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Background: Acute myocardial infarction (AMI) is one of the most deleterious conditions leading to cardiovascular diseases and mortality. The importance of an early and accurate diagnosis assures immediate medical treatments, which are fundamental to reduce mortality and improve prognoses. AMI is associated to an inflammatory response which includes the increase of circulating inflammatory cytokines, chemokines and immune cell activation. This study aimed to identify which are the very early immune-related biomarkers that may be used as predictors of myocardial infarction severity. In order to mimic the pathophysiological events involved in human myocardial infarction, a temporary occlusion (90 min) of the mid-left anterior descending coronary artery was performed in a swine animal model., Results: Lymphocyte subsets analysis in peripheral blood revealed significant alterations in CD4+/CD8+ ratio and naïve and effector/memory T cell percentages at 1 h post-myocardial infarction. Changes in TH1/TH2-related cytokine, monocyte and neutrophil markers gene expression were observed in peripheral blood lymphocytes, as well. Additionally, significant correlations between cardiac parameters (cardiac enzymes, left ventricular ejection fraction and % infarct) and blood-derived parameters (cytokine expression and lymphocyte subset distribution) were found., Conclusions: Peripheral blood lymphocyte alterations are easily and swiftly detectable, so they may be good biomarkers for a very early prognosis and to predict myocardial infarction severity.

Classical swine fever virus (CSFV) is one of the most important pathogens affecting swine. After infection with a moderate virulence strain at 8 hours after birth, CSFV is able to induce viral persistence. These animals may appear clinically healthy or showed unspecific clinical signs despite the permanent viremia and high viral shedding, in absence of immune response to the virus. Given the role played by this infection in disease control, we aimed to evaluate the capacity of CSFV to induce postnatal persistent infection at 3 weeks after birth. Nine pigs were CSFV infected and sampled weekly during 6 weeks and viral, clinical, pathological and immunological tests were carried out. Also, the CD4/CD8 ratio was calculated with the purpose to relate this marker with the CSFV persistent infection. The IFN-α response was detected mainly 1 week after infection, being similar in all the infected animals. However, 44.4% of animals were CSFV persistently infected, 33.3% died and 22.2% developed specific antibody response. Interestingly, in persistently infected pigs, the T-CD8 population was increased, the T-CD4 subset was decreased and lower CD4/CD8 ratios were detected. This is the first report of CSFV capacity to confer postnatal persistent infection in pigs infected at 3 weeks after birth, an age in which the weaning could be carried out in some swine production systems. This type of infected animals shed high amounts of virus and are difficult to evaluate from the clinical and anatomopathological point of view. Therefore, the detection of this type of infection and its elimination in endemic areas will be relevant for global CSF eradication. Finally, the low CD4/CD8 ratios found in persistently infected animals may be implicated in maintaining high CSFV replication during persistence and further studies will be performed to decipher the role of these cells in CSFV immunopathogenesis., (© 2018 The Authors. Transboundary and Emerging Diseases published by Blackwell Verlag GmbH.)

We previously showed that cats that were infected with non-pathogenic Puma lentivirus (PLV) and then infected with pathogenic feline immunodeficiency virus (FIV) (co-infection with the host adapted/pathogenic virus) had delayed FIV proviral and RNA viral loads in blood, with viral set-points that were lower than cats infected solely with FIV. This difference was associated with global CD4⁺ T cell preservation, greater interferon gamma (IFN-γ) mRNA expression, and no cytotoxic T lymphocyte responses in co-infected cats relative to cats with a single FIV infection. In this study, we reinforced previous observations that prior exposure to an apathogenic lentivirus infection can diminish the effects of acute infection with a second, more virulent, viral exposure. In addition, we investigated whether the viral load differences that were observed between PLV/FIV and FIV infected cats were associated with different immunocyte phenotypes and cytokines. We found that the immune landscape at the time of FIV infection influences the infection outcome. The novel findings in this study advance our knowledge about early immune correlates and documents an immune state that is associated with PLV/FIV co-infection that has positive outcomes for lentiviral diseases.

Canine lymphoma is a heterogeneous disease with many different subtypes. Lymphoma of T cell type in particular is variable in outcome, and includes subtypes with non-progressive, slowly- and rapidly-progressive disease course. Association of immunotype with disease course is incompletely defined. Here, results of flow cytometric immunotyping of 127 canine T cell lymphomas were analyzed in relation to survival and progression free interval. Samples originated from 101 multicentric, 8 mediastinal, 6 cutaneous, 5 hepatosplenic, 5 gastrointestinal and 2 other anatomic subtypes of T cell lymphoma. Compared to multicentric T cell lymphoma, gastrointestinal lymphoma had shorter survival and progression free interval, and hepatosplenic lymphoma had shorter survival. Among dogs with multicentric T cell lymphoma, immunotypes of CD4 + /CD8 - /MHCII + , CD4 - /CD8 + /MHCII + and CD4 - /CD8 + /MHCII - were associated with longer survival times than the immunotype of CD4 + /CD8 - /MHCII - , and immunotypes of CD4 + /CD8 - /MHCII + , CD4 - /CD8 + /MHCII - , and CD4 - /CD8 - /MHCII + were associated with longer progression free intervals. Dogs with multicentric T cell lymphoma and concurrent leukemia had shorter survival but similar progression free interval compared to those without leukemia. Body weight, sex, hypercalcemia, cell size, expression of CD3 and use of combination or single agent chemotherapy did not significantly affect outcome of multicentric TCL., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Three separate experiments were conducted to study the effects of whole yeast cell product supplementation in pullets and layer hens. Body weight gain, fecal and intestinal coccidial oocyst counts, cecal microflora species, cytokine mRNA amounts, and CD4+ and CD8+ T-cell populations in the cecal tonsils were analyzed following an experimental coccidial infection. In Experiment I, day-old Leghorn layer chicks were fed 3 experimental diets with 0, 0.1, or 0.2% whole yeast cell product (CitriStim®, ADM, Decatur, IL). At 21 d of age, birds were challenged with 1 × 105 live coccidial oocysts. Supplementation with whole yeast cell product decreased the fecal coccidial oocyst count at 7 (P = 0.05) and 8 (P < 0.01) d post-challenge. In Experiment II, 27-week old Leghorn layer hens were fed 3 experimental diets with 0, 0.05 or 0.1% whole yeast cell product and challenged with 1 × 105 live coccidial oocysts on d 25 of whole yeast cell product feeding. Supplementation with whole yeast cell product decreased the coccidial oocyst count in the intestinal content (P < 0.01) at 5, 13, and 38 d post-coccidial challenge. Supplementation with whole yeast cell product increased relative proportion of Lactobacillus (P < 0.01) in the cecal tonsils 13 d post-coccidial challenge. Supplementation with whole yeast cell product decreased CD8+ T cell percentages (P < 0.05) in the cecal tonsils at 5 d post-coccidial challenge. In Experiment III, 32-week-old Leghorn layer hens were fed 3 experimental diets with 0, 0.1, or 0.2% whole yeast cell product and challenged with 1 × 105 live coccidial oocysts on d 66 of whole yeast cell product feeding. At 5 d post-coccidial challenge, whole yeast cell product supplementation down-regulated (P = 0.01) IL-10 mRNA amount. It could be concluded that supplementing whole yeast cell product can help minimize coccidial infection in both growing pullets and layer chickens., (© 2017 Poultry Science Association Inc.)

