Maud Brandely

Activity of intrapleural recombinant gamma‐interferon in malignant mesothelioma

Twenty‐two consecutive patients with malignant diffuse pleural mesothelioma were treated with recombinant gamma‐interferon by the intrapleural route. Diagnosis was made by thoracoscopic examination and all cases were confirmed by the French Mesothelioma Panel of Pathologists. Patients were staged based on thoracoscopic examination and computed tomography (CT) scan: 12 patients were classified as Stage I and 10 were Stage II. A solution of gamma‐interferon (40 × 106 U) was infused twice a week over 2 months. Every patient experienced fever. One patient had a Grade 2 leukopenia and one patient suffered from pleural empyema. Response evaluation was based on the following: (1) CT scan performed 2 weeks after treatment ended, and (2) repeat thoracoscopic examination with histopathologic verification in nine patients who had demonstrated a stabilization or a regression of the disease on CT scan. From the original group, 19 patients could be evaluated. Four complete thoracoscopic histopathologic responses and one partial response were observed in Stage I patients (56%). One partial response was observed in Stage II patients.

A PHASE II STUDY OF INTERLEUKIN-2 IN 49 PATIENTS WITH RELAPSED OR REFRACTORY ACUTE LEUKEMIA

In this report we present the results of a multicenter phase II study of high-dose recombinant Interleukin-2 (rIL-2) in patients with refractory or relapsed acute leukemia. Forty-nine patients with acute myeloid leukemia (AML: 30 patients) or acute lymphoblastic leukemia (ALL: 19 patients) were included. Median age was 30 years (range: 4-71). Four patients were treated for primary refractory disease and 45 for relapsed disease (16 patients > 2nd relapse). Twenty-four patients (49%) had previously received bone marrow transplantation (allogeneic: 5, autologous: 19). Patients were scheduled to receive three 5-day cycles of rIL-2 given every other week. rIL-2 was administered as bolus I.V. infusion of 8 x 10(6) UI/m2 every 8 hours during cycle I and every 12 hours during cycles 2 and 3. Patients received a mean of 76% of rIL-2 planned dose. Main toxicity was hematologic (grade IV thrombopenia: 84%). Hemodynamic and metabolic toxicities lead to treatment discontinuation in 10 patients (20%). Strong immune activation was achieved including a significant increase in activated T-cells and Lymphokine-Activating-Killer cell (LAK) activity. Twenty-seven out of 30 AML patients could be evaluated for response: 2(7%) achieved complete remission (CR) which lasted 3 and 4 months. No response was observed in the 18 assessable ALL patients, most of whom (77 %) presented absolute drug resistance. These results show that this high dose rIL-2 regimen induces significant biological effects and provides some anti-leukemic activity in patients with advanced leukemia. Considering the severe toxicity observed and the limited remission rate achieved here, rIL-2 does not appear to be a valuable therapeutic option for such patients. However, the undoubted anti-leukemic activity of this cytokine invites further investigation especially in the minimal residual disease situation.

The use of a sequential high dose recombinant interleukin 2 regimen after autologous bone marrow transplantation does not improve the disease free survival of patients with acute leukemia transplanted in first complete remission.

We report the outcome of 50 consecutive patients with CR1 acute leukemia (AML = 22; ALL = 28) treated with autologous BMT, after cyclophosphamide and TBI, followed with a sequential high dose rIL2 regimen. rIL-2 (RU 49637 from Roussel-Uclaf, Romainville, France) was started after hematological reconstitution an average of 72 +/- 22 days post transplant. The schedule consisted of a continuous infusion over 5 cycles (Cycle 1: 5 days starting on day 1; cycle 2-5: 2 days starting on day 15, 29, 43 and 57). Patients were treated at 4 different dosages (12 (N = 40), 16 (N = 3), 20 (N = 2), 24 (N = 5) x 10(6) IU/m2/day). Toxicities were mainly related to capillary leak syndrome and thrombocytopenia. Patients received an average of 122 +/- 49 10(6) IU/m2. Two patients with AML died from toxicity. rIL-2 infusion was associated with very a high level of immune stimu-lation of both T-cells (P < 0.05) and natural killer (NK) cells (P < 0.05) and associated cytolytic functions (P < 0.05). With a minimal and median follow-up of 21 and 46 months, 3 year leukemia free survival is 41 +/- 6% overall, 39 +/- 10% and 43 +/- 8% for AML and ALL respectively. Relapse probabilities at 3 years are 59 +/- 11% for AML and 57 +/- 8% for ALL. We conclude that this short infusion of rIL-2 over 2 months, resulting in an increased immune stimulation, is not associated with a better leukemic control for patients with acute leukemia transplanted early after reaching first complete remission.

