T. N. Harris

Effect of Cortisone on Some Reactions of Hypersensitivity in Laboratory Animals.

Summary In rabbits and guinea pigs sensitized with tubercle bacilli (BCG) dermal reactivity to Old Tuberculin and to cytoplasmic particles from tubercle bacilli and systemic shock produced by intraperitoneal injection of OT could be suppressed by treatment with cortisone. Four days after cessation of treatment the skin test again became positive.

SEROLOGIC RESPONSE TO STREPTOCOCCAL HEMOLYSIN AND HYALURONIDASE IN STREPTOCOCCAL AND RHEUMATIC INFECTION

Although it has as yet not been possible to demonstrate the specific role of a causative organism in rheumatic fever, there has been considerable epi-demiologic (1) and serologic (2) evidence of some relationship between the hemolytic streptococcus and this disease. Studies of both cellular and extracellular substances of the hemolytic strepto-coccus have with rare exceptions shown that groups of patients with acute rheumatic fever were indistinguishable, by serologic reactions, from streptococcal convalescents. These studies have included those of reactions to cellular antigens, such as agglutination, precipitation with the M protein, and, more recently , complement fixation with somatic nucleo-protein fractions (3, 4); and, among the reactions to extracellular antigens, antistreptolysin, anti-fibrinolysin, and, most recently, antihyaluronidase. A number of reports have shown that the sera of patients with acute rheumatic fever contained antibodies to streptococcal hyaluronidase in equal or greater concentration than those of patients convalescing from acute infection with the hemo-lytic streptococcus (5-10). This difference was manifest in both a higher geometric mean titer among the rheumatic fever patients and by a distribution of these titers which extended beyond the range of antihyaluronidase titers found after acute streptococcal infection. (The antihyaluronidase referred to here is the neutralizing antibody specific for hyaluronidase of streptococcal origin, and is stable at 560 C. for 30 minutes. This point is made because in addition to the one or more thermolabile inhibitors of testicular hyaluronidase which have been described in human sera [11-14], thermolabile inhibitors of streptococcal hyaluroni-dase have been described which can be reactivated by complement [15, 16]. The latter are distinct from the specific, thermostable inhibitor which has been measured in these studies.) This difference between the antihyaluronidase titers in rheumatic and streptococcal infection has been discussed elsewhere (7). First, the factor of time was considered as a possible cause for this difference, since the streptococcal convalescents included in our series were studied three weeks after the onset of the disease, and it was felt that perhaps the longer contact with the streptococcus which some of the rheumatic patients may have had might have given rise to a greater production of this antibody. Other serologic circumstances which might give rise to such a difference were considered. Finally it was pointed out that only after thorough exploration of such possibilities could the question be raised as to whether the relatively higher antihyaluronidase titers in rheumatic fever might have any implication of some role of …

Gel-Precipitation of Streptococcal Culture Supernates with Sera of Patients With Rheumatic Fever and Streptococcal Infection.∗

Summary 1. Precipitin tests in semi-solid agar (gel-precipitin) have been carried out between concentrates of streptococcal culture supernates and sera of various sources. Two technics of diffusion in 1 dimension (tube) were used: the single-diffusion technic of Oudin, and a double-diffusion technic. 2. Sera of rabbits previously injected with streptococcal culture supernates showed increasing numbers of bands as the course of injections progressed. Sera of such rabbits showed as many as 5 bands when tested with such concentrates, indicating that this was the minimum number of antigen-antibody systems present. 3. Sera of patients with acute rheumatic fever and of patients convalescent from an acute streptococcal infection (scarlet fever) were also tested by this technic against concentrates of streptococcal culture supernates. The group with rheumatic fever showed a greater number of bands (a range of 2-7) than did the group with scarlet fever (0-4), when tested by the double-diffusion technic. The corresponding ranges found with the single-diffusion technic were 1-5 and 0-2, respectively. In a sampling of sera from convalescents from scarlet fever the number of bands was found to be no greater 6 weeks after the onset of the infection than 3 weeks after the onset.

Solubilization of h-2 histocompatibility antigens of the mouse by triton x-100 and butanol.

Histocompatibility antigens (HCA) have been solubilized by the use of Triton X-100 and n-butanol from cell membrane fragments of spleens and livers of BALB/c and CBA mice. These soluble preparations could inhibit or neutralize allogeneic antibody (CBA anti-BALB or BALB anti-CBA) in two biological tests, cytotoxicity and the suppression of antibody formation by appropriately stimulated spleen cells of the same strain. The neutralizing activities of the spleen-derived preparations, per milligram, were about 10 times those derived from liver, and preparations extracted by butanol or by papain were of substantially higher specific activity than those extracted by Triton X-100. All preparations contained active molecular species excluded by Sephadex G-200 and others, nonexcluded, in various proportions. Both Sephadex fractions could be adsorbed to sheep red blood cells (SRBC) but only the adsorption of the G-200-excluded material rendered the SRBC agglutinable by CBA anti-BALB antibodies; the non-excluded fraction could compete with the excluded fraction to prevent agglutination of the coated SRBC by such antibody. The soluble antigen could inhibit the allogeneic antibody in two serological tests, the adsorption hemagglutination (AH) reaction referred to, and the standard hemagglutination test of BALB RBC by CBA anti-BALB antibody.

