CD3 x CD19 bispecific antibodies and CD28 costimulation for locoregional treatment of low-malignancy non-Hodgkin's lymphoma.

In advance of using bispecific antibodies for the treatment of B cell lymphoma in humans, we analysed CD3 x CD19 bispecific antibodies for their capacity to induce T cell activation in cell suspensions from follicular lymphoma lymph nodes. Here, we demonstrate that the lack of costimulatory molecules, such as members of the B7 family, on the tumour cells resulted in insufficient activation of autologous T lymphocytes. However, stimulation and proliferation of T cells could be induced by addition of monospecific CD28 antibodies.

Single-step purification of bispecific monoclonal antibodies for immunotherapeutic use by hydrophobic interaction chromatography.

A method for large scale production and single-step purification of bispecific antibodies is described. Hybrid-hybridomas were grown in hollow-fibre bioreactors with an average yield of 8 to 12 g of immunoglobulin per month. Bispecific antibodies were purified from the bioreactor supernatant by hydrophobic interaction chromatography which resolves bispecific antibodies, monospecific immunoglobulins, and culture medium supplements in one single chromatographic step.

Classical Hodgkin's disease. Clinical impact of the immunophenotype.

Antibodies against CD15, -30, and -20 are often used to support morphological diagnosis of Hodgkin's Disease (HD). The classical HD, i.e.

Expression of p56lck in B-cell neoplasias.

The protooncogene p56lck is considered to participate in malignant transformation of lymphoid cells. In order to evaluate the role of this tyrosine kinase in B cell neoplasias, we investigated the expression of p56lck by Western blot analysis. In 12/16 Burkitt's lymphoma derived cell lines, 3/3 lymphoblastoid cell lines, 1/6 Hodgkin's disease derived cell lines, and 10/10 freshly isolated chronic lymphocytic leukemia cells constitutive expression of the protein was detected.

[31P-MR spectroscopy of the human liver--the spectral indications of lymphoma infiltration].

Purpose: To evaluate whether phosphorus magnetic resonance spectroscopy (31P-MRS) enables a non-invasive detection of liver involvement in systemic diseases like Hodgkin's lymphoma.

Materials And Methods: Using a clinical 1.5 Tesla whole-body MR system image-guided localised phosphorus MR spectra from the anatomically defined volumes of interests were measured.

Dose-response relationship of complementary radiotherapy following four cycles of combination chemotherapy in intermediate-stage Hodgkin's disease.

Purpose: To determine the appropriate irradiation dose after four cycles of modern combination chemotherapy in nonbulky involved field (IF/BF) and noninvolved extended-field (EF/IF) sites in patients with intermediate-stage Hodgkin's disease (HD).

Materials And Methods: HD patients in stage I to IIIA with a large mediastinal mass, E stage, or massive spleen involvement were treated with two double cycles of alternating cyclophosphamide, vincristine, procarbazine, and prednisone (COPP) plus doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) followed by EF irradiation in two successive trials (HD1 and HD5). In the HD1 trial (1983 to 1988), 146 patients who responded to chemotherapy were randomized to receive 20 Gy (70 patients) or 40 Gy (76 patients) of EF irradiation in all fields outside bulky disease sites.

Prevention of Epstein-Barr virus-induced human B-cell lymphoma in severe combined immunodeficient mice treated with CD3xCD19 bispecific antibodies, CD28 monospecific antibodies, and autologous T cells.

Bispecific CD3 x antitumor antibodies in combination with coactivating CD28 antibodies can induce resting T cells to proliferate and to lyse syngeneic tumor cells (M. Azuma et al., J.

BEACOPP: an intensified chemotherapy regimen in advanced Hodgkin's disease. The German Hodgkin's Lymphoma Study Group.

Purpose: At present, treatment results for patients with advanced-stage Hodgkin's disease remain unsatisfactory. Standard chemotherapy M(C)OPP (nitrogen mustard (cyclophosphamide). vincristine, procabazine, and prednisone).

A phase-II study with idarubicin, ifosfamide and VP-16 (IIVP-16) in patients with refractory or relapsed aggressive and high grade non-Hodgkin's lymphoma.

We report a phase-II study of idarubicin, ifosphamide and etoposide (IIVP-16) in heavily pretreated patients with relapsed or refractory aggressive non-Hodgkin's lymphoma. The IIVP-16 regimen consisted of idarubicin (10 mg/m2 i.v.

A phase-I study of an anti-CD25 ricin A-chain immunotoxin (RFT5-SMPT-dgA) in patients with refractory Hodgkin's lymphoma.

The anti-CD25 immunotoxin (IT), RFT5-SMPT-dgA, was used in a phase I dose escalation trial in patients with refractory Hodgkin's lymphoma. The IT was constructed by linking the monoclonal antibody RFT5 via a sterically hindered disulfide linker to deglycosylated ricin-A. All patients in this trial were heavily pretreated with a mean of 5 (range, 2 to 8) different prior therapies, including autologous bone marrow transplantation in 8 of 15.

Mapping of the human IL10 gene and further characterization of the 5' flanking sequence.

Interleukin-10 (IL-10) is an important regulatory cytokine whose involvement extends into diverse areas of the human immune system. Recent characterization of the promoter and 5' flanking regions has demonstrated the presence of positive and negative regulatory segments in the promoter. In addition, the characterization of two dinucleotide repeat elements immediately upstream of the gene has shown that there is considerable polymorphism directly associated with the human IL10 gene.

Idiotype vaccination strategies against a murine B-cell lymphoma: dendritic cells loaded with idiotype and bispecific idiotype x anti-class II antibodies can protect against tumor growth.

