FURTHER STUDIES OF THE STIMULATION OF DNA SYNTHESIS IN CULTURES OF SPLEEN CELL SUSPENSIONS BY HOMOLOGOUS CELLS IN INBRED STRAINS OF MICE AND RATS

The early proliferative response previously demonstrated in rabbits has now been shown to follow the mixing of spleen cell suspensions from 2 inbred strains of mice or rats. The size of the response is comparable to that seen in cells from hyperimmune animals exposed to antigen in vitro. Autoradiographs of cells from stimulated cultures showed 1 to 4 per cent of the total population had incorporated thymidine. Modifications in the conditions necessary for the culture of mice and rat spleen cell suspensions and the measurement of thymidine incorporation are described. No responses were observed in isologous mixes. The responses obtained on mixing individual pairs of spleens from different strains showed relatively little variation. Responses were obtained in all of the 21 possible combinations between 7 inbred strains of mice. Responses were obtained when parental cells were mixed with their F1 hybrids. Analysis of these responses showed that, in every case, parental-F1 hybrid responses were less intense than those between the 2 parents. It was shown that there was no inherent defect in the ability of the hybrid cells to respond when mixed with an unrelated strain. The results suggested that the hybrid cells made no response to the parent cells although this was not conclusively established. This has been taken as circumstantial evidence that the response is immunological in nature. The significance of the vigor of the response and the large fraction of the immunologically competent cells that take part is discussed

SPLEEN CELL PROLIFERATION IN RESPONSE TO HOMOLOGOUS ANTIGENS STUDIED IN CONGENIC RESISTANT STRAINS OF MICE

The proliferative responses obtained when spleen cell suspensions from two different inbred strains of mice were mixed were investigated further using congenic resistant strain pairs. Strong responses were obtained in 9 cases out of 12 where the two strains differed at a single gene locus controlling an H-2 histocompatibility antigen. No responses were obtained where the difference occurred at loci controlling weak histocompatibility antigens. These findings have been taken to provide additional circumstantial evidence that the response represents an in vitro homograft reaction to homologous tissue antigens

REGULATION OF THE IMMUNE RESPONSE : I. DIFFERENTIAL EFFECT OF PASSIVELY ADMINISTERED ANTIBODY ON THE THYMUS-DERIVED AND BONE MARROW-DERIVED LYMPHOCYTES

The effect of passively transfered antiserum against sheep erythrocytes (SRBC) on the antigen stimulated increase of SRBC-specific plaque-forming cells (anti-SRBC-PFC) and SRBC-specific thymus-derived lymphocytes (SRBC-specific T-cells) in the mouse spleen was examined. A dose of antiserum which severely suppressed the development of anti-SRBC-PFC did not prevent the increase in SRBC-specific T-cells, as measured by their ability to cooperate in the in vitro response to trinitrophenylated (TNP) SRBC. It was shown that the insensitivity of these T-cells to antiserum could not be explained by their low antigen requirement as compared to that of PFC. In the in vivo response of mice to TNP-SRBC, antibody specific for TNP suppressed the appearance of both anti-TNP- and anti-SRBC-PFC. The presence of free SRBC specifically prevented the suppression of the anti-SRBC-PFC. These observations are consistent with opsonization by phagocytic cells as the primary means of the observed suppression of PFC development by antibody

CELLULAR EVENTS IN THE IMMUNE RESPONSE : ANALYSIS AND IN VITRO RESPONSE OF MOUSE SPLEEN CELL POPULATIONS SEPARATED BY DIFFERENTIAL FLOTATION IN ALBUMIN GRADIENTS

Cell suspensions from the spleens of normal mice or mice injected with sheep erythrocytes were separated on a discontinous bovine serum albumin density gradient. Four bands or subpopulations were obtained and were assayed for antibody-forming cells, and for antigen-sensitive precursor cells. The antibody-forming cells were assayed by the hemolytic plaque assay and the antigen-sensitive precursors were assayed by the number of plaque-forming cells which developed after 3 or 5 day's culture with antigen. It was found that both antibody-forming cells and their precursors were present in the denser region of the gradient when spleen cell suspensions were taken from unimmunized mice. In contrast, both antibody-forming cells and precursors floated to the top in cell suspensions from mice sacrificed 1, 2, or 3 days after antigen injection. The change in density was detectable as early as 12 hr and was complete by 18 hr. The cell which changed in density was specific for the antigen that brought about that change. The significance of these findings is discussed