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Biomedical subjects

H M Johnson

Publications and source records attributed to H M Johnson.

At least 145 records · Page 8Linked to original sources

The role of a gamma interferon-like lymphokine in the activation of T cells for expression of interleukin 2 receptors.

Human peripheral T cells, but not non-T cells, expressed receptors for interleukin 2 when treated with partially purified human gamma interferon (IFN-gamma). The expression of the receptors was evidenced by proliferation of IFN-gamma-treated cells in the presence of IL 2 and absorption of IL 2 by treated cells. The IFN-gamma-induced expression of IL 2 receptors was associated with partially purified IFN-gamma (approximately 2000-fold purified) and was blocked by treatment of IFN-gamma with specific antibodies or destruction of IFN-gamma by acid pH. IFN-gamma induced expression of receptors in a manner similar to that of concanavalin A (Con A). Further, Con A induction of expression of receptors was blocked by simultaneous treatment of cultures with anti-IFN-gamma sera, suggesting that IFN-gamma was involved in Con A induction of IL 2 receptors. T cells in culture are continuously exposed to various forms of antigen stimulation, and thus the function of IFN-gamma may be the enhancement of expression of antigen- or mitogen-induced IL 2 receptors in a manner similar to its enhancement of expression of antigens of the major histocompatibility complex.

Adult↗

Recombinant mouse interferon-gamma regulation of antibody production.

Interferon-gamma produced in monkey cells by transfection with mouse interferon-gamma cDNA suppressed the mouse in vitro antibody response in a manner similar to that of natural mouse interferon-gamma. Significant suppression was obtained with as little as 1 U of interferon. Recombinant human interferon-gamma produced by cloning in a similar fashion was not suppressive. Both the suppressive and the antiviral activities of recombinant interferon-gamma were neutralized by antibodies to mouse natural interferon-gamma. Thus, interferon-gamma was responsible for the immunosuppression. At the cellular level, the recombinant interferon-gamma was capable of activating macrophages to suppress antibody production. The data provide clear-cut evidence that interferon-gamma plays an important role in regulation of immunological processes.

Animals↗

Interleukin 2 enhances natural killer cell activity through induction of gamma interferon.

Highly purified interleukin 2 (IL 2), free of interferon activity, enhanced natural killer (NK) cell activity against tumor cells in mouse spleen cell cultures and in human peripheral lymphocyte cultures in a manner similar to that of interferon (IFN). We determined that IL 2 enhanced NK activity indirectly in a cascade manner by the induction of gamma IFN (IFN-gamma) in the cultures, which actually mediated the enhanced killing. Accordingly, lymphocyte cultures treated with IL 2 alone produced 10 to 100 U of IFN per ml in 6 to 24 h of culture. The IFN was typed as IFN-gamma by specific antibodies. Specific antibodies either to natural IFN-gamma or to a synthetic peptide corresponding to the human IFN-gamma N-terminal amino acids, when added to cultures treated with IL 2, completely blocked IL 2 enhancement of NK cell activity for both the mouse and human systems. IL 2-induced proliferation was not affected by the antibodies. Thus, the enhancement of NK cell activity by IL 2 is completely mediated by IL 2-induced IFN-gamma. The findings clearly indicate a cascade effect whereby one lymphokine (IL 2) induces the production of another. The latter lymphokine (IFN-gamma) then mediates an important biological effect (natural killing).

Animals↗

Staphylococcus aureus protein A induces the production of interferon-alpha in human lymphocytes and interferon-alpha/beta in mouse spleen cells.

Human and mouse lymphocytes produced IFN-alpha and IFN-alpha/beta, respectively, when incubated with highly purified Staphylococcus aureus protein A (SPA). Maximum IFN induction occurred when lymphocytes were incubated with SPA at 250 to 500 micrograms/ml for 24 hr. The SPA induced-IFN were identified by neutralization of antiviral activity with specific antisera. A commercial preparation of SPA was found to induce both HuIFN-alpha and HuIFN-gamma, with the latter comprising over 90% of the antiviral activity. The SPA preparation that induced IFN-gamma was shown by SDS-polyacrylamide gel electrophoresis and an enzyme-linked immunosorbent assay to be contaminated with 5% S. aureus enterotoxin A (SEA), a potent T lymphocyte mitogen and IFN-gamma inducer. Preincubation of the commercial SPA with a specific antiserum to SEA blocked the induction of IFN-gamma and had no effect on IFN-alpha production. SEA is a very potent IFN-gamma inducer, effective at concentrations of 1 ng/ml or less. Thus, very minor contamination of SPA by SEA or other IFN-gamma inducers could be responsible for previous reports that SPA induces IFN-gamma in lymphocytes or that SPA behaves as a T cell mitogen.