Cutaneous side effects associated with interleukin 2 administration for metastatic melanoma

BACKGROUND Cutaneous changes associated with recombinant interleukin 2 (r IL-2) administration are frequent but have been rarely studied in a large series. OBJECTIVE We analyzed these clinical, microscopic, and immunologic changes. METHODS Patients with metastatic melanoma treated with r IL-2 were studied. The eruption was scored as mild or severe. Biopsy specimens were obtained for histopathology, ultrastructural analysis, and immunophenotyping. Chi-square and t tests were used for statistics. RESULTS Twenty-five patients were included. Eruptions were observed in 56 of 78 cycles (72%); 53 were mild with a burning pruriginous erythema, and 3 were severe with urticaria, necrotic lesions, and blisters. Regression was constant without sequelae. Pathologic changes were mild with a mononuclear cell infiltrate of activated helper T phenotype, expressing LFA-1. Keratinocytes and endothelial cells displayed intercellular adhesion molecule-1 and HLA-DR. Cells rarely expressed CD25. CONCLUSION Administration of r IL-2 triggers non-treatment-limiting cutaneous inflammation.

Intrapleural immunotherapy with escalating doses of interleukin-2 in metastatic pleural effusions

Background. The authors assessed the tolerance and efficacy of intrapleural interleukin‐2 (IL‐2) in patients with malignant effusion.

A Pilot Study of Autologous Bone Marrow Transplantation Followed by Recombinant Interleukin-2 in Malignant Lymphomas

In this study, we investigated the impact of recombinant interleukin-2 (rIL-2) after high dose chemotherapy and autologous bone marrow transplantation (ABMT) in 25 patients with refractory or relapsed Hodgkins disease (HD) (11 patients) and non Hodgkins lymphoma (NHL) (14 patients). 48% of patients had resistant disease, 84% achieved complete remission after ABMT. rIL-2 was started at a median of 54 days post-transplant and consisted of a first cycle of 5 days followed by 4 cycles of 2 days every other week. Patients received a mean of 160 x 10(6) IU/m2 rIL-2 and hematological toxicity was moderate and transient. None of the 5 evaluable patients with measurable disease responded to rIL-2. After a 5 year median follow-up, the probability of survival and DFS is 72% (HD: 73% and NHL: 70%, p = NS) and 45% (HD: 36% and NHL: 48%, p = NS) respectively. These somewhat encouraging results warrant further evaluation of rIL-2 after ABMT in controlled studies, especially in NHL patients stratified for previous chemosensitivity.

Phase I trial with recombinant interleukin-2 (rIL-2): immune activation by rIL-2 alone or following pretreatment with recombinant interferon-gamma.

Alterations of imniunological parameters were analysed in patients with advanced malignancies during a phase I trial with rIL‐2. Five‐day infusions of rIL‐2 at doses from 1 × 106 to 24 × 106 biological response modifiers program (BRMP) U/m2 per day were given to 29 patients, with a minimum of three patients per dose. The dose of 24 × 106 U/m2 per day was the maximal tolerated dose (MTD). Immunological parameters were analysed at days 0, 8 and 11 of the rIL‐2 courses. Following a leucopenia during rlL‐2 infusion, a lymphocytosis was found in all patients except one. The lymphocylosis peaked at day 8 and was detected at doses of rIL‐2 as low as 1 × 106 U/m2 per day, reaching a plateau at a dose of 16 × 106 U/m2 per day. Although all lymphocyte subsets were increased in patients receiving rIL‐2. some patients had predominant T cells (CD3+, NKH1(CD56)‐), others had predominant natural killer (NK) cells (CD3‐. NKH1(CD56)+), and yet others showed a mixed profile. A strong induction of cells cytotoxic for K.562 targets was found in all patients at days 8 and 11. Eighteen patients received, 1 month later, a second treatment in which infusion of rIL‐2 was preceded by a course of 5 days infusion of 2 × 106 U/m2 per day recombinant interferon‐gamma (rIFN‐γ)‐ The infusion of rIFN‐γ prior to rIL‐2 had no effect on the rIL‐2‐induced alterations of immunological parameters. Taken together, our results suggest that immune stimulation by rIL‐2 occurs even at low doses and is maximal at a dose below the MTD; and that pretreatment with low‐dose rlFN‐γ does not modify the immune stimulation by rIL‐2.

