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Journal of Dairy Science Sep 2022It is necessary for the dairy industry to reduce calf morbidity and mortality, and the reliance on antibiotics to treat sick calves, to address the growing concern...
It is necessary for the dairy industry to reduce calf morbidity and mortality, and the reliance on antibiotics to treat sick calves, to address the growing concern regarding antibiotic resistant bacteria. The primary objective of this study was to evaluate the effect that feeding dairy calves medium-chain fatty acids (MCFA) has on growth performance and health, and the secondary objective was to evaluate the effect of MCFA on energy status around weaning and the adaptive immune response following a vaccine challenge. Thirty-three Holstein bull calves (5 ± 1.6 d of age) were randomly assigned to 1 of 2 treatments. Control (CON) calves were fed milk replacer with no C8:0 or C10:0 oil added and MCFA calves were fed milk replacer with 0.5% of a combination of C8:0 or C10:0 oil added. Body weight and average daily gain were measured weekly. Feed efficiency (gain/feed) and the change in body condition score, hip width, hip height, heart girth, and paunch girth were calculated for the duration of the study. Fecal scores were recorded daily and all medical treatments were documented for the duration of the trial. On d 42, 49, and 56 of the study, a serum sample was collected from each calf and used to measure nonesterified fatty acids, β-hydroxybutyric acid, insulin, and glucose concentrations to evaluate energy status around weaning. A subset of 11 calves per treatment were enrolled in a vaccine challenge. At 21 ± 1.9 d of age (mean ± standard deviation) calves were vaccinated intramuscularly with 1 mL of endotoxin-free ovalbumin (OVA) mixed with aluminum hydroxide adjuvant. At 42 d of age (±1.9 d), blood samples were collected and used to analyze OVA-specific IgG and IgG, and calves were vaccinated a second time. At 56 d of age (±1.9 d), blood samples were collected to analyze IgG and IgG as well as IFN-γ and IL-4 secreted from peripheral blood mononuclear cells (PBMC) treated with OVA or phytohemagglutinin. Data were analyzed as a completely randomized design with repeated measures when applicable. A tendency for greater daily fecal score was observed for MCFA calves compared with CON. At d 42 of the study, nonesterified fatty acid concentrations were greater in CON calves compared with MCFA. At 42 and 56 d of age, anti-OVA IgG concentrations for CON and MCFA calves were greater than prevaccination samples. This study suggests that feeding MCFA to calves affects the energy status of calves around weaning and vaccinating dairy calves with ovalbumin combined with an aluminum hydroxide adjuvant is an effective way to evaluate the adaptive immune responses.
Topics: Aluminum Hydroxide; Animal Feed; Animals; Body Weight; Cattle; Diet; Fatty Acids; Fatty Acids, Nonesterified; Immunity; Immunoglobulin G; Leukocytes, Mononuclear; Male; Ovalbumin; Weaning
PubMed: 35940917
DOI: 10.3168/jds.2021-21567 -
Allergy Nov 2022Antibody-based tests are available for measuring SARS-CoV-2-specific immune responses but fast T-cell assays remain scarce. Robust T cell-based tests are needed to...
Combined assessment of S- and N-specific IL-2 and IL-13 secretion and CD69 neo-expression for discrimination of post-infection and post-vaccination cellular SARS-CoV-2-specific immune response.
BACKGROUND
Antibody-based tests are available for measuring SARS-CoV-2-specific immune responses but fast T-cell assays remain scarce. Robust T cell-based tests are needed to differentiate specific cellular immune responses after infection from those after vaccination.
METHODS
One hundred seventeen individuals (COVID-19 convalescent patients: n = 40; SARS-CoV-2 vaccinees: n = 41; healthy controls: n = 36) were evaluated for SARS-CoV-2-specific cellular immune responses (proliferation, Th1, Th2, Th17, and inflammatory cytokines, activation-induced marker [AIM] expression) by incubating purified peripheral blood mononuclear cells (PBMC) or whole blood (WB) with SARS-CoV-2 peptides (S, N, or M), vaccine antigens (tetanus toxoid, tick borne encephalitis virus) or polyclonal stimuli (Staphylococcal enterotoxin, phytohemagglutinin).
