Comparison of ATP production in whole blood and lymphocyte proliferation in response to phytohemagglutinin.
Lymphocyte proliferation in response to mitogens, phytohemagglutinin (PHA), concanavalin A, pokeweed, and/or specific antigens has been the method of choice for in vitro diagnosis of cell-mediated immune dysfunction. Recently, an assay to measure intracellular adenosine triphosphate (ATP) production in response to PHA has been developed that requires a shorter, overnight incubation. We compared a standard 5- to 7-day lymphocyte mitogen stimulation assay utilizing tritiated thymidine (3H-thy) incorporation to one in which ATP production in response to PHA by CD4-positive cells is measured in a luminometer that requires only 18-24 hr. A total of 20 patient samples suspected of having decreased cell-mediated immunity submitted for mitogen induced lymphocyte proliferation and 21 normal controls were tested in both assays. A comparison of these two methods has demonstrated that the screening ATP assay has a sensitivity at 24 hr of 100% in detecting decreased PHA induced lymphocyte proliferation at 5 days and a specificity of 85% in the samples obtained from normal controls. The data indicate that the ATP assay may be a useful screening tool for more rapid detection of blood samples with decreased cell-mediated immune responses. However, a positive screen should always be confirmed by 3H-thy uptake using mitogens and recall antigens like candida and tetanus.
Augustine
Nancy H
NH
Department of Pathology, University of Utah School of Medicine, Salt Lake City, Utah 84132, USA. nancy.augustine@path.utah.edu
Pasi
Brian M
BM
Hill
Harry R
HR
D000255:Adenosine Triphosphate / Q000097:blood*; D000293:Adolescent; D000328:Adult; D049109:Cell Proliferation / Q000187:drug effects; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007111:Immunity, Cellular / Q000502:physiology; D007153:Immunologic Deficiency Syndromes / Q000175:diagnosis* / Q000276:immunology; D007159:Immunologic Tests / Q000379:methods; D007223:Infant; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects; D008297:Male; D008875:Middle Aged; D008934:Mitogens / Q000494:pharmacology*; D010835:Phytohemagglutinins / Q000494:pharmacology*; D012680:Sensitivity and Specificity
D008934:Mitogens; D010835:Phytohemagglutinins; D000255:Adenosine Triphosphate