Background: The pathogenesis of canine generalized demodicosis is poorly understood but is thought to involve dysfunction of the immune system. Previous studies showed diminished CD4+ T lymphocyte counts in affected dogs, but none has evaluated this subpopulation through resolution of the disease., Hypothesis/objectives: In this longitudinal study, we tested whether quantification of CD4+ cells, CD8+ cells and the ratio of CD4+ to CD8+ cells are good indicators of immunological status and could be used as biomarkers of treatment efficacy and prognosis., Animals: Sixteen dogs of several breeds with diagnoses of generalized demodicosis, plus 30 age/breedmatched healthy dogs., Methods: Total lymphocytes, CD4+, CD8+ and CD4+:CD8+ ratio were quantified at four time points: at diagnosis, 30 days after diagnosis (during treatment), at first negative parasitological examination and at clinical cure., Results: Absolute numbers of CD4+ cells were significantly lower in affected dogs at the time of diagnosis. Absolute numbers of CD4+ and CD8+ cells were significantly augmented in affected animals compared with control dogs after treatment was established, and this persisted until the first negative parasitological examination, at which time the CD4+ counts equalled those of the control group., Conclusions and Clinical Importance: Our findings suggest that longitudinal quantification of CD4+ and CD8+ T lymphocytes is a useful indicator of the efficacy of demodicosis treatment., (© 2014 ESVD and ACVD.)

Antibody testing based on individual risk assessments is recommended to determine feline immunodeficiency virus (FIV) status, but ELISA and Western blot tests cannot distinguish between anti-FIV antibodies produced in response to natural infection and those produced in response to FIV vaccination. The aim of this cross-sectional study was to test the hypothesis that FIV-infected cats could be differentiated from FIV-vaccinated uninfected cats using lymphocyte subset results, specifically the CD4%:CD8(low)% T-lymphocyte ratio. Comparisons of the CD4%:CD8(low)% T-lymphocyte ratio were made among the following four groups: Group 1 - FIV-infected cats (n=61; FIV-antibody positive by ELISA and FIV PCR positive); Group 2 - FIV-uninfected cats (n=96; FIV-antibody negative by ELISA); Group 3 - FIV-vaccinated uninfected cats (n=31; FIV-antibody negative by ELISA before being vaccinated against FIV, after which they tested FIV ELISA positive); and Group 4 - FIV-uninfected but under chronic/active antigenic stimulation (n=16; FIV-antibody negative by ELISA; all had active clinical signs of either upper respiratory tract disease or gingival disease for ≥ 21 days). The median CD4%:CD8(low)% T-lymphocyte ratio was lower in Group 1 (1.39) than in each of the other three groups (Group 2 - 9.77, Group 3 - 9.72, Group 4 - 5.64; P<0.05). The CD4%:CD8(low)% T-lymphocyte ratio was also the most effective discriminator between FIV-infected cats and the other three groups, and areas under ROC curves ranged from 0.91 (compared with Group 4) to 0.96 (compared with Group 3). CD4%:CD8(low)% shows promise as an effective test to differentiate between FIV-infected cats and FIV-vaccinated uninfected cats., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: Endogamy increases the risk of manifestation of deleterious recessive genes. Mitochondrial DNA allows the separation of American Zebu (Bos indicus and Bos taurus) and evaluate the effect of mitochondrial DNA on productive traits of cattle. However, the effect of endogamy and mitochondrial DNA (mtDNA) on the immune system remains unclear. The aim of this study was to evaluate the association between endogamy, mtDNA and immune parameters., Results: A total of 86 cattle (43 cows and 43 calves) were used in this study. Age, endogamy, milk yield, and origin of mtDNA were measured and their influence on immunological parameters was evaluated. Older cows had increased CD4+ T cells, decreased CD21+ and γδhigh T cells as well as increased CD4+/CD8+ and T/B ratio. Multiple regression analysis indicated that endogamy in calves was associated with increased CD8+ T and CD21+ B lymphocytes, and decreased γδhigh T cells in peripheral blood. Cows with medium and lower endogamy had a lower percentage of B lymphocytes and γδlow T cells and cows with lower endogamy had higher levels of γδ T cells and γδhigh T cells, as well as the CD4+/CD48+ cell ratio. Calves with higher endogamy had higher levels of CD8+ T lymphocytes, whereas calves with lower endogamy had lower levels of γδlow T cells., Conclusions: These results demonstrated for the first time that endogamy influences the immune system of cattle.

Detailed biological analyses (e.g. epidemiological, genetic) of animal health and fitness in the field are limited by the lack of large-scale recording of individual animals. An alternative approach is to identify immune traits that are associated with these important functions and can be subsequently used in more detailed studies. We have used an experimental dairy herd with uniquely dense phenotypic data to identify a range of potentially useful immune traits correlated with enhanced (or depressed) health and fitness. Blood samples from 248 dairy cows were collected at two-monthly intervals over a 10-month period and analysed for a number of immune traits, including levels of serum proteins associated with the innate immune response and circulating leukocyte populations. Immune measures were matched to individual cow records related to productivity, fertility and disease. Correlations between traits were calculated using bivariate analyses based on animal repeatability and random regression models with a Bonferroni correction to account for multiple testing. A number of significant correlations were found between immune traits and other recorded traits including: CD4(+):CD8(+) T lymphocyte ratio and subclinical mastitis; % CD8(+) lymphocytes and fertility; % CD335(+) natural killer cells and lameness episodes; and serum haptoglobin levels and clinical mastitis. Importantly these traits were not associated with reduced productivity and, in the case of cellular immune traits, were highly repeatable. Moreover these immune traits displayed significant between-animal variation suggesting that they may be altered by genetic selection. This study represents the largest simultaneous analysis of multiple immune traits in dairy cattle to-date and demonstrates that a number of immune traits are associated with health events. These traits represent useful selection markers for future programmes aimed at improving animal health and fitness.

It has been considered that drinking oxygenated water improves oxygen availability, which may increase vitality and improve immune functions. The present study evaluated the effects of oxygenated drinking water on immune function in pigs. Continuous drinking of oxygenated water markedly increased peripheral blood mononuclear cell proliferation, interleukin-1β expression level and the CD4(+):CD8(+) cell ratio in pigs. During Salmonella Typhimurium infection, total leukocytes and relative cytokines expression levels were significantly increased in pigs consuming oxygenated water compared with pigs consuming tap water. These findings suggest that oxygenated drinking water enhances immune activity in pigs and increases immune responses of pigs during S. Typhimurium Infection.

Jembrana disease virus (JDV) is an unusual bovine lentivirus that causes an acute and sometimes fatal disease after a short incubation period in Bali cattle (Bos javanicus). The pathological changes occur primarily in lymphoid tissues, which feature proliferating lymphoblastoid-like cells predominantly throughout parafollicular (T-cell) areas, and atrophy of follicles (B-cell) areas. Five Bali cattle were experimentally infected with JDV and all developed typical clinical signs of Jembrana disease characterised by a transient febrile response, enlargement of superficial lymph nodes and a significant leukopenia. Flow cytometric analysis of PBMC during the acute (febrile) disease phase showed that the reduced number of lymphocytes was due to a significant decrease in both the proportion and absolute numbers of CD4(+) T cells, but not CD8(+) T-cells or CD21(+) B-cells. At the end of the febrile phase, total numbers of both CD8(+) T-cells and CD21(+) B-cells increased significantly, while CD4(+) T-cell numbers remained below normal values, resulting in a significantly reduced CD4(+):CD8(+) ratio. We speculate that the persistent depletion of CD4(+) T cells following JDV infection, through lack of CD4(+) T cell help to B cells, may explain the lack of production of JDV-specific antibodies for several weeks after recovery despite an increase in CD21(+) B cell numbers. Further, our previous data showing that IgG(+) plasma cells are targets for JDV infection, correlated with our current data demonstrating an increase in CD8(+) T cell numbers, supports the suggestion that anti-viral cytotoxic T cell or other cell-mediated immune responses may be critical in the recovery process, although this remains to be formally demonstrated for JDV., (Copyright © 2012 Elsevier B.V. All rights reserved.)

