Characteristics of the adaptive response in cultured salmon cells exposed to ionizing radiation

The aim of this study was to investigate the influence of "priming" doses of ionizing irradiation on salmon cell survival in vitro prior to being challenged with subsequent higher doses. A radiation-induced adaptive response (AR) was examined in the Chinook salmon embryo cell line (CHSE-214). Cells were initially irradiated with a range of priming (conditioning) doses of (60)Co gamma (gamma) rays (0.25-0.75 Gy), followed by a challenge dose of 7.50 Gy at intervals of 24, 48, and 72 hr. The AR was assessed using a colony-forming assay. Cell survival was determined by counting the number of colonies (viable clones) after 40 days of culture. This study revealed that cells that received a priming dose of 0.50 Gy before delivering the higher challenge dose became more radiation resistant with an increase in cell survival of 29% over cells receiving the challenge dose alone. The cells showed maximum resistance to ionizing radiation when the priming dose was given 72 hr prior to the higher challenge dose. This study is one of the first to demonstrate an AR using an in vitro piscine system, and is generally consistent with other studies of both in vitro and in vivo systems across the taxa.     

Modulation of the immune response by a methanol-insoluble fraction of attenuated tubercle bacilli (BCG). II. Relationship of antigen dose to heightened primary and secondary immune responses to sheep red blood cells

A methanol extraction residue (MER) of BCG has previously been shown to heighten host resistance to a subsequent challenge with microorganisms or syngeneic tumour grafts and to stimulate the antibody response to sheep red blood cells (SRBC) and phage T₂. Present investigations on the effect of antigen dose on MER stimulation of the primary and secondary response to SRBC indicated that:

(1) Pretreatment by MER stimulates the early primary response most effectively when the immunizing dose of SRBC is less than maximum, and this is due to a greater increase by MER of 19S antibody with low doses of antigen than with high doses of SRBC. Maximum 7S antibody production is higher in MER treated animals at all antigen doses.

(2) MER administered after low doses of SRBC but not high doses stimulates the ongoing primary haemagglutinin response.

(3) Maximum secondary responses of mice treated with MER before priming are considerably elevated above the corresponding controls and the secondary responses in treated mice are not inhibited by high priming doses of antigen.

These results are discussed in relation to the locus of MER activity.

     

Hormesis and Ginkgo biloba (GB): Numerous biological effects of GB are mediated via hormesis

Ginkgo biloba (GB) extracts have been shown to commonly induce biphasic dose responses in a range of cell types and endpoints (e.g., cochlea neural stem cells, cell viability, cell proliferation). The magnitude and width of the low dose stimulation of these biphasic dose responses are similar to those reported for hormetic dose responses. These hormetic dose responses occur within direct stimulatory responses as well as in preconditioning experimental protocols, displaying acquired resistance within an adaptive homeodynamic and temporal framework and repeated measurement protocols. The demonstrated GB dose responses further reflect the general occurrence of hormetic dose responses that consistently appear to be independent of the biological model, endpoint, inducing agent, and/or mechanism. These findings have important implications for consideration(s) of study designs involving dose selection, dose spacing, sample size, and statistical power. This illustrates and strengthens the need to characterize the low dose stimulatory response range and optimal dose in order to explore potential public health and clinical applications of plant-derived agents, such as GB.     

Capability of adriamycin and busulfan to induce adaptive response in vitro.

The capability to induce an adaptive response by low doses of busulfan (BS) or adriamycin (ADR) was studied in two kinds of mammalian cells with acquired or inherent resistance to ADR (ME18/R and V3) cultured in vitro. The results indicate the presence of an adaptive response to ADR in both kinds of used cells pretreated with a low priming dose of ADR. In the same kind of cells no adaptive response to BS after priming with a low dose of this drug was found.     

Adaptive response after X-irradiation of human lymphocytes?

The induction of an adaptive response of human lymphocytes fromthree donors exposed to 0.01 or 0.05 Gy of X-rays at 32 h ofculture and to 1.5 Gy at 48 h was studied. There was no evidencefor a significant reduction of the incidences of chromosomalaberrations in cultures pre-treated with low doses and subsequentlyexposed to the higher dose compared with cultures exposed tothe high dose alone.     

