Zebrafish ISG15 exerts a strong antiviral activity against RNA and DNA viruses and regulates the interferon response.

ISG15, a 15-kDa interferon-induced protein that participates in antiviral defenses of mammals, is highly conserved among vertebrates. In fish, as in mammals, viral infection and interferon treatment induce isg15 expression. The two ubiquitin-like domains of ISG15 and the presence of a consensus LRLRGG sequence in the C-terminal region, which is required for the covalent conjugation to a substrate protein, are also conserved in fish. Our data demonstrate that overexpression of zebrafish ISG15 (zf-ISG15) in EPC cells is sufficient to inhibit viral infection by RNA viruses belonging to the genera Novirhabdovirus and Birnavirus and by DNA viruses of the genus Iridovirus. In coexpression experiments with IHNV proteins, we demonstrate specific ISGylation of phosphoprotein and nonvirion protein. Mutation of the glycine residues in the consensus LRLRGG motif abolishes zf-ISG15 conjugation to these proteins and the cellular protection against viral infection, thus connecting ISGylation and ISG15-dependent viral restriction. Additionally, zf-ISG15 overexpression triggers induction of the rig-I and viperin genes as well as, to a lesser extent, the IFN gene. Overall, our data demonstrate the antiviral effect of a fish ISG15 protein, revealing the conservation among vertebrates of an ISGylation mechanism likely directed against viruses. Furthermore, our findings indicate that zf-ISG15 affects the IFN system at several levels, and its study shall shed further light on the evolution of the complex regulation of the innate antiviral response in vertebrate cells.

[Evolution of food supply (apart from school catering) between 2004/2005 and 2009/2010 in middle- and high-schools of Aquitaine, France]. / Évolution de l'offre alimentaire hors restauration scolaire entre 2004-2005 et 2009-2010 dans les établissements du second degré d'Aquitaine.

BACKGROUND: The aim of this study was to assess the evolution of food supply (apart from school catering) between school years 2004/2005 and 2009/2010, in middle- and high-schools from the Aquitaine region (southwest France), in order to evaluate the impact of actions conducted within the framework and the program Nutrition, Prevention and Health of children and adolescents in Aquitaine (southwest France). METHODS: Two surveys were carried out among all middle- and high-schools of the Aquitaine region in 2004/2005 (n=536) and 2009/2010 (n=539) within the framework of a regional multidisciplinary public health program "Nutrition, prevention and health of children and teenagers in Aquitaine". For both 2004/2005 and 2009/2010, data were collected using the same questionnaire and dealt with school characteristics and modalities of food supply (apart from school catering). RESULTS: Response rate was 84.1% in 2004/2005 and 79.6% in 2009/2010. The proportion of schools offering food to pupils (apart from school catering) significantly decreased in 5 years (from 80.1% to 50.1%, P<0.001). Between 2004/2005 and 2009/2010, we observed a stabilization in the proportion of schools offering free food (from 19.7 to 17%, P=0.3), a significant decrease of those selling food (from 62.8 to 37.1, P<0.001), offering vending machines (from 43.5 to 7.2, P<0.001) and a significant increase of those offering water supply (from 8.2 to 44%, P<0.001). The composition of each modality of food supply (apart from school catering) has also been improved: less sweet and fat food, more bread and fruits. CONCLUSION: This study shows an overall improvement of food supply apart from school catering (food sale, free food and vending machines) in middle- and high-schools from the Aquitaine region (southwest France) between 2004/2005 and 2009/2010. This improvement is related to the proportion of schools offering food (quantitative improvement), as well as to the composition of food supply (qualitative improvement). These results show an improvement of food supply (apart from school catering), suggesting that actions implemented in the framework of the program "Nutrition, prevention and health of children and adolescents in Aquitaine" may have led to these improvements.

Cosmetic surgery volume and its correlation with the major US stock market indices.

