The Journal of Experimental Medicine

Cantor, Harvey; Asofsky, Richard; Talal, Norman

doi: 10.1084/jem.131.2.223pmid: 4392945

The ability of spleen cells from young (3 month) and old (1 yr) NZB mice to induce GVH reactions in newborn C57BL/6N mice was compared quantitatively using the Simonsen spleen assay. Young NZB cells were five times more reactive than cells from older mice. The minimum number of cells producing detectable reactions was 2 x 10 6 for the young and 10 x 10 6 for the old. Young and old cells combined and injected together produced GVH reactions quantitatively similar to those obtained with inocula composed of young cells alone. Mixtures of two cell populations producing no detectable reactions when injected separately into different recipients (1 x 10 6 young cells and 4 x 10 6 old cells) produced reactions approximately equal to those obtained with 5 x 10 6 young cells. As few as 0.25 x 10 6 young cells were sufficient to effect a reaction when combined with 4.75 x 10 6 old unreactive cells. Viability of both cell populations was essential for GVH reactivity. This evidence of synergy in GVH reactions indicates that old NZB spleen cells can be rendered immunologically more reactive in the presence of a normally reactive population. Footnotes Submitted: 28 August 1969

Cantor, Harvey; Asofsky, Richard

doi: 10.1084/jem.131.2.235pmid: 4392946

The capacity of cells from different lymphoid tissues obtained from Balb/c mice to produce graft-vs.-host (GVH) reactions was quantitatively determined in C57BL/6N by Balb/c F 1 hybrid recipients. Synergistic responses were observed when small numbers of cells from lymphoid tissues that were rich in GVH activity such as spleen and femoral lymph node were combined with weakly reactive thymus cells. Thymus and spleen cells obtained from 1-wk old mice were separately inactive but produced moderate GVH reactions when combined in equal proportions. GVH activity of spleen cells from mice thymectomized at 3 days of age was partially restored by the addition of small numbers of spleen or thymus cells from adult mice. Changes in ratio between the two cell populations markedly affected the degree of synergy. Synergy was not observed when Balb/c cells were combined with Balb/c x C57BL/6N F 1 hybrid cells and inoculated into C57BL/6N recipients, but was demonstrated when Balb/c and C57BL/6N cells were combined and inoculated into F 1 recipients, indicating that a genetic disposition to mount GVH reactions in both populations is required to produce synergy. The data indicate that at least two cell types are necessary for GVH reactions, and that synergy between cell populations results from favorable adjustments in the ratio between these two cell types. Footnotes Submitted: 28 August 1969

Feldmann, Marc; Diener, Erwin

doi: 10.1084/jem.131.2.247pmid: 5463248

Antibody-mediated suppression of the in vitro immune response to polymerized flagellin of Salmonella adelaide and to sheep erythrocytes was studied at the cellular level. Normal mouse spleen cells, preincubated in vitro with mixtures of antigen and antibody for short periods of time before being washed, did not respond to an optimal antigenic challenge in vitro, whereas similar cells treated with antibody alone gave a normal response. The degree of immune suppression was found to depend on the time of preincubation. Significant immune suppression could be induced in as short a time as 15 min, whereas profound suppression (90%) required the incubation of cells with mixtures of antigen and antibody for 4–6 hr. Mouse spleen cells treated similarly were also unable to respond subsequently to the antigen upon transfer to lethally irradiated hosts, as measured at both the level of the antigen-reactive cell and that of serum antibody production. These results were taken as evidence that in vitro an effect of antibody-mediated suppression occurred at the level of the immunocompetent cell. Similarities between immune tolerance and antibody-mediated suppression in vitro were described, and the significance of the findings discussed in the light of current concepts of the mechanism of antibody-mediated suppression. Footnotes Submitted: 2 September 1969

Aisenberg, Alan C.

doi: 10.1084/jem.131.2.275pmid: 4392947

Irradiated and thymectomized CBA mice are markedly depressed in several immunological parameters (skin homograft rejection, graft-vs.-host activity and hemolytic plaque-forming cells of the spleen, hemolysin and hemagglutinin formation, and peripheral lymphocyte counts). In the present experiments the ability of homografts of neonatal thymus placed beneath the kidney capsule to restore immunological capacity of such animals was studied. Thymus homografts which share the same H-2 locus with the CBA mouse were permanently tolerated and immunological restoration was complete. Skin from the thymus donor was specifically retained, but third party skin with even minor (non-H-2) incompatibility was normally rejected and hemolytic plaque-forming cells of the spleen were restored. Thymus homografts which differ at the H-2 locus were promptly rejected and led to accelerated rejection of skin subsequently grafted from the thymus donor. With such H-2 incompatible thymus grafts, third party skin with minor histo-incompatibility was retained while there was slight to moderate restoration of rejection of skin with major (H-2) incompatibility. Graft-vs.-host activity was restored, but there was no return of plaque-forming spleen cells, hemolysins, hemagglutinins, or peripheral lymphocyte counts. In view of the cross-reactivity at the H-2 locus in CBA mice between thymus and third party skin donors, it was felt that restoration of skin rejection and graft-vs.-host activity could be adequately explained on the basis of immunization by the thymus graft and did not require the postulation of true immune restoration or a thymus hormone. Footnotes Submitted: 5 September 1969

Henson, P. M.

