Volume 33, Issue 3,
, Pages 344-352
Author links open overlay panel, , , ,
The TH1–TH2 paradigm provides an elegant model of directed response to infectious pathogens. Developed in the mouse, the model has provided a framework for systematic and mechanistic studies of immune regulation, protective immunity, and vaccine development in swine. Interleukin-4 (IL-4) plays a central role in the paradigm as a regulatory molecule directing development of the TH2 phenotype, as a developmental cytokine essential for antibody production, and as a soluble diagnostic marker of the TH2 cell type. In contrast, while characterizing the biological properties of porcine IL-4, we discovered that it was not a stimulatory factor for porcine B cells. Rather, it blocked antibody and IL-6 secretion and suppressed antigen-stimulated proliferation of B cells. Inhibition was not reversed by treatment with IL-2 and IL-6 treatment. IL-4 did not stimulate T lymphocyte proliferation, but induced cell growth in lymphoblasts in a dose-dependent fashion. These results suggest that IL-4 plays a different role in pigs than in mice and humans, in which it stimulates B cells and is essential for antibody production. Furthermore, the functions of IL-4 in swine cannot be inferred from results in model systems such as the mouse. General models of disease resistance show substantial variation between pigs and mice at the cellular and molecular level. Advances in somatic cell technologies and animal engineering to enable gene knockouts in pigs, in combination with a continuously expanding immunological toolkit, promise an exciting future for pig immunology, detailed mechanistic elucidation of the TH1–TH2 paradigm, and an improved understanding of the role of IL-4 in porcine immunity to infectious disease.
In our view, the immune system evolved to resist rapidly proliferating pathogens that, in the absence of an immediate response, had the potential to overwhelm and kill a host. Slowly replicating pathogens, by comparison, posed a less immediate threat and so required less of an immediate response. The cells and molecules responsible for immunological function are most easily explained as arising at single points in evolutionary history. The sensing receptors of innate immunity are ancient, and lymphocytes first appeared in the cartilaginous fishes. Because similar cells and molecules of immunity can be recognized through evolutionary time and across vertebrate species, knowledge obtained in one species is widely disseminated and assumed to be relevant to other species. In this way, comparative immunology has been employed widely to guide vaccine development in food and companion animals since the tools and resources for direct interrogation of basic immunology in these species often is limited or lacking.
Pioneering studies at DNAX by the Mosmann and Coffman laboratories laid the foundation of the TH1–TH2 paradigm, a simple concept summarized in Fig. 1. It is based on patterns of cytokine secretion in activated helper T cells that provided a conceptual and mechanistic basis for development of immune responses against intracellular and extracellular pathogens , . TH1 cells secreted IL-2 and IFNγ that activated cytotoxic T cells and macrophages, cell types actively involved in the control of infection by intracellular pathogens. TH2 T cells secreted IL-4, which stimulated antibody production, and controlled infections by extracellular pathogens. IFNγ and IL-4 were not only activating molecules of cellular and humoral immunity, they also were key negative regulatory molecules suppressing TH2 and TH1 cells, respectively.
The relevance of a murine immunoregulatory paradigm in pigs is related to the degree that the underlying assumptions are valid. Little is known about IL-4 in swine. Here, we introduce the basic function of IL-4 in the TH1–TH2 paradigm and provide new data on its genomic organization, regulation of expression and its biological properties. The purpose is to address what, to us, is a fundamental contradiction in IL-4 function that we observed in the pig when trying to replicate the original observation that IL-4 stimulated B-cell growth. IL-4 can easily be a central character in reviews of anti-parasite immunity, asthma, allergy, T-cell differentiation, and cytokine signaling. Here, we focused on its regulatory role in B-cell differentiation and function. The findings indicate that the gene and protein are conserved through mammalian evolution, but that its role in regulation of adaptive immune responses in the pig is different than in mouse or human. One consequence of these differences is that the key TH2-type immunoregulatory molecule may be different in pigs or the pathways of signal transduction may be different from those in mouse and human.