BACKGROUND: As a consumer-driven industry, cosmetic plastic surgery is subject to ebbs and flows as the economy changes. There have been many predictions about the short, intermediate, and long-term impact on cosmetic plastic surgery as a result of difficulties in the current economic climate, but no studies published in the literature have quantified a direct correlation. OBJECTIVES: The authors investigate a possible correlation between cosmetic surgery volume and the economic trends of the three major US stock market indices. METHODS: A volume analysis for the time period from January 1992 to October 2008 was performed (n = 7360 patients, n = 8205 procedures). Four cosmetic procedures-forehead lift (FL), rhytidectomy (Rh), breast augmentation (BA), and liposuction (Li)-were chosen; breast reduction (BRd), breast reconstruction (BRc), and carpal tunnel release (CTR) were selected for comparison. Case volumes for each procedure and fiscal quarter were compared to the trends of the S&P 500, Dow Jones (DOW), and NASDAQ (NASD) indices. Pearson correlation statistics were used to evaluate a relationship between the market index trends and surgical volume. P values <.05 were considered statistically significant. RESULTS: Three of the four cosmetic surgery procedures investigated (Rh, n = 1540; Li, n = 1291; BA, n = 1959) demonstrated a direct (ie, positive) statistical correlation to all three major market indices. FL (n =312) only correlated to the NASD (P = .021) and did not reach significance with the S&P 500 (P = .077) or DOW (P = .14). BRd and BRc demonstrated a direct correlation to two of the three stock market indices, whereas CTR showed an inverse (ie, negative) correlation to two of the three indices. CONCLUSIONS: This study, to our knowledge, is the first to suggest a direct correlation of four cosmetic and two reconstructive plastic surgery procedures to the three major US stock market indices and further emphasizes the importance of a broad-based plastic surgery practice in times of economic recession.

Lysyl-tRNA synthetase produces diadenosine tetraphosphate to curb STING-dependent inflammation.

Inflammation is an essential part of immunity against pathogens and tumors but can promote disease if not tightly regulated. Self and non-self-nucleic acids can trigger inflammation, through recognition by the cyclic GMP-AMP (cGAMP) synthetase (cGAS) and subsequent activation of the stimulator of interferon genes (STING) protein. Here, we show that RNA:DNA hybrids can be detected by cGAS and that the Lysyl-tRNA synthetase (LysRS) inhibits STING activation through two complementary mechanisms. First, LysRS interacts with RNA:DNA hybrids, delaying recognition by cGAS and impeding cGAMP production. Second, RNA:DNA hybrids stimulate LysRS-dependent production of diadenosine tetraphosphate (Ap4A) that in turn attenuates STING-dependent signaling. We propose a model whereby these mechanisms cooperate to buffer STING activation. Consequently, modulation of the LysRS-Ap4A axis in vitro or in vivo interferes with inflammatory responses. Thus, altogether, we establish LysRS and Ap4A as pharmacological targets to control STING signaling and treat inflammatory diseases.

Bi-directional cell trafficking between mother and fetus in mouse placenta.

It is now well established that cells are exchanged between mother and fetus during gestation. It has been proposed that some of these exchanges take place in the placenta, but it has never been demonstrated. Here, we made use of EGFP (Enhanced Green Fluorescent Protein) transgenic mice to precisely visualize the juxtaposition of maternal and fetal tissues at the implantation site, as well as to describe the bi-directional cell trafficking between mother and fetus at different stages of gestation. The influence of genetic differences between mother and fetus on the cell migration was also addressed by studying various types of matings: syngeneic, allogeneic and outbred. The frequency of maternal-fetal cell exchanges within the placenta is much higher in syngeneic and allogeneic gestations than in outbred ones. Maternal cells were mainly localized in the labyrinth where they were scattered or sometimes grouped in or near blood spaces. Groups of maternal cells could also be observed in maternal blood sinuses of the spongiotrophoblast. Conversely, fetal cells were organized in rings surrounding maternal blood sinuses in the decidua at 10-12 days of gestation. After day 13, they invaded the decidua. Fetal cells could also be detected in maternal peripheral blood and organs by nested PCR and fluorescence microscopy on cryosections, respectively. This suggests a role in the establishment and maintenance of the maternal tolerance to the fetus.

