title: Depletion of alloreactive T cells by a specific anti-interleukin-2 receptor p55 chain immunotoxin does not impair in vitro antileukemia and antiviral activity.
abstract: The success of bone marrow transplantation (BMT) from HLA-disparate donors depends on the development of new strategies able, on one hand, to efficiently prevent graft-versus-host disease (GVHD) and, on the other hand, to protect leukemic patients from relapse and infections. Using an immunotoxin (IT) directed against the alpha chain (p55) of the human interleukin-2 receptor (RFT5-SMPT-dgA), we previously showed that it is possible to kill mature T cells activated against a specific HLA complex by a one-way mixed lymphocyte culture (MLC). The present study was performed to investigate whether this protocol of allodepletion affects the capacity of residual T cells to display antileukemia and antiviral activity evaluated by limiting dilution assays (LDA), measuring the frequency of cytotoxic T-lymphocyte precursors (CTLp) directed against autologous leukemic blasts (LB) and cytomegalovirus (CMV)- and Epstein-Barr virus (EBV)-infected target cells. Antileukemia activity was evaluated in peripheral blood mononuclear cells (PBMC) of 3 patients treated for acute myeloid leukemia who had developed a high frequency of LB-reactive CTLp after either autologous or allogeneic BMT. Results demonstrate that (1) depletion with RFT5-SMPT-dgA efficiently inhibited MLC; (2) fresh PBMC of patients yielded a high frequency of LB-reactive CTLp comparable to that of the mock-treated PBMC; and (3) effector cells obtained after allodepletion fully retained the capacity to lyse pretransplant LB. By contrast, the frequency of CTLp directed against patient's pretransplant BM remission cells was always undetectable. Data obtained in 4 healthy donors showed that specifically allodepleted T cells recognized and killed autologous CMV-infected fibroblasts and autologous EBV-B-lymphoblastoid cell lines. In conclusion, our data indicate that allodepletion using RFT5-SMPT-dgA efficiently removed alloreactive cells, while sparing in vitro antileukemic and antiviral cytotoxic responses.
authors.affiliation: Laboratorio di Immunologia e Unità di Trapianto di Midollo Osseo, Dipartimento di Scienze Pediatriche, Università degli Studi di Pavia, IRCCS Policlinico San Matteo, Pavia, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Montagna
authors.forename: E
authors.initials: E
authors.lastname: Yvon
authors.forename: V
authors.initials: V
authors.lastname: Calcaterra
authors.forename: P
authors.initials: P
authors.lastname: Comoli
authors.forename: F
authors.initials: F
authors.lastname: Locatelli
authors.forename: R
authors.initials: R
authors.lastname: Maccario
authors.forename: A
authors.initials: A
authors.lastname: Fisher
authors.forename: M
authors.initials: M
authors.lastname: Cavazzana-Calvo
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology* / Q000473:pathology; D016026:Bone Marrow Transplantation; D002460:Cell Line; D002648:Child; D003587:Cytomegalovirus / Q000276:immunology; D006680:HLA Antigens / Q000276:immunology; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D000922:Immunotoxins / Q000276:immunology*; D015470:Leukemia, Myeloid, Acute / Q000276:immunology* / Q000628:therapy*; D019520:Living Donors; D008213:Lymphocyte Activation; D008212:Lymphocyte Depletion / Q000379:methods*; D015375:Receptors, Interleukin-2 / Q000276:immunology*; D012016:Reference Values; D013601:T-Lymphocytes / Q000276:immunology* / Q000821:virology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014182:Transplantation, Autologous; D014184:Transplantation, Homologous
chemical_list: D006680:HLA Antigens; D000922:Immunotoxins; D015375:Receptors, Interleukin-2