Interleukin-4 (IL-4) was originally recognized as a co-stimulatory factor for the anti-IgM antibody-induced proliferation of resting B cells . It plays an important role in promoting humoral immune responses against extracellular pathogens, and its expression is characteristic of TH2 cells, where it is primarily produced , . Gene knockout experiments demonstrated in mice that IL-4 is essential for TH2 cytokine responses  and antibody production . In addition to its B-cell
Effects of IL-4 on B-cell proliferation in the pig
IL-4 was discovered by its potentiation of anti-IgM induced B-cell proliferation . Thus, we examined this effect of IL-4 in swine using the same approach. In contrast to the marked stimulation of B-cell proliferation observed in mice, neither porcine nor human IL-4 stimulated B-cell proliferation in the pig (Fig. 2). Instead, there appeared to be a reduced response to antigen receptor engagement by anti-porcine IgM antibodies, and there was no proliferation in response to IL-4 alone. The
Effects of IL-4 on B-cell antibody and cytokine secretion in the pig
It was possible that IL-4 was acting in a stimulatory manner on B-cells in other stages of differentiation that we could not detect by staining for the B-cell antigen receptor, or that responding cells were resident in lymphoid tissue rather than in the circulation. To address these possibilities, we determined its effects on immunoglobulin and IL-6 secretion, which are commensurate with a TH2-type immune response, in B cells isolated from the spleen in a model of antigen-specific recall. Pigs
Effect of IL-4 on T cell proliferation
The negative effects of IL-4 on B-cell function did not appear to be due to non-specific toxicity insofar as human recombinant IL-4 also suppressed B-cell proliferation, as shown in Fig. 2. To further demonstrate that the findings were biologically relevant and not due to a nonspecific toxicity, we examined the effect of IL-4 on T lymphocyte proliferation. Alone, it had no effect on cell proliferation in PBMC. However, mitogenic stimulation of PBMC for 5 day to produce lymphoblasts made both
Porcine IL-4 and the TH1–TH2 paradigm
Our findings and published data show that in swine, IL-4 suppresses macrophage inflammatory responses , , suppresses natural killer cell activity , is involved in anti-parasite responses , helps to differentiate macrophages and monocytes into dendritic cells , , and stimulates T lymphoblast proliferation (Fig. 7). In dendritic cell differentiation, IL-4 appears to facilitate cell survival but not differentiation , and in B cells it promotes survival of activated B
Re-evaluation of the TH1–TH2 paradigm in porcine immunoregulation
Pigs have a number of immunological characteristics that are different from mice in addition to T lymphocyte subpopulations. For example, lymph node architecture is inverted relative to most other species ; development of antibody diversity involves gene conversion ; and γδ receptor features are unusual . Pigs are comprised of outbred populations with diverse MHC class I and class II compositions, in contrast to inbred mouse strains widely used in immunological research. Due to
The authors thank Csaba Pasmany and Mark Moody, Pierce Endogen, Inc., Woburn, MA for providing recombinant porcine IL-4. Supplementary Fig. 1 was prepared by Geoff Hirsch and Juan Abrahante, University of Minnesota. Special thanks go to Reinhard Pabst, Medical School of Hannover, Germany, whose encouragement was a significant motivation, and to Artur Summerfield, Institute of Virology and Immunoprophylaxis, Mittelhäusern, Switzerland, for insightful comments and suggestions.
- A. O’Garra
Cytokines induce the development of functionally heterogeneous T helper cell subsets
- A.A. te Velde et al.
Interleukin-4 (IL-4) inhibits secretion of IL-1 beta, tumor necrosis factor alpha, and IL-6 by human monocytes
- A. Nuntaprasert et al.
Characterization of specific antibodies and the establishment of sandwich ELISA and ELISPOT systems for swine IL-4
Comp Immunol Microbiol Infect Dis
- I. Diaz et al.
Use of ELISPOT and ELISA to evaluate IFN-gamma, IL-10 and IL-4 responses in conventional pigs
Vet Immunol Immunopathol
- E.M. Bautista et al.
IL-13 replaces IL-4 in development of monocyte derived dendritic cells (MoDC) of swineSee AlsoSpecies specialization in cytokine biology: Is interleukin-4 central to the TH1–TH2 paradigm in swine?Cytokine upregulation of the antigen presenting function of acute myeloid leukemia cells - DocumentExposure to particulate matter 2.5 (PM2.5) induced macrophage-dependent inflammation, characterized by increased Th1/Th17 cytokine secretion and cytotoxicity
Vet Immunol Immunopathol
- Y. Zhou et al.
