Lymphocyte proliferation in vitro induced by soluble protein antigens. II. Cellular requirements

Abstract

Immune guinea pig lymph node cells were fractionated on Ig anti-Ig or HSA anti-HSA affinity columns or on plastic surface in medium containing carbonyl iron. These techniques selectively removed B lymphocytes, K lymphocytes or adherent cells. The residual cells (Fc receptor-negative T lymphocytes) responded to soluble antigen in vitro in the same way or even better compared with nonfractionated cells. In addition, there was no indication that antigen-antibody complexes were superior to antigen in triggering lymph node cells or purified lymph code T lymphocytes into DNA synthesis. The results obtained suggested that memory T lymphocytes can be stimulated by antigen autonomously.

Rat Memory T Lymphocytes

“…3) to recognize antigen and to proliferate in vitro is in agreement with results of T-lymphocyte cultures from guineapig [18,22,23,24] and humans [3], in whom the response was macrophage-dependent. In contrast, Hertel-Wulff & Rubin [13] showed that soluble protein antigens activated primed guinea-pig T cells independently of macrophages. This discrepancy might be explained by the fact that macrophage precursors were not detectable by the acridine-orange or the latex particle ingestion techniques used [13], but were inactivated by preculture in the presence of silica (Fig.…”

“…Thus, viability of the phagocytic cells was not necessary. The help that T cells required for the anamnestic in vitro response was presumably provided by lymphocyte-like macrophage precursors [9,30] passing through the Degalan Ig-anti-IgG columns, because the mature phagocytes were retained under the conditions used [13]. The interaction of antigen with Ir-gene products on the surfaces of such precursor cells or with soluble molecules released by these cells could possibly deliver the signals necessary to trigger T cell proliferation [14,15,18].…”

“…Activation of primed T lymphocytes by antigen in vitro has been reported to require the help of macrophages in the mouse, guineapig, and man [3,10,18,[22][23][24], as shown for the activation of T cells by mitogens [19,22]. However, contrasting data have been reported which point to an autonomous mode of stimulation by antigens [13] or mitogens [17,20]. It is known that the removal of adherent and/or phagocytic cells from rat spleen cell suspensions enhanced the subsequent stimulation by phytohaemagglutinin (PHA) or concanavalin A (Con A) [11,12], the generation of plaque-forming cells [27], or the induction of cytotoxic T lymphocytes in mixed leucocyte cultures [26].…”

Effects of Basic Proteins on Spontaneous Rosette Formation between Human Lymphocytes and Xenogeneic Erythrocytes

“…By identifying monocytes and granulocytes under fluorescence microscopy after acridine orange staining [10] the suspensions were found to consist of 80% lymphocytes (range 67-93%), 18% monocytes (range 4-31%) and 2% granulocytes (range 0-6%).…”

Two Small Lymphocyte Subpopulations in Human Peripheral Blood

“…Monocytes were identified in cell suspensions by their ability to phagocytoze latex or fluolite particles [Arvilommi, 1975], In rosette suspensions morphological criteria after acridine-orange stain ing were used [Hertel-Wulff et al, 1976]. In the aforementioned isolations the mean percentage of monocytes was 18.5 (range 9-25).…”