RESULTS
N-peptide mix stimulation of WB identified the combination of IL-2 and IL-13 secretion as superior to IFN-γ secretion to discriminate between COVID-19-convalescent patients and healthy controls (p < .0001). Comparable results were obtained with M- or S-peptides, the latter almost comparably recalled IL-2, IFN-γ, and IL-13 responses in WB of vaccinees. Analysis 10 months as opposed to 10 weeks after COVID-19, but not allergic disease status, positively correlated with IL-13 recall responses. WB cytokine responses correlated with cytokine and proliferation responses of PBMC. Antigen-induced neo-expression of the C-type lectin CD69 on CD4 (p < .0001) and CD8 (p = .0002) T cells informed best about the SARS-CoV-2 exposure status with additional benefit coming from CD25 upregulation.
CONCLUSION
Along with N- and S-peptide-induced IL-2 and CD69 neo-expression, we suggest to include the type 2 cytokine IL-13 as T-cellular recall marker for SARS-CoV-2 specific T-cellular immune responses after infection and vaccination.
Topics: Humans; COVID-19; Cytokines; Immunity, Cellular; Interleukin-13; Interleukin-2; Leukocytes, Mononuclear; SARS-CoV-2; Vaccination
PubMed: 35690994
DOI: 10.1111/all.15406 -
Brain, Behavior, & Immunity - Health May 2021In the last decades, it is growing the idea that stress-induced immunomodulation is bimodal: with acute stress associated with enhancing effects while chronic stress...
In the last decades, it is growing the idea that stress-induced immunomodulation is bimodal: with acute stress associated with enhancing effects while chronic stress with suppressive effects. However, the immune-endocrine interactions and its implications are often overlooked in ectotherms. We investigated the impact of corticosterone (CORT) treatment and short-term stressors on CORT plasma levels and the immunity of male toads (), using three distinct protocols: restraint, immune challenge (with lipopolysaccharide, LPS), and CORT transdermal application (TA). Our results showed increased CORT and neutrophil: lymphocyte ratio (NLR) regardless of the stress input (restraint, LPS challenge) or CORT TA. In the meantime, the bacterial killing ability (BKA) was not affected by any treatment, suggesting this immune parameter might be a more constitutive and robust response. Interestingly, the cellular immune response showed distinct patterns. Increased phagocytosis of blood leukocytes and phytohemagglutinin edema followed LPS and CORT TA (15 μg), respectively. In contrast, the phagocytosis of peritoneal leukocytes decreased after CORT TA (1 and 10 μg), indicating that short-term increases in CORT levels might impair local immune function. Such differences in cellular immunity might also be associated with CORT doses or the interaction between CORT and other immune mediators, such as melatonin, testosterone, and cytokines. Overall, our results highlight the immune-enhancing effects of the acute stress response and CORT TA, and the complexity of the immune-endocrine interaction in anurans. It also highlights the relevance of investigating distinct contexts for CORT increase arising from different situations, as well as diverse immune components for a better understanding of the stress-induced immunomodulation.
PubMed: 34589745
DOI: 10.1016/j.bbih.2021.100230 -
Journal of Investigative Medicine : the... Jan 2021Genital inflammation is an established risk factor for increased HIV acquisition risk. Certain HIV-exposed seronegative populations, who are naturally resistant to HIV...