Interleukin-4 suppresses the expression of macrophage NADPH oxidase heavy chain subunit (gp91-phox)
Biochim Biophys Acta
- D.L. Foss et al.
Differentiation of porcine dendritic cells by granulocyte-macrophage colony-stimulating factor expressed in Pichia pastoris
Vet Immunol Immunopathol
- A. Nuntaprasert et al.
The effect of recombinant swine interleukin-4 on swine immune cells and on pro-inflammatory cytokine productions in pigs
Comp Immunol Microbiol Infect Dis
- T.V. Nguyen et al.
Transfer of maternal cytokines to suckling piglets: in vivo and in vitro models with implications for immunomodulation of neonatal immunity
Vet Immunol Immunopathol
- P. Matzinger
An innate sense of danger
Specific versus cooperative regulatory mechanisms of the cytokine genes that are clustered on the same chromosome
J Allergy Clin Immunol
Mucosal immunogenicity and adjuvanticity of cholera toxin in swine
Isolation and characterization of gamma delta T lymphocyte cell lines from Sinclair swine peripheral blood
Vet Immunol Immunopathol
Establishment and characterization of porcine cytolytic cell lines and clones
Vet Immunol Immunopathol
Phenotypic and functional characterisation of porcine CD4(+)CD25(high) regulatory T cells
Vet Immunol Immunopathol
Characterization of porcine T lymphocytes and their immune response against viral antigens
Organisation of the lymphoreticular system and lymphocyte markers in the pig
Vet Immunol Immunopathol
Fc receptors in livestock species
Vet Immunol Immunopathol
Identification of a conserved GATA3 response element upstream proximal from the interleukin-13 gene locus
J Biol Chem
TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional properties
Annu Rev Immunol
Two types of murine helper T cell clone. Part I. Definition according to profiles of lymphokine activities and secreted proteins
Identification of a T cell-derived B cell growth factor distinct from interleukin 2
J Exp Med
Disruption of the murine IL-4 gene blocks TH2 cytokine responses
Generation and analysis of interleukin-4 deficient mice
IL-4 inhibits the expression of IL-8 from stimulated human monocytes
Interleukin-4 suppression of monocyte tumour necrosis factor-alpha production. Dependence on protein synthesis but not on cyclic AMP production
IL-4 suppresses cytokine gene expression induced by IFN-gamma and/or IL-2 in murine peritoneal macrophages
IL-4 down-regulates IL-1 and TNF gene expression in human monocytes
Differential regulation of IL-1 production in human monocytes by IFN-gamma and IL-4
Isolation and characterization of a human interleukin cDNA clone, homologous to mouse B-cell stimulatory factor 1, that expresses B-cell- and T-cell-stimulating activities
Proc Natl Acad Sci USA
Structural comparison and chromosomal localization of the human and mouse IL-13 genes
Cited by (55)
Characterization of immune development of fetal and early-life of minipigs
2023, International Immunopharmacology
Fetal and child’s immune systems differ from those of adults. Developing immune systems exhibit increased or decreased sensitivity to drugs, infection, or toxicants compared to adult immune systems. Understanding fetal and neonatal immune systems will help predict toxicity or the pathogenesis or prognosis of diseases. In this study, we evaluated whether the innate and adaptive immune system of fetal and young minipigs could respond to external stimuli compared to a medium-treated group and analyzed several immunological parameters for developmental immunotoxicity according to developmental stages. We performed a hematological analysis of fetal cord bloods and the bloods of neonatal and 4-week-old piglets. Splenocytes were isolated at each developmental stage and treated with lipopolysaccharide (LPS), R848, and concanavalin A (ConA). Various cytokines were measured in the cell supernatants. Total antibody production was also evaluated in serum. The percentage of lymphocytes was dominant in gestational weeks (GW) 10 and 12 and started to decrease from postnatal day (PND) 0. From PND0, the percentage of neutrophils increased. Interleukin (IL)-1β, IL-6, and interferon (IFN)-α were induced from GW10 in response to LPS and R848 stimulation. Th1 cytokine induction was detected from PND0 upon ConA stimulation, whereas Th2 cytokine release was observed from GW10. IgM and IgG production was sustained at low levels at fetal stages and was significantly increased after birth. This study reconfirmed that the fetal immune system could respond to external stimuli and that hematological analysis, cytokine evaluation, and antibody subclass measurement can be useful parameters for developmental immunotoxicity using minipigs.
