Interaction of Mouse Spleen Cells in Diffusion Chambers

Abstract

IT has been suggested that the interaction which occurs between lymphoid cells of two individuals in mixed cell cultures provides a measure of histocompatibility difference^1–4. This suggestion is based on the assumption that the interaction, which takes the form of a proliferative response, is immunological in nature. While most work has been performed with human cells, it is obviously desirable that this assumption is tested in situations where histocompatibility difference can be precisely defined, as in inbred strains of the rat and mouse, Wilson^5,6 has shown that in mixed cultures of parental strain and F₁ hybrid rat cells most of the response is parental in type, as would be expected for an interaction with an immune basis. Dutton^7,8 has demonstrated a proliferative response in short term mixed spleen cell cultures of mouse origin. Murine cells, however, have proved refractory to growth in vitro during long periods and, to facilitate their survival and function, Nettescheim et al.^9,10 have cultured them in diffusion chambers in vivo. We have followed this method to examine the interaction between mixed suspensions of mouse spleen cells paying particular attention to the comparative proliferation of cells from C57Bl and CBA H-T₆T₆ to each other and to their F₁ hybrid. The presence of two T⁶ marker chromosomes in CBA H-T₆T₆ cells and of a single T⁶ chromosome in F₁ cells enables the direction of the response to be determined and so, in parental type/F₁ hybrid mixtures, provides evidence for the immunological specificity of the reaction.