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.

Study by cathode-ray oscillography of some innocent and pathologic cardiac murmurs of children☆☆☆

Abstract Cardiac murmurs of several types found in children and adolescents have been recorded on magnetic tape and examined by cathode-ray oscillography. The types of murmurs studied have included two presumably innocent types of murmurs, the precordial vibratory (“twanging-string” murmur of Still) and the pulmonic-systolic murmur, which comprise the great majority of nonpathologic murmurs of childhood, and several types of organic murmurs, those due to aortic and mitral insufficiency and to a variety of congenital malformations. In terms of wave form, the precordial vibratory murmurs examined showed a discernible basic wave of frequency ranging from 90 to 180 cycles per second, among various subjects, with a small and variable degree of distortion. In oscillograms of pulmonic-systolic murmurs a basic wave in the range of frequency given above could also be discerned, but the degree of distortion was noticeably greater than that found in the precordial vibratory murmurs. In the case of the specimens examined in all the groups of organic murmurs mentioned above the wave forms were highly complex, with no detectable basic wave or recognizable pattern. A possible implication of these differences in wave form is discussed.

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.

STREPTOCOCCAL ANTIHYALURONIDASE TITERS IN THE SERA OF PATIENTS WITH RHEUMATOID ARTHRITIS AND GLOMERULONEPHRITIS

Excerpt Recent observations on streptococcal antihyaluronidase in human sera have shown that the concentration of that antibody is elevated above the normal in acute rheumatic and streptococcal inf...

SUPPRESSION OF TRANSFERRED LYMPH NODE CELLS IN NEONATAL RABBITS BY THE INJECTION OF LEUKOCYTES

Although tissue transplantation immunity has been most widely studied i n t he area of skin homografting, aspects of transplantation immunity have also been studied in a number of other systems, including recently the homotransfer of lymph node cells. Whereas in skin homografting the timing of rejection of the graft affords the measure of immunological response of the host, in the studies of lymph node cell transfer the measure of transplantation reaction of the recipient’s tissues has been provided by the degree of interference with the functional activity of transferred cells in the production of antibody. The system of lymph node cell transfer in which tissue transplantation effects have been studied in this laboratory is summarized schematically in FIGURE 1. On the left side of the figure are shown the experimental situation and the kind of data obtained on simple transfer of lymph node cells in this system. Lymph node cells obtained from rabbits not previously injected with Shigella paradysenteriae are incubated in vitro with a Shigella-trypsin filtrate and transferred to homologous recipient animals. In the sera of the latter, agglutinins to Shigella can be found subsequently in a typical pattern of concentration versus time.’ It was shown recently that, if the transfer of such cells was preceded, by about a week, by the injection of the prospective recipient animals with leukocytes obtained from the donors of the lymph node cells, agglutinins failed to appear in the sera of the recipients or did so in low concentration, as shown in the central section of FIGURE 1. This indicated that the injection of leukocytes prior to transfer had induced an immunological response to the transplantation antigens of the donor, and that this reaction had caused suppression of the production of agglutinins on subsequent transfer of lymph node cells? It was shown later that this suppressive effect could be transferred passively by serum. If leukocytes were obtained from the prospective donors, pooled, and injected into other adult rabbits, the pooled sera of the latter, if injected into recipients together with lymph node cells, could cause suppression of the transferred cells, as indicated by the low concentration of agglutinin in the sera of the recipient animal^.^ This experimental design is shown in the last panel of FIGURE 1. The present study grew out of an attempt to demonstrate in this system another effect that has been observed in tissue transplantation, namely, that of immunological tolerance. It was considered that, if a state of tolerance could be established in neonatal rabbits by the injection of rabbit leukocytes, this state might be reflected on subsequent transfer of lymph node cells from the adult to the young rabbits by agglutinin levels in the sera of the young recipient animals higher than those usually found after transfer. Such an effect

Suppressive antibody in heterologous anti-lymphocytic serum estimated in vitro by the hemolytic antibody plaque test, species-specific and strain-specific antibodies in rabbit-anti-mouse anti-lymphocytic sera.

Summary: Heterologous anti-lyniphocytic sera (ALS) produced by injection of rabbits with spleen cells from CBA or BALB/c mice were found to suppress hemolytic antibody plaque formation by antigenically stimulated spleen cells following prior incubation of such cells with ALS and complement. Cytotoxicity of such antisera for mouse spleen cells was also shown. Both antibody effects were demonstrable in high titer in both anti-CBA and anti-BALB ALS tested against cells of both strains. The bulk of the antibody was species-specific. However, the presence of significant amounts of strain-specific antibody was indicated, first, by higher titers in some classes of sera tested against cells of the immunizing strain than against those of the other strain; second, by absorption of the ALS with cell fragments of the other strain before testing against cells of the immunizing strain; and, third, by sequential absorption of ALS by cell fragments of the other strain and then those of the immunizing strain and elution of antibodies from these absorptions. Hemolytic antibodies appearing in such sera were more preponderantly species-specific. If strainspecific hemolysins were present, they could have constituted at most a substantially smaller fraction. The strain-specific component of an anti-CBA ALS was shown in vivo to cause prolonged retention of BALB skin grafts by CBA mice and reduction in the production of agglutinins to Sltigella by such mice