Miriam B. Farber

Effect of injection of rabbit leucocytes into neonatal rabbits on subsequent lymph node cell transfer.

Summary 1. Newborn rabbits were injected with leucocytes obtained from adult rabbits which were prospective donors of lymph node cells. When antigen-incubated lymph node cells from these donors were transferred to young rabbits 1-2 1/2 months later, agglutinins failed to appear, or did so in low titer. This suppression of transferred lymph node cells indicated that tissues of newborn rabbits had reacted to antigens of leucocytes of donor animals. 2. Transfer of lymph node cells incubated in vitro with Shigella-trypsin filtrate to uninjected newborn rabbits (1-11 days of age) resulted in the appearance of anti-Shigella agglutinins.

Transfer to x-irradiated rabbits of lymph node cells incubated in vitro with Shigella paradysenteriae.

Summary Cells were teased from the popliteal lymph nodes of rabbits which had not been injected with dysentery bacilli. These cells were incubated with dysentery bacilli in vitro and transferred to X-irradiated recipients. Agglutinins were detected in the sera of such recipients on the fourth day after transfer. When the cells were heated prior to transfer agglutinins did not appear during the first week after transfer. Similar results were obtained when suspensions of lymph node cells and suspensions of dysentery organisms were injected separately into irradiated rabbits.

Effect of antigraft strain antibodies on skin allografts in immunologically tolerant mice.

SUMMARY Alloantibody-containing globulins of the mouse (CBA or C3H versus BALB/c), selected for relatively high titer of anti-BALB/c suppressive antibody, have been examined for their effect on BALB/c skin grafts in CBA or C3H mice rendered immunologically tolerant of BALB/c grafts by perinatal injections of (CBA × BALB/c) F1h spleen cells. Such globulin pools, obtained from ascitic fluid collected 11-13 days after secondary injection of BALB/c spleen cells and injected at the time of grafting, were found to cause complete rejection of the grafts between 10 and 20 days after grafting, the rejection by individual pools occurring largely within a relatively narrow range of 3-6 days. Globulins obtained from the same groups of mice later than 11-13 days caused rejection a few days later. It was also possible to cause rejection of 15 of 17 established BALB/c skin grafts after 2 months in situ on immunologically tolerant hosts, more than one-half of the 15 grafts rejected being distinctly smaller within 10 days after the beginning of the injections, and completely rejected within 16 days. Mice whose tolerated grafts had been rejected following the injections of the alloantibodies were then given second allografts of BALB/c skin to test for residual tolerance. All of these were retained for periods between 21 and 45 days.

PREPARATION BY ELUTION FROM SPECIFIC AGGREGATES OF THE ISOANTIBODY CAUSING REJECTION OF TRANSFERRED RABBIT LYMPH NODE CELLS.

A fraction of rabbit lymph node cells and splenic cells has been prepared by differential centrifugation of sonically disrupted cells: a high‐speed centrifugal sediment from the supernate of a lower speed centrifugation. This cell fraction gives complement fixation with rabbit anti‐rabbit‐leucocyte serum and gives evidence of containing nearly all the complement‐fixing isoantigen of such cells. The antibody in these sera which causes the rejection or suppression of homotransferred lymph node cells can be removed from such sera by absorption with the cell fraction. From the specific aggregates thus obtained the suppressive antibody can be eluted with recovery of 25% of the antibody present in the original serum. The suppressive antibody can be absorbed from such eluates by the cell‐fraction. The antibody thus eluted shows properties of a pseudoglobulin.

Accelerated rejection of allogeneic skin grafts in the mouse after injections of histocompatibility antigens solubilized by Triton, butanol, and papain.

BALB/e histocompatibility antigens (HCA), solubilized from cell fragments of spleens and livers by Triton X-100, butanol, and papain, were tested for immunogenicity as transplantation antigens by injection into normal CBA mice and later observation of the time of rejection of BALB skin grafts in these mice. Each type of preparation, injected 3 times in saline or in oil adjuvant, caused accelerated rejection of such grafts. Preparations solubilized by butanol or papain showed higher specific activity than the Triton-solubilized, and preparations from spleen showed higher activity than those from liver. These differences were similar to the differences in specific activity shown by the respective preparations in the inhibition of 2 effects of CBA anti-BALB antibody, cytotoxicity, and the suppression of hemolytic antibody plaque production by BALB spleen cells. CBA antigen solubilized by butanol from liver cell fragments similarly caused accelerated rejection of CBA skin grafts in BALB mice following injection on the same schedule.

Various Effects Of Alloantibody-containing Globulins On The Time Of Retention Of Skin Allografts In The Mouse

A study of the effect of alloantibody-containing ascitic fluid globulin of mice on the retention of skin allografts has been carried out using, as a tentative measure of the relevant antibody, the titer of the antibody in such globulins which can suppress the production of hemolytic antibody plaques by donor strain spleen cells. When globulins with sufficiently high suppressive titer were available in sufficient quantities, pools of such globulins were injected into normal mice which were given skin allografts in the same strain combination as that used for production of the alloantibodies. Among globulins, all of which were above a given threshold of suppressive titer and all within a given range, four kinds of effect were found on the time of retention of the allograft: accelerated rejection in almost all of the grafted mice (relative to control mice injected with normal ascitic globulin), or accelerated rejection in some of the grafted mice, or no apparent effect, or, finally, prolonged retention of the graft. There was no relation between the suppressive titer of the globulin pool and its effect on the time of rejection of the skin graft. The fact that anti-H-2 antibody-containing globulin pools could cause, in given instances, these opposite effects of accelerated rejection and prolonged retention of the allogeneic skin grafts suggested the participation of at least two alloantibodies with the same specificity but with differing biological effect, the fate of the graft depending on the relative concentrations of the two. The possible roles of two such competing antibodies are being explored.

Suppression of growth of a mouse plasmacytoma by the igg2 fraction of syngeneic antitumor globulin.

SummaryThe effects of syngeneic antitumor antibody on transplanted plasmacytoma cells have been examined. Globulin was prepared from ascites fluids produced in Balb/c mice injected with MOPC 315 tumor cells and bearing the solid tumor. Normal Balb/c mice were given inoculations of tumor cells that had been incubated with the antitumor globulin obtained at various intervals after immunization, or with portions of such globulins. These materials were prepared to express the IgG2 class antibody by three procedures: precipitation with heterologous anti-mouse IgG1, passage through columns of Sepharose anti-IgG1, or adsorption to and elution from heat- and formalin-killed protein A-bearing staphylococci. The original antitumor globulins showed differences with time relative to the second injection of the immunizing tumor, in that a number of the earlier pools led to some suppression of tumor growth, and a number of the later pools led to some enhancement. Of the globulins obtained later, preparations expressing the IgG2 class of antibody by precipitation with anti-IgG1 serum caused some suppression of tumor growth. Pronounced suppression of growth was consistently obtained with anti-MOPC 315 globulin freed of IgG1 by passing it through an anti-IgG1 immunoadsorbent, and with IgG2a preparations obtained by elution from the protein A-bearing staphylococci. Of the anti-IgG1 column-treated preparations, the suppressive effect was maximal in globulin obtained 15–26 days after the second immunization. The suppressive effect of these preparations could be removed by absorption with MOPC 315 cells, but not by cells of another Balb/c tumor nor by two other plasmacytomas.