dinitrobenzenes and Lymphoma

T cells were isolated from spleens of C57BL/Ka/Thy 1.1/Lb mice immunized with 2,4-dinitrophenylated keyhole limpet hemocyanin (DNP-KLH). The mice were infected in vitro with radiation leukemia virus and injected intrathymically into congenic C57BL/Ka (Thy 1.2) recipients. Within 3-4 months, seven thymic lymphomas developed, five of which were of donor type. The donor lymphomas were explanted and permanent cell lines were established in vitro. These lymphoma cells are capable of providing antigen-specific carrier (KLH)-primed help to DNP-primed B lymphocytes in secondary antibody production to DNP-KLH. They also enhance the secondary antibody response of whole period spleen cell populations to DNP-KLH. The availability of these immortal clonable populations of immunofunctional neoplastic T lymphocytes should facilitate biological and biochemical investigations of lymphocyte interactions during synthesis of antibody to thymus-dependent antigens.

Topics: Animals; Antibody Formation; Carrier Proteins; Cell Line; Cell Transformation, Viral; Dinitrobenzenes; Epitopes; Immunologic Memory; Leukemia Virus, Murine; Leukemia, Experimental; Lymphocyte Cooperation; Lymphoma; Mice; T-Lymphocytes

A cell fusion technique was used to produce hybridomas between the T lymphoma cell line, EL-4, derived from C57BL (H-2(b)), and an enriched population of human gamma globulin (HGG)-specific suppressor T cells prepared from the spleens of HGG-tolerant CBA mice (H-2(k)). Membrane fluorescence analysis of the hybridoma cells within 6 wk of cell fusion revealed expression of H-2(k) and I-J(k) gene products as well as H-2(b) antigens. Sonicates prepared from hybridomas which contained I-J(k) cells were tested for suppressive activity in vivo in irradiated mice given HGG-primed cells, dinitrophenyl (DNP)-primed cells, HGG-DNP, and horse erythrocytes. Among 18 such hybridoma lines, 6 showed specific suppressive activity, 5 nonspecific suppression, and 7 no suppression. Most lines progressively lost, with time, those properties derived from the normal parent cell. By about 3 mo after fusion few cells expressed CBA markers and only one cell line (number 77) retained some specific suppressive activity. In parallel with the losses was an alteration in chromosome number from near-tetraploid, soon after cell fusion, to near- diploid. Preliminary results with the T lymphoma-sensitive hypoxanthine aminopterin thymidine cell line, L5178, indicate retention of the expression of surface markers derived from the normal parent for 18 wk after hybidization. This suggests that T lymphoma cell lines may have to be screened for their capacity to produce hybridomas with stable properties.

Topics: Antibody Formation; Cell Line; Cell Separation; Dinitrobenzenes; H-2 Antigens; Hybrid Cells; Immune Tolerance; Immunosuppression Therapy; Lymphoma; Major Histocompatibility Complex; Neoplasms, Experimental; T-Lymphocytes