Different macrophage populations distinguished by means of fluorescent polysaccharides. Recognition and properties of marginal‐zone macrophages

Abstract

Fluorescent conjugates of hydroxyethyl (OEt) starch or Ficoll are selectively ingested and retained in vivo by spleen marginal‐zone (MZ) and lymph node marginal‐sinus macrophages of mice, whereas similar conjugates of type 3 pneumococcal capsular polysaccharide (SIII) are generally retained by macrophages (Kupffer cells, histiocytes, macrophages of spleen, lymph nodes, bone marrow and peritoneal cavity). MZ and other macrophages are readily identifiable by fluorescence after injection in vivo of OEt starch and SIII labeled with tetraethylrhodamine isothiocyanate and fluorescein isothiocyanate. Collagenase digestion was required for recovery of intact MZ macrophages from spleen in single cell suspensions and for maximum yields of other macrophages. MZ macrophages are larger and morphologically distinct from other macrophages, but resemble them in respect of EA, EAC receptors and acid phosphates and nonspecific esterase content and are equally radio‐resistant. They appear normal in CBA/N nude and in beige mice. Freshly isolated MZ macrophages in suspension have adherent lymphocytes, dispersible by EDTA treatment, with B but not T cell markers. It is suggested that selective adherence to MZ macrophages is a factor in determining B cell traffic. MZ macrophages did not have demonstrable surface I‐A or I‐EC antigens. Only 4–8% of other spleen macrophages freshly isolated by collagenase treatment expressed I‐A in the same preparation, whereas 35% of other cells (lymphocytes and blasts) reacted with monoclonal anti‐I‐A and anti‐I‐EC. After adherence to glass or plastic, 40% or more red‐pulp, but not MZ macrophages, became I‐A‐positive. When taken from mice recently restimulated with sheep erythrocytes, half the red‐pulp macrophages expressed I‐A even before adherence. The relation of MZ to other macrophages is not known. However, their properties are consistent with the demonstrated ability of thymus‐independent antigens selectively taken up by these cells to elicit long‐lasting IgM antibody responses by direct interaction with B cells. The unexpected observation that only a small proportion of spleen macrophages freshly isolated from unstimulated mice had detectable surface I‐A, but that this proportion was much increased after attachment to plastic, is discussed in relation to the possibility that macrophages do not express surface Ia antigens unless they have been stimulated.