CD154 Expression Indicates T Cell Activation Following Tetanus Toxoid Vaccination of Horses (original) (raw)
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OBTENTION OF TETANUS ANTITOXIN HYPERIMMUNE SERA IN EQUINES
2016
The study aimed to obtain a potent antitoxin for tetanus as the available antiserum is not routine in Cuba. The researchers emulsified different amounts of immunogen in Immune and carried out immunizations using several multisite protocols. The antitoxin antibody titers varied significantly among the protocols, with the fourth group of animals showing significantly higher titers. The study concluded that immunization protocols that are more prolonged produce better antitoxin outputs. The obtained potent antitoxin could be used as internal reference material in the tetanus toxoid antigen.
Subpopulations of equine blood lymphocytes expressing regulatory T cell markers
Veterinary Immunology and Immunopathology, 2010
Several distinct T lymphocyte subpopulations with immunoregulatory activity have been described in a number of mammalian species. This study performed a phenotypic analysis of cells expressing regulatory T cell (Treg) markers in the peripheral blood of a cohort of 18 horses aged 6 months to 23 years, using antibodies to both intracellular and cell surface markers, including Forkhead box P3 (FOXP3), CD4, CD8, CD25, interferon gamma (IFN␥) and interleukin 10 (IL-10). In peripheral blood, a mean of 2.2 ± 0.2% CD4+ and 0.5 ± 0.1% CD8+ lymphocytes expressed FOXP3. The mean percentage of CD4+FOXP3+ cells was found to be significantly decreased in horses 15 years and older (1.5%) as compared to horses 6 years and younger (2.7%), but did not differ between females and males and ponies and horses. Activation of peripheral blood mononuclear cells by pokeweed mitogen resulted in induction of CD25 and FOXP3 expression by CD4+ cells, with peak expression noted after 48 and 72 h in culture respectively. Activated CD4+FOXP3+ cells expressed IFN␥ (35% of FOXP3+ cells) or IL-10 (9% FOXP3+ cells). Cell sorting was performed to determine FOXP3 expression by CD4 + CD25 − , CD4 + CD25 dim and CD4 + CD25 high subpopulations. Immediately following sorting, the percentage of CD4+FOXP3+ cells was higher within the CD4 + CD25 high population (22.7-26.3%) compared with the CD4 + CD25 dim (17% cells) but was similar within the CD4 + CD25 dim and CD4 + CD25 high cells after resting in IL-2 (9-14%). Fewer than 2% of cells in the CD4 + CD25 − population expressed FOXP3. These results demonstrate heterogeneity in equine lymphocyte subsets that express molecules associated with regulatory T cells. CD4+FOXP3+ cells are likely to represent natural Tregs, with CD4+FOXP3+IL-10+ cells representing either activated natural Tregs or inducible Tregs, and CD4+FOXP3+IFN␥+ cells likely to represent activated Th1 cells.
Immunology, 2007
cd T-cell receptor + T lymphocytes are an important element of the innate immune system. Early production of interferon (IFN)-c by cd T cells may have a role in linking innate and adaptive immune responses and contribute to T helper-1 bias. We investigated the role of cytokines in the activation and induction of IFN-c secretion by bovine workshop cluster 1 + (WC1 +) cd T cells. The effects of culture with interleukin (IL)-12, IL-18, IL-15 and IL-2 were investigated; these cytokines are known to influence murine and human cd T cells. We report that bovine WC1 + cd T cells are synergistically stimulated by IL-12 and IL-18 to secrete large quantities of IFN-c. Neonatal calves were shown to have significantly higher numbers of circulating WC1 + cd T cells than adult animals. In addition, the response of peripheral blood WC1 + cd T cells was significantly higher in neonatal calves compared with adult animals. However, in adult animals the response of lymph node WC1 + cd T cells to IL-12/IL-18 was more pronounced than that of peripheral blood WC1 + cd T cells. We hypothesize that the induction of IFN-c secretion from WC1 + cd T cells by IL-12 and IL-18 is likely to be an important element of the innate response to pathogens such as Mycobacterium bovis. The high numbers of WC1 + cd T cells in neonatal calves, and their inherent ability to respond to inflammatory cytokines, could be a key factor in the enhanced responses seen in calves to BCG vaccination.
