mismatched allografts could provide a much more effective tool for eradication of tumor cells resistant to chemotherapy. We have pioneered a new approach for treatment of patients with resistant hematological malignancies (AML/MDS 5; ALL 1; Biphenotype 2; NHL 3; HD 1) using matched siblings (n=4), matched unrelated donor (n=1) or haploidentically mismatched donors (n=7). Prevention of rejection of mismatched allografts was accomplished by combination of fludarabine and deletion of donor reactive host lymphocytes by infusion of donor mononuclear blood cells and elimination of alloreactive lymphocytes susceptible to high-dose cyclosphosphamide (60mg/kgx3) one day later. Prevention of GVHD following infusion of G-CSF mobilized, haploidentically mismatched blood stem cells was accomplished using Miltenyi’s immunomagentic beads coupled with anti-AC133 (n=6) or using anti-CD3 (n=1). No other anti-GVHD prophylaxis was used. Following transplantation, patients were treated with rIL-2 activated donor peripheral blood lymphocytes activated for 4 days at 37°C in 5% C02 in air incubator with rIL-2 6,000 IU/ml. T cell depletion was accomplished either by positive selection of CD56+ (n=10) or negative selection of CD3 (n=2) for optimal induction of graft vs leukemia (GVL) effects by mismatched and fully activated NK cells. One patient with resistant leukemia became disease free for 8 months but died of resistant aspergilosis which was evident prior to transplantation. Five out of 12 patients with intractable and fully resistant leukemia are alive with no GVHD and no evidence of disease 1–18 (median 13) months post transplantation. Based on our ongoing preliminary study we conclude that patients with resistant hematological malignancies may benefit from cell therapy mediated by rIL-2 activated donor lymphocytes, and most likely from intentionally mismatched haploidentical allografts following elimination of host anti-donor alloreactive lymphocytes and prevention of GVHD by positively or negatively selected stem cells, followed by immunotherapy with rIL-2 activated CD3 depleted NK cells. Intentionally mismatched rIL-2 activated NK cells represents a safe approach for elimination of residual tumor cells, aiming for induction of GVL while avoiding GVHD.
The conditioning prior to allogeneic stem cell transplantation was originally designed as a myelo... more The conditioning prior to allogeneic stem cell transplantation was originally designed as a myeloablative conditioning, designed to eliminate malignant or genetically abnormal cells and then use the transplant procedure for rescue of the patients or to replace missing bone marrow products. However, allografts can induce effective graft vs. malignancy effects and can also eliminate undesirable hematopoietic stem cells in patients with genetic disorders and autoimmune diseases, thus documenting that alloreactive effects mediated by donor lymphocytes post-grafting can play a major role in eliminating hematopoietic cell of host origin, as well as provide effective immunotherapy for the treatment of disease recurrence. The efficacy of donor lymphocyte infusion (DLI) could be improved by activation with rIL-2 or by donor immunization. The cumulative experience over the years suggesting that alloreactive donor lymphocytes were most effective in eliminating tumor cells of host origin resulted in an attempt to reduce the intensity of the conditioning in preparation for the transplant procedure used for the treatment of hematological and other malignancies as well as life-threatening non-malignant disorders for which allogeneic stem cell transplantation may be indicated. Our working hypothesis proposed that the myeloablative conditioning which is hazardous and may be associated with early and late side effects, may not be required for treatment of patients with any indication for allogeneic stem cell transplantation. Instead, nonmyeloablative conditioning based on the use of reduced intensive preparatory regimen, also known as nonmyeloablative stem cell transplantation, may be sufficient for engraftment of donor stem cells while avoiding procedure-related toxicity and mortality, followed by elimination of undesirable cells of host origin by post-transplant effects mediated by alloreactive donor lymphocytes infused along with donor stem cells or administered subsequently as DLI. Improvement of the immediate outcome of stem cell transplantation using NST due to a significant decrease in transplant related mortality has broadened the spectrum of patients eligible for allogeneic stem cell transplantation, including elderly patients and other patients with less than optimal performance status. Likewise, the safer use of stem cell transplantation prompted expanding the scope of potential indications for allogeneic stem cell transplantation, such as metastatic solid tumors and autoimmune disorders, which now are slowly becoming much more acceptable. Current strategies focus on the need to improve the capacity of donor lymphocytes to eliminate undesirable malignant and non-malignant hematopoietic cells of host origin, replacing abnormal or malignant stem cells or their products with normal hematopoietic stem cells of donor origin, while minimizing procedure-related toxicity and mortality and improving the quality of life by reducing the incidence and severity of hazardous acute and chronic GVHD.