Three strategies were used to evaluate 38C13 B-cell lymphoma-specific idiotype immunization to protect against subsequent lymphoma challenge in C3H/He mice. It was observed that tumor-specific immunity could be induced by immunization with (i) KLH-conjugated 38C13 B-cell lymphoma idiotype in complete Freund's adjuvants (survival rate 80%), (ii) dendritic cells pulsed in vitro with native idiotype protein (survival rate 80%), and (iii) bispecific antibodies composed of B-lymphoma-related idiotype and an MHC class II binding moiety (survival rate 40%). Presentation of idiotype determinants by dendritic cells or bispecific antibody resulted in lymphoma-specific immunity and obviated the requirement for carrier protein or adjuvant.

A deletion mutant of Pseudomonas exotoxin-A fused to recombinant human interleukin-9 (rhIL-9-ETA') shows specific cytotoxicity against IL-9-receptor-expressing cell lines.

The receptor for human interleukin-9 (hIL-9) might be a target for selective immunotherapy. It is expressed on a variety of malignant cells, including Hodgkin's lymphoma, non-Hodgkin lymphoma and acute myeloid leukemia (AML). We therefore constructed a new chimeric toxin by fusing hIL-9-cDNA to modified Pseudomonas aeruginosa exotoxin A (ETA').

Hodgkin's disease. Treatment of intermediate stages.

Intermediate stage Hodgkin's disease (HD) is defined as nodal disease with certain clinical risk factors. Patients with intermediate stage HD qualify for a combination of chemo- and radiotherapy, since radiotherapy alone results in a high relapse rate. The precise definition of this group varies between trial groups.

Promotion of IL8, IL10, TNF alpha and TNF beta production by EBV infection.

Burkitt's lymphoma (BL) represents a high malignant B cell tumour. It has been proposed that cytokines are responsible for some of the characteristics of BL. We have analysed a panel of different BL and lymphoblastoid cell lines (LCLs) for the expression of cytokines, including: IL 1 alpha, IL 1 beta, IL2, IL3, IL4, IL6, IL8, IL10, TNF alpha and TNF beta and for the soluble cytokine receptor for IL2 (slL2R).

Hodgkin's disease: from basic science to clinical application.

Although the pathogenesis of Hodgkin's disease is not clear, molecular analyses revealed characteristic features which proved the clonal origin of the disease. EBV injection can be demonstrated in more than 50% of cases at the DNA or protein level. Recently, immunoglobulin gone rearrangements were found in single Hodgkin and Reed-Sternberg cells.

Interleukin 10 inhibits cytokine production of human AML cells.

Background: Unlike normal hematopoietic cells leukemic blasts from patients with AML constitutively express cytokines like IL1, GM-CSF and TNF alpha and it has been suggested that these cytokines may regulate growth and differentiation of the malignant cells. IL10 inhibits cytokine production of activated macrophages and T-helper 1 cells. We analysed whether IL10 can also suppress cytokine production and may inhibit growth of AML cells.

[Clinical aspects and primary therapy of Hodgkin's lymphomas].

In spite of intensive research, the etiology and the pathogenesis of Hodgkin's lymphoma are still unclear. Hodgkin's lymphoma is clinically characterized by the appearance of enlarged lymph nodes and systemic symptoms. The diagnosis is exclusively based on the identification of the typical Reed-Sternberg cells surrounded by a mixture of reactive cells.

Current clinical trials in Hodgkin's disease.

With the advent of modern polychemotherapy and radiation techniques, cure rates in Hodgkin's disease (HD) have improved substantially. This article summarises current concepts of treatment for early, intermediate and advanced stage HD. Current strategies focus on the balance between cure rate and toxicity.

Further chemotherapy versus low-dose involved-field radiotherapy as consolidation of complete remission after six cycles of alternating chemotherapy in patients with advance Hodgkin's disease. German Hodgkins' Study Group (GHSG).

Objective: It was the aim of this prospective randomized multicenter study to compare chemotherapy and radiotherapy as consolidation treatments in patients achieving complete remission (CR) after 6 cycles of doxorubicin-containing chemotherapy in advanced-stage Hodgkin's disease (HD).

Methods: A total of 288 previously untreated patients aged 18-60 years with stage IIIB or IV HD received induction chemotherapy with 3 X (COPP + ABVD). Patients achieving CR were eligible for randomisation to either 20 Gy radiotherapy to initially involved fields (RT-arm) or to an additional 1 X (Copp + ABVD) (CT-arm ).

Isolation and characterization of allergen-binding cells from normal and allergic donors.

Background: Flow cytometry of the immune system so far has been limited to the analysis of subpopulations according to lineage markers. The cells involved in a particular immune response could not be assayed due to their low frequency. Here we show the potential of antigen-specific high gradient magnetic cell sorting to enrich cells for visualisation in multiparameter cytometry, functional studies and immortalization.

IL6 and IL6 receptor expression in Burkitt's lymphoma and lymphoblastoid cell lines: promotion of IL6 receptor expression by EBV.

We have analysed a panel of different Burkitt's lymphoma (BL) and lymphoblastoid cell lines (LCLs) for the expression of IL6 and IL6 receptor (IL6R). Epstein-Barr-Virus (EBV) positive or negative BL cell lines and the corresponding lymphoblastoid cell lines (LCL), derived from EBV immortalized mononuclear cells of the BL patients, were tested for the expression of IL6 mRNA and protein by Northern blot experiments and ELISA, and for the expression of the IL6R mRNA and protein by Northern blot Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and flow cytometry. Our results demonstrate that six out of 19 Burkitt's lymphoma cell lines produced IL6 constitutively.