To evaluate the relationship among immune status and increased morbidity and mortality, peripheral blood lymphocytes (CD3(+), CD4(+), CD8(+) and CD21(+) cells) from 32 healthy dogs over 8 years of age were analyzed. Twenty-five of the 32 dogs were followed-up for 3 years after the analysis; and 14 dogs were found to be diseased, and nine dogs died. There was no notable difference between the ages of the dogs that died compared with the ones that survived. The relative percentage of CD4(+) and the CD4(+):CD8(+) ratio decreased notably in dogs falling ill compared with healthy dogs. The relative percentage of CD3(+) lymphocytes showed a notable decrease in dogs that died within 3 years in comparison with dogs that survived. In a discriminant analysis of morbidity and mortality, most patients were correctly classified as diseased or not and surviving or dead, respectively. These results indicate that the immunophenotypes of peripheral blood lymphocytes in older dogs offer promise as parameters for evaluating mortality and morbidity.

A total of 128 1-week-old chicks were classified into four groups; T-2 toxin fed (T-2), IBV infected (IBV), T-2 toxin fed and co-infected with IBV (T-2+IBV), and untreated (control) for a period of 6 weeks. Within their respective groups, the birds belonged to T-2 and T-2+IBV were exposed to 2 ppm of T-2 toxin contaminated feed for 6 weeks, and 0.2 ml of 10 EID(50) (10(5.69)/0.2 ml) inoculums of IBV isolate (India/LKW/56/IVRI/08) was used to challenge the chicks belonged to IBV alone and T-2+IBV groups after 3 weeks of the experiment. To study immunopathological effects, parameters such as lymphocyte stimulation indices (SI), haemagglutination inhibition, enzyme linked immunosorbant assay (ELISA), peripheral lymphocytes CD(4)(+) and CD(8)(+) analysis, and histopathological examination of lymphoid organs were done. Accordingly, SI values were significantly (P<0.05) lower in all the treatment groups as compared to control, however, the SI values of IBV infected group were significantly higher than the values in toxin fed groups. The mean HI titres to ND vaccine was significantly (P<0.05) lower in the toxin groups at all the intervals, and the antibody titres in IBV infected group were significantly (P<0.05) higher than that of T-2 toxin fed and co-infected with IBV group but were significantly (P<0.05) lower than the control at 21 (3) and 28 (10) days of toxin feeding (DTF) (days post infection (DPI)). Similarly, the mean IBV ELISA antibody titres in the toxin fed groups were significantly (P<0.01) reduced as compared with the IBV ELISA antibody titres of IBV infected but not toxin fed group, at all intervals. Peripheral CD(4)(+):CD(8)(+) ratios in T-2+IBV group and number of CD(4)(+) and CD(8)(+) peripheral lymphocytes in all treatment groups were significantly reduced as compared to the values in control birds. However, CD(4)(+):CD(8)(+) ratios of IBV infected group at 42 (21) DTF (DPI) were found significantly (P<0.05) higher than the values in control birds. Histopathologically, lymphoid organs (bursa of Fabricius, spleen, thymus, caecal tonsils and Harderian glands) showed moderate to severe necrosis (lymphocytolysis) and extensive lymphocyte depletion in all the toxin groups (T-2 and T-2+IBV groups) where the severity and extent of the lesions were more in T-2+IBV group. The findings of the present experiment revealed immunosuppressive effects of T-2 toxin and aggravated the pathology and pathogenesis of IBV infection., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Background: Feline immunodeficiency virus (FIV) and human immunodeficiency virus (HIV) infection cause similar clinical syndromes of immune dysregulation, opportunistic infections, inflammatory diseases, and neoplasia. Renal disease is the 4th most common cause of death associated with HIV infection., Objective: To investigate the association between FIV infection and renal disease in cats., Animals: Client-owned cats (153 FIV-infected, 306 FIV-noninfected) and specific-pathogen-free (SPF) research colony cats (95 FIV-infected, 98 FIV-noninfected)., Methods: A mixed retrospective/prospective cross-sectional study. Blood urea nitrogen (BUN), serum creatinine, urine specific gravity (USG), and urine protein:creatinine ratio (UPC) data were compared between FIV-infected and FIV-noninfected cats. In FIV-infected cats, total CD4+ and CD8+ T lymphocytes were measured using flow cytometry, and CD4+:CD8+ T lymphocyte ratio was calculated. Renal azotemia was defined as a serum creatinine ≥ 1.9 mg/dL with USG ≤ 1.035. Proteinuria was defined as a UPC > 0.4 with an inactive urine sediment., Results: Among the client-owned cats, no association was detected between FIV infection and renal azotemia (P = .24); however, a greater proportion of FIV-infected cats were proteinuric (25.0%, 16 of 64 cats) compared to FIV-noninfected cats (10.3%, 20 of 195 cats) (P < .01). Neither neuter status nor health status were risk factors for proteinuria in FIV-infected cats, but UPC was positively correlated with the CD4+:CD8+ T lymphocyte ratio (Spearman's rho = 0.37, P = .01). Among the SPF research colony cats, no association was detected between FIV infection and renal azotemia (P = .21) or proteinuria (P = .25)., Conclusions and Clinical Importance: Proteinuria but not azotemia was associated with natural FIV infection., (Copyright © 2012 by the American College of Veterinary Internal Medicine.)

