Macrophage‐Lymphocyte Clusters in the Immune Response to Soluble Protein Antigen in Vitro

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Abstract
T-cell populations from guinea-pigs sensitized to the protein antigens purified protein derivative of Mycobacterium tuberculosis, ovalbumin, or horseradish peroxidase can be selectively depleted of cells capable of initiating antigen-specific macrophage-lymphocyte clusters in vitro. The depletion is achieved by incubating the T cells on a monolayer of antigen-pulsed macrophages in a Petri dish for some hours and then gently aspirating the cells not adhering to the bottom of the dish. When subsequently assayed, the aspirated cells were found to be depleted of cluster-initiating lymphocytes. To remove the cluster-initiating lymphocytes committed to ovalbumin, monolayers of ovalbumin-pulsed macrophages must be used as immunoabsorbent. To remove cluster-initiating lymphocytes committed to horseradish peroxidase, monolayers of macrophages pulsed with that antigen must be used. The optimum time for incubation on the absorbing monolayer appears to be 4 h, and two successive incubations are more effective than one. The cell density of the absorbing monolayer and the handling of the Petri dish may be critical for effective removal of the cluster-initiating lymphocytes. With optimum procedure we have achieved up to 90% depletion of specific cells with no depletion of cells committed to a control antigen.