Genital inflammation is an established risk factor for increased HIV acquisition risk. Certain HIV-exposed seronegative populations, who are naturally resistant to HIV infection, have an immune quiescent phenotype defined by reduced immune activation and inflammatory cytokines at the genital tract. Therefore, the aim of this study was to create an immune quiescent environment using immunomodulatory drugs to mitigate HIV infection. Using an peripheral blood mononuclear cell (PBMC) model, we found that inflammation was induced using phytohemagglutinin and Toll-like receptor (TLR) agonists Pam3CSK4 (TLR1/2), lipopolysaccharide (LPS) (TLR4) and R848 (TLR7/8). After treatment with anti-inflammatory drugs, ibuprofen (IBF) and betamethasone (BMS), PBMCs were exposed to HIV NL4-3 AD8. Multiplexed ELISA was used to measure 28 cytokines to assess inflammation. Flow cytometry was used to measure immune activation (CD38, HLA-DR and CCR5) and HIV infection (p24 production) of CD4+ T cells. BMS potently suppressed inflammation (soluble cytokines, p<0.05) and immune activation (CD4+ T cells, p<0.05). BMS significantly reduced HIV infection of CD4+ T cells only in the LPS (0.98%) and unstimulated (1.7%) conditions (p<0.02). In contrast, IBF had minimal anti-inflammatory and immunosuppressive but no anti-HIV effects. BMS demonstrated potent anti-inflammatory effects, regardless of stimulation condition. Despite uniform immunosuppression, BMS differentially affected HIV infection according to the stimulation conditions, highlighting the complex nature of these interactions. Together, these data underscore the importance of interrogating inflammatory signaling pathways to identify novel drug targets to mitigate HIV infection.
Topics: Anti-Inflammatory Agents; Anti-Inflammatory Agents, Non-Steroidal; Betamethasone; CD4-Positive T-Lymphocytes; Cells, Cultured; Disease Transmission, Infectious; Female; HIV; HIV Infections; Humans; Ibuprofen; Immunosuppression Therapy; In Vitro Techniques; Leukocytes, Mononuclear; Phytohemagglutinins; Toll-Like Receptors
PubMed: 33004468
DOI: 10.1136/jim-2020-001424 -
Viruses Apr 2021The multimammate mouse () has been identified as a major reservoir for multiple human pathogens including Lassa virus (LASV), spp., spp., and spp. Although are...
The multimammate mouse () has been identified as a major reservoir for multiple human pathogens including Lassa virus (LASV), spp., spp., and spp. Although are related to well-characterized mouse and rat species commonly used in laboratory models, there is an absence of established assays and reagents to study the host immune responses of . As a result, there are major limitations to our understanding of immunopathology and mechanisms of immunological pathogen control in this increasingly important rodent species. In the current study, a large panel of commercially available rodent reagents were screened to identify their cross-reactivity with Using these reagents, ex vivo assays were established and optimized to evaluate lymphocyte proliferation and cytokine production by lymphocytes. In contrast to C57BL/6J mice, lymphocytes from were relatively non-responsive to common stimuli such as phytohaemagglutinin P and lipopolysaccharide. However, they readily responded to concanavalin A stimulation as indicated by proliferation and cytokine production. In summary, we describe lymphoproliferative and cytokine assays demonstrating that the cellular immune responses in to commonly used mitogens differ from a laboratory-bred mouse strain.
Topics: Animals; Biomarkers; Cytokines; Host-Pathogen Interactions; Humans; Immunity, Cellular; Immunophenotyping; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Murinae; Rats; Rodent Diseases; Species Specificity; T-Lymphocyte Subsets
PubMed: 33922222
DOI: 10.3390/v13050729 -
Cell Calcium Jan 2021Gain-of-function RyR1-p.R163C mutation in ryanodine receptors type 1 (RyR1) deregulates Ca signaling and mitochondrial function in skeletal muscle and causes malignant...