Animals↗

Recombinant mouse gamma interferon induces the priming step in macrophage activation for tumor cell killing.

Mouse macrophages become activated to kill tumor cells by traversing a series of steps (1-3). The first of these does not cause the expression of cytolytic activity; instead, it primes macrophages to respond to a second signal(s) that then triggers the onset of killing (4-7). The mediator that is responsible for priming is contained, along with other lymphokines, in the culture supernatants of concanavalin A-stimulated spleen cells (5-7) cloned T lymphocytes (8), or some T cell hybridomas (9). Close association has been noted between macrophage priming activity and antiviral activity that is attributable to gamma interferon (10-12). However, unequivocal evidence that the two activities are products of the same molecule has not been available. We now how conclusively by using mouse gamma interferon (MulFN-gamma)3 produced by recombinant DNA technology that, in addition to antiviral activity, this lymphokine has the capacity to induce the priming step in the process of macrophage activation for tumor cell killing.

Animals↗

Regulation of murine macrophage Ia antigen expression by a lymphokine with immune interferon activity.

A culture supernatant of concanavalin A-activated spleen cells (Con A supernatant) induced murine macrophages to express Ia antigens in vitro. Biochemical characterization of the Con A supernatant indicated that the macrophage Ia antigen regulatory activity shares molecular weight, pI, and hydrophobic and affinity characteristics with immune interferon (IFN-gamma). Antiserum to mouse IFN-gamma neutralized both the macrophage Ia antigen regulatory and IFN-gamma bioactivities of the Con A supernatant. Furthermore, both partially purified murine IFN-gamma (10(7) U/mg protein sp act) and IFN-containing culture supernatants of the murine BFS T cell line-induced macrophage Ia antigen expression in vitro. Culture supernatants containing colony-stimulating factor, interleukin 1, interleukin 2, macrophage migration inhibitory factor, and a macrophage-activating activity that were distinct from IFN-gamma did not induce macrophage Ia antigen expression. Taken together, the data indicate that the in vitro expression of Ia antigens on macrophages is regulated by an activity that has the characteristics of interferon.

Animals↗

Human natural killer-like activity against mouse spleen cells.

Unstimulated human peripheral lymphocytes were cytotoxic for normal mouse spleen cells and suppressed the in vitro antibody plaque-forming cell (PFC) response of these cells to sheep red blood cells and dinitrophenylated Ficoll. The cells in the lymphocyte population that were responsible for the immunosuppression had properties of natural killer (NK) or NK-like cells in that they were: (a) non-E-rosetting, (b) nonadherent, (c) unaffected by treatment with anti-human immunoglobulin plus complement, (d) cytotoxic against an established human NK target, K562 leukemic cells, and (e) partially inactivated by mitomycin C. Addition of the human NK-like cells to mouse spleen cell cultures at the time of antigen addition and at an effector cell to target cell ratio as low as 0.67:1 resulted in 85 to 96% suppression of the PFC response. Addition of NK-like cells to cultures 18 h before harvesting in 5-day cultures required higher concentrations and ratios (2.7:1) of effector to target cells to significantly suppress the PFC response. The data suggest that human NK-like activity in suppression of the mouse PFC response is due to killing of the targets. The mouse spleen cell PFC system represents a potential model for assessment of human NK activity that is quite dramatic in its effect and can be used in addition to the well known 51Cr-release assay. Also, since the mouse spleen cell is a normal cell, it provides a model in the PFC system for studying the mechanism of NK regulation of normal cellular function. An additional finding of this study was the observation that E-rosetting T cells significantly enhanced the mouse PFC response. Thus, human peripheral lymphocytes contain discrete cellular population that either enhance or suppress the mouse PFC response.

Animals↗

Regulation of the in vitro antibody response by neuroendocrine hormones.

