High‐frequency establishment of human immunoglobulin‐producing lymphoblastoid lines from normal and malignant lymphoid tissue and peripheral blood

Abstract

Human hematopoietic tissue and lymphocytes separated from 10‐20 ml samples of peripheral blood have been grown in vitro in a lens‐paper and a gelatin foam (Spongostan) grid organ culture. Lymphoblastoid cell lines were established from the lymph nodes, and in one case from the spleen, of 22/23 consecutive, unselected adult individuals without manifest malignancy or infectious mononucleosis. Biopsies from 5/8 patients with malignancy were successful. The blood tines were derived from 5/10 patients with and 4/10 donors without malignancy. The very high frequency of success from normal tissue confirms the assumption made before that the spontaneous establishment of lymphoblastoid cell lines is unrelated to manifest malignancy of the donor. The results indicate that lymphoid cells with a potential for infinite proliferation (“lymphoblastoid transformation”) are present in almost all adult individuals. The Spongostan grid culture is a superior instrument to select and/or adapt these cells in vitro. All lymphoblastoid lines produced immunoglobulins. The majority started with a “polyclonal” pattern of immunoglobulin production but changed towards stable “ monoclonality “ during the course of long‐term cultivation. It is suggested that lymphoblastoid lines have a polyclonal origin and that the reason for development of a monoclonal line is a selection of one cell clone either in the organ culture during establishment or in long‐term culture.

Regulation of Human Cell Engraftment and Development of EBV-Related Lymphoproliferative Disorders in Hu-PBL-scidMice

“…B lymphoblastoid cell lines (B-LCL) were initiated from donor peripheral blood mononuclear cells by induction with supernatant from the EBVproducing cell line B95.8 as previously described (22). B-LCL lines were maintained in culture for use as targets or stimulators for EBV-specific T cell analyses.…”

Bone marrow-derived lymphoid cell lines from patients with agammaglobulinemia.

“…In cultures stimulated by PWM and LMF, some of these lymphoblasts mature into plasma cells (15). In cultures stimulated by EBV, some of the lymphoblasts are immortalized, giving rise to long-term lymphoid cell lines that secrete immunoglobulin, usually of several classes (16)(17)(18). These cells have surface receptors for C3, Fc, and EBV and bear surface immunoglobulin, usually of several heavy chain classes.…”

“…Prolonged cultivation of a LCL may result in phenotypic restriction to production of a single class of immunoglobulin (16,17,45). All of the agammaglobulinemic cell lines were examined during the 1st mo after they were established and at least once again after 6-12 mo of culture.…”

Continuous Lymphoid Cell Lines with Characteristics of B Cells (Bone-Marrow-Derived), Lacking the Epstein-Barr Virus Genome and Derived from Three Human Lymphomas

“…The few exceptions where the derived, EBVcarrying cells had discordant markers were probably due to the presence of allogeneic lymphocytes in the tumor, derived from transfused blood (9). Explantation of leukemia and myeloma cells usually does not lead to the outgrowth of the leukemia or the myeloma cells; instead, a small minority of EBV-genome-carrying B-lymphoid cells will grow, as a rule, that are thus nonrepresentative for the neoplastic cell in these conditions (10)(11)(12)(13). In two very exceptional cases, the myeloma cell itself grew out, however.…”

“…It was therefore of great interest to determine whether this cell line was representative for the virus-genome-negative biopsy or was merely the outgrowth of an admixed, nonrepresentative EBV-genome-carrying lymphoid cell-type. U-698M and U-715M lines were established by the Spongostan grid organ culture technique previously described (13). Fragmented tissue from a lymphosarcoma tonsil (U-698M) and a lymphoreticuloma lymph node (U-715M) was explanted on the grids, which were placed in 60-mm Falcon petri dishes.…”