Cytotoxic T-cell clones derived from pluripotent stem cells (CFU-GEMM) of patients with Hodgkin's lymphoma.

Pluripotent stem cells (CFU-GEMM) give rise to multilineage hemopoietic colonies in culture. The cellular composition revealed that mixed colonies contain cells of different myeloid lineages and mononuclear cells with T-cell surface antigens. T lymphocytes of primary colonies and replated secondary clones from 5 patients with Hodgkin's lymphoma (stage I--II) were identified by their reaction with the monoclonal antibody OKT-8. Replated secondary clones do act functionally as cytotoxic cells using K562 as target cells. Evidence for a common progenitor of myeloid and lymphoid cells is provided by analysis of individual secondary colonies with the use of OKT-3, OKT-4, OKT-8, VIM-D5, and IgM + D antibodies for each individual clone. Primary mixed and replated secondary colonies revealed OKT-8-positive cells. No reaction with OKT-3, OKT-4, VIM-D 5, or IgM + D was observed. In mixed colonies grown from putative bone marrow transplant donors, only OKT-3-positive cells could be observed. Secondary replated colonies did not stain for OKT-8 and failed to lyse 51Cr-labeled K562 cells.

Identification of human megakaryocytes derived from pure megakaryocytic colonies (CFU-M), megakaryocytic-erythroid colonies (CFU-M/E), and mixed hemopoietic colonies (CFU-GEMM) by antibodies against platelet associated antigens.

Pure megakaryocytic colonies, megakaryocytic-erythroid colonies and mixed hemopoietic colonies can be cultured from human bone marrow under appropriate culture conditions. Human plasma and mercaptoethanol support the growth for these different types of hemopoietic colonies. However, the addition of medium conditioned by leucocytes in the presence of phytohemagglutinin (PHA-LCM), as a source of thrombopoietin, is required for the formation of megakaryocytic colonies or megakaryocytes within mixed colonies. Megakaryocytes were identified by their typical morphological appearance in culture. Pure megakaryocytic colonies, megakaryocytic-erythroid colonies and mixed colonies were plucked by micropipette and analysed by the PAP-slide technique using antibodies to human factor VIII-related protein or serum derived from a patient with posttransfusion purpura; this particular serum demonstrated anti-P1A1 antibody activity. These antibodies might provide an excellent probe to identify megakaryocytic progeny from committed and non-committed hemopoietic progenitors, facilitating studies of early events in megakaryopoiesis.

Suppressor T-cell clones derived from pluripotent stem cells (CFU-GEMM) of a patient with Hodgkin's lymphoma.

Pluripotent stem cells (CFU-GEMM) give rise to multilineage hemopoietic colonies in culture. The cellular composition revealed that mixed colonies contain cells of different myeloid lineages and mononuclear cells with T-cell surface antigens. T-lymphocytes of primary colonies, replated secondary and tertiary colonies from a patient with Hodgkin's Lymphoma were identified by their reaction with the monoclonal antibody OKT 8. Evidence for a common progenitor of myeloid and lymphoid cells is provided by analysis of individual secondary and tertiary colonies using OKT 3, OKT 4, OKT 8, VIM-D 5, and Ig M + D antibodies for each individual colony. Primary mixed, replated secondary and tertiary colonies revealed OKT 8 positive cells. No reaction with OKT 3, OKT 4, VIM-D 5, or Ig M + D was observed.