Treatment of lymphocytes with inducers of interferon alpha (IFN-alpha) results in the production of corticotropin (ACTH) and endorphin-like activities. The pro-opiomelanocortin-derived hormones ACTH and alpha-, beta-, and gamma-endorphin and the structurally related hormones [Leu]- and [Met]enkephalin were therefore tested for their effects on the in vitro antibody response of mouse spleen cells. ACTH and alpha-endorphin were potent inhibitors (>/=80% suppression) of the antibody response to the T-cell-dependent antigen sheep erythrocytes at a concentration of 0.5 muM. [Met]- and [Leu]enkephalin were moderate inhibitors (approximately 60% suppression) at 0.2-2 muM, and beta- and gamma-endorphin were minimal inhibitors (approximately 20% suppression) at 5-6 muM. At higher concentrations ACTH also inhibited the antibody response to the T-cell-independent antigen dinitrophenyl-Ficoll, suggesting that T-cell function was more sensitive to blockage by these hormones than was B-cell function. ACTH and IFN had similar suppression properties; thus, the hormone-like activities associated with IFN-alpha may play a role in IFN-induced immunosuppression. alpha-Endorphin immunosuppression was blocked by naloxone, which suggested that alpha-endorphin exerted its effects through binding to opiate-like receptors on the spleen cells. The failure of beta-endorphin to suppress the immune response significantly was not due to its failure to bind to the opiate-like receptors because it blocked alpha-endorphin-induced suppression. Direct evidence for both opiate and ACTH receptors on the spleen cells was obtained in binding studies with labeled enkephalin and ACTH. Such studies revealed the presence of both high- and low-affinity receptors. The data show that neuroendocrine polypeptide hormones can regulate the immune response.

Adrenocorticotropic Hormone↗

Long-term mortality profile of heavily-exposed lead smelter workers.

Of 241 male smelter workers diagnosed as having lead poisoning during 1928 to 1959, 140 are known to have subsequently died. Their cause-of-death profile has been compared with that of 695 other male decedents (predominantly nonoffice production workers) from the same smelter and with that of the Australian male population. Age-standardized proportional mortality analysis shows a substantial excess in the numbers of deaths from chronic renal disease and cerebral hemorrhage, particularly prior to 1965. A moderate excess was also apparent for the other smelter workers. In recent years, these mortality excesses in lead-exposed workers have largely dissipated.

Adult↗

Cellular regulation of gamma interferon production: Lyt phenotype of the suppressor cell.

Suppressor cell activity for in vitro gamma interferon (IFNgamma) production can be induced in C57BL/6 mice with the T-cell mitogens staphylococcal enterotoxin A and concanavalin A. The suppressor cell lacked mitogen specificity in that injection of mice with either staphylococcal enterotoxin A or concanavalin A induced potent suppressor cell activity that inhibited the in vitro induction of IFNgamma by either mitogen. The suppressor cell is a T cell with a relatively high density of Thy 1 antigen. The IFNgamma-producing cell has a low density of Thy 1 and appears to have evolved from cells with a high density of Thy 1.2 or to have required such cells as helpers in the production of IFNgamma. Monoclonal anti-Lyt 2.2 antibody completely eliminated the suppressor cell activity, whereas anti-Lyt 1.2 partially, but significantly, reduced the suppressor cell activity. The suppressor cell, then, may be or may require a Lyt 1,2,3(+) cell with a relatively low density of the Lyt 1 antigen for activity. The Lyt phenotype of the IFNgamma- producing cells was examined for both virgin and suppressor cell populations. Treatment of cells with either anti-Lyt 1.2 or anti-Lyt 2.2 completely eliminated IFNgamma production in both virgin and suppressor cell cultures. Cocultivation of Lyt 1(+) and Lyt 2(+) cells in both virgin and suppressor cell cultures significantly restored the ability to produce IFNgamma when cells were stimulated with staphylococcal enterotoxin A. For the virgin cells, this seems to indicate clearly that both Lyt 1(+) and Lyt 2(+) cells are required for IFN production, but that Lyt 1,2,3(+) cells are not required. For the suppressor cell population this type of treatment resulted in IFNgamma production comparable to that of the virgin cell population. The treatment eliminated or depleted Lyt 1,2,3(+) cells from the suppressor cell population, thus providing additional evidence that a Lyt 1,2,3(+) cell is required for suppressor cell activity. Treatment of spleen cells with anti-Lyt antibodies at the time that they were involved in ongoing IFNgamma production showed that the IFNgamma-producing cell is Lyt 2(+). At least a 1:1 ratio of virgin to suppressor cells was required for optimal suppression of IFNgamma production by virgin spleen cells. This suggests that suppression involved cell-to-cell contact. Preliminary data showed that the suppressor cell activity could also be induced in in vitro cultures incubated with mitogen for 4 days. This is evidence that the suppressor cell is inducible and not necessarily recruited to the spleen from other lymphoid organs.