Ex Vivo Monitoring of Antigen-Specific CD4+ T Cells after Recall Immunization with Tetanus Toxoid
Clinical and Vaccine Immunology, 2007
To monitor antigen-specific CD4 ؉ T cells during a recall immune response to tetanus toxoid (TT), a sequential analysis including ex vivo phenotyping and cytokine flow cytometry, followed by cloning and T-cell-receptor (TCR) spectratyping of cytokine-positive CD4 ؉ T cells, was performed. Grossly, twice as many TT-specific CD4 ؉ T-cell clones, ex vivo derived from the CCR7 ؉/؊ CD69 ؉ interleukin-2-positive (IL-2 ؉ ) CD4 ؉ subsets, belonged to the central memory (T CM ; CD62L ؉ CD27 ؉ CCR7 ؉ ) compared to the effector memory population (T EM ; CD62L ؊ CD27 ؊ CCR7 ؊ ). After the boost, a predominant expansion of the T CM population was observed with more limited variations of the T EM population. TCR beta-chain-variable region (BV) spectratyping and sequencing confirmed a large concordance between most frequently expressed BV TCR-CDR3 from ex vivo-sorted CCR7 ؉/؊ CD69 ؉ IL-2 ؉ CD4 ؉ subsets and BV usage of in vitro-derived TT-specific CD4 ؉ T-cell clones, further demonstrating the highly polyclonal but stable character of the specific recall response to TT. Taken together, ex vivo flow cytometry analysis focused on the CCR7 ؉/؊ CD69 ؉ IL-2 ؉ CD4 ؉ subsets appears to target the bulk of antigen-specific T cells and to reach an analytical power sufficient to adequately delineate in field trials the profile of the antigen-specific response to vaccine.
Macrophage effector responses of horses are influenced by expression of CD154
Veterinary Immunology and Immunopathology, 2016
Reactive intermediates contribute to innate immunity by providing phagocytes with a mechanism of defense against bacteria, viruses and parasites. To better characterize the role of CD154 in the production of reactive intermediates, we cloned and expressed recombinant equine CD154 (reqCD154) in Chinese Hamster Ovary (CHO). In co-culture experiments, CHO cells ectopically expressing reqCD154 elicited superoxide production in monocyte-derived macrophages (MDM). Collectively, our results indicate that regulation of CD154 expression plays a role in innate host defenses.
Journal of Immunological Methods, 2005
One of the challenges for immunomonitoring in clinical trials is to detect an antigen specific T cell-mediated immune response. In an attempt to define the most suitable assay, tetanus toxoid was used to compare the capacity of 4 different methods to detect cytokine responses, before and after recall vaccination, in peripheral blood mononuclear cells (PBMC) of 14 healthy volunteers. ELISA, ELISPOT, intracytoplasmic detection and real-time RT-PCR were chosen to measure IFN-g production before and after vaccination. As far as the detection of memory T cell status (before vaccination) was concerned, we found that ELISPOT was the most sensitive method to discriminate TT-induced from spontaneous responses. On the other hand, intracytoplasmic cytokine detection was the most efficient method to detect the restimulating effect of TT vaccination. D
Transcriptomic response of porcine PBMCs to vaccination with tetanus toxoid as a model antigen
2013
The aim of the present study was to characterize in vivo genome-wide transcriptional responses to immune stimulation in order to get insight into the resulting changes of allocation of resources. Vaccination with tetanus toxoid was used as a model for a mixed Th1 and Th2 immune response in pig. Expression profiles of PBMCs (peripheral blood mononuclear cells) before and at 12 time points over a period of four weeks after initial and booster vaccination at day 14 were studied by use of Affymetrix GeneChip microarrays and Ingenuity Pathway Analysis (IPA). The transcriptome data in total comprised more than 5000 genes with different transcript abundances (DE-genes). Within the single time stages the numbers of DEgenes were between several hundred and more than 1000. Ingenuity Pathway Analysis mainly revealed canonical pathways of cellular immune response and cytokine signaling as well as a broad range of processes in cellular and organismal growth, proliferation and development, cell signaling, biosynthesis and metabolism. Significant changes in the expression profiles of PBMCs already occurred very early after immune stimulation. At two hours after the first vaccination 679 DE-genes corresponding to 110 canonical pathways of cytokine signaling, cellular immune response and other multiple cellular functions were found. Immune competence and global disease resistance are heritable but difficult to measure and to address by breeding. Besides QTL mapping of immune traits gene expression profiling facilitates the detection of functional gene networks and thus functional candidate genes. Citation: Adler M, Murani E, Brunner R, Ponsuksili S, Wimmers K (2013) Transcriptomic Response of Porcine PBMCs to Vaccination with Tetanus Toxoid as a Model Antigen. PLoS ONE 8(3): e58306.