Glutamine (Gln) is an important substrate for the innate immune cells including lymphocytes and macrophages. In this report, the effects of alanyl-glutamine dipeptide (Ala-Gln) on the naïve immune system, intestinal integrity and gain performance of early-weaned calves were investigated. Early-weaned Chinese Holstein calves were intravenously administered different dosages of Ala-Gln. The effects of Ala-Gln administration were ascertained by evaluating the blood for naïve T lymphocyte subpopulations, the concentrations of serum IgG, serum IgA and intestinal mucosal secretory IgA (s-IgA), the intestinal integrity, as well as the gain performance. Results demonstrated that intravenous administration of Ala-Gln dipeptide (1.01 g/kg×d(-1)) for 7 days had a positive effect on gain performance, intestinal integrity and the immune system. Calves administered doses of Ala-Gln displayed an improvement in gain performance and health status concurrent with increases in blood CD2(+) and CD4(+) lymphocytes, the ratio of CD4(+)/CD8(+), serum IgA and IgG, intestinal mucosal s-IgA while decreasing the occurrence of diarrhea. Moreover, we found that animals given the effective dose (1.01 g/kg×d(-1)) of Ala-Gln resulted in improved immune status and intestinal integrity relative to those given a lower (0.49 g/kg×d(-1)) or higher dose (1.99 g/kg×d(-1)) of Ala-Gln. These findings suggest that maintaining a certain concentration of plasma and/or tissue glutamine in the early stages of weaning is an effective alternative approach for improvement of growth performance in early-wean calves., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Molecular analysis, serology and immunophenotyping for T lymphocytes and their subsets, B lymphocytes and monocytes were performed on dogs naturally infected with Leishmania infantum. Animals were categorised as asymptomatic dogs I (AD-I), with negative serology and positive molecular results, and asymptomatic dogs II (AD-II), with positive serology and positive molecular results, and these were compared to symptomatic dogs (SD) and control dogs (CD). AD-I exhibited immunophenotypic features similar to those of CD, including isotype profiles and concentrations of monocytes. Similar biomarkers were found in AD-II and SD, such as, higher levels of immunoglobulins IgG, IgG2, IgM and IgA and higher concentrations of eosinophils. High frequencies of T lymphocytes and CD4(+) T cells were observed in both AD-I and AD-II compared to SD, whereas CD8(+) T cells were higher only in AD-II compared with SD. Analysis of B lymphocytes revealed an increased frequency of this cell type only in AD-II animals compared with SD. Asymptomatic dogs appear to have a dichotomous infection spectrum that can influence the humoral and cellular immunological status during canine visceral leishmaniasis., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Type-I interferons (IFNs) are cytokines that have non-specific antiviral activity, participating mostly in innate defense mechanisms. Their administration has been proposed to treat several viral and immunomediated diseases as an immunomodulatory therapy. Due to its availability, recombinant human interferon-alpha (rHuIFN-α) has been studied in relation to feline retrovirosis, both in vitro and in vivo. However, IFNs are species-specific and antibodies have been shown to develop in response to the high rHuIFN-α doses necessary for an effective therapy. A recombinant feline IFN has been developed, which has been characterized as interferon-omega (rFeIFN-ω), designed to overcome these problems. Nonetheless, very few studies have been undertaken to evaluate its efficacy in cats naturally infected with FIV or FeLV. In an initial study, we here demonstrated that rFeIFN-ω can dramatically improve the clinical condition of infected cats, and induce improvement of hematologic parameters. Minor changes or no change was observed for hypergammaglobulinemia, CD4/CD8 ratio, proviral load, viremia and RT activity, suggesting that the overall effect of IFN was on innate immunity. More studies are needed in order to better understand its in vivo mechanisms., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Changes in an individual's immune status are considered major contributing factors towards the morbidity of cancer and mortality of aging. To evaluate age-related changes in the immune status of dogs, the immunophenotypes (CD3, CD4, CD8 and CD21) of peripheral blood lymphocytes were measured in 160 healthy dogs aged from 1 to 17 years, and in 365 dogs with various tumors and at various stages. In healthy dogs, the absolute numbers of white blood cells, lymphocytes, and CD3(+), CD4(+) and CD21(+) lymphocytes decreased significantly with age. The relative percentages of lymphocytes and CD4(+) cells decreased significantly, while CD8(+) cells increased significantly with age. The CD4:CD8 ratio showed a significant age-related decrease. In contrast, dogs with tumors possessed significantly lower absolute numbers and relative percentages of all lymphocyte phenotypes, while the CD4:CD8 ratio was significantly higher than in age-matched controls. The relative percentages of CD3(+) and CD8(+) lymphocytes were significantly lower in dogs with distant metastases compared with dogs without metastases, and the CD4:CD8 ratio increased with advanced stage. These observations illustrate the significant changes in immune status with age and the presence of marked immunological defects in a large-scale study of dogs with advanced tumors., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The objective of this study was to determine the effects of dietary n-6:n-3 polyunsaturated fatty acid (PUFA) ratios on heterophil:lymphocyte (H:L) ratios and T cell subsets in the peripheral blood of the growing Yangzhou goose. A total of 160 healthy Yangzhou geese (0.407 ± 0.023 kg), aged 21 d, were randomly divided into 4 groups consisting of diets with different n-6:n-3 PUFA ratios: 12:1, 9:1, 6:1, and 3:1. White blood cell counts and H:L ratios were measured at 42, 56, and 70 d of age; T lymphocyte subsets were also analyzed using monoclonal antibodies and flow cytometry simultaneously. The results showed that differences were found in white blood cells between groups (P < 0.05) but not within groups (P > 0.05), and white blood cells of 70-d-old geese were significantly higher than those of other ages within the same group. The H:L ratios of all groups decreased as age increased, and differences were found in 6:1 and 3:1 groups between ages (P < 0.05). As for the comparison of H:L ratio between treatments, 6:1 and 3:1 were lower than the other 2 groups at 42 or 70 d of age (P < 0.05), and 3:1 was lower than the other 3 groups at 56 d of age (P < 0.05). Moreover, H:L ratios had a tendency to decrease with the decrease of n-6:n-3 PUFA ratios. Furthermore, cluster of differentiation (CD) 3(+) and CD8(+) increased gradually, whereas CD4(+) and CD4(+):CD8(+) ratios first increased and then decreased as age increased. Differences were detected in T cell subsets among ages (P < 0.05). In addition, CD4(+) and CD4(+):CD8(+) ratios were affected remarkably by n-6:n-3 PUFA ratios but CD3(+) and CD8(+) were not, and 6:1 had much higher CD4(+) percentage and CD4(+):CD8(+) ratio compared with the other groups (P < 0.05). Taken together, diets containing low n-6:n-3 PUFA ratios could decrease H:L ratios in the peripheral blood. Additionally, CD4(+) percentage and CD4(+):CD8(+) ratio were much higher in the 6:1 group, which could be relevant for improving positive immune responses for Yangzhou goslings from 42 to 70 d of age.

Canine leishmaniosis (CL) is a common systemic parasitic disease that is endemic in many Mediterranean countries including Greece. The immune reaction to the parasite is critical to the outcome of the infection and the response to treatment. Some studies have shown a reduction of circulating CD4+ T cells and of the CD4+/CD8+ ratio in dogs with CL and these changes normalised following treatment with meglumine antimoniate or amphotericin B. Allopurinol is used as a monotherapy for the chronic treatment of CL. The aim of the present study was to determine the circulating CD4+ and CD8+ T lymphocyte numbers and the CD4+/CD8+ ratio in 19 dogs diagnosed with CL before and after prolonged allopurinol monotherapy (18 months). A significant decrease in circulating CD4+ T cells was observed in dogs with CL before treatment. Prolonged allopurinol monotherapy improved the number of circulating CD4+ T cells, but did not restore their number to within the normal range., (Copyright © 2009 Elsevier Ltd. All rights reserved.)

The duration of immunity of the dual-subtype feline immunodeficiency virus (FIV) vaccine, Fel-O-Vax FIV, for protection against subtype-B FIV was assessed in this study. Vaccinated cats along with controls were challenged with FIV(FC1), a subtype-B FIV strain, 54 weeks after the final vaccination, and monitored for 46-48 weeks for provirus and viral RNA in peripheral blood, provirus in lymphoid organs, and CD4:CD8 ratios. Results of provirus detection in peripheral blood and lymphoid organs and plasma viral RNA loads showed that 10/14 vaccinated cats were fully protected for 48 weeks against infection with FIV(FC1) whereas 5/5 controls were persistently infected with FIV(FC1). CD4:CD8 inversions were noted in association with FIV infection and viral loads were not significantly different between FIV infected controls and the unprotected vaccinated animals., (Copyright 2009 ISFM and AAFP. Published by Elsevier Ltd. All rights reserved.)