Domestic pig prioritized in one health action against fascioliasis in human endemic areas: Experimental assessment of transmission capacity and epidemiological evaluation of reservoir role
2021, One Health
The Northern Bolivian Altiplano is the human fascioliasis hyperendemic area where the highest prevalences and intensities in humans have been reported. Preventive chemotherapy was implemented in the last ten years. Surveillance showed high human infection and re-infection rates in between the annual triclabendazole monodose treatments. A complementary One Health control action was launched to decrease the infection risk. Among the multidisciplinary axes, there is the need to establish animal reservoir species priorities for a more efficient control. Laboratory and field studies were performed for the first time to assess the Fasciola hepatica transmission capacity of the pig and its potential reservoir role. The experimental follow-up of altiplanic pig isolates through altiplanic Galba truncatula snail vector isolates were performed at different miracidial doses and different day/night temperatures. Experiments included egg embryonation, miracidial infectivity, lymnaeid snail infection, intramolluscan larval development, cercarial production, chronobiology of the cercarial shedding, vector survival to infection, metacercarial infectivity of mammal host, and adult stage development. Surveys included the assessment of prevalence, intensity, egg measurements and egg shedding rates in nature. Pig contribution was evaluated by comparing with the main altiplanic reservoirs sheep and cattle. Results demonstrated that the pig assures the whole F. hepatica life cycle and participates in its transmission in this area. The fast egg embryonation, high cercarial production, long multi-wave shedding chronobiological pattern in monomiracidial infections at permanent 20°C temperature, and the high daily egg outputs per pig are worth mentioning. The high infection risk suggests early infection of freely running piglets and evolutionary long-term adaptation of the liver fluke to this omnivorous mammal, despite its previously evoked resistance or non-suitability. Genetic, physiological and immune similarities with humans may also underlie the parasite adaptation to humans in this area. The pig should be accordingly included for appropriate control measures within a One Health action against human fascioliasis. The pig should henceforth be considered in epidemiological studies and control initiatives not only in fascioliasis endemic areas with human infection risk on other Andean countries, but also in rural areas of Latin America, Africa and Asia where domestic pigs are allowed to run freely.
The pig as a medical model for acquired respiratory diseases and dysfunctions: An immunological perspective
2021, Molecular Immunology
By definition no model is perfect, and this also holds for biology and health sciences. In medicine, murine models are, and will be indispensable for long, thanks to their reasonable cost and huge choice of transgenic strains and molecular tools. On the other side, non-human primates remain the best animal models although their use is limited because of financial and obvious ethical reasons. In the field of respiratory diseases, specific clinical models such as sheep and cotton rat for bronchiolitis, or ferret and Syrian hamster for influenza and Covid-19, have been successfully developed, however, in these species, the toolbox for biological analysis remains scarce. In this view the porcine medical model is appearing as the third, intermediate, choice, between murine and primate. Herein we would like to present the pros and cons of pig as a model for acquired respiratory conditions, through an immunological point of view. Indeed, important progresses have been made in pig immunology during the last decade that allowed the precise description of immune molecules and cell phenotypes and functions. These progresses might allow the use of pig as clinical model of human respiratory diseases but also as a species of interest to perform basic research explorations.
Swine dysentery disease mechanism: Brachyspira hampsonii impairs the colonic immune and epithelial repair responses to induce lesions
2020, Microbial Pathogenesis
Swine dysentery (SD) is a global, production-limiting disease of pigs in commercial farms. It is associated with infection by Brachyspira hyodysenteriae and B. hampsonii, and characterized by mucohaemorrhagic diarrhea and colitis, SD prevention, treatment or control relies heavily on antimicrobials as no commercial vaccines are available. This is linked to our poor understanding of the disease pathogenesis. Our goal was to characterize the host-pathogen interactions during the early stage of infection. We employed dual RNA-seq to profile mRNA and miRNA following 1-h incubation of colonic explants with a pathogenic or a non-pathogenic B. hampsonii strain. Our results suggest that the pathogenic strain more efficiently interfered with the host's ability to activate and build a humoral response (through IL-4/CCR6/KLHL6 interactions), epithelial wound repair mechanisms (associated with LSECtin impairment of macrophages), induced mitochondrial dysfunction (linked to MDR1), and loss of microbiome homeostasis. The pathogenic strain also up-regulated the expression of stress-associated genes, when compared to the non-pathogenic strain. These results shed a light on the pathophysiological mechanisms that lead to SD and will contribute to the development of novel disease control tools.
