Detection of HSV-1-induced lymphokine production in human cells by a direct indicator cell addition assay.
These studies present an efficient and sensitive method for detection of T cell growth factor (TCGF) activity in human lymphocyte cultures and illustrate that T cell growth factors are associated with T lymphocyte-mediated anti-HSV-1 responses. Secretion of TCGF is induced after stimulation of human peripheral blood mononuclear cells ( PBMNC ) with herpes simplex virus type 1 (HSV-1). Lymphokine activity is detected in a simple, sensitive method by studying [3H]thymidine incorporation after the addition of murine CTLL -20 cells to cultures of gamma-irradiated (4000 R), virus-stimulated PBMNC . By using this assay, we find that PBMNC from seropositive but not seronegative individuals produce detectable TCGF activity in a dose-dependent manner after incubation with HSV-1. Maximum activity is detected between 24 to 48 hr of incubation and correlates with in vitro proliferation of nonirradiated PBMNC in response to the virus. In addition, gamma-irradiated (1000 to 3000 R) PBMNC , which are frequently used as a source of antigen-presenting cells (APC), can secrete TCGF after contact with HSV-1. Lymphokine production by the APC-containing population is eliminated by gamma-irradiation (5000 R); such APC can still present UV-inactivated HSV-1 to HSV-1-responsive lymphoblasts, indicating that lymphokine production by T cells residing in the APC population is not essential for antigen presentation.
D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis*; D008213:Lymphocyte Activation; D008211:Lymphocyte Cooperation; D007959:Lymphocyte Culture Test, Mixed; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000528:radiation effects; D051379:Mice; D018139:Simplexvirus / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
D007376:Interleukin-2