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The etiologic relationship of certain infections of the upper respiratory tract to group A hemolytic streptococci is well established and the further relationship of these infections to rheumatic fever and acute glomerulonephritis is highly probable, hence it seems important to learn more about host-parasite relations. Host-parasite relations at the cellular level are difficult to study in the patient and although tissue culture techniques at best are not a true counterpart of what happens in the patient, these techniques have some application in the study of these relationships. In this paper are reported experiments designed to study the spacial relationship and effect of group A streptococci on human tissue cells growing in tissue culture, and the effect of the tissue cells on the bacteria.Methods and materiais. Tissue cel1s.t Human tonsils were the source of lymphocytes. The tonsils were cut into fine pieces within 60 minutes of removal. The fibrous tissue was separated and the lymphoid tissue minced as fine as possible with scissors. The mince was filtered through sterile gauze. The filtrate, which contained the lymphocytes suspended in about 20 ml of medium, was composed of equal parts of mixture 199 and Eagle's basal medium containing 5% human serum, 40 pg of neomycin/ml, 1000 units of mycostatin/ml, and 100 units each of peni-*These studies were aided by a contract between the Office of Naira1 Research, Dept. of the Navy, and Vanderbilt University. cillin and streptomycin/ml. The cells were separated by centrifuging at 2500 r.p.m. for 15 minutes, resuspended in 3 ml of the above medium, then incubated for 2 hours at 37OC. Two-tenths to 0.3 ml of packed lymphocytes were the usual yield from each pair of tonsils. The lymphocytes were usually bacteriologically sterile after this treatment. They then were washed 3 times in the culture medium, taken up in 3 ml of medium without antibiotics and distributed in tubes, 1 ml per tube.Cell preparations of human tonsil,$ human liver and human monocytes were made by sub-culturing cells in antibiotic-free medium in Leighton tubes containing coverslips of No. 1 thickness. Each tube contained 0.8 ml t Cell lines used in these experiments and growth medium for each type of cell were human tonsil epithelial-like cells and mixture 199 -k 10% calf serum; human liver cells and Eagle's Basal medium + 10 % calf serum + 0.01 mg L-glutamineJml; human monocytes and Eagle's Basal medium 4-10% calf serum f 0.01 mg L-glutamindml; monkey heart and minimal essential medium -l-10% calf serum; monkey kidney and mixture 199 -k 10% calf serum; human heart (Girardj.) and minimall esentdal medium + 10% human serum -/ -0.01 mg L-glutamine/ml; human lymphocytes and equal parts of mixture 199 and Eagle's Basal medium -k 5% human serum. Unless antibio'tic-free medium was required, 1 mg each of penicillin and streptomydn was added to each 10 ml of medium. $

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