doi: 10.1084/jem.131.2.287pmid: 5419850

The immunological release of vasoactive amines from rabbit platelets by a mechanism requiring blood leukocytes has been described. The reaction involved leukocytes from an immunized rabbit, antigen, and platelets and did not require the complement system. The leukocytes appeared to be mononuclear, had the ability to adhere to glass, and were found in greater numbers in the blood than in spleen, lymph nodes, thoracic duct lymph, bone marrow, and peritoneal cavity washings. Part or all of the effect on the platelets appeared to result from active release of a soluble factor from the leukocytes, which induced vasoactive amine release from the platelets. The platelets were not lysed during this reaction and the release of vasoactive amines required the active participation of metabolic pathways in the platelet. Footnotes Submitted: 8 September 1969

Coyne(Zavatone), Veronica E.; Millman, Irving; Cerda, James; Gerstley, B. J. S.; London, Thomas; Sutnick, Alton; Blumberg, Baruch S.

doi: 10.1084/jem.131.2.307pmid: 4911697

We have studied the localization of Australia antigen, a particulate substance associated with hepatitis, by means of the fluorescent antibody technique. Preparations were made from 61 liver biopsy specimens taken from patients with infectious hepatitis, serum hepatitis, and a variety of other diseases. When tested with fluorescein-conjugated rabbit anti-Au(1) antisera all 26 patients who had Au(1) in their serum had specific fluorescence in their liver cells. The fluorescence appeared in three forms: as discrete particles within the nucleus, diffuse fluorescence of the entire nucleus, and fluorescence of the nuclear rim. Occasionally there were also fluorescent particles in the cytoplasm. Other specimens were tested with the fluorescent antibody including a variety of human tissues, buffy coat smears, peripheral lymphocyte cultures, and cells obtained from bile and duodenal drainage. Among these specimens, fluorescence was found in the cytoplasm of a few cells in the bone marrow of two patients with hepatitis and Au(1) in their serum, and in the liver, spleen, mesentery, and testis of one patient with leukemia, chronic hepatitis, and Au(1) in his serum. We have shown that the presence of fluorescent particles in the liver cells is strongly associated with the presence of Au(1) in the serum and the diagnosis of viral hepatitis. We believe that this study adds support to the hypothesis that Australia antigen is an antigenic determinant of a virus capable of causing hepatitis. Footnotes Submitted: 8 September 1969

Huggins, Charles; Grand, Lorraine; Oka, Hisao

doi: 10.1084/jem.131.2.321pmid: 5419851

A series of pulse-doses of 7, 8, 12-trimethylbenz(a)anthracene-induced leukemia rapidly and consistently in very high yield in rats of Long-Evans (L-E) strain. The predominant type was a diffuse hepatic leukemia of erythroblastic stem cells. Progressive hypothermia and a decline in pituitary function are newly recognized signs of advanced leukemia in rat. Footnotes Submitted: 12 September 1969

Sugiyama, Taketoshi; Brillantes, Filomena P.

doi: 10.1084/jem.131.2.331pmid: 5419852

Cytogenetic studies on 64 rat leukemias induced with 7,8,12- and 6,8,12-trimethylbenz(a)anthracene were performed. Highly distinctive changes were found repeatedly in one special pair of chromosomes. 10 leukemias (15.6%) showed the presence of stemline(s) with trisomy of the largest telocentric chromosome (C-1 trisomy). Another chromosome abnormality, elongation of one of the pairs of the same chromosome (long C-1), was found in 5 leukemias (7.8%) as the predominant stemline and in 7 other cases as a small cell population. This chromosome abnormality, despite slight differences in their elongation rate, was defined as a new specific chromosome abnormality. Other chromosome abnormalities not related with C-1 chromosome were found in 8 cases (12.5%). The remaining 41 leukemias (64.1%) had the predominant stemline with normal karyotype. From this study it appears that three structurally different hydrocarbon carcinogens, 7,12-dimethylbenz(a)anthracene and 7,8,12- and 6,8,12-trimethylbenz(a)anthracene act on blood-forming cells by a common specific mechanism. Footnotes Submitted: 12 September 1969

Kindt, Thomas J.; Todd, Charles W.; Eichmann, Klaus; Krause, Richard M.

doi: 10.1084/jem.131.2.343pmid: 5419853

Rabbit antibodies to streptococcal polysaccharide are described which show selectivity of expression of the allotypic specificities on both the heavy (H) and light (L) chains. One of these antibodies binds weakly to Sephadex. A purification method based on this binding has yielded antibody completely lacking any group a allotypic marker on its H chains. Footnotes Submitted: 16 September 1969

Andersson, J.; Killander, D.; Möller, E.; Möller, G.

doi: 10.1084/jem.131.2.355pmid: 5419854

The mechanism of growth stimulation in allogeneic lymphocytes mixed in vitro was studied at the cell level by means of cytophotometric techniques. A pronounced increase in fluorescence intensity of fixed and acridine orange (AO) stained lymphocytes was observed as soon as after 1–3 hr in mixed culture. No increase in the amount of DNA took place during this time. The higher fluorescence intensity was due to an increased accessibility of AO binding sites in the deoxyribonucleoprotein (DNP) complex, most probably as a result of weakened bonds between the DNA and the protein moiety in the DNP complex. Similar DNP changes have been found in other systems of growth stimulation and may be one prerequisite for later induction of cellular synthetic processes. Increased AO binding only occurred when the lymphocyte donors were incompatible at the major histocompatibility locus (HL-A); there was no change in AO binding in cases of HL-A identity. The AO binding reaction probably reflects a specific recognition of HL-A antigens, whereas other antigenic discrepancies between the individuals do not seem to cause an analogous response. Footnotes Submitted: 19 September 1969