The Journal of Experimental Medicine

Jose, David G.; Good, Robert A.

doi: 10.1084/jem.137.1.1pmid: 4688318

Mice were fed diets deficient in a single essential amino acid, and the primary immune responses to inoculation of allogenic tumor cells was measured by in vitro assay of cellular immunity. Moderate reduction of the amino acids phenylalanine-tyrosine, valine, threonine, methionine-cystine, isoleucine, and tryptophane in the diet produced profound depression of hemagglutinating and blocking antibody responses, although cytotoxic cell-mediated immunity remained intact. These diets had previously been shown to result in a selective depression of tumor growth in mice. Limitation of the amino acids arginine, histidine, and lysine in the diets gave rise to only slight depression of the immune responses. These diets had previously been shown to produce a proportional decrease in both tumor growth and host body weight. Moderate leucine restriction resulted in a paradoxical depression of cytotoxic cell-mediated immunity with little effect on serum blocking activity. Slight increases had previously been noted in the weight of tumors in mice fed leucine-restricted diets. Deficiency or imbalance of essential amino acids in the diet may produce profound depression of immune responses and apparent, marked changes in the immune resistance of the host animal to tumors. Footnotes Submitted: 4 August 1972

Thompson, Jan; van Furth, Ralph

doi: 10.1084/jem.137.1.10pmid: 4688316

To elucidate mechanisms underlying the prolonged monocytopenia induced in the peripheral blood of mice by injection of a subcutaneous depot of hydrocortisone acetate, the effect of this compound on the production of monocytes and their release from the bone marrow was studied. Hydrocortisone was found to cause a rapid reduction of the bone marrow promonocytes to about 65% of their initial number. The number of monocytes in the bone marrow decreased gradually, over a period of 96 h, to 75% of the initial value. The mitotic activity of the promonocytes was not diminished, as judged from the labeling in vitro with 3 Hthymidine and the DNA-synthesis and cell-cycle times of these cells. The production of monocytes was only moderately diminished, i.e., to about 80% of the normal amount. The release of monocytes from the bone marrow was found to be influenced by hydrocortisone. After in vivo labeling with 3 Hthymidine the monocyte-labeling indices were initially significantly higher in hydrocortisone-treated than in normal mice. It is concluded that a decreased production of monocytes in the bone marrow cannot account for the prolonged monocytopenia in the peripheral blood after hydrocortisone administration. However, hydrocortisone interferes with the release of newly formed monocytes from the bone marrow, resulting in a prolonged sojourn of these cells in this compartment. Footnotes Submitted: 31 August 1972

Pawlak, Laura L.; Mushinski, Elizabeth B.; Nisonoff, Alfred; Potter, Michael

doi: 10.1084/jem.137.1.22pmid: 4120094

Anti- p -azophenylarsonate (anti-Ar) antibodies elicited in all strain A/J mice tested share one or more idiotypic specificities. These specificities are also found in the anti-Ar antibodies of mice of the closely related strain, AL/N, but not in those of BALB/c mice. Anti-Ar antibodies were elicited in congenic mice in which the IgC H locus of AL/N mice, which controls allotypic markers in the constant regions of heavy chains, had been introgressively backcrossed for nine generations onto a BALB/c background; the mice were then rendered homozygous for the AL/N allotypic determinant. On the average, these antibodies were quantitatively equivalent, with respect to content of the cross-reactive idiotype, to those of AL/N mice. This indicates that the gene controlling the idiotype is closely linked to the IgC H locus. Since idiotype must be a function of V region sequences, the results suggest close linkage of V H and C H genes. The cross-reactive idiotype was found in nearly all F 1 mice (C57/BL x A/J or BALB/c x A/J) tested. Footnotes Submitted: 11 September 1972

Dwyer, J. M.; Kantor, F. S.