Zebrafish Plzf transcription factors enhance early type I IFN response induced by two non-enveloped RNA viruses.

The BTB-POZ transcription factor Promyelocytic Leukemia Zinc Finger (PLZF, or ZBTB16) has been recently identified as a major factor regulating the induction of a subset of Interferon stimulated genes in human and mouse. We show that the two co-orthologues of PLZF found in zebrafish show distinct expression patterns, especially in larvae. Although zbtb16a/plzfa and zbtb16b/plzfb are not modulated by IFN produced during viral infection, their over-expression increases the level of the early type I IFN response, at a critical phase in the race between the virus and the host response. The effect of Plzfb on IFN induction was also detectable after cell infection by different non-enveloped RNA viruses, but not after infection by the rhabdovirus SVCV. Our findings indicate that plzf implication in the regulation of type I IFN responses is conserved across vertebrates, but at multiple levels of the pathway and through different mechanisms.

Scripting MODFLOW Model Development Using Python and FloPy.

Graphical user interfaces (GUIs) are commonly used to construct and postprocess numerical groundwater flow and transport models. Scripting model development with the programming language Python is presented here as an alternative approach. One advantage of Python is that there are many packages available to facilitate the model development process, including packages for plotting, array manipulation, optimization, and data analysis. For MODFLOW-based models, the FloPy package was developed by the authors to construct model input files, run the model, and read and plot simulation results. Use of Python with the available scientific packages and FloPy facilitates data exploration, alternative model evaluations, and model analyses that can be difficult to perform with GUIs. Furthermore, Python scripts are a complete, transparent, and repeatable record of the modeling process. The approach is introduced with a simple FloPy example to create and postprocess a MODFLOW model. A more complicated capture-fraction analysis with a real-world model is presented to demonstrate the types of analyses that can be performed using Python and FloPy.

A mouse monoclonal antibody specific for the V beta 5.3 chain of the human TcR recognizes a subgroup of the mouse TcR V beta 8.2 chains.

We have analyzed the reactivity of a mouse monoclonal antibody directed against the human T cell receptor for antigen (TCR). This antibody (111-427) of immunoglobulin G1 isotype has been produced in a BALB/c mouse immunized with HPB-ALL cells and normal human peripheral blood leukocytes. It reacts specifically with the HPB-ALL lymphoma and 2 to 7% of normal human blood lymphocytes, on which it has a mitogenic effect in vitro. We have shown that it immunoprecipitates the alpha beta TCR of HPB-ALL and that it is specific for the V beta 5.3 chain of the human TCR. In addition, we have observed that this antibody stains a minor fraction of T lymphocytes in different strains of mice. We have screened a number of murine T cell clones or hybridomas and have found that the T cell hybrid line DO.11.10.S4.4 is positive. We have been unable to immunoprecipitate reproducibly the molecule recognized by 111-427 after 125I cell surface labeling and cell lysis in NP-40 or digitonin, probably because of low-affinity binding. On Western blotting, 111-427 revealed one band that has an apparent molecular mass of 89 kDa in nonreducing conditions and disappears after reduction. Similar results were obtained in parallel with the F23.1 and F23.2 antibodies. Thus, this antibody appears to recognize an epitope present primarily on the V beta 8.2 chain of the mouse TCR. We have assayed its capacity to stimulate splenic T lymphocytes in vitro. We have observed that it is capable of triggering, to a minor degree in soluble form and very effectively when coupled to Sepharose beads, the proliferation of spleen T lymphocytes from mice chronically infected with the blood parasite Trypanosoma cruzi.

Effects of noninherited maternal antigens on allotransplant rejection in a transgenic mouse model.