Gain-of-function RyR1-p.R163C mutation in ryanodine receptors type 1 (RyR1) deregulates Ca signaling and mitochondrial function in skeletal muscle and causes malignant hyperthermia in humans and mice under triggering conditions. We investigated whether T lymphocytes from heterozygous RyR1-p.R163C knock-in mutant mice (HET T cells) display measurable aberrations in resting cytosolic Ca concentration ([Ca]), Ca release from the store, store-operated Ca entry (SOCE), and mitochondrial inner membrane potential (ΔΨ) compared with T lymphocytes from wild-type mice (WT T cells). We explored whether these variables can be used to distinguish between T cells with normal and altered RyR1 genotype. HET and WT T cells were isolated from spleen and lymph nodes and activated in vitro using phytohemagglutinin P. [Ca] and ΔΨ dynamics were examined using Fura 2 and tetramethylrhodamine methyl ester fluorescent dyes, respectively. Activated HET T cells displayed elevated resting [Ca], diminished responses to Ca mobilization with thapsigargin, and decreased rate of [Ca] elevation in response to SOCE compared with WT T cells. Pretreatment of HET T cells with ryanodine or dantrolene sodium reduced disparities in the resting [Ca] and ability of thapsigargin to mobilize Ca between HET and WT T cells. While SOCE elicited dissipation of the ΔΨ in WT T cells, it produced ΔΨ hyperpolarization in HET T cells. When used as the classification variable, the amplitude of thapsigargin-induced Ca transient showed the best promise in predicting the presence of RyR1-p.R163C mutation. Other significant variables identified by machine learning analysis were the ratio of resting cytosolic Ca level to the amplitude of thapsigargin-induced Ca transient and an integral of changes in ΔΨ in response to SOCE. Our study demonstrated that gain-of-function mutation in RyR1 significantly affects Ca signaling and mitochondrial fiction in T lymphocytes, which suggests that this mutation may cause altered immune responses in its carrier. Our data link the RyR1-p.R163C mutation, which causes inherited skeletal muscle diseases, to deregulation of Ca signaling and mitochondrial function in immune T cells and establish proof-of-principle for in vitro T cell-based diagnostic assay for hereditary RyR1 hyperfunction.
Topics: Animals; Calcium; Calcium Signaling; Cell Proliferation; Genotype; Intracellular Space; Lymphocyte Activation; Machine Learning; Malignant Hyperthermia; Membrane Potential, Mitochondrial; Mice; Mitochondria; Mutant Proteins; T-Lymphocytes; Thapsigargin
PubMed: 33310301
DOI: 10.1016/j.ceca.2020.102325 -
Poultry Science Dec 2023This study aimed to evaluate the immunity of chickens up to 35 d subjected to posthatch fasting and supplementation with conjugated linoleic acid (CLA). A total of 320...
This study aimed to evaluate the immunity of chickens up to 35 d subjected to posthatch fasting and supplementation with conjugated linoleic acid (CLA). A total of 320 chicks were housed in a completely randomized design with a 2 × 2 factorial arrangement (0 or 12 h of fasting × 0.000 or 0.025% CLA in a prestarter diet), totaling 4 treatments (No-F-12 h; F-12 h; No-CLA; CLA) with 8 replicates of 10 birds each. The relative weights (% body weight) of the spleen and bursa were determined 12 h posthatch (Post-12 h) and then weekly. Immunoglobulin Y (IgY) titers against Newcastle disease virus (NDV) were measured by ELISA in the yolk sac contents Post-12 h and in the serum weekly. Hypersensitivity to phytohemagglutinin (PHA) inoculation was evaluated by toe-web swelling response on d 13 and 34, 4 times a day (after 3 h, 6 h, 12 h, and 24 h inoculation, respectively, PHA-3 h, PHA-6 h, PHA-12 h, and PHA-24 h). The data were subjected to analysis of variance (P < 0.05). F-12h reduced the Post-12 h relative weight of the spleen, and CLA reduced the relative weight of the bursa at this stage and at 28 d. At 13 d, F-12 h reduced PHA-3 h, whereas PHA-12 h was increased by CLA. At 34 d, CLA reduced PHA-3 h. A greater reaction was observed in the No-F-12 h-CLA chicks, for the PHA-24 h. In the Post-12 h evaluation, F-12h reduced, whereas CLA increased NDV-specific IgY titers in the yolk sac. No-F-12 h-No-CLA chicks had the lowest serum titers. At 21 d, F-12 h-CLA chicks exhibited the highest serum titers. Titers were higher in the F-12 h-No-CLA chicks, when compared to other treatments. At 28 d, fasting reduced the titers. In conclusion, F-12 h and CLA accelerated the transfer of immunoglobulins from the yolk sac to the serum. F-12 h impairs cellular immunity, whereas CLA favors it.