doi: 10.1084/jem.137.1.32pmid: 4540056

Two potential mechanisms for terminating delayed hypersensitivity (DH) reactions have been examined in desensitized guinea pigs. Lack of macrophage responsiveness to lymphokines was sought as an explanation for the reduced ability of these animals to express delayed hypersensitivity. Skin-reactive factor was injected into the skin of desensitized guinea pigs and a control group of similarly immunized animals. The resulting inflammatory reactions were similar in size and intensity in both groups indicating normal macrophage responsiveness in the desensitized state. Passive cellular transfer of DH responses to desensitized animals was markedly less successful than transfer to normal animals. However, cells from desensitized guinea pigs did transfer DH responsiveness to normal animals. These data support the concept of a humoral suppressant of cellular immunity, perhaps acting as a feedback inhibitor, produced when guinea pigs are desensitized. Footnotes Submitted: 5 September 1972

Fidler, John M.; Golub, Edward S.

doi: 10.1084/jem.137.1.42pmid: 4120095

Treatment of mice with a nonimmunogenic preparation of free reactive hapten, trinitrobenzene sulfonic acid (TNBS), leads to the induction of a state of tolerance to the hapten, 2,4,6-trinitrophenyl (TNP). This is determined by the lack of response to the haptenic moiety in an immunogenic hapten-carrier conjugate (TNP-SRBC) as assayed both by serum antibody titrations and the hemolytic plaque assay. The tolerance produced is specific for the hapten, since the anticarrier responses are essentially unaltered compared with the control values. The unresponsiveness induced by TNBS treatment is a dose-dependent phenomenon, becoming less complete at lower doses of TNBS. The tolerance is of a definite length, both in its induction phase and in the duration of the established unresponsive state. Tolerance can be maintained and extended, and may also be reentered once escape has been initiated. Footnotes Submitted: 11 September 1972

Ferrone, S.; Cooper, N. R.; Pellegrino, M. A.; Reisfeld, R. A.

doi: 10.1084/jem.137.1.55pmid: 4734592

The interaction of histocompatibility (HL-A) antibodies and complement with synchronized human lymphoid cells in continuous culture has been investigated. The sensitivity of cultured lymphoid cells to HL-A antibody-mediated lysis in the cytotoxic test, the extent of activation of the complement system, the degree to which labeled complement components are bound, and the ability of these cells to absorb HL-A alloantibodies do not vary significantly during the cell growth cycle. The constancy of histocompatibility antigen expression throughout the growth cycle of cultured cells suggests that these cell surface markers are an essential part of membrane cytoarchitecture and could well play a critical role in determining the normal function of the cell membrane. Footnotes Submitted: 17 August 1972

Ashman, Robert F.; Raff, Martin C.

doi: 10.1084/jem.137.1.69pmid: 4569470

Anti-θAKR antibody conjugated to fluorescein has been used in direct immunofluorescence tests to identify spleen θ + (T) sheep erythrocyte rosette-forming cells in AKR mice. Specificity studies involving A and cogenic A/θAKR mice clearly demonstrated that the cell surface fluorescence and cytotoxicity produced by the antiserum is directed solely toward the θAKR alloantigen. Approximately ⅜ of rosette-forming and non-rosette-forming spleen cells were found to be θ + . The tendency for T cells to bind less antigen and the tendency for antigen-binding T cells to bear less θ than other spleen T cells, first suggested by other studies involving rosette-elimination by anti-θC3H plus complement, were confirmed by direct immunofluorescence. All AKR rosettes are specifically inhibitable by anti-immunoglobulin, including T rosettes. Antigen-induced redistribution of T cell receptors, analogous to that previously described for B cell receptors (16), occurs as readily in θ + RFC as in θ - RFC, without altering the symmetrical ring distribution of θAKR antigen. Footnotes Submitted: 2 August 1972

Unkeless, J. C.; Tobia, A.; Ossowski, L.; Quigley, J. P.; Rifkin, D. B.; Reich, E.