We studied the influence of noninherited maternal antigen (NIMA) on allotransplant rejection using a mouse transgenic model. CBK transgenic (CBA [H-2k] expressing K(b) MHC class I transgene) mice were used as donors in heart transplantation experiments. Offspring of BM3.3 (CBA anti-K(b) TCR transgenic) male mice and (CBA x CBK)F1 females were used as NIMA (offspring that did not inherit K(b)) and IMA (offspring that inherited K(b) maternal antigen) recipient mice. Survival of allografts was monitored and the alloimmune response evaluated using an ELISPOT assay. IMA mice accepted CBK heart allografts and displayed no alloresponse to K(b+) cells. In contrast, mice never exposed to K(b) (offspring of BM3.3 males and CBA females) acutely rejected their grafts within 18 days posttransplantation and exhibited potent inflammatory alloresponses to K(b+) cells. NIMA mice displayed prolonged survival of allotransplants (MST >60 days). Although no deletion of anti-K(b) TCR transgenic cells was detected in these mice, they had a marked reduction in the frequency of activated alloreactive T cells producing type 1 (IFN-gamma and IL-2) cytokines and concomitant expansion of type 2 (IL-4) cytokine-secreting cells. Finally, depletion of CD4+ T cells from NIMA mice restored acute rejection of CBK hearts. This study is the first demonstration of the tolerogenic effects of NIMA on alloimmunity and allotransplant rejection in a transgenic model. It is shown that, although the NIMA tolerogenic effect is not due to deletion of alloreactive T cells, it is mediated by CD4+ T cells producing type 2 cytokines.

MODFLOW-Based Coupled Surface Water Routing and Groundwater-Flow Simulation.

In this paper, we present a flexible approach for simulating one- and two-dimensional routing of surface water using a numerical surface water routing (SWR) code implicitly coupled to the groundwater-flow process in MODFLOW. Surface water routing in SWR can be simulated using a diffusive-wave approximation of the Saint-Venant equations and/or a simplified level-pool approach. SWR can account for surface water flow controlled by backwater conditions caused by small water-surface gradients or surface water control structures. A number of typical surface water control structures, such as culverts, weirs, and gates, can be represented, and it is possible to implement operational rules to manage surface water stages and streamflow. The nonlinear system of surface water flow equations formulated in SWR is solved by using Newton methods and direct or iterative solvers. SWR was tested by simulating the (1) Lal axisymmetric overland flow, (2) V-catchment, and (3) modified Pinder-Sauer problems. Simulated results for these problems compare well with other published results and indicate that SWR provides accurate results for surface water-only and coupled surface water/groundwater problems. Results for an application of SWR and MODFLOW to the Snapper Creek area of Miami-Dade County, Florida, USA are also presented and demonstrate the value of coupled surface water and groundwater simulation in managed, low-relief coastal settings.

[Impact of an intervention improving the food supply (excluding school meals) with educational support in middle and high schools]. / Impact d'une intervention sur l'offre alimentaire et sur les comportements alimentaires des adolescents (hors restauration scolaire).

Within the Nutrition, Prevention, and Health Program for children and teenagers in Aquitaine, an experimental intervention was implemented in 2007-2008 in the middle and high schools in Aquitaine (southwest France). This intervention aimed to improve the eating habits of adolescents, combining actions to improve the food supply sold during recreational times (remove/limit fat and sugar products sold and promote the sale of fruits and bread) and health education actions to make adolescents aware of the concept of nutritional balance and steer their choice towards recommended products. The aim of this paper was to evaluate the impact of the intervention on the eating behavior of adolescents and the food supply sold during recreational times in middle and high schools in Aquitaine. A survey was conducted before and after the implementation of the intervention in seven middle and high schools that have implemented actions (intervention group) and eight middle and high schools that have not implemented actions (control group). In these schools, 1602 adolescents answered the survey before and 1050 after the intervention (samples were independent because of the anonymity of responses). The impact of the intervention on the dietary behavior of teenagers was modeled using logistic regression adjusted on potential confounding variables (sex, age, and educational status). In multivariate analyses, the intervention was associated with more frequent daily intake of breakfast (OR=2.63; 95% CI [1.89; 3.66]) and lower intake of morning snacks (OR=0.66; 95% CI [0.48; 0.90]), higher consumption of starchy foods (OR=1.77; 95% CI [1.30; 2.42]), bread at breakfast, morning snacks, and a light afternoon meal (OR=1.43; 95% CI [1.07; 1.90]), and the food supply sold at recreational times (OR=1.34 95% CI [1.01; 1.78]). These results show that the "Improving food supply in middle and high schools associated with educational support actions" project led to the sales of recommended foods during recreational times and improved students' eating behavior. These results encourage partners to pursue these actions in all volunteer middle and high schools.