Pulmonary inflammatory response to influenza virus infection in pigs is regulated by DAP12 and macrophage M1 and M2 phenotypes
2020, Cellular Immunology
We delineated the expression of DAP12 (DNAX-Activating Protein) and its associated receptors, TREM-1, TREM-2 and MDL-1 in pig alveolar monocyte/macrophages (AMM) that have attained M1 or M2 phenotypes. Pig AMM stimulated in vitro with IFN-γ and IL-4 induced the expression of M1 (TNFα and iNOS) and M2 (ARG1 and no MMR) phenotypic markers, respectively. In influenza virus infected pigs at seven days post-infection, in addition to substantial modulations in the M1 and M2 markers expression, DAP12, TREM-1 and MDL-1 were downregulated in AMM. Thus, DAP12 signaling promoted the anti-inflammatory pathway in AMM of influenza virus infected pigs.
The regulatory actions of retinoic acid on M2 polarization of porcine macrophages
2019, Developmental and Comparative Immunology
We previously demonstrated that the most bioactive vitamin A metabolite, all-trans retinoic acid (ATRA), increased T helper 2-associated responses induced in pigs by infection with the parasitic nematode Ascaris suum We also showed that ATRA potentiated the mRNA expression of several IL-4 induced chemokines (chemokine (CC motif) ligand 11 [(CCL11), CCL17, CCL22 and CCL26] associated with alternative activation (M2a) in porcine macrophages in vitro. Herein, several mechanisms whereby ATRA affects IL-4 signaling are profiled using large-scale real time PCR and RNA-Seq analysis. Twenty-three genes associated with M2a markers in other species were independently upregulated by both IL-4 and ATRA, including the adenosine receptor A2B (ADORA2B), cysteinyl leukotriene receptor 2 (CYSLTR2) and the vitamin D receptor (VDR). ATRA synergistically enhanced IL-4 up-regulation of Hepatitis A virus cellular receptor 2 (HAVCR2) and transglutaminase 2 (TGM2) and further repressed IL-4 down-regulated CD163 and Cytochrome b-245, beta polypeptide (CYBB) mRNA. Macrophages treated with ATRA exhibited a dose-dependent reduction in phagocytosis of opsonized Staphylococcus aureus. In addition, the combination of IL-4 and ATRA up-regulated the anti-inflammatory protein, IL-1R antagonist (IL1RN) and TGM2. These data indicate that ATRA induces a state of partial alternative activation in porcine macrophages, and amplifies certain aspects of M2a activation induced by IL-4. Given the prevalence of allergic and parasitic diseases worldwide and the close similarities in the porcine and human immune responses, these findings have important implications for the nutritional regulation of allergic inflammation at mucosal surfaces.
Recommended articles (6)
Polyanhydride nanovaccine against swine influenza virus in pigs
Vaccine, Volume 35, Issue 8, 2017, pp. 1124-1131
We have recently demonstrated the effectiveness of an influenza A virus (IAV) subunit vaccine based on biodegradable polyanhydride nanoparticles delivery in mice. In the present study, we evaluated the efficacy of ∼200nm polyanhydride nanoparticles encapsulating inactivated swine influenza A virus (SwIAV) as a vaccine to induce protective immunity against a heterologous IAV challenge in pigs. Nursery pigs were vaccinated intranasally twice with inactivated SwIAV H1N2 (KAg) or polyanhydride nanoparticle-encapsulated KAg (KAg nanovaccine), and efficacy was evaluated against a heterologous zoonotic virulent SwIAV H1N1 challenge. Pigs were monitored for fever daily. Local and systemic antibody responses, antigen-specific proliferation of peripheral blood mononuclear cells, gross and microscopic lung lesions, and virus load in the respiratory tract were compared among the groups of animals. Our pre-challenge results indicated that KAg nanovaccine induced virus-specific lymphocyte proliferation and increased the frequency of CD4+CD8αα+ T helper and CD8+ cytotoxic T cells in peripheral blood mononuclear cells. KAg nanovaccine-immunized pigs were protected from fever following SwIAV challenge. In addition, pigs immunized with the KAg nanovaccine presented with lower viral antigens in lung sections and had 6 to 8-fold reduction in nasal shedding of SwIAV four days post-challenge compared to control animals. Immunologically, increased IFN-γ secreting T lymphocyte populations against both the vaccine and challenge viruses were detected in KAg nanovaccine-immunized pigs compared to the animals immunized with KAg alone. However, in the KAg nanovaccine-immunized pigs, hemagglutination inhibition, IgG and IgA antibody responses, and virus neutralization titers were comparable to that in the animals immunized with KAg alone. Overall, our data indicated that intranasal delivery of polyanhydride-based SwIAV nanovaccine augmented antigen-specific cellular immune response in pigs, with promise to induce cross-protective immunity.