The existence of a relationship between cortisol levels, after an acute stress, and behavioral activities, immunological profile, and production performance in sheep was studied. An initial flock of 30 Comisana ewes was involved in the experiment, and each of the 30 ewes was individually subjected to an isolation test in a novel environment. Subsequently, from the initial flock, 2 groups of 8 Comisana ewes were each retrospectively selected, and the animals were divided, according to their cortisol concentration 10 min after the isolation test, into high cortisol (HC) ewes, having a peak of cortisol concentration >90 ng/mL (average: 119.3 ng/mL +/- 11.8), and low cortisol (LC) ewes having a peak of cortisol concentration <80 ng/mL (average: 52.4+/-11.8). During the isolation test, the behavior of each animal was video-recorded and behavioral activities were registered. Blood samples were collected before the isolation test, immediately after the test (10 min), and at 60, 120, 300 min, 24 h, and 48 h after the test to evaluate percentages of T-helper (CD4(+)) and T-cytotoxic (CD8(+)) cells, CD4(+)/CD8(+) ratio, and IL-1beta and IL-6 levels. The ewes were milked for 3 d after the isolation test to determine cortisol levels and IL-1beta and IL-6 concentrations in whey. Milk yield was recorded at each milking, and milk samples were analyzed for pH, nutritional parameters, renneting properties, and somatic cell count. During the isolation test, HC ewes exhibited a shorter duration of movement and fewer bleats than LC ewes. The average plasma IL-1beta concentration was higher in HC than in LC ewes. The average whey IL-1beta and IL-6 concentrations were higher in whey from HC ewes than in LC ewes. A positive correlation emerged between plasma and whey IL-1beta concentrations. The average CD4(+)/CD8(+) ratio in blood was lower in HC than in LC ewes. Time from isolation affected the CD4(+)/CD8(+) ratio: at 120 min, the CD4(+)/CD8(+) ratio increased compared with that at 10 min after isolation and then decreased until 300 min after isolation. On average, ewes with low cortisol concentrations showed higher milk production and lower SCC than ewes with high cortisol concentrations. Results suggest that plasma cortisol concentration is connected to the behavioral response and immune competence of dairy ewes and cytokine concentrations. Both whey IL-1beta and IL-6 can be considered reliable indicators of the magnitude of hypothalamic-pituitary-adrenal axis activation. The stress-induced changes in CD4(+)/CD8(+) ratio are critical for controlling disease incidence and planning appropriate vaccination programs. High reactivity of the hypothalamic-pituitary-adrenal axis is also associated with a reduction in milk production and an increased predisposition to develop intramammary inflammatory processes., (2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study was designed to compare cutaneous mycoflora isolation and CD4+:CD8+ ratio in feline immunodeficiency virus (FIV)-infected cats with that in FIV-uninfected cats. Sixty cats were examined. Twenty-five were FIV-infected cats and 35 were FIV-uninfected cats. All 60 cats were FeLV-negative. Fungi were speciated and immunophenotyping of peripheral CD4+ and CD8+ T lymphocytes was performed. At least one fungal colony was isolated from 22/25 (88%) FIV-infected cats. Among the FIV-uninfected cats fungal colonies were recovered from 13/35 (37%) specimens. Dermatophytes were recovered from 2/25 (8%) FIV-infected cats (one Microsporum gypseum, one Microsporum canis) and 3/35 (8.5%) FIV-uninfected cats (M gypseum). Malassezia species was the most commonly isolated organism from both groups of cats (51.6%). Malassezia species was more commonly isolated from FIV-infected cats than FIV-uninfected cats (84% vs 28.6%). The CD4+ to CD8+ lymphocyte ratio for FIV-infected cats was significantly lower than the CD4+ to CD8+ ratio in the FIV-uninfected cats. The CD4+ to CD8+ lymphocyte ratio for FIV-infected cats with cutaneous overall fungal isolation was significantly lower than the CD4:CD8 lymphocyte ratio in the FIV-infected cats but without cutaneous fungal isolation. We can conclude that immunologic depletion due to retroviral infection might represent a risk factor to cutaneous fungal colonization in cats., (Copyright 2010 ISFM and AAFP. Published by Elsevier Ltd. All rights reserved.)

Feline immunodeficiency virus (FIV), an immunosuppressive lentivirus found in cats worldwide, is studied to illuminate mechanisms of lentiviral pathogenesis and to identify key components of protective immunity. During replication, lentiviruses accumulate errors of nucleotide mis-incorporation due to the low-fidelity of reverse transcriptase and recombination between viral variants, resulting in the emergence of a complex viral "quasispecies". In patients infected with HIV-1, env sequences may vary by up to 10% and the detection of quasispecies with greater heterogeneity is associated with higher viral loads and reduced CD4+ T cell numbers [1], indicating that transmission of more complex quasispecies may lead to disease progression. However, little is known about how FIV evolves as disease progresses, or why some cats develop AIDS rapidly while disease progression is slow in others. The aim of this study was to determine whether disease progression may be governed by viral evolution and to examine the diversity of viral variants emerging following infection with an infectious molecular clone. The FIV env gene encoding the envelope glycoprotein (Env) was examined at early (12 weeks) and late (322 weeks) stages of FIV infection in two groups of cats infected experimentally with the FIV-GL8 molecular clone. Viral variants were detected within quasispecies in cats in the late stages of FIV infection that contained differing amino acid compositions in several variable loops of Env, some of which were identified as determinants of receptor usage and resistance to neutralization. Therefore these results indicate that the FIV env gene evolves during the course of infection, giving rise to variants that resist neutralization and likely lead to disease progression., (Copyright 2009 Elsevier B.V. All rights reserved.)

Steroid-responsive Meningitis-Arteritis (SRMA) is a systemic inflammatory disease of juvenile to young adult dogs with a relapsing course and most prominent manifestation in the cervical meninges. Immunophenotyping and flow cytometric measurement of lymphocytes in peripheral blood (PB) and CSF was performed in the acute phase of SRMA (n=12) and during glucocorticosteroid treatment (n=10). Values were compared to those from dogs with other neurologic diseases (n=63) and healthy individuals (n=7). Dogs with SRMA had high CD4:CD8alpha ratios in PB and low T:B cell ratios in PB and CSF suggesting that a T(H)2-mediated immune response occurs. The T:B cell ratio in CSF was markedly lower than that in PB indicating that either a selective recruitment of B cells or, alternatively, their strong intrathecal proliferation takes place. SRMA appears to be a valuable animal model for the investigation of compartmentalization of immune responses and for studies on differences in local central nervous system and systemic immune responses.

This study was designed to test the effect of antioxidant supplementation on feline immunodeficiency virus (FIV)-infected felines. Six acutely FIV-infected cats (> or =16 weeks post-inoculation) were given a propriety oral superoxide dismutase (SOD) supplement (Oxstrin; Nutramax Laboratories) for 30 days. Following supplementation, the erythrocyte SOD enzyme concentration was significantly greater in the supplemented FIV-infected group than the uninfected control group or the unsupplemented FIV-infected group. The CD4+ to CD8+ ratio increased significantly (0.66-0.88) in the SOD supplemented FIV-infected cats but not in the unsupplemented FIV-infected cats. Proviral load and reduced glutathione (GSH) levels in leukocyte cell types did not change significantly following supplementation. Antioxidant supplementation resulted in an increase in SOD levels, confirming the oral bioavailability of the compound in FIV-infected cats. This result warrants further investigation with trials of antioxidant therapy in FIV-infected cats that are showing clinical manifestations of their disease, as well as in other feline patients where oxidative stress likely contributes to disease pathogenesis, such as diabetes mellitus and chronic renal failure.