A mouse placental immunoregulatory factor different from transforming growth factor beta.

Of the growth-promoting factors, transforming growth factor beta (TGF-beta) has been most clearly shown to act as a potent regulator of inflammation and immunity. It is highly suppressive for T and B lymphocyte proliferation, cytotoxic T lymphocyte generation, and lymphokine-activated killer cell development, as well as natural killer cell activity. Moreover, there is accumulating evidence that TGF-beta also may contribute to impaired immune surveillance of tumor development. In previous work, we isolated and described a 40 kD glycoprotein extracted from mouse placenta. This placental factor (PF) is also a potent immune modulator in vivo: it is highly inhibitory of secondary antibody responses as well as cellular responses, such as local graft-versus-host reactions. Because placenta has been shown to be a major source of TGF-beta and several reports have indicated an important role for TGF-beta in the immunosuppressive mechanisms taking place during the course of mammalian gestation, we have looked for the presence of TGF-beta in our placental factor preparations. Our results clearly indicate that they do not contain TGF-beta or TGF-beta-like molecules by the following criteria: (1) no inhibition of Mv-1 Lu cell proliferation at any dose tested; (2) no band detected by immunoblotting using different polyclonal reagents specific for TGF-beta 1; and (3) no activity retained on or eluted from an affinity column made of immobilized monoclonal antibody against TGF-beta 2. Aliquots of the same preparations retained their full immune inhibitory capacity in vivo throughout the various assays.(ABSTRACT TRUNCATED AT 250 WORDS)

A quick method for the preparation of stable DNP red cell conjugates.

Another procedure for preparing dinitrophenyl (DNP) sheep erythrocytes (SRBC) using the DNP-alanylglycylglycyl hapten has been developed. These DNP-SRBC also are stable for as long as 1 month, but the conjugation procedure is much simpler to carry out than the previously reported method.

Fibrosing cholehepatitis in broiler chickens induced by bile duct ligations or inoculation of Clostridium perfringens.

The pathogenesis of fibrosing hepatitis causing condemnations in broiler chickens was investigated. Three to four week old broilers were inoculated via the hepatoenteric bile duct with saline washed suspensions of Clostridium perfringens (10(7) and 10(8) organisms). In another group of broilers, both bile ducts were ligated. The sequential development of liver and gall bladder lesions was studied at intervals ranging from 1-28 days postsurgery. The lesions were similar in both experiments in that the liver became mottled and swollen by five to seven days. Fibrinoid necrosis, heterophil and lymphocyte infiltration, bile duct hyperplasia and fibrosis with reticulin fiber proliferation occurred. By 14-17 days, the liver was enlarged, tan colored and firm with red and white foci. By 28 days, bile duct proliferation and fibrosis were massive with only a few hepatocytes remaining. The liver capsule was not involved. Jaundice was not present but the birds with ligated bile ducts excreted intensely yellow stained droppings after six to seven days. The gall bladder in inoculated birds was edematous and distended with flocculent or inspissated material. Clostridium perfringens was reisolated from gall bladder and/or liver of inoculated birds up to 28 days postsurgery. It is suggested that this organism plays a role in the pathogenesis of fibrosing cholehepatitis by inducing septic intrahepatic cholestasis.