A novel adjuvanted capsule based strategy for oral vaccination against infectious diarrhoeal pathogens
Journal of Controlled Release, Volume 233, 2016, pp. 162-173
Diarrhoeal infections are a major cause of morbidity and mortality with enterotoxigenic Escherichia coli (ETEC) and cholera imposing a significant global burden. There is currently no licensed vaccine for ETEC. Development of new nonliving oral vaccines has proven difficult due to the physicochemical and immunological challenges associated with the oral route. This demands innovative delivery solutions to protect antigens, control their release and build in immune-stimulatory activity. We describe the Single Multiple Pill® (SmPill®) vaccine formulation which combines the benefits of enteric polymer coating to protect against low gastric pH, a dispersed phase to control release and aid the solubility of non-polar components and an optimized combination of adjuvant and antigen to promote mucosal immunity. We demonstrate the effectiveness of this system with whole cell killed E. coli overexpressing colonization factor antigen I (CFA/I), JT-49. Alpha-galactosylceramide was identified as a potent adjuvant within SmPill® that enhanced the immunogenicity of JT-49. The bacteria associated with the dispersed phase were retained within the capsules at gastric pH but released at intestinal pH. Vaccination with an optimized SmPill® formulation promoted CFA/I-specific immunoglobulin A (IgA) responses in the intestinal mucosa in addition to serum IgG and a solubilized adjuvant was indispensable for efficacy.
Ex vivo activated CD4+ T cells from young calves exhibit Th2-biased effector function with distinct metabolic reprogramming compared to adult cows
Veterinary Immunology and Immunopathology, Volume 248, 2022, Article 110418
As maternal passive immunity wanes at 6–8 weeks, young calves must rely on their own naïve and developing immune system for protection against pathogens. Typically, an infection in the young induces a T cell-mediated response, which skews towards a Th2 phenotype and results in a reduced effector response. Our study examines the implications this transitional period of immunocompetency has on cellular metabolism in young calves, focusing on effector function of CD4+ T cells in comparison to those from adult cows. Results from sorted CD4+ T cells from young calves and adult cows activated by α-CD3:α-CD28, show that young calves exhibit a significantly greater propensity to produce the Th2 cytokine, IL-4, in comparison to IFN-γ. Concomitantly, cells from young calves and adult cows exhibit no statistical difference in cell surface marker expression induced by α-CD3:α-CD28 stimulation. Metabolically, activated CD4+ T cells from young calves show significantly greater utilization of mitochondrial respiration, measured by oxygen consumption rate (OCR), and greater glycolytic reserve, measured by extracellular acidification rate (ECAR). However, adult cows have a significantly higher change in glycolytic rate after α-CD3:α-CD28 stimulation compared to young calves. Further, CD4+ T cells from young calves have an increased mRNA expression signature associated with glycolytic metabolism (GAPDH, HK2, FBP1, HIF1A) and Th2-associated metabolic signaling (RPTOR) in comparison to adult cows. The distinct metabolic phenotype and associated gene expression in activated CD4+ T cells may be intrinsic drivers of the Th2-biased response by young calves. Additionally, CD4+ recent thymic emigrant cells (RTEs) may further contribute to altered effector function, as they are preferential precursors to Tregs, and based on the microenvironment, have the propensity to polarize toward Th2. Evaluation of T cell master transcription regulators, as well as measuring signal joint T cell receptor excision circles between young calves and adult cows, we observed a significantly increased proportion of RTEs from sorted CD4+ T cells. In this study, we show a unique metabolic profile exhibited by activated CD4+ T cells from young calves in which mitochondrial respiration and glycolytic capacity is significantly increased compared to adult cows.