Topics: Animals; Chickens; Linoleic Acids, Conjugated; Immunity, Humoral; Diet; Fasting; Animal Feed
PubMed: 37926012
DOI: 10.1016/j.psj.2023.103167 -
Comparative Medicine Apr 2020Olive baboons () have provided a useful model of human diseases and conditions, including cardiac, respiratory, and infectious diseases; diabetes; and involving... (Comparative Study)
Comparative Study
Olive baboons () have provided a useful model of human diseases and conditions, including cardiac, respiratory, and infectious diseases; diabetes; and involving genetics, immunology, aging, and xenotransplantation. The development of a immunologically defined SPF baboons has advanced research further, especially for studies involving the immune system and immunosuppression. In this study, we compare normal immunologic changes of PBMC subsets, and their function in age-matched conventional and SPF baboons. Our results revealed that both groups have comparable numbers of different lymphocyte subsets, but phenotypic differences in central and effector memory T-cell subsets are more pronounced in CD4+ T cells. Despite equal proportions of CD3+ T cells among the conventional and SPF baboons, PBMC from the conventional group showed greater proliferative responses to phytohemagglutinin and pokeweed mitogen and higher numbers of IFNγ-producing cells after stimulation with concanavalin A or pokeweed mitogen, whereas plasma levels of the inflammatory cytokine TNFα were significantly higher in SPF baboons. Exposure of PBMC from conventional baboons to various Toll-like (TLR) ligands, including TLR3, TLR4, and TLR8, yielded increased numbers of IFNγ producing cells, whereas PBMC from SPF baboons stimulated with TLR5 or TLR6 ligand had more IFNγ-producing cells. These findings suggest that although lymphocyte subsets share many phenotypic and functional similarities in conventional and SPF baboons, specific differences in the immune function of lymphocytes could differentially influence the quality and quantity of their innate and adaptive immune responses. These differences should be considered in interpreting experimental outcomes, specifically in studies measuring immunologic endpoints.
Topics: Animals; Female; Immunity, Cellular; Male; Monkey Diseases; Papio; Papio anubis; T-Lymphocytes
PubMed: 32014083
DOI: 10.30802/AALAS-CM-19-000035 -
Journal of Animal Science Aug 2021Disease resilience refers to the productivity of an animal under disease. Given the high biosecurity of pig nucleus herds, traits that can be measured on healthy pigs...