[Morning snack offer in kindergarten: outlook and practices in Aquitaine, France, between 2004 and 2008]. / La collation à l'école maternelle: évolution des perceptions et pratiques des enseignants d'Aquitaine entre 2004 et 2008.

The Nutrition, Prevention and Health for Children and Teenagers in Aquitaine program is a regional implementation of the French National Program of Nutrition and Health (PNNS). The first of two surveys of a representative sample of Aquitaine preschools was conducted in 2004-2005. This survey showed that more than 2/3 of teachers offered a morning snack to their pupils at around 10 o'clock in the morning, mainly composed of sweet cakes, cookies, or candies. Following this initial survey, actions were implemented starting in September 2005, aiming to stop systematic morning snacks or improve their composition. The same survey was repeated during 2007-2008 in order to analyze the changes in morning snack practices in preschools between 2004-2005 and 2007-2008. Data were collected from teachers of Aquitaine's preschools who filled out questionnaires. Schools were randomly chosen according to their size and whether or not they belonged to a priority education zone (ZEP). The results show that 57.9% of teachers organized a morning snack in 2007-2008, versus 68.7% in 2004-2005. In 2004-2005, 22% of teachers considered the morning snack as "unjustified" compared to 44% in 2007-2008. The composition of morning snacks improved: 17.7% of teachers offered fruit and/or milk in 2007-2008 versus 8.5% in 2004-2005. Morning snacks composed of other foods (such as sweets, chocolate pieces, cookies, pastries, bread, fruit juice) decreased from 60.2% in 2004-2005 to 40.2% in 2007-2008. In 2007-2008, 19% of the teachers reported that children had snacks in their schoolbag versus 34% in 2004-2005. The proportion of teachers reporting children having snacks in their schoolbag decreased from 34% in 2004-2005 to 19% in 2007-2008. Comparison between these two surveys is encouraging as it shows an improvement of the perception and practices of teachers regarding morning snacks. These results encourage the partners of this program to continue the fieldwork actions.

High performance liquid chromatographic determination of azelastine and desmethylazelastine in guinea pig plasma and lung tissue.

Analytical methods have been developed for the simultaneous quantitation of azelastine (AZ) and desmethylazelastine (DAZ) in guinea pig plasma and lung tissue. The methods require a 1.00 mL plasma sample and a minimum 0.100 g lung sample. Both methods employ liquid/liquid organic extraction and back-extraction into dilute acid. Quantitation is performed by high performance liquid chromatography on a 2 x 250 mm 5 microns Hypersil CPS column using fluorescence detection. The linear quantitative ranges for AZ.HCl and DAZ.HBr in plasma are 0.156-160 ng/mL and 0.313-160 ng/mL, respectively. The linear quantitative ranges for AZ.HCl and DAZ.HBr in lung tissue are 0.039-20 micrograms/g for both.

Antigen-specific modulation of graft-versus-host reactions by two distinct placental factors.

We have previously shown that two distinct mouse placental fractions (PF) are potent immunomodulators in vivo. A 40 kDa PF induces a marked decrease of plaque forming cell (PFC) responses, while a 60 kDa PF increases them. Both effects are specific for the priming antigen. In the present study, these two PF are assayed on a cell-mediated response to allogeneic cells, i.e. in a local graft-versus-host reaction (LGVHR). Mice were primed with allogeneic cells in the presence of various amounts of the 40 kDa or 60 kDa PF, or liver extract (LE) as control. Six days later, their spleen cells were injected into the footpads of F1 recipients. Precise dose-response curves were established and the kinetics of the GVH response were carefully followed. Parallel with the modulation of PFC responses, the 40 kDa PF caused a potent inhibition of the LGVHR, while the 60 kDa PF greatly enhanced it. Both effects were specific for the alloantigens injected with the PF. Furthermore, we showed that these modulations were observed whatever the intensity of the GVH reaction, which varied according to the number of primed spleen cells transferred. This report also demonstrates that these PF can be greatly enriched by passage over affinity columns made of insolubilized lectins. The 40 kDa PF is retained on and can be eluted from columns of insolubilized concanavalin A (Con A) or wheat germ agglutinin (WGA), which indicates that it is a glycoprotein. Conversely, the 60 kDa PF does not bind to any of the above lectins and is probably not a glycoprotein. This biochemical purification step is also a good procedure for obtaining an even cleaner separation of the two fractions from each other. Thus, this paper demonstrates that both PF have important regulatory properties on specific cellular immune responses.