Animals↗

Phorbol ester replacement of helper cell and interleukin 2 requirements in gamma interferon production.

Immune interferon (IFN-gamma) production by mitogen-stimulated C57BL/6 mouse spleen cells required both Lyt 1+,2- and Lyt 1-,2+ lymphocytes. The Lyt 1-,2+ cells were the IFN-gamma producers, whereas the Lyt 1+,2- cells were the helpers. Interleukin 2 mediated the Lyt 1+,2- cell help. Phorbol myristic acetate (10 ng/ml) completely replaced Lyt 1+,2- helper cell or interleukin 2 requirements in IFN-gamma production. The phorbol ester exerted its helper effects by direct interaction with IFN-gamma-producing Lyt 1-,2+ cells and not through stimulation of production of interleukin 2 by Lyt 1+, 2- cells. Suppressor cell effects in IFN-gamma production were also completely abrogated by phorbol myristic acetate. The data suggest a mechanism for phorbol myristic acetate regulation of IFN-gamma production at the cellular level.

Animals↗

Interleukin 2 regulates immune interferon (IFN gamma) production by normal and suppressor cell cultures.

A Lyt-2+ lymphocyte is responsible for immune interferon (IFN gamma) production in mitogen-stimulated mouse spleen cell cultures. A Lyt-1+ helper cell is required for the induction of IFN gamma. Interleukin 2 (IL 2) can specifically replace the Lyt 1 cell requirement. IL 2 will also abrogate suppressor cell inhibition of IFN gamma production. IFN gamma production appears to be regulated by a dynamic interaction between helper cells (source of IL 2), suppressor cells (absorption of IL 2?) and IFN gammna-producer cells.

Animals↗

Vasopressin replacement of interleukin 2 requirement in gamma interferon production: lymphokine activity of a neuroendocrine hormone.

The neuroendocrine antidiuretic hormone arginine vasopressin (AVP) was capable of replacing the interleukin 2 (IL 2) requirement for gamma-interferon (IFN gamma) production by Lyt-2+ cells from C57BL/6 mouse spleen cells. The AVP replacement did not stimulate DNA synthesis in the target lymphocytes. This suggested that AVP was capable of replacing an IL 2 function that did not involve stimulation of cellular proliferation or DNA synthesis. This was confirmed by the demonstration that mitomycin C inhibition of IFN gamma production was reversed by IL 2 or AVP without concomitant reversal of blockage of DNA synthesis. Oxytocin, which is structurally related to AVP, was also capable of replacing IL 2 requirement for IFN gamma production, whereas insulin was ineffective. The data show that the neuroendocrine hormones AVP and oxytocin are capable of lymphokine-like activity. This activity may involve the induction of a second messenger such as cyclic GMP.

Animals↗

Neutralization of native human gamma interferon (HuIFN gamma) by antibodies to a synthetic peptide encoded by the 5' end of HuIFN gamma cDNA.

A synthetic peptide corresponding to the N-terminal amino acid sequence of human gamma-interferon (HuIFN gamma), based on the cDNA sequence, was used to produce antibodies in rabbits that were reactive with native HuIFN gamma. Antibodies from all immunized rabbits neutralized the antiviral activity of HuIFN gamma. Significant neutralization of other HuIFN and mouse IFN was not observed. The peptide had the sequence Cys-Tyr-Cys-Gln-Asp-Pro-Tyr-Val-Lys-Glu-Ala-Glu-Asn-Leu-Lys-Lys-Tyr-Phe-Asn-Ala ,and was coupled to keyhole limpet hemocyanin by disulfide linkage with the use of cystamine. The specificity of the antibodies produced to the peptide was compared to that of antibodies produced to native HuIFN gamma by neutralization of HuIFN gamma and by reactivity with peptide in the enzyme-linked immunosorbent assay (ELISA). The ratio of anti-peptide antibody neutralization of HuIFN gamma vs reactivity with peptide in the ELISA was at least 28-fold lower than for anti-HuIFN gamma antibody. Thus the antibodies to peptide and to HuIFN gamma were directed primarily against different determinants on native HuIFN gamma or the anti-HuIFN gamma antiserum probably contained antibodies to additional determinants. The anti-peptide antibodies should be useful for further characterization and purification of HuIFN gamma.

Amino Acid Sequence↗