Introduction: Prolonged treatment of graft vs. host disease (GVHD) is extremely immunosuppressive... more Introduction: Prolonged treatment of graft vs. host disease (GVHD) is extremely immunosuppressive. Local therapy with intra-arterial (IA) injection of steroids may induce remission with lower extent of systemic immune suppression. Here, we present our experience with IA treatment of gastrointestinal (GI) and/or hepatic steroid resistant/dependent GVHD with 2 consecutive protocols. Patients and methods: Thirty five patients (37 GVHD events (hepatic, n=15), (GI, n=16), (combined, n=6)) were treated with 53 IA sessions. Most side effects were minor. Results: We found that IA steroid therapy was associated with partial and complete remission among patients with steroid resistant/dependent hepatic or GI GVHD. Hepatic partial response was observed in 14 (66.6%) patients among whom 7 (33.3%) reached complete response. GI partial response was observed in 19 (86.4%) patients among whom 12 (54.4%) reached complete response. Early administration of the local therapy, female gender, myeloid basic disease, and a non-active status of the basic disease at the day of transplantation were found related for predicting a better response for the intra-arterial treatment. The use of high dose steroids in the hepatic IA protocol from was at least as good as intermediate dose steroids with methotrexate (table 1, figure 1) and may be safer. Conclusions: Intra-arterial catheter guided steroid therapy is safe and effective in steroid resistant/dependent GVHD. Hepatic artery treatment with methotrexate can be safely substituted with high dose IA methylprednisolone. Further research is warranted characterizing the patients benefit most. Table 1 - comparison between 1st and 2nd hepatic IA treatment protocols 1st protocol 2nd protocol Significance Median age (range) 25 years (7–42) 32 years (18–59) P=0.09 Sex (M:F) 6:1 8:5 NS family donor vs MUD 6:1 9:4 NS Median time in days SCT-GVHD (range) 27 (13–133) 45 (13–248) NS Median time in days GVHD-IA (range) 15 (6–218) 190 (12–2615) P=0.09 Median peak GVHD grade (range) 3 (3–4) 3 (2–4) NS Highest pre-IA treatment bilirubin level (in mmol/L; normal&amp;amp;amp;amp;lt;17) (range) 186 (138–321) 225.5 (83–672) NS Median time to initial response in days (range) 14.5 (4–100) 8 (1–31) P=0.073 Median time to complete response in days (range) 130.5 (35–226) 49 (17–80) Figure 1A, Figure 1B Figure 1A, Figure 1B.
Background and objectives: Busulfan is the most commonly used myeloablative alkylating agent, but... more Background and objectives: Busulfan is the most commonly used myeloablative alkylating agent, but is considered a poor anti-lymphocyte agent. Since engraftment of allogeneic stem cells depends not only on adequate immunosuppression but also on successful hematopoietic competition, and considering the fact that residual lymphocytes of host origin may play a beneficial role in preventing graft-versus-host disease (GVHD), we used low doses of oral busulfan as a single agent for conditioning prior to stem cell transplantation (SCT) in recipients of transplants from a variety of donors. Design and methods: Fifteen heavily pretreated high-risk patients (age 25-66, median 42 years) with hematologic malignancies were conditioned with busulfan alone, 4mg/kg/day for 2, 3, or 4 consecutive days. No additional pre- or post-transplant immunosuppressive agents were used in order to exploit the capacity of donor lymphocytes to induce graft-versus-malignancy (GVM) effects. Results: Conditioning was well tolerated, trilineage engraftment was documented in all patients and none exhibited immune-mediated rejection. Time to recovery of absolute neutrophil count >0.5x10(9)/L and 1.0x10(9)/L was 12 - 38 (median 15) days and 12 - 41 (median 15) days, respectively. The time to platelet recovery >or=20 and >or=50x10(9)/L ranged from 0 to 26 (median 11) days, and from 0 to 83 (median 14) days, respectively. Acute GVHD (<or=grade I) occurred in 13/15 patients. Three patients benefited from long-term survival. Interpretation and conclusions: We suggest that using busulfan alone for the preparation of patients for SCT may be sufficient for engraftment, in very high-risk heavily pre-treated patients.