IFN-gamma plays an important role in cellular immunity contributing to microorganism elimination. We have previously reported that bovine colostrum contains high levels of IFN-gamma as well as immunoglobulin. Lymphocytes are potent producers of IFN-gamma, so establishing the lymphocyte population in colostrum will help to identify the source of colostral cytokines. In this study, we used flow cytometric analysis to quantify the population of three types of lymphocytes found in colostrum; namely, CD4 (Th) cells, CD8 (cytotoxic T) cells, and gammadelta-T cells. We also quantified the concentration of colostral IFN-gamma using ELISA. IFN-gamma concentrations were measured in colostrum obtained from 96 healthy Holstein cows; the levels tended to decrease on the first day post-parturition. Flow cytometric analysis showed that many gammadelta-T- and CD8-positive cells were present in the colostrum, and that the CD4/CD8 ratio was low. The ratios of CD8- and gammadelta-T-positive cells to cells of other types decreased during the 5 days after parturition, but that of CD4-positive cells showed no change during the observation period. To identify IFN-gamma-expressing colostral lymphocytes, we used magnetic separation technology to separate the lymphocytes (CD4, CD8 and gammadelta-T) from colostral cells, then examined them for IFN-gamma mRNA expression with real-time PCR (RT-PCR). RT-PCR analysis revealed potent expression of the IFN-gamma gene in CD8-positive cells, reaching higher levels than in CD4- or gammadelta-T-positive cells. These results suggest that the CD8-positive T cells in colostrum play a role as producers of IFN-gamma.

Immune responses against polymorphic host molecules incorporated into lentiviral envelopes during cell budding have induced protection against primate immunodeficiency virus infection. Dendritic cells (DCs) express high levels of MHC molecules and are infectable by lentiviruses. Therefore, in this pilot study we addressed the hypothesis that immunization of cats with allogeneic DC would induce immune responses that protect against challenge with the feline immunodeficiency virus. Two groups of 3 cats each received 3 subcutaneous injections of allogeneic or autologous DC, and were then challenged with viruses propagated in the immunizing DC. Infection status and lymphocyte parameters of cats were assessed during 6 weeks after challenge. MHC II antigens were incorporated into viral particles as identified by Western blot; and antibodies reactive with MHC class II antigens were detected in the serum of cats immunized with allogeneic but not autologous DC. After challenge, all cats had proviral DNA in blood leukocytes from 2 weeks post-challenge onward and seroconverted. Cats immunized with allogeneic DC maintained higher total and CD21(+) lymphocyte concentrations, and higher CD4(+)/CD8(+) lymphocyte ratios; however, these differences were not significantly different from cats that received autologous DC immunizations. Plasma viral load was not significantly different between groups of cats (p=0.204). These results suggest that immunization of cats with allogeneic DC does not induce protective immunity against FIV infection.

Johne's disease (JD) is characterized by a protracted period of subclinical infection. Infected cows may remain in the subclinical state until stressors such as parturition and lactation invoke more clinical signs of disease. The objective of this study was to evaluate changes in the percentages of CD4(+), CD8(+), and gammadelta T-cells, B-cells, monocytes, as well as the expression of the activation marker, CD5, on these cell subpopulations in the peripheral blood of dairy cows naturally infected with Mycobacterium avium subsp. paratuberculosis (MAP) during the periparturient period. Peripheral blood mononuclear cells (PBMCs) were collected from 3 wk pre- to 4 wk post-calving and freshly isolated or cultured for 7d. Day 7 cultures were infected with live MAP at a 10:1 MOI (bacteria to adherent PBMC), and cultures were incubated for an additional 24h. Fluorescent antibody labeling of lymphocyte subsets and monocytes was conducted and analyzed with flow cytometry. Freshly isolated PBMCs from subclinical cows expressed a greater (P<0.05) percentage of CD8(+) and gammadelta T-cells compared with clinical cows. The percentage of CD4(+) T-cells increased (P<0.08) in clinical cows as parturition approached. During the postpartum period, clinical cows had greater (P<0.05) CD4:CD8 ratios compared with subclinical and control cows. After 8d, uninfected PBMCs from clinical cows had greater (P<0.05) percentages of CD14(+) cells compared with subclinical cows. When infected with live MAP, there was no effect of infection group or parturition on cell subpopulations. In fresh PBMCs, clinical cows expressed lower percentages of CD4(+)CD5(bright) and CD8(+)CD5(bright) compared with control cows, but greater percentages of CD5(dim) cells for all lymphocyte subsets. These results suggest changes in the percentages of lymphocyte subsets, monocytes, and CD5 markers are modulated by both infection status and the periparturient period.

Feline immunodeficiency virus (FIV) infection of cats is an animal model for the pathogenesis of CD4+ lymphopenia and thymus dysfunction in HIV-infected humans. Recently, a monoclonal antibody (755) was reported to recognize the feline homologue to CD45RA, allowing the enumeration of naïve T cells in cats. We tested the hypothesis that pediatric FIV infection would be associated with a selective loss of naïve CD4+ lymphocytes by inoculating newborn cats with a pathogenic clone of FIV (JSY3) or a related clone with an inactive ORF-A gene (JSY3-DeltaORFA), and compared the data to age-matched uninfected control cats. Both FIV inocula were associated with a reduction in the CD4-CD8 ratio (p=0.01), which was attributable to a disproportionate loss of naïve CD4+ cells (p=0.01) vs. naïve CD8+ cells. Therefore, the reduced CD4:CD8 ratio in FIV-infected juvenile cats is associated with a selective depletion of naïve CD4+ cells from the blood.

Although migration of leukocytes into the mammary gland is pivotal for a cow's response against intramammary invading pathogens, the contribution of lymphocyte subsets to this response remains unclear. To investigate the dynamics of lymphocyte populations during Escherichia coli mastitis, T-lymphocyte subsets, CD4+/CD8+ ratio, CD21+ cells, and lymphoproliferation were studied in blood and milk of primiparous cows exposed to different quantities of bacteria. The cows were intramammarily inoculated with 10(4) cfu of E. coli (group A) and 10(6) cfu (group B). Compared with group A, a much greater number of lymphocytes migrated into the infected quarters at postinfection hour (PIH) 6 to 24 in group B, and the CD8+ cells were the first-recruited T cells in the milk. There was a significant decline in the CD4+/CD8+ ratios at PIH 6 to 24 in group B. The decrease of CD4+/CD8+ ratios at PIH 6 to 24 resulted mainly from greater CD8+ cell concentrations in milk. In contrast, at PIH 72, CD4+/CD8+ ratios increased about 2-fold in both groups. This increase was mainly due to the increase in CD4+ cell concentration. The increased concentration of CD4+ cells coincided with an increase in the CD21+ cell population in the milk. In blood, the increase of CD8+ cells appeared much faster in group B (PIH 6 and 12) than in group A. The results from lymphoproliferation also indicated a greater increase in the proliferative response in both blood and milk lymphocytes of group B. Our study demonstrates for the first time that an increase of E. coli inoculum dose accelerates the trafficking of CD8+ cells during initiation of E. coli mastitis, and these cells are the predominant T cells in milk during the early hours of bovine E. coli mastitis.