No radioadaptive response to micronucleated polychromatic erythrocyte (MN-PCE) induction in murine peripheral blood in vivo

The effect of conditioning pretreatment with 0.025 Gy of gamma rays on micronucleated polychromatic erythrocyte (MN-PCE) induction by 1.0 or 0.1 Gy of gamma rays was determined in murine peripheral blood. The adaptive and challenge doses as well as the timing of their administration were taken from a previously reported experiment [Far-ooqi and Kesavan (1992): Mutat Res 302:83–89]. The response was determined by the strategy of measuring the area below the curve (ABC) of MN-PCE induction vs. time. This strategy permits one to determine an index of total damage and to establishif conditioning exposure affects the timing of MN-PCE appearance in the blood stream, which in turn could cause an apparent difference in response between the conditioned and the unconditioned groups at specific times. The results indicate that low dose gamma ray pretreatment does not protect against MN-PCE induction by the challenge gamma ray dose, and that there was no change on the kinetics of MN-PCE appearance in peripheral blood. Environ. Mol. Mutagen. 29:289-295, 1997 ® 1997 Wiley-Liss, Inc.     

Suppression of anti-hapten antibody response in vitro by hapten-carrier conjugates

Production of antibodies was stimulated or suppressed arbitrarily by antigen treatment in vitro of spleens cultured at various time intervals after in vivo immunization. Spleens of mice immunized to the 2,4-dinitrophenyl or (4-hydroxy-3-iodo-5-nitrophenyl)acetyl haptenic determinants produced antibodies in culture when no antigen was applied in vitro. When a conjugate of the hapten to the same carrier employed for priming was given in vitro, an initial reduction of the response was observed, the level of which was dependent on antigen dose. Subsequently, increased amounts of antibodies were measured. In contrast, in vitro exposure to the hapten conjugated to an unrelated carrier resulted in significant reduction of the response for the entire period of the test. This suppressive effect manifested with various carrier proteins (ovalbumin, bovine IgG, bovine and rabbit serum albumin and keyhole limpet hemocy-anin), when applied to cultures in doses which were potentially immunogenic.     

Antibody synthesis by chicken spleen cells in vitro. I. Requirements of B cells at various stages after immunization for T cells,macrophages and antigen

The requirements for T cells, macrophages and antigen during the induction of in vitro antibody responses were ascertained with chicken spleen cells obtained at various times after immunization with sheep red blood cells (SRBC). The IgM plaque-forming cell (PFC) response was T cell independent exclusively in cultures initiated 3 days after priming, but macrophage dependent at all time intervals tested. In cultures started 4 to 10 days after priming the IgG response was both T cell and macrophage independent and PFC numbers remained at a high plateau level throughout the culture period. In contrast, IgG responses initiated more than 15 days after priming showed a reversal to complete T cell and macrophage dependence and were characterized by a sharp increase in PFC numbers between days 2 and 4 of culture. Formaldehyde-fixed SRBC were immunogenic for IgG PFC 4 to 10 days after priming but failed to stimulate later IgG memory and all IgM responses. Contrasting antigen dose requirements for IgM and IgG responses were found in cultures initiated at various periods after priming. The results suggest that direct contact with fixed antigen was sufficient to maintain IgG antibody synthesizing PFC in vitro, while native antigen and cell co-operation were required for late secondary IgG and all IgM responses. These results are interpreted in terms of separate pathways of differentiation for IgM and IgG antibody-producing cells. A distinct 3rd day stage of T cell-independent but macrophage-dependent responsiveness for both classes of antibody was also defined.     

Dietary factors, hormesis and health

The impact of dietary factors on health and longevity is increasingly appreciated. The most prominent dietary factor that affects the risk of many different chronic diseases is energy intake -- excessive calorie intake increases the risk. Reducing energy intake by controlled caloric restriction or intermittent fasting increases lifespan and protects various tissues against disease, in part, by hormesis mechanisms that increase cellular stress resistance. Some specific dietary components may also exert health benefits by inducing adaptive cellular stress responses. Indeed, recent findings suggest that several heavily studied phytochemicals exhibit biphasic dose responses on cells with low doses activating signaling pathways that result in increased expression of genes encoding cytoprotective proteins including antioxidant enzymes, protein chaperones, growth factors and mitochondrial proteins. Examples include: activation of the Nrf-2 -- ARE pathway by sulforaphane and curcumin; activation of TRP ion channels by allicin and capsaicin; and activation of sirtuin-1 by resveratrol. Research that establishes dose response and kinetic characteristics of the effects of dietary factors on cells, animals and humans will lead to a better understanding of hormesis and to improvements in dietary interventions for disease prevention and treatment.     

