Characterization of the Blood Mononuclear Leucocytes Producing Alpha Interferon after Stimulation with Herpes Simplex Virus in Vitro, by Means of Combined Immunohistochemical Staining and in Situ RNA‐RNA Hybridization

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Abstract
A procedure is described for combined immunohistochemical staining and in situ RNA‐RNA hybridization of human peripheral blood mononuclear leucocytes (PBMC). These cells were first stimulated in vitro by herpes simplex virus (HSV)‐infected fibroblasts and after 6 h fixed and stained with a panel of antibodies against differentiation antigens. Alpha interferon (IFN‐α)‐producing cells (IPC) were identified by in situ hybridization by means of a 35S‐labelled IFN‐α2 cRNA probe. The IPC were infrequent, one in 200–5000 PBMC, but heavily labelled with the cRNA probe. They lacked antigens typical of T and B lymphocytes, and were also essentially negative for the Leu‐M5 antigen, present on a majority of monocytes. However, 50% of IPC expressed OKM5 antigens, corresponding to the thrombospondin receptor. The IPC lacked the antigens present on null lymphocytes detected by OKT16, but most of them expressed HLA‐DR, ‐DP and ‐DQ antigens. The IPC may represent a small subpopulation in the monocyte/macrophage lineage, resembling cells described as antigen presenting and stimulators of autologous mixed lymphocyte reactions. Alternatively, they constitute a subpopulation among the null lymphocytes.

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