Increased expression of perforin and granzyme B genes in patients with metastatic melanoma treated with recombinant interleukin-2

The frequency of peripheral blood cells expressing the perforin gene or the granzyme B gene was evaluated by in situ hybridization in nine patients suffering from metastatic melanoma and treated with recombinant interleukin-2 (rIL-2). A spontaneous expression of both genes was detected in five to seven patients, rIL-2 administration increased the frequency of positive cells in all patients (P<0.03 for each gene), the highest frequency being reached in the patients who already expressed these genes prior to rIL-2 treatment (P<0.02). Expressions of the granzyme B gene and of the perforin gene were strongly correlated before IL-2 treatment and they were similarly affected by rIL-2 administration. In contrast, their modification under treatment did not correlate with that of CD56+ cell counts, of natural killer activity and of sCD8 release. This indicates that perforin and granzyme B gene expressions are markers of cytotoxic cell activation independent of those previously described, and that they should be further evaluated in patients with malignancies to delineate their potential value in predicting clinical outcome.

Cell surface expression of ICAM-1 (CD54) and LFA-3 (CD58), two adhesion molecules, is up-regulated on bone marrow leukemic blasts after in vivo administration of high-dose recombinant interleukin-2.

High-dose recombinant interleukin-2 (rIL-2) therapy can induce long-term remission in patients with melanoma and renal and colon cancer. More recently, in vivo IL-2 therapy was shown to induce complete or partial remission in some cases of relapsed chemotherapy-resistant acute myeloid leukemia. We have investigated the phenotypic modifications of bone marrow cells obtained from five patients with acute myeloid leukemia in relapse receiving high-dose i.v. rIL-2. We found that, in three of five patients, IL-2 could induce, in vivo, an increase in the expression of CD54/ICAM-1 and to a lesser extent of CD58/LFA-3 on bone marrow leukemic blasts. This demonstrates that rIL-2 modifies directly or indirectly the expression of the cell surface molecules of the tumor cells themselves. Upregulation of such adhesion molecules could account for the enhancement of cell interactions between the tumor and effector cells such as T, natural killer, and phagocytic cells as well as being indicators of differentiation signaling.

Interferon-γ (IFN-γ) and interleukin-2 in the generation of lymphokine-activated killer cell cytotoxicity — IFN-γ-induced suppressive activity

SummaryIncubation of human lymphocytes with recombinant interleukin-2 (rIL-2) results in the generation of lymphokine-activated killer (LAK) cells capable of lysing a wide variety of tumor cells. The present study was undertaken to examine the effect of recombinant γ interferon (rIFN-γ) on LAK cell cytotoxicity generated from different peripheral blood mononuclear cell (PBMC) subpopulations. When unseparated PBMC were stimulated by rIL-2 and rIFN-γ, the latter induced a transient enhancement after 2 days followed by a suppression of LAK cell cytotoxicity at day 6. Enhancement of LAK cell cytotoxicity was moderate and inconstant, whereas the inhibition was strong and observed with all the donors tested. This suppression was not associated with a decrease in the [3H]thymidine uptake. PBMC depleted of adherent cells were more sensitive to the stimulation by rIL-2 and the induced cytotoxicity was not modified by rIFN-γ. Monocyte-enriched plastic-adherent cells, when incubated with rIL-2 and rIFN-γ, became cytotoxic after 2–3 days of culture and inhibited LAK cell activity after 5–6 days. Collectively, our results suggest that rIFN-γ affects LAK cell cytotoxicity through the activation of plastic-adherent, monocyte-rich, cells which modulate natural killer cells, first in a positive, then in a negative way.