doi: 10.1084/jem.137.1.85pmid: 4347290

Chick embryo fibroblast cultures develop fibrinolytic activity after transformation by Rous sarcoma virus (RSV). This fibrinolytic activity is not present in normal cultures, and it does not appear after infection with either nontransforming strains of avian leukosis viruses or cytocidal RNA and DNA viruses. In cultures infected with a temperature sensitive mutant of RSV the onset of fibrinolysis appears after exposure to permissive temperatures and precedes by a short interval the appearance of morphological evidence of transformation. See PDF for Structure The rate of fibrinolysis in transformed cultures depends on the nature of the serum that is present in the growth medium: some sera (e.g., monkey or chicken serum) promote high enzymatic activity, while others (calf, fetal bovine) do not. Some sera contain inhibitors of the fibrinolysin. Based on the effect of a small number of known inhibitors, at least one step of the fibrinolytic process shows specificity resembling that of trypsin. The sera of sarcoma-bearing chickens contain an inhibitor of the fibrinolysin, whereas normal chicken sera do not. For general discussion, conclusions, and summary see the accompanying paper, part II, ( J. Exp. Med . 137 :112). Footnotes Submitted: 27 September 1972

Ossowski, L.; Unkeless, J. C.; Tobia, A.; Quigley, J. P.; Rifkin, D. B.; Reich, E.

doi: 10.1084/jem.137.1.112pmid: 4347288

Chick, hamster, mouse, and rat embryo fibroblast cultures, transformed by either DNA or RNA viruses, show fibrinolytic activity under suitable conditions of growth and in appropriate media; normal counterpart cultures do not. The fibrinolysin is produced by the interaction of two protein factors: one of these, a cell factor, is released by transformed cells and accumulates in the medium when cultures are incubated in the absence of scrum. The second factor, the serum factor, is a specific protein that is present in sera of many avian and mammalian species, including man. Not all sera yield fibrinolysin on interaction with any given transformed cell factor, and the spectrum of activating sera is distinctive for each cell factor. This pattern appears to be determined by the cell type, rather than by the transforming virus. An important role for the fibrinolysin in oncogenic transformation is suggested by the following correlations. ( a ) The initial appearance of fibrinolysin precedes the morphological change after the transfer to permissive temperatures of chick fibroblast cultures infected with a temperature-sensitive mutant of RSV. ( b ) The initiation of fibrinolysis and of morphological change both require the synthesis of new protein, but not the synthesis of either DNA or rRNA. ( c ) The activity of the fibrinolysin is correlated with the retention of abnormal morphology in hamster cells transformed by SV-40. ( d ) The sera of normal chicks effectively activate fibrinolysis with the cell factor from transformed chick cells. In contrast the sera of chicks with RSV tumors do not; these contain an inhibitor of the fibrinolytic activity. Footnotes Submitted: 27 September 1972

Nilsson, Bengt S.; Sultzer, Barnet M.; Bullock, Wesley W.

doi: 10.1084/jem.137.1.127pmid: 4120093

Purified protein derivative (PPD) tuberculin induced immunoglobulin production in cultures of nonimmune mouse spleen cells, as measured by plaque-forming cells (PFC) against sheep erythrocytes (SRBC), horse erythrocytes, and 4-hydroxy-3,5-dinitrophenacetyl-SRBC. The increase started between 15 and 20 h of culture and reached a peak at 48–72 h. Higher PPD concentrations resulted in earlier peak responses than low concentrations. The Ig produced was mainly 19S and of very low avidity. The response elicited by PPD was of the same type as that caused by lipopolysaccharide of bacterial origin. Mitomycin treatment of cells before culture did not change the numbers of PFC/10 6 recovered cells but the cell recovery was considerably lower. Also injection of PPD in vivo resulted in increased numbers of PFC. On the basis of these results it is suggested that PPD nonspecifically activates a majority of the B cell population to proliferation and immunoglobulin synthesis. Footnotes Submitted: 15 September 1972