Role of the Lymphocyte in Antibody Formation 83 



us for use with the soluble foreign protein immunologic techniques in the 



rabbit. 



MATERIALS AND METHODS 



Complete descriptions of the materials and methods used in these cell 

 transfer studies have been published previously. 44 - 48 - 51 Table 7-1 outlines a 

 typical experiment. 



Table 7-1. Cell Transfer Schema 



Donors 



Days 1-30 BSA (antigen) injections, total =•= 250 nig. 



Day 53 Lymph node cells harvested 



Recipients 



Day 51 400 r whole body x-radiation 



Day 53 Injection of donor cells, S.C. and I.M. injection of I* BSA 



Days 54-62 Sacrificed at various intervals, after bleedings for immunologic studies 



Adult rabbits were immunized by repeated intravenous and subcutaneous 

 injections of bovine serum albumin (BSA) totaling over 250 mg. for a period 

 of about a month, and those making good antibody responses were selected 

 as donors. Approximately three weeks after the last antigen injection, when 

 the production of antibody was Low, the donors were sacrificed, and mes- 

 enteric and popliteal lymph nodes pooled. The lymph node pools were 

 minced, and free cell suspensions were obtained by filtering the cold mince 

 through fine-mesh tantalum gauze. Over 90 per cent of these cells did not 

 stain with trypan blue at the time of injection and were considered to be 

 viable. 



After washing with macromolecular suspending solutions, usually buf- 

 fered polyvinylpyrrolidone (PYP-Macrose, Schenley), about half a billion 

 of the donor cells were injected subcutaneouslv and intramuscularly into 

 the anterior abdominal walls of recipient rabbits. 



The recipient rabbits had received 400 r whole body x-radiation two days 

 prior to the transfer of cells and had never been exposed to any foreign 

 protein antigens. This dosage of x-radiation to rabbits will ablate tem- 

 porarily the immune response to foreign proteins. ■"'- 



Immediately after the injection of cells, 1.5 mg. of I ,3, -trace labeled bovine 

 serum albumin (I* BSA) were injected intravenously into the recipient. In 

 relation to the number of transferred cells, this antigen dosage is compar- 

 able to 150-300 mo. in an intact adult rabbit, since the transferred cells 



O 



equalled about 0.5 to 1 per cent of the total lymphoid tissue of a normal 

 o- to 8-pound rabbit. 53 The radioactive label was employed for ease in 

 antigen determinations. The immune response to the antigen bovine serum 