Converging concepts: adaptive response, preconditioning, and the Yerkes-Dodson Law are manifestations of hormesis

The adaptive response in toxicology and environmental mutagenesis, preconditioning in biomedicine and the Yerkes-Dodson Law in psychology have dominating research themes with widespread and significant scientific and societal implications. This paper suggests that these apparently independent biological dose-response phenomena are manifestations of the common and more general biphasic dose-response relationship concept called hormesis. These three types of dose-response, as well as the hormesis concept, may represent the same general type of adaptation, which were discovered independently in different biological disciplines, amongst which there has been little communication. This intellectual isolation, due principally to progressively greater disciplinary specialization, resulted in the evolution of different terminologies for dose-response phenomena with strikingly similar quantitative features. This lack of recognition of converging dose-response concepts across disciplines has important implications since it limits the recognition of a common and basic biological concept while minimizing collaborations by investigators in related areas. The paper concludes that the broadly recognized biological adaptive responses, as described by the concepts of adaptive response, preconditioning and the Yerkes-Dodson Law, are special cases of the more general hormesis dose-response concept.     

Priming with high and low respiratory allergen dose induces differential CD4+ T helper type 2 cells and IgE/IgG1 antibody responses in mice

Sensitization of allergic patients normally takes place over several years and is the result of repeated exposure to low levels of allergen. Most mouse asthma models use a high dose of allergen administered over a short period. We have investigated the role of dose in the immune response to an inhaled respiratory allergen (Blomia tropicalis). We observed the effect of priming dose on the allergic response in mice intranasally immunized with low (0·5 μg) and high (50 μg) doses of B. tropicalis extract and killed 1 day after the last challenge. For both doses of allergen, T helper type 2 (Th2) cells and Th2 cytokines were evident as well as eosinophilic inflammation accompanied by mucus hyper‐secretion. By contrast, IgE and IgG1 antibody responses were normally only detected at high‐dose priming. To investigate the mechanism for these effects, we found group 2 innate lymphoid cells (ILC2s) were increased 48 hr after challenge in the low‐dose‐treated but not the high‐dose‐treated mice. Furthermore, we determined whether repeated low‐dose exposure with different priming protocols could induce an antibody response. Repeated low‐dose exposure to 0·5 μg three times weekly for 4 weeks (cumulative 6 μg) had the same effect as a shorter high‐dose exposure (cumulative 80 μg) and increasing cumulative dose induced antibody responses. These data indicate that low doses of allergen are sufficient to prime Th2 cells and ILC2s, but insufficient to induce antibody responses. Cumulative exposure to small amounts of allergen induces both Th2 and antibody responses and may better reflect natural sensitization.     

Photodynamic therapy induces an immune response against a bacterial pathogen

Photodynamic therapy (PDT) employs the triple combination of photosensitizers, visible light and ambient oxygen. When PDT is used for cancer, it has been observed that both arms of the host immune system (innate and adaptive) are activated. When PDT is used for infectious disease, however, it has been assumed that the direct antimicrobial PDT effect dominates. Murine arthritis caused by methicillin-resistant Staphylococcus aureus in the knee failed to respond to PDT with intravenously injected Photofrin^®. PDT with intra-articular Photofrin produced a biphasic dose response that killed bacteria without destroying host neutrophils. Methylene blue was the optimum photosensitizer to kill bacteria while preserving neutrophils. We used bioluminescence imaging to noninvasively monitor murine bacterial arthritis and found that PDT with intra-articular methylene blue was not only effective, but when used before infection, could protect the mice against a subsequent bacterial challenge. The data emphasize the importance of considering the host immune response in PDT for infectious disease.     