We examined the effects of vitamin E (VE) and Arg on the subpopulations of T lymphocytes in peripheral blood after an infectious bursal disease virus vaccination (20 d old, n = 54). Broiler chickens were fed diets with normal levels of Arg (NARG, 1.2%) or high levels of Arg (HARG, 2.2%) and 3 levels of VE (40, 80, or 200 IU/kg of feed) in a factorial arrangement of treatments. The percentages of CD4+ and CD8+ cells 9 d after vaccination were not different in birds fed the HARG or NARG feed, but they were higher in birds fed the VE80 diet than in birds fed the VE40 diet. Birds fed the VE200 feed had similar levels of CD4+ and CD8+ cells as birds fed the VE40 diet. However, 19 d after vaccination, the percentage of CD4+ cells was higher in birds fed the HARG (43.93 +/- 1.05) diet than in birds fed the NARG diet (41.14 +/- 1.05) and in birds fed the VE80 and VE200 (44.1 +/- 1.28 and 44.42 +/- 1.28, respectively) diets compared with birds fed the VE40 diet (39.04 +/- 1.28). On the other hand, the percentage of CD8+ cells was highest in birds fed the HARG-VE80 feed compared with all the other Arg and VE combinations. The percentages of CD3+ cells and BU-1+ cells were higher in birds fed the HARG (81.35 +/- 1.46 and 22.3 +/- 0.89, respectively) diet than in birds fed the NARG diets (77.82 +/- 1.46 and 18.93 +/- 0.89, respectively). Birds fed the VE40 had a higher amount of BU-1+ (24.35 +/- 1.073) but lower amounts of CD3+ cells (75.76 +/- 1.76) than birds fed the VE80 (18.46 +/- 1.073 and 81.67 +/- 1.76) and VE200 (19.03 +/- 1.073 and 81.19 +/- 1.76, respectively). Neither Arg nor VE had an effect on the CD4+:CD8+ cell ratio and on the percentage of immature (CD4+CD8+) T lymphocytes 9 d after vaccination. These results suggest that Arg and VE have complimentary effects on cellular and humoral immune function and may enhance the resistance of broilers to infectious diseases.

Fifteen piglets were used to determine the effect of vitamin E supplementation on the number of CD4-immunoreactive (CD4+) T-lymphocytes, CD8-immunoreactive (CD8+) T-lymphocytes and IgA-immunoreactive (IgA+) B-lymphocytes per follicle in the Peyer's patch of distal ileum and the mesenteric lymph nodes of weaned piglets. Piglets, following a 3-day adaptation period after weaning, were assigned to one of three experimental groups: control (no vitamin E supplementation), vitamin E supplementation of 100 mg/kg of diet and vitamin E supplementation of 300 mg/kg of diet. Supplementation of vitamin E lasted for a period of 36 days. The basal diet contained 80 mg alpha-tocopherol/kg of diet. All piglets were killed at day 39 after weaning and samples of the distal ileum and adjacent mesenteric lymph nodes were collected. The number of cells for each lymphocyte subset was counted in the Peyer's patch and the mesenteric lymph nodes follicles, in cryostat sections processed for immunohistochemistry. Results showed that vitamin E supplementation (300 mg/kg diet) of piglets caused an increase (P < 0.05) in the number of IgA+ B-lymphocytes in the Peyer's patch, but not in the mesenteric lymph nodes, compared with the corresponding values in control animals. Vitamin E supplementation had no effect (P > 0.05) on the number of CD4+ and CD8+ T-lymphocytes in the follicles of the Peyer's patch and the adjacent mesenteric lymph nodes. Thus, vitamin E had relatively minor effects on distribution of the major immunocompetent cells in the gut. The numbers of CD4+ and CD8+ T-lymphocytes as well as IgA+ B-lymphocytes per follicle were higher by 26-77% (P < 0.05) in the mesenteric lymph nodes than the corresponding values in the Peyer's patch.

We previously demonstrated that stimulation of bovine peripheral blood mononuclear cells (PBMCs) with staphylococcal enterotoxin C (SEC), led to an inversion of the CD4(+):CD8(+)T cell ratio and generation of an atypical CD8(+)T cell subpopulation expressing CD26. In the present study, we examined T cell apoptosis and proliferation profiles of PBMC subpopulations in cultures stimulated with SEC. Unlike when stimulated with concanavalin A, nucleic acid synthesis in bovine PBMC cultures stimulated with SEC was low during the first four days but increased greatly on day 5. In contrast, nucleic acid synthesis in human PBMC cultures stimulated with SEC increased continuously. To investigate the mechanism of delayed bovine T cell proliferation, various cell phenotypes were monitored. The inversion of the bovine CD4(+):CD8(+)T cell ratio in PBMC cultures stimulated by SEC was associated with higher proliferation and lower apoptosis of CD8(+)T cells compared to CD4(+)T cells. The mRNA levels for interleukin (IL)-4 and IL-13 were sustained over 4 days but IL-12 mRNA levels dropped to background on day 2. These data suggest that SEC induces a prolonged Th-2- biased microenvironment, and together with the inversion of the bovine CD4(+):CD8(+)T cell ratios in bovine PBMC cultures with SEC, may in part explain the inability of the mammary immune system to establish an effective response to Staphylococcus aureus infections.

Reasons for Performing Study: Ageing appears to affect immune and neuroendocirne function in horses and response to acute exercise. No studies have examined the combined effects of training and ageing on immune and neuroendocirne function in horses., Hypothesis: To ascertain whether training and age would affect the plasma beta-endorphin (BE) and cortisol (C) as well as immune function responses to acute exercise in Standardbred mares., Methods: Graded exercise tests (GXT) and simulated race tests (SRT) were performed before and after 12 weeks training at 60 % HRmax. BE and C were measured at rest and at 5, 10, 20, 40, 60 and 120 min post GXT. Leucocyte cell number, CD4+ and CD8+ lymphocyte subsets, and mitogen stimulated lymphoproliferative response (LPR), were measured in jugular blood before and after the SRTs., Results: Cortisol rose by 5 min post GXT in young (Y) and middle-age (MA) mares (P<0.05) and remained elevated until 40 and 60 min post GXT, respectively during both pre- and post training GXT. There was no rise in C in old (0) mares after either GXT (P>0.05). Pretraining BE rose (P<0.05) by 5 min post GXT in all mares. After training, BE was higher in Y and O vs. MA (P<0.05) at 5 min post GXT. Post training BE was higher at 5 min post GXT in Y and O vs. pretraining (P<0.05). After SRT, lymphocyte number rose in all mares (P<0.05); however, lower lymphocyte numbers (P<0.05) were seen in MA vs. Y and O vs. MA (P<0.05). The O had reduced LPR to Con A and PHA stimulation (P<0.05) compared to Y and MA after the SRT after both pre- and post training SRT. LPR to PWM was lower (P<0.05) in O vs. Y and MA after the pretraining SRT. Training caused an increase in resting LPR to PWM in MA only (P<0.05)., Conclusion: Both age and training altered the plasma beta-endorphin and cortisol responses as well as and immune responses to acute exercise., Potential Relevance: This study provides important information on the effects of ageing and training that will aid in the management and care of an increasing number of active older horses.

