Primary immune responses in vitro general considerations

Mononuclear cell cultures derived from secondary lymphoid organs are preferable, in general, to blood cell cultures. Within the former, the cell populations required for an immune response are correctly represented, whereas blood mononuclear cells have a greater dominance of T lymphocytes, and an important population of suppressor cells. The latter can be impaired, unless they are the subject of interest, using agents such as leucyl methyl esters, although not all species possess cells with equal sensitivity to such treatments.

The expected concentration of antigen-reactive lymphocytes within a primary immune response in vitro will be not more than 1 in 10h. Considering the required cellular interactions, and that conventional cultures employ between 2 x 106 and 2 x 10 leukocytes ml"1, this low concentration of potentially reactive cells does not lend itself to success. Enrichment procedures have been particularly useful in circumventing this problem. The potential also exists to apply hollow fiber or similar capillary culture vessels, wherein cell concentrations can be maintained at 10-1000 times that in conventional culture systems. Nevertheless, the methods which have found the widest application are the use of exogenous cytokines and the preprocessing of antigen. These techniques, which will be dealt with in the next section, have the advantage over enrichment procedures in that the balance of leukocyte populations within a mononuclear cell preparation has not been modified.

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