Lymphocyte transformation and interferon production in human mononuclear cell microcultures for assay of cellular immunity to herpes simplex virus

Abstract

Interferon production and transformation in respowse to herpes simplex virus antigen were studied in microcultures of human mononuclear cells. Mononuclear cells consisting of monocytes and both T and B lymphocytes were purified by Ficoll-Hypaque gradients. Lymphocytes, predominantly T with 5% B, were obtained by passage of buffy-coat cells through nylon fiber columns. For some experiments, autochthonous macrophages and column-purified lymphocytes were stimulated with herpesvirus antigen. The effect of specific antibody and cell concentration on reactivity is described. Crude and purified antigens were compared as cell culture stimulants. Significant differences in transformation and interferon were observed between donors with a history of herpes labialis and donors with no detectable antibody, both in cultures prepared by Ficoll-Hypaque gradients and by column purification of lymphocytes. Cultures from seronegative donors prepared by Ficoll-Hypaque gradients produced interferon but did not transform when stimulated by herpes simplex antigen. "Immune" interferon production, that is, type II as opposed to type I, occurred only with autochthonous macrophage and columm-purified lymphocyte cultures. Interferon produced by Ficoll-Hypaque-purified mononuclear cultures was type I, and its production was unrelated to immune status. Similarly, column-purified lymphocytes responded to herpes simplex virus antigen with type I interferon if obtained from a seropositive donor.

Interferon Production, Cellular and Humoral Immunity in Splenectomized Patients

“…The serum interferon assay was performed on serum obtained from heparin stabilized blood. The assay was performed by using a micromethod, as previously described (Haahr et al 1976). The interferon titer was the the highest dilution that reduced the vesicular stomatitis virus cytophatic effect by 50%.…”

Immune Interferon Produced In Vitro as a Quantitative Indicator of Cell‐Mediated Immunity

“…Interferon has been found to be one of the mediators of cell-mediated immunity (7,10,20). It is produced in vitro upon exposure of human or mouse leukocytes to mitogens and to viral or nonviral antigens (4,6,9,18…”

“…In 1966, Glasgow provided the first direct evidence of the fact that immunity and immune mechanisms are intimately associated with interferon production (10). Subsequent experiments demonstrated that nonviral antigens stimulated interferon production in cultures of lymphocytes from sensitized donors but not in cultures from donors who lack sensitivity to those antigens (4,6,9).…”

Sequential Changes in T and B Lymphocyte Responses to Herpes Simplex Virus in Man

“…These findings were unexpected as it seemed probable that the responding cell in the UF population was a T cell. However, Haahr et al [6] have noted a reduced response to HSV of nylon wool purified T cells as compared to UF cells 1-12 weeks after a recurrent infection although the response was not related to the time since the onset of the infection.…”

“…The cell type responsible for each response has not been previously determined in man. Immune specific interferon is released in response to HSV antigens by sensitized human T lymphocytes and autologous macrophages [6]. Depletion of T cells in mice increases their susceptibility to herpetic infection, but resistance can be restored by transfer to T cells [13,15].…”