Construction and preclinical evaluation of mmCT, a novel mutant cholera toxin adjuvant that can be efficiently produced in genetically manipulated Vibrio cholerae
Vaccine, Volume 34, Issue 18, 2016, pp. 2121-2128
There is an urgent need for new adjuvants that are effective with mucosally administered vaccines. Cholera toxin (CT) is the most powerful known mucosal adjuvant but is much too toxic for human use. In an effort to develop a useful mucosal adjuvant we have generated a novel non-toxic mutant CT molecule that retains much of the adjuvant activity of native CT. This was achieved by making the enzymatically active A subunit (CTA) recalcitrant to the site-specific proteolytic cleavage (“nicking”) required for toxicity, which was found to require mutations not only in the two residues rendering the molecule resistant to trypsin but also in neighboring sites protecting against cleavage by Vibrio cholerae proteases. This multiple-mutated CT (mmCT) adjuvant protein could be efficiently produced in and purified from the extracellular medium of CT-deleted V. cholerae. The mmCT completely lacked detectable enterotoxicity in an infant mouse model and had >1000-fold reduced cAMP inducing activity compared to native CT in a sensitive mammalian target cell system. It nonetheless proved to have potent adjuvant activity on mucosal and systemic antibody as well as cellular immune responses to mucosally co-administered antigens including oral cholera and intranasal influenza vaccines. We conclude that mmCT is an attractive novel non-toxic mucosal adjuvant for enhancing immune responses to co-administered mucosal vaccines.
iNKT cell activation plus T-cell transfer establishes complete chimerism in a murine sublethal bone marrow transplant model
American Journal of Transplantation, Volume 18, Issue 2, 2018, pp. 328-340
Transplant tolerance induction makes it possible to preserve functional grafts for a lifetime without immunosuppressants. One powerful method is to generate mixed hematopoietic chimeras in recipients by adoptive transfer of donor-derived bone marrow cells (BMCs). In our murine transplantation model, we established a novel method for mixed chimera generation using sublethal irradiation, CD40-CD40L blockade, and invariant natural killer T-cell activation. However, numerous BMCs that are required to achieve stable chimerism makes it difficult to apply this model for human transplantation. Here, we show that donor-derived splenic T cells could contribute to not only the reduction of BMC usage but also the establishment of complete chimerism in model mice. By cotransfer of T cells together even with one-fourth of the BMCs used in our original method, the recipient mice yielded complete chimerism and could acquire donor-specific skin-allograft tolerance. The complete chimeric mice did not show any remarks of graft versus host reaction in vivo and in vitro. Inhibition of the apoptotic signal resulted in increase in host-derived CD8+ T cells and chimerism brake. These results suggest that donor-derived splenic T cells having veto activity play a role in the depletion of host-derived CD8+ T cells and the facilitation of complete chimerism.
Progress in the use of swine in developmental immunology of B and T lymphocytes
Developmental & Comparative Immunology, Volume 58, 2016, pp. 1-17
The adaptive immune system of higher vertebrates is believed to have evolved to counter the ability of pathogens to avoid expulsion because their high rate of germline mutations. Vertebrates developed this adaptive immune response through the evolution of lymphocytes capable of somatic generation of a diverse repertoire of their antigenic receptors without the need to increase the frequency of germline mutation. The focus of our research and this article is on the ontogenetic development of the lymphocytes, and the repertoires they generate in swine. Several features are discussed including (a) the “closed” porcine placenta means that de novo fetal development can be studied for 114 days without passive influence from the mother, (b) newborn piglets are precocial permitting them to be reared without their mothers in germ-free isolators, (c) swine are members of the γδ−high group of mammals and thus provides a greater opportunity to characterize the role of γδ T cells and (d) because swine have a simplified variable heavy and light chain genome they offer a convenient system to study antibody repertoire development.
Current address: Center for Immunology, Department of Laboratory Medicine and Pathology, MMC 334, University of Minnesota, Minneapolis, MN 55455, USA.
Current address: Boehringer Ingelheim Vetmedica Inc., 2621 North Belt Highway, St. Joseph, MO 64506, USA.
Current address: Department of Clinical Investigation, Walter Reed Army Medical Center, Washington, DC 20307, USA.
Copyright © 2008 Elsevier Ltd. Published by Elsevier Ltd. All rights reserved.