N-acetylcysteine (NAC) is a known antioxidant and induces modulation of glutathione cellular cont... more N-acetylcysteine (NAC) is a known antioxidant and induces modulation of glutathione cellular content effects. It has been suggested that in the context of stem cell transplantation (SCT), NAC can prevent and treat graft-vs.-host disease, veno-occlusive disease and idiopathic pneumonia syndrome. We investigated the possible effect of NAC on graft-vs.-leukemia effect (GVL) and lymphokine activated cells (LAK) activity in murine models. After 10 days of NAC treatment, the cytotoxic activity of the LAK cells did not significantly differ from LAK activity generated from spleen cells obtained from untreated controls. However, NAC mildly suppressed GVL (appearance of leukemia in 8/36 animals treated with NAC as compared to 0/20 in the SCT control group, p=0.023). In spite of this mild suppression of GVL, no negative effect on achievement of donor chimerism was seen. We conclude that NAC usage in SCT may be relatively safe with regard to the GVL effect, yet further clinical studies are warranted.
Persistent influenza virus replication during antiviral therapy in patients undergoing hematopoie... more Persistent influenza virus replication during antiviral therapy in patients undergoing hematopoietic stem cell transplantation (HSCT) could promote the emergence of antiviral drug resistance. To follow the viral genotypic and drug susceptibility changes in a patient who developed progressive influenza A/H3N2 pneumonia despite oseltamivir therapy after haploidentical HSCT. Direct genotypic analysis of the neuraminidase (NA) and hemagglutinin (HA) genes in successive bronchoalveolar lavage specimens was employed in combination with hemagglutination and NA enzymatic activity assays of the corresponding viral isolates. The emergence of NA oseltamivir-resistance mutation R292K was detected by 12 days of oseltamivir treatment with 44,286-fold increase in oseltamivir IC50. Resurgence of wild type viral population was identified by 7 days after cessation of oseltamivir. Sequential HA mutations R228S and A138S were identified and associated with a shift in the HA receptor binding pattern reflected by loss of the ability to agglutinate chicken erythrocytes. These rapid evolutionary changes warrant close virologic monitoring of immunocompromised patients treated for influenza infection, and raise concern about the efficacy of mono-drug therapy for influenza-associated disease in HSCT recipients.
Critical Reviews in Oncology Hematology, Oct 1, 2007
The development of reduced intensity or non-myeloablative conditioning (NST) in preparation for a... more The development of reduced intensity or non-myeloablative conditioning (NST) in preparation for allogeneic stem cell transplantation (SCT) revolutionized the field and led to reconsideration of the dogma of upper age limit that was set up by the transplant centers as an eligibility parameter. Analysis of the literature data showed that NST regimens are associated with decreased transplant related mortality, and graft-versus-host disease, in comparison with standard myeloablative conditioning, in patients above the age of 50-55 years, or in younger patients with significant comorbidities. However we have to mention, that our considerations are based on the retrospective analysis of the literature data, and that well controlled prospective randomized studies are needed in order to definitely assess the role of NST. Comorbidity indices might be proved as the most important parameters for the choice of the most proper regimen for each patient in need and should be included in future trials.
Recent studies suggest that myocardial administration of stem cells improves perfusion and functi... more Recent studies suggest that myocardial administration of stem cells improves perfusion and function of ischemic myocardium. The present study evaluated the safety and efficacy of simple intracoronary administration of mononuclear autologous bone marrow (BM) cells in patients with ischemic cardiomyopathy without revascularization option. We enrolled 6 consecutive patients with ischemic cardiomyopathy, who were in New York Heart Association classes III to IV despite optimal medical treatment without revascularization options and who, on dobutamine stress echocardiograph (DSE), were found to have left ventricular ejection fraction &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 35% with significant hibernation and ischemia in at least 2 myocardial segments. BM cell suspension was collected, and on the next day, during coronary angiography, mild ischemia was induced by a short balloon inflation in each coronary conduit with a TIMI flow of &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt; or = 2 followed by slow infusion of up to 50 mL of BM cells suspension to each conduit. At baseline and 4 months&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; follow-up, patients underwent clinical evaluation, Holter monitoring, and DSE. BM infusion was successful in all patients. One patient developed postprocedure hypotension and troponin increase. At 4 months&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; follow-up New York Heart Association class improved from 3.5 +/- 0.5 to 2.3 +/- 1.0, P = .04, and resting ejection fraction improved from 25% +/- 7% to 28% +/- 8%, P = .055. We observed improvement in resting wall motion score only in the segments with hibernation in baseline DSE (2.3 +/- 0.5 to 2.0 +/- 0.6, P = .03) and improvement in high-dose dobutamine wall motion score, only in segments showing significant ischemia at baseline DSE (2.5 +/- 0.5 to 2.0 +/- 0.6, P = .001). There were no clinical arrhythmias or increased arrhythmia burden by Holter monitoring. In patients with severe symptomatic ischemic cardiomyopathy, mild induction of ischemia followed by intracoronary infusion of unmanipulated autologous BM is feasible and safe and may improve hibernation and ischemia.