Functional and biochemical properties of a mouse placental immunoregulatory factor.

The authors have a long standing interest in immune regulations which control the absence of rejection of a semi-allogeneic fetus by the mother. A previous work described a soluble 40 kDa factor extracted from mouse placenta and capable of inhibiting secondary immune responses in vitro. The present paper reports the following on its mode of action in vivo: (1) it is active even in a fully allogeneic host; (2) it can be administered i.v. or i.p. along with antigen; and (3) the injections of factor and antigen must not be more than 2 days apart for maximum efficacy. Moreover, the results of the study described here indicate also that this factor is a concanavalin A-binding glycoprotein, sensitive to heat and pronase, and different from interleukin 10 (IL-10). Thus, this placental factor appears to be different from previously described immune regulators such as IL-10 and could contribute significantly to immune regulations at the level of the placenta.

T cell tolerization in vitro: modulation of helper and suppressor cell activities.

This paper analyzes the conditions for in vitro tolerization of purified whole T cell populations and the consequences on helper and suppressor T cell functions. Highly purified splenic T cells from adult DBA/2 mice were incubated in vitro for 24 hr with high doses of trinitrophenyl coupled to human gamma-globulins (TNP-HGG). A profound inhibition of the TNP-specific helper function of these T lymphocytes was observed in a cooperative culture with normal purified splenic B cells and TNP-SRBC as antigen. This state of specific unresponsiveness was maintained after trypsin treatment of the cells, at the end of the 24-hr incubation with the tolerogen. We checked that this procedure removed the vast majority of F23.1 T cell receptor determinants from the cells. This result indicates that T cell receptors for antigen were not merely blocked by the tolerogen. In addition, B cells preincubated with tolerized T cells for 24 hr remained as responsive to TNP as B cells mixed with normal T cells in similar conditions. This demonstrates that the decreased response is not the result of secondary B cell tolerization. In addition, anti-Ia monoclonal antibodies were shown to block the induction of tolerance. We also showed that tolerized T cells significantly decreased the anti-TNP response of normal T and B cells in vitro, whereas the anti-SRBC response in the same cultures was unaffected. When tolerized T cells were separated into Lyt-2- and Lyt-2+ cells, it was found that tolerized Lyt-2- cells had lost about 75% of their helper activity and that Lyt-2+ cells suppressed 70% of the response of a normal T and B cell culture. Thus, in vitro induction of T cell tolerance results in a specific T cell unresponsiveness which is due to both helper T cell inactivation and induction of specific suppressor T cells.

Role of suppressor cells in antibody-induced inhibition of mixed lymphocyte reactions.

Facilitating immune sera (FIS) have both in vivo enhancing and in vitro blocking properties. They inhibit the mixed lymphocyte reaction (MLR) in the mouse. In the present report, we studied the inhibitory capacity of these sera after removal of populations of suppressor cells both in vivo by adult thymectomy and in vitro by depletion of histamine-receptor-bearing cells (HRC). A significant decrease of the MLR inhibition by FIS after either of these depletions was observed. It appeared therfore that facilitating antibodies and thymus-dependant and/or HRC could cooperate in the antibody-induced inhibition of the MLR in the mouse.