Peripheral blood lymphocytes of nonleukemic lymphoma patients exhibit aberrant expression of T-cell activation markers after polyclonal stimulation in vitro

Abstract

The authors evaluated suppressed in vitro functions of peripheral blood lymphocytes (PBL) as a possible tool in the early diagnosis of human lymphoma. In 13 of 22 patients with recent onset of various types of nonleukemic lymphomas (Mb. Hodgkin and non-Hodgkin's lymphomas of B-cell and T-cell origin) the mitogen response of PBL against phytohemagglutinin (PHA) and concanavalin A (Con A), as measured by 3H-thymidine (3HTdR) uptake, was found to be significantly suppressed, whereas the response to pokeweed mitogen (PWM) was normal in 18 cases. In parallel, cytofluorimetric analysis was done with PBL after 72 hours in culture with and without PHA, using antibodies against the differentiation antigens: CD3, CD8, CD4, CD19, and CDw14 and the activation antigens: interleukin-2 (IL-2) receptor (IL-2R, CD25), human leukocyte antigen DR (HLA-DR), and transferrin receptor (TR). Compared with healthy controls and patients with other diseases, a significant reduction of the total T-cell blast response, i.e., the percentage of large T-cells bearing activation markers, was found in all lymphoma cases including those with a normal 3HTdR uptake. Furthermore, a pronounced inhibition in the expression of the activation markers Il-2R and TR, but not of HLA-DR, was detected on CD3+ cells in PHA-stimulated PBL of all lymphoma cases. Thus, polyclonal activation combined with activation antigens seems to give more accurate information about the functional defect(s) of PBL in an early state of lymphoma; these parameters may therefore be valuable diagnostically. The abnormal pattern in the expression of T-cell activation antigens after polyclonal stimulation may help in the understanding the cellular immune defects associated with lymphoma.

Immune reconstitution after purified autologous and allogeneic blood stem cell transplantation compared with unmanipulated bone marrow transplantation in children

“…A whole blood stimulation assay was used (Mangge et al , 1990). Briefly, various volumes (5, 10 and 20 μ l) of fresh, heparinised blood were diluted in microtitre plates to a final volume of 200 μ l with medium (RPMI‐1640 medium supplemented with antibiotics and 10 −5 mol/l 2‐mercaptoethanol) containing phytohaemagglutinin (PHA 20, 10, 5, 2·5, 0·2 mg/ml), concanavalin A (ConA 40, 20, 10, 5 mg/ml), or pokeweed mitogen (PWM 160, 80, 40, 20 mg/ml) (all from Sigma, St Louis, MO, USA).…”

Unrelated peripheral blood stem cell transplantation with ‘megadoses’ of purified CD34+ cells in three children with refractory severe aplastic anemia

“…9 Briefly, various volumes (5, 10 and 20 l) of fresh, heparinized blood were diluted in microtiter plates to a final volume of 200 l with medium (RPMI-1640 with antibiotics and 10 −5 m 2-mercaptoethanol) containing the mitogens (all from Sigma, St Louis, MO, USA) phytohemagglutinin (PHA 20, 10, 5, 2.5 0.2 g/ml), concanavalin A (ConA 40, 20, 10, 5 g/ml), or pokeweed mitogen (PWM 160, 80, 40, 20 g/ml). Each culture was incubated for 72 h at 37°C in 5% CO 2 humidified air.…”

Unrelated partially matched peripheral blood stem cell transplantation with highly purified CD34+ cells in a child with Wiskott–Aldrich syndrome

“…T lymphocytes showed a normal proliferative response to polyclonal activators (phytohemagglutinin, concanavalin A, pokeweed mitogen) within 12 months after transplantation (Table 1). 17 Repeated chimerism analysis of peripheral blood revealed permanent 100% donor hematopoiesis and the underlying gene defect (deletion of the Cnucleotide of exon 10, position 1065) could not be detected in the peripheral blood or the bone marrow cells. Eczema disappeared 2 weeks after starting conditioning chemotherapy, but reappeared within 5 weeks after transplantation due to an allergic reaction against yeast and milk protein.…”