Although allogenenic stem cell transplantation may provide a cure for a growing number of patient... more Although allogenenic stem cell transplantation may provide a cure for a growing number of patients with hematologic malignancies and several metastatic solid tumors, several problems remain to be solved. In routine medical practice transplant can be offered for patients with a matched donor available whereas the large majority of patients in need have no matched donor available. Although alloreactive lymphocytes may eliminate residual malignant cells, such an effect is accompanied by acute and chronic GVHD which may be hazardous even in recipients with perfectly matched allografts, and prohibitive in recipients treated with haploidentically mismatched allografts. On the other hand immunotherapy with intentionally mismatched allografts could provide a much more effective tool for eradication of tumor cells resistant to chemotherapy. We have pioneered a new approach for treatment of patients with resistant hematological malignancies (AML/MDS 5; ALL 1; Biphenotype 2; NHL 3; HD 1) using matched siblings (n=4), matched unrelated donor (n=1) or haploidentically mismatched donors (n=7). Prevention of rejection of mismatched allografts was accomplished by combination of fludarabine and deletion of donor reactive host lymphocytes by infusion of donor mononuclear blood cells and elimination of alloreactive lymphocytes susceptible to high-dose cyclosphosphamide (60mg/kgx3) one day later. Prevention of GVHD following infusion of G-CSF mobilized, haploidentically mismatched blood stem cells was accomplished using Miltenyi’s immunomagentic beads coupled with anti-AC133 (n=6) or using anti-CD3 (n=1). No other anti-GVHD prophylaxis was used. Following transplantation, patients were treated with rIL-2 activated donor peripheral blood lymphocytes activated for 4 days at 37°C in 5% C02 in air incubator with rIL-2 6,000 IU/ml. T cell depletion was accomplished either by positive selection of CD56+ (n=10) or negative selection of CD3 (n=2) for optimal induction of graft vs leukemia (GVL) effects by mismatched and fully activated NK cells. One patient with resistant leukemia became disease free for 8 months but died of resistant aspergilosis which was evident prior to transplantation. Five out of 12 patients with intractable and fully resistant leukemia are alive with no GVHD and no evidence of disease 1–18 (median 13) months post transplantation. Based on our ongoing preliminary study we conclude that patients with resistant hematological malignancies may benefit from cell therapy mediated by rIL-2 activated donor lymphocytes, and most likely from intentionally mismatched haploidentical allografts following elimination of host anti-donor alloreactive lymphocytes and prevention of GVHD by positively or negatively selected stem cells, followed by immunotherapy with rIL-2 activated CD3 depleted NK cells. Intentionally mismatched rIL-2 activated NK cells represents a safe approach for elimination of residual tumor cells, aiming for induction of GVL while avoiding GVHD.
