DNA SYNTHESIS IN MIXED CULTURES OF DOG LEUCOCYTES: DIFFERENTIAL EFFECT OF X-RADIATION AND FREEZE-THAWING ON CELLULAR ISOANTIGENICITY.,
Abstract
A marked induction of DNA synthesis occurred in 50-50 mixed cultures of peripheral blood leucocytes derived from sets of allogeneic (Beagle and mongrel) normal dog donors. DNA-synthetic activity at selected times was determined by liquid scintillation counting following pulse labeling of cell populations with 3H-thymidine. In a typical experiment, DNA synthesis is mixed cultures attained a maximum value at the 6th day (61,000 CPM) followed by a steady decline thereafter (to 7,500 CPM by day 9), as contrasted with control values (65-86 CPM). In leucocyte cultures of cells isolated and mixed on the same day, considerable DNA labeling (1900-3000 CPM) occurred at time 0. These values declined, (600 CPM at 18 hours; 230 CPM at day 2), and then rose later (2400 CPM at day 4). Incubation of individual cell suspensions for 18 hours prior to mixing eliminated the elevated zero time label. X-irradiation (2500 rads) of one (or the other) of the cell components of the mixture changed the reation from a 'two way' immunological reaction. When one (or the other) of a pair was frozen (-196C) and thawed (37C) three times and momentarily homogenized before mixing with intact allogeneic leucocytes, no induction of DNA synthesis occurred. These results suggest that leucocyte isoantigenic factors are not inactivated or destroyed by X-irradiation and can induce DNA synthesis in allogeneic leucocytes; whereas, they are labile to freeze-thawing. The results are compatible with the known chemical and immunological properties of transplantation antigens. (Author)
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 30, 1966
- Accession Number
- AD0640261
Entities
People
- Henry M. Haire
- Leonard J. Cole
- Marjorie J. Jones
- Raymond K. Main
Organizations
- Naval Radiological Defense Laboratory