Disease resilience refers to the productivity of an animal under disease. Given the high biosecurity of pig nucleus herds, traits that can be measured on healthy pigs and that are genetically correlated with disease resilience, that is, genetic indicator traits, offer a strategy to select for disease resilience. Our objective was to evaluate mitogen stimulation assays (MSAs) on peripheral blood mononuclear cells (PBMCs) from young healthy pigs as genetic indicators for disease resilience. Data were from a natural disease challenge in which batches of 60 or 75 naïve Yorkshire × Landrace piglets were introduced every 3 wk into a continuous flow barn that was seeded with multiple diseases. In this environment, disease resilience traits, including growth, treatment, and mortality rates, were recorded on 3,136 pigs that were genotyped with a high-density marker panel. PBMCs from 882 of these pigs from 19 batches were isolated from whole blood collected prior to the disease challenge and stimulated with five mitogens: concanavalin A (ConA), phytohemagglutinin (PHA), pokeweed mitogen (PWM), lipopolysaccharide (LPS), and phorbol myristate acetate (PMA). The proliferation of cells was evaluated at 48, 72, and 96 h and compared with unstimulated samples (rest count). Heritabilities of cell proliferation were estimated using a model with batch as a fixed effect and covariates of entry age; rest count; complete blood count proportions of lymphocytes, monocytes, eosinophils, and basophils; and pen, litter, and animal genetics as random effects. Heritability estimates were highest for response to ConA (0.30 ± 0.09, 0.28 ± 0.10, 0.17 ± 0.10, and 0.25 ±0.10 at 48, 72, and 96 h after stimulation and for area under the curve across the three time points, respectively). Estimates were in a similar range for response to PHA and PMA but low for PWM and LPS. Responses to ConA, PHA, and PMA were moderately genetically correlated with several disease resilience traits and in the expected direction, but individual estimates were not significantly different from zero due to large SEs. In conclusion, although validation is needed, MSAss, in particular based on ConA, show promise as genetic indicator traits for disease resilience.
Topics: Animals; Cell Proliferation; Leukocytes, Mononuclear; Lymphocyte Activation; Mitogens; Phytohemagglutinins; Pokeweed Mitogens; Swine
PubMed: 33944943
DOI: 10.1093/jas/skab084 -
AIDS (London, England) Feb 2020Opioid-use disorders (OUD) and hepatitis C or B co-infection (HEP) are common among people living with HIV (PLHIV). The impact of OUD on innate and adaptive immunity...
BACKGROUND
Opioid-use disorders (OUD) and hepatitis C or B co-infection (HEP) are common among people living with HIV (PLHIV). The impact of OUD on innate and adaptive immunity among PLHIV with and without HEP is unknown.
OBJECTIVES
To investigate the impact of OUD on monocyte and T-cell phenotypes, cytokine responses to lipopolysaccharide (LPS) and phytohemagglutinin (PHA), and plasma inflammatory markers, among PLHIV with and without HEP.
METHODS
Cross-sectional study enrolling PLHIV receiving ART, with and without OUD. Flow cytometry determined monocyte and T-cell phenotypes; LPS and PHA-induced cytokine production was assessed following LPS and PHA stimulation by multiplex cytokine array; plasma IL-6, soluble CD163, and soluble CD14 were measured by ELISA.
RESULTS
Twenty-two PLHIV with OUD and 37 PLHIV without OUD were included. PLHIV with OUD exhibited higher frequencies of intermediate (CD14CD16) and nonclassical (CD14CD16) monocytes when compared with PLHIV without OUD (P = 0.0025; P = 0.0001, respectively), regardless of HEP co-infection. Soluble CD163 and monocyte cell surface CD163 expression was increased among PLHIV with OUD and HEP, specifically. Regardless of HEP co-infection, PLHIV with OUD exhibited reduced production of IL-10, IL-8, IL-6, IL-1alpha, and TNF-alpha in response to LPS when compared with PLHIV without OUD; PHA-induced production of IL-10, IL-1alpha, IL-1beta, IL-6, and TNF-alpha were also reduced among individuals with OUD.
CONCLUSION
OUD among PLHIV are associated with altered monocyte phenotypes and a dysregulated innate cytokine response. Defining underlying mechanisms of opioid-associated innate immune dysregulation among PLHIV should be prioritized to identify optimal OUD treatment strategies.
Topics: Adult; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Biomarkers; Cross-Sectional Studies; Cytokines; Female; Flow Cytometry; HIV Infections; Humans; Interleukin-10; Interleukin-1beta; Male; Middle Aged; Monocytes; Opioid-Related Disorders; Randomized Controlled Trials as Topic; Receptors, Cell Surface; T-Lymphocytes; Tumor Necrosis Factor-alpha
PubMed: 31687981
DOI: 10.1097/QAD.0000000000002416