Genetics of the immune response. I. The immune response to the phage fd in high and low responding inbred strains of mice

The immune response to the bacteriophage fd has been analyzed in several inbred strains of mice. The inactivation curve of bacteriophage fd by specific antisera follows initially first order kinetics and finally levels off gradually. The strength of the antisera has been characterized primarily by the velocity constant K of the inactivation kinetics. Two other methods, endpoint and plateau determination, have been used for the analysis of selected problems. The threshold dose for priming for a secondary response varies widely among the strains tested. Low responding strains have a hundred- to thousandfold higher threshold dose than high responding strains. High responsiveness is dominant over low responsiveness in genetical analysis and inherited as a single dominant trait. The dose for an optimal response in high and low responders lies within the same range at about 1 × 10⁹ fd/g body weight. Further increase of the priming dose leads in all except one strain to a suppression of the response probably due to partial high zone paralysis. One strain of mice, CBA/MH, shows a dose response curve different from the others. No suppression of the response with high priming doses is found. The cooperating role of thymus derived cells and of macrophages has been tested. The previous uptake of antigen by macrophages probably prevents induction of high zone paralysis. No evidence has been found for macrophages being responsible for high and low responsiveness. The immune response to phage fd is strongly thymus dependent. The response of animals lacking thymic influence is below the response of low responders.     

Impact of the magnitude of sensitization dose on the incidence and intensity of CHS to dinitrochlorobenzene (DNCB): Insight from ear swelling and challenged-skin draining lymph node response in rats

Abstract

Allergic contact dermatitis (ACD) is a common skin inflammatory disease that develops in hosts sensitized with contact allergens. Elucidation of dose–response relationships represents one of the approaches in studying the type of ACD in humans/animal models, termed as contact hyper-sensitivity reaction (CHS). Such studies have demonstrated that the intensity of sensitization determines the response to elicitation with a contact allergen, but underlying mechanisms are unclear. The aim of this study is to explore the impact of the sensitization on contact hypersensitivity to dinitrochlorobenzene (DNCB) in rats by measuring the incidence and intensity of a challenge response in hosts sensitized with two different doses (i.e. low and high) of this hapten. Assumptions concerning the contribution from the magnitude of sensitization doses were drawn on the basis of effects from the two doses on the measured reaction parameters. Ear swelling and activity of lymph nodes that drain challenged skin (cdLN), including cellularity, proliferation, and effector cytokine IFNγ and IL-17 production was measured in rats sensitized with 0.4% or 4% DNCB and challenged with a non-irritant (0.13%) dose. Sensitization with 4% DNCB resulted in a greater proportion of rats who responded more intensely (than unsensitized challenged rats) to challenge in terms of ear swelling and increases in cdLN activity (except for IFNγ). The intensity of cdLN responses was higher in these hosts as well. Among the high-dose-sensitized rats, greater cellularity/proliferation of cells from lymph nodes (sdLN) that drain the high-dose-sensitized skin, as well as higher IL-17 production, was noted compared to what was seen in rats that received low-dose sensitization. In contrast, unchanged spontaneous and even decreased hapten-stimulated IFNγ production after the high DNCB dose was observed. Based on the data, it seems the impact of magnitude of sensitization dose on CHS might be related to the rise in the proportion of rats that responded to challenge with an increase of dLN activity. Coincidental higher production of IL-17 by dLN cells from the high-dose-sensitized rats and following challenge of these hosts underscored the significance of IL-17 for a CHS to DNCB.     

Airway responses to histamine, acetylcholine, and propranolol in anaphylactic hypersensitivity in guinea pigs

Using changes in tidal volume and dynamic lung compliance, airway responses to histamine, acetylcholine, and propranolol were investigated in different groups of guinea pigs before and after active or passive sensitization. The threshold doses to inhaled histamine, acetylcholine, and propranolol were identical before and after sensitization. Similarly, the slope of dose response curves to these drugs and the time course of recovery from airway constriction were comparable in control and sensitized animals. The slopes of dose response curves to histamine and acetylcholine were parallel. Threshold doses to acetylcholine (T_ACH) and histamine (T_H) were proportional. The average ratio T_ACH/T_H was 7.7. There was no correlation between threshold doses and slopes of dose response curves for propranolol and those for histamine or acetylcholine. Antigen challenge resulted in decreases of dynamic lung compliance and in increases of airway resistance and frequency, which were maximal 60 to 130 seconds after challenge. When functional parameters after antigen challenge had returned to normal, bronchial responses to histamine, acetylcholine, and propranolol were exaggerated. These exaggerated responses were reproducible, transient, variable from animal to animal, and related to the antigen dose. Guinea pig anaphylaxis does not lead to the prolonged hypersensitivity to chemical mediators, which characterizes human reaginic asthma, but to a temporary enhancement of responses to acetylcholine, histamine, and propranolol.     

