The requirement for macrophages in the secondary immune response to antigens of small and large size in vitro.

Tissue culture techniques were combined with cell separation procedures, antimacrophage serum and soluble and particulate forms of sheep red cell antigen to investigate the cellular requirements for a secondary antibody response. By using the highly efficient active adherence column separation method of Shortman which almost completely removes phagocytic cells, it was found that the secondary response to SRC was macrophage dependent. This conclusion was verified by the use of specific antimacrophage serum and by a combination of both methods. Critical tests were used to verify that these separation methods acted on phagocytic and not on other cells, for example, thymus or bone marrow derived lymphocytes. In contrast, the secondary immune response to POL and solubilized SRC antigen was not dependent on the presence of phagocytic cells, as highly purified lymphocytes responded normally to these antigens. Antimacrophage serum did not depress the immune response to these soluble antigens. These results indicate that the requirement for macrophage activity depends on the physical size of the antigen. These findings, obtained in the secondary response in vitro, were closely analogous to previous findings from this laboratory on the cellular basis of the primary response to SRC and POL. The reasons for the different cellular bases of the secondary immune response to various molecular forms of antigen are discussed. The similar cellular basis of the primary and secondary response in vitro suggests that the basic mechanisms of immunization in these responses are the same.