Typhoid vaccine and sheep erythrocytes were injected subcutaneously into the feet of rabbits, and the subsequent formation of agglutinins and hemolysins in the popliteal lymph node was compared with the output of lymphocytes through the efferent lymph and with changes in the lymph node. Antibodies began to appear in the efferent lymph 2 to 4 days after the injection of the antigen and reached their highest titer after 6 days. This was preceded by a sharp rise in the output of lymphocytes through the efferent lymph, while in the lymph node there was lymphatic hyperplasia after preliminary infiltration of granulocytes and monocytes. This hyperplasia was first of a diffuse type, but was later superseded by large so called germinal centers, the latter lagging somewhat behind the rise in antibody titer. The fact that the tissue response accompanying the formation of antibodies was chiefly a lymphocytic one points to the lymphocyte as a factor in the formation of antibodies.
In a previous paper(l) the production of antibodies in the p~pliteal lymph node of the rabbit was compared with the output of lymphocytes in the efferent lymph vessel of the node and with the morphological changes which took place within the node itself. Antibodies first appeared in the efferent lymph 2 to 4 days after the injection of the antigen and reached their highest titer after 6 days. This antibody formation was preceded and accompanied by a rise in the output of lymphocytes in the efferent lymph which ranged from 15,000 to 20,000 per c. ram. to 60,000 to 80,000 per c. ram. or more. At the same time hyperplasia of the lymphatic tissue within the node occurred, resulting in some experiments in a weight increase of the node of 0.2 gm. to 1.0 grn. or more. The fact that the cellular response during antibody formation was chiefly lymphocytic suggested that the lymphocyte may play an important r61e in antibody formation.The present investigation deals with the r61e in antibody formation played by the lymphocyte itself. Instead of comparing antibody production and cellular response in the pad of the hind foot of the rabbit (which was the site of injection of antigen), the lymph contained in the afferent lymph vessel, the popliteal lymph node (the only node regional to the site of injection), the efferent lymph, and the serum, we now determined antibodies within the lymphocytes and the supernatant fluid of the efferent lymph collected during the period of antibody formation in the lymph node. The results of this study not only stregthen the conclusions drawn in the previous paper, but appear to show for the first time that the lymphocyte is engaged in antibody formation. MethodsThe antigens were prepared as follows: Typhoid bacilli, strain tt 901, were taken from an 18 hour nutrient broth culture, washed 3 times by centrifugafion in saline, heated in a water bath at 56°C. for 1 hour, and then diluted with an equal volume of saline to make a suspension of approximately 50 per cent. Sheep erythrocytes, obtained from blood drawn within the week, were washed 3 times and diluted with saline to make an approximately 50 per cent suspension. In all cases, 0.2 cc. of antigen was injected subcutaneously into the plantar surface of the hind feet of 2000 gin. Chinchilla rabbits, and the puncture wounds were closed with collodion.
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