Opsonic Adherence of Sensitized Ox Red Cells to Human Lymphocytes as Measured by Rosette Formation

Abstract

Ox red blood cells sensitized with rabbit anti-ox red cell antibody adhered opsonically to freshly prepared human peripheral blood lymphocytes. When tested by a centrifugation technique 24–59% of live lymphocytes in normal individuals formed rosettes with the indicator red cells while 41–87% of the lymphocytes in chronic lymphatic leukaemia reacted. After storage at + 4 °C the lymphocytes gradually lost the operative receptor; killing the cells by freezing and thawing, by sodium azide or by fixation in formaldehyde also resulted in loss of capacity to bind the indicator cells. Pretreatment of the lymphocytes with high doses of anti-human IgM in diluted serum resulted in a definite, although incomplete, inhibition. Also, soluble antigen-antibody complexes were inhibitory, although strong inhibition was difficult to show. Undiluted, heated normal human or rabbit serum was highly inhibitory. The high incidence of lymphocytes showing antibody opsonic adherence in patients with chronic lymphatic leukaemia, together with the finding that most lymphocytes binding sensitized ox red cells did not bind unsensitized sheep red cells (a possible T-cell marker) suggests that lymphocytes showing antibody opsonic adherence represent a subpopulation of peripheral blood lymphocytes. This subpopulation might be an analogue of the B-cell population in mice.

A subpopulation of normal human peripheral B lymphcytes that bind IgE.

“…with washed ox erythrocytes (E0) (14). Antisera having an agglutination titer of 1:32-1:64 were pooled and the IgG fraction was isolated by DEAE-cellulose chromatography in 0.015 M phosphate buffer at pH 8.0.…”

“…IgE rosette assay. Eo, which do not form spontaneous rosettes with human lymphocytes (14), were used as indicator red cells in the other rosette assays. To facilitate adsorption of proteins and allow their use for up to 5 wk when stored at 4°C, Eo were pretreated with trypsin, then fixed with pyruvic aldehyde and formaldehyde in a modification of the method described by Hirata and Brandriss (19).…”

Suppression of human T-cell mitogenesis by prostaglandin. Existence of a prostaglandin-producing suppressor cell.

“…The preparation of such columns has been previously described (12). The effluent contained > 90% lymphocytes which formed E-rosettes with sheep erythrocytes (13) and < 1% of cells with surface immunoglobulin or receptors for IgG (14).…”

Robin Coombs: his life and contribution to haematology and transfusion medicine

“…In the early 1970s Robin Coombs and colleagues began to use rosetting reactions to define the lymphocyte subpopulations present in peripheral blood (PBL) and other tissues (Hallberg et al, 1973). By incubating PBL with sheep red blood cells (SRBC) or ox red blood cells (ORBC) coated with either immunoglobulin or complement in the mixed antiglobulin rosetting reaction they were able to determine the proportion of T -and B-cells in the blood of normal individuals.…”