The conditioning prior to allogeneic stem cell transplantation was originally designed as a myelo... more The conditioning prior to allogeneic stem cell transplantation was originally designed as a myeloablative conditioning, designed to eliminate malignant or genetically abnormal cells and then use the transplant procedure for rescue of the patients or to replace missing bone marrow products. However, allografts can induce effective graft vs. malignancy effects and can also eliminate undesirable hematopoietic stem cells in patients with genetic disorders and autoimmune diseases, thus documenting that alloreactive effects mediated by donor lymphocytes post-grafting can play a major role in eliminating hematopoietic cell of host origin, as well as provide effective immunotherapy for the treatment of disease recurrence. The efficacy of donor lymphocyte infusion (DLI) could be improved by activation with rIL-2 or by donor immunization. The cumulative experience over the years suggesting that alloreactive donor lymphocytes were most effective in eliminating tumor cells of host origin resulted in an attempt to reduce the intensity of the conditioning in preparation for the transplant procedure used for the treatment of hematological and other malignancies as well as life-threatening non-malignant disorders for which allogeneic stem cell transplantation may be indicated. Our working hypothesis proposed that the myeloablative conditioning which is hazardous and may be associated with early and late side effects, may not be required for treatment of patients with any indication for allogeneic stem cell transplantation. Instead, nonmyeloablative conditioning based on the use of reduced intensive preparatory regimen, also known as nonmyeloablative stem cell transplantation, may be sufficient for engraftment of donor stem cells while avoiding procedure-related toxicity and mortality, followed by elimination of undesirable cells of host origin by post-transplant effects mediated by alloreactive donor lymphocytes infused along with donor stem cells or administered subsequently as DLI. Improvement of the immediate outcome of stem cell transplantation using NST due to a significant decrease in transplant related mortality has broadened the spectrum of patients eligible for allogeneic stem cell transplantation, including elderly patients and other patients with less than optimal performance status. Likewise, the safer use of stem cell transplantation prompted expanding the scope of potential indications for allogeneic stem cell transplantation, such as metastatic solid tumors and autoimmune disorders, which now are slowly becoming much more acceptable. Current strategies focus on the need to improve the capacity of donor lymphocytes to eliminate undesirable malignant and non-malignant hematopoietic cells of host origin, replacing abnormal or malignant stem cells or their products with normal hematopoietic stem cells of donor origin, while minimizing procedure-related toxicity and mortality and improving the quality of life by reducing the incidence and severity of hazardous acute and chronic GVHD.
Introduction: Prolonged treatment of graft vs. host disease (GVHD) is extremely immunosuppressive... more Introduction: Prolonged treatment of graft vs. host disease (GVHD) is extremely immunosuppressive. Local therapy with intra-arterial (IA) injection of steroids may induce remission with lower extent of systemic immune suppression. Here, we present our experience with IA treatment of gastrointestinal (GI) and/or hepatic steroid resistant/dependent GVHD with 2 consecutive protocols. Patients and methods: Thirty five patients (37 GVHD events (hepatic, n=15), (GI, n=16), (combined, n=6)) were treated with 53 IA sessions. Most side effects were minor. Results: We found that IA steroid therapy was associated with partial and complete remission among patients with steroid resistant/dependent hepatic or GI GVHD. Hepatic partial response was observed in 14 (66.6%) patients among whom 7 (33.3%) reached complete response. GI partial response was observed in 19 (86.4%) patients among whom 12 (54.4%) reached complete response. Early administration of the local therapy, female gender, myeloid basic disease, and a non-active status of the basic disease at the day of transplantation were found related for predicting a better response for the intra-arterial treatment. The use of high dose steroids in the hepatic IA protocol from was at least as good as intermediate dose steroids with methotrexate (table 1, figure 1) and may be safer. Conclusions: Intra-arterial catheter guided steroid therapy is safe and effective in steroid resistant/dependent GVHD. Hepatic artery treatment with methotrexate can be safely substituted with high dose IA methylprednisolone. Further research is warranted characterizing the patients benefit most. Table 1 - comparison between 1st and 2nd hepatic IA treatment protocols 1st protocol 2nd protocol Significance Median age (range) 25 years (7–42) 32 years (18–59) P=0.09 Sex (M:F) 6:1 8:5 NS family donor vs MUD 6:1 9:4 NS Median time in days SCT-GVHD (range) 27 (13–133) 45 (13–248) NS Median time in days GVHD-IA (range) 15 (6–218) 190 (12–2615) P=0.09 Median peak GVHD grade (range) 3 (3–4) 3 (2–4) NS Highest pre-IA treatment bilirubin level (in mmol/L; normal&amp;amp;amp;amp;lt;17) (range) 186 (138–321) 225.5 (83–672) NS Median time to initial response in days (range) 14.5 (4–100) 8 (1–31) P=0.073 Median time to complete response in days (range) 130.5 (35–226) 49 (17–80) Figure 1A, Figure 1B Figure 1A, Figure 1B.
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Papers by Igor Resnick