Immunoresponses to Neisseria meningitidis epitopes: in vivo analysis of immunocompetent cells involved in suppression of secondary response to phosphorylcholine

The immune response to phosphorylcholine (PC) antigens has been extensively studied in recent years. Neisseria meningitidis serogroup B M986 (NMB) was recently reported to induce a PC-specific plaque-forming cell (PFC) immunoresponse in mice, a characteristic useful for the study of immunomodulating properties of N. meningitidis. With this technique, priming mice with low doses of NMB has been shown greatly to impair their ability, one month after priming, to mount an anti-PC response induced by NMB; this suppression is permanent, does not involve switching from IgM to another immunoglobulin class, transiently affects the T15 idiotype expression and is carrier specific. We report, based on an analysis of spleen cells from NMB-primed mice in an adoptive transfer model, that this suppression does not appear to be mediated by B lymphocytes nor does it seem to be under the direct control of T lymphocytes; rather, it involves radio-resistant cells. Additionally, our results show that NMB modulates the idiotype composition of the anti-phosphorylcholine response, probably by enhancing the expression of so called hapten-augmentable PFC. These results demonstrate that NMB can interfere effectively with the immune response in a variety of ways.Abbreviations BNF1 (BALB/c × CBA/N)F₁ hybrid mice - HI heterogeneity index - IFA incomplete Freund's adjuvant - KLH keyhole limpet hemocyanin - NBF1 (CBA/N × BALB/c)F₁ hybrid mice - NMB Neisseria meningitidis serogroup B M986 - PC phosphorylcholine - PC ₅₀ amount of PC-chloride that suppresses 50% of response - PC-KLH PC coupled to KLH - PC-(NMB)HI PC coupled to heat inactivated NMB - PFC plaque forming cell - TNP-Ficoll trinitrophenylated Ficoll     

Airways of allergic rhinitics are 'primed' to repeated allergen inhalation challenge

The hypothesis that repeated exposure to a specific allergen will further increase bronchial responsiveness to that allergen is supported by indirect evidence. However, it has not been tested as intensely in the laboratory setting, and in some cases, conflicting results are presented. In order to test the hypothesis in the atopic subjects, allergen inhalation challenge tests were performed in 29 house dust mite (Dermatophagoides pternyssinus) sensitive subjects with allergic rhinitis. Nine subjects displayed early asthmatic responses (EARs) to the first challenge (Group 1). Twenty subjects with no significant airway response were submitted to the second challenge 24 h later. Thirteen subjects showed EARs (Group FI) and two of these showed late asthmatic responses (LARs)aswell. In Group II, there were significant changes between the first and second challenge in post-allergen early phase FEV₁, (88.1 ± 4.2 vs 71.7 ± 4.2%, baseline, P < 0.05) and in post-allergen late phase FEV₁, (93.1 ± 3.4 vs 86.6 ± 7.8. P < 0.05). After the second challenge. PD20 (provocative dose of methacholine required to produce a 20% fall in FEV¹) decreased significantly from the baseline values. When challenged separately with twofold dose of allergen, only three and one of the Group II showed EAR and LAR respectively. PD20 did not change significantly after this challenge. These results indicated that two repeated exposure to allergen dose, which is not enough to cause significant airway responses at a time, may provoke asthmatic airway responses in the subjects with allergic rhinitis and that this effect of priming is not attributed to the cumulative dose but to the consequent effect of repeated allergen exposure.     

Carrier inhibition in the in vitor response to hapten-erythrocyte conjugate

The in vitro induction of 4-hydroxy-5-iodo-3-nitrophenacetyl (NIP)-specific plaque-forming cells by NIP-SRC (sheep red blood cells) can be specifically inhibited by the presence of free SRC in the cultures, but only when low doses of hapten-erythrocyte conjugate and of free SRC are used. At high concentrations of free SRC, the NIP response is specifically enhanced. Thus, only at low antigen concentrations can the in vitro system be seen as a model of physiological T-B interactions.