To evaluate immunocompetence in commercially raised chickens, we immunophenotyped Dekalb Delta and H&N White Leghorn (WLH) hybrids, 20 chickens in each of 3 age groups (9 wk [juvenile], 25 wk [young adult], and 79 or 80 wk [adult]), for circulating CD3+, CD4+, CD8+, TCR1+, TCR2+, and TCR3+ lymphocytes. The proportion of CD3+ T cells, including CD4+ and CD8+ subsets, was increased in the hybrids as compared with published values for laboratory-raised outbred WLH chickens. The proportion of the TCR2+ (Vbeta1) T cell subpopulation was also increased. An age-related decrease in the proportion of TCR1+ (gammasigma) T cells was noted in both hybrids. Further, a remarkably low CD4:CD8 ratio was evident in all age groups of both hybrids, indicating decreased immunocompetence. Overall, these experiments provide age-related proportions of various peripheral-blood T lymphocyte subpopulations in commercially raised Dekalb Delta and H&N chickens that diverge from the proportions in laboratory-raised outbred WLH chickens and suggest reduced immunocompetence. Such a decline in immunocompetence, including humoral immune capacity, could be attributed to genetic selection for production traits, environmental factors associated with commercial operations, and intense immunization.

The study was undertaken to evaluate adverse effects of larvicidal treatment in horses naturally infected with cyathostomins. Out of 24 ponies kept on pasture, four animals were housed in September and anthelmintically cured to serve as worm-free controls (group C-0). The others were housed in December. Eight animals each were treated 8 weeks later with 5 x 7.5mg/kg fenbendazole (FBZ) or 1 x 0.4 mg/kg moxidectin (MOX). Four animals remained untreated (group C-i). Two, 4, 6 and 14 days after the end of treatment two animals of each of the treated groups were necropsied together with group C-0 and C-i animals. Infected animals before treatment showed weight loss, eosinophilia, increased plasma protein and globulin contents. Treatment was followed by weight gain and temporal plasma protein and globulin increase. Proportions of CD4+ and CD8+ T lymphocytes in the peripheral blood did not differ between the groups before treatment but dropped significantly temporally after FBZ treatment. Group C-0 was worm-free at necropsy. Group C-i animals contained variable numbers of luminal and tissue cyathostomins. Histological sections showed larval stages in the lamina propria und submucosa surrounded by macrophages. Either treatment was effective against luminal parasites and reduced the number of larvae in the bowel wall beginning 4-6 days after FBZ and 6-14 days after MOX treatment. Histologically, as a first reaction after FBZ application T lymphocytes accumulated around morphologically intact L4 in the submucosa. Subsequently T lymphocytes associated with eosinophils infiltrated the submucosa. Parasites became enclosed by granulomas with eosinophils adhering to and invading the larvae which started to disintegrate on day 4. Later on, particularly on day 14 inflammation extended into the mucosa and was frequently associated with ulcerations. Third stage larvae in general and L4 in the lamina propria, however, seemed not to be affected until day 14 and even then, parasites did usually not generate extensive inflammation. After MOX treatment severe morphologically detectable alterations of tissue larvae could not be observed earlier than day 14. Different from FBZ treatment, larvae disintegrated and were obviously resorbed without causing severe inflammation in the gut wall. In conclusion treatment with either drug was efficacious against tissue larvae of cyathostomins but there may be different clinical consequences: in contrast to MOX effects, killing of larvae due to FBZ was associated with severe tissue damage, which clinically may correspond to reactions caused by synchronous mass emergence of fourth stage larvae, i.e., may mimic larval cyathostominosis.

Feline immunodeficiency virus (FIV) is a lentivirus related to human immunodeficiency virus (HIV) that causes feline AIDS in the domestic cat (Felis catus). Serological surveys indicate that at least 25 other species of cat possess antibodies that cross-react with domestic cat FIV. Most infected nondomestic cat species are without major symptoms of disease. Long-term studies of FIV genome variation and pathogenesis reveal patterns consistent with coadaptation of virus and host in free-ranging FIV-Ple-infected African lions (Panthera leo) and FIV-Pco-infected pumas (Puma concolor) populations. This report examined correlates of immunodeficiency in wild and captive lions and pumas by quantifying CD5(+), CD4(+), and CD8(+) T-cell subsets. Free-ranging FIV-Ple-infected lions had immunofluorescence flow cytometry (IFC) profiles marked by a dramatic decline in CD4(+) subsets, a reduction of the CD4(+)/CD8(+) ratio, reduction of CD8(+)beta(high) cells, and expansion of the CD8(+)beta(low) subset relative to uninfected lions. An overall significant depletion in CD5(+) T-cells in seropositive lions was linked with a compensatory increase in total CD5(-) lymphocytes. The IFC profiles were altered significantly in 50% of the seropositive individuals examined. The FIV-Pco-infected pumas had a more generalized response of lymphopenia expressed as a significant decline in total lymphocytes, CD5(+) T-cells, and CD5(-) lymphocytes as well as a significant reduction in CD4(+) T-cells. Like lions, seropositive pumas had a significant decline in CD8(+)beta(high) cells but differed by not having compensatory expansion of CD8(+)beta(low) cells relative to controls. Results from FIV-infected lions and pumas parallel human and Asian monkey CD4(+) diminution in HIV and SIV infection, respectively, and suggest there may be unrecognized immunological consequences of FIV infection in these two species of large cats.

In this study, we investigated whether ozonated autohemoadministration (OAHA) influences leukocyte populations in cows with clinical inflammatory disease. Eleven cows with inflammatory disease (Inflammatory Group) and three healthy cows (Control Group) were used for this study. The CD4(+)/CD8(+) ratio in the Inflammatory Group increased significantly compared to that in the Control Group 3 to 4 days after OAHA treatment. In the Inflammatory Group, the number of CD14(+) cells decreased gradually after OAHA, but CD14(+) levels remained stable in the Control Group. The number of MHC class-II(+) cells decreased gradually in the Inflammatory Group, but increased gradually in the Control Group, and the difference between the groups was significant on day 14 after OAHA. These findings suggest a possible difference in the activation of immune response after OAHA in infected cows compared to healthy cows.

The study sought to evaluate whether truck-transportation had an impact on the respiratory immune system of cattle. Six castrated 6-10-month-old Holstein calves were shipped approximately 100 km by road for 4 h. Plasma and bronchoalveolar lavage (BAL) fluid samples, collected immediately before transportation, at 4 h (soon after transportation), and on days 3 and 7 after transportation, were examined. A marked elevation of plasma cortisol concentration was observed at 4 h, but this level was unchanged in controls. The chemiluminescence (CL) response of phagocytes in BAL fluid cells, composed mainly of alveolar macrophage, decreased significantly after transportation (P<0.05). Transportation increased the CD3+ T cell population significantly (P<0.05), and a significant increase (P<0.05) in the ratio of CD4+ to CD8+ cells in BAL fluid was evident. We conclude that short-term road transportation alters pulmonary cells and their function, which may engender bovine respiratory disorders.

The effectiveness of flow cytometric assays for canine use is still requiring standardization. Despite several studies using purified mononuclear cells, no methodology or reference ranges are available for immunophenotyping of whole blood leukocytes (WBL). Fresh and pre-fixed WBL were used to identify cell-subsets, (Thy-1(+)/CD5(+)/CD4(+)/CD8(+)/CD21(+) and CD14(+)) and measure MHC-II, CD45RA/CD45RB expression. We described here an efficient method for fast quantification of canine-WBL, using pre-fix in a microplate assay, which allows long-term sample storage prior to phenotyping. Decreased percentage of CD5(+)-T-cells within the lymphocyte-gate and increased percentage of CD21(+)-B-cells were observed in young animals, which led to higher T/B cell ratios in middle-aged dogs. Lower numerical counts of Thy-1(+), CD4(+), CD8(+) and CD21(+) lymphocyte were observed when compared to young animals. In addition, we identified an age-related decline of MHC-II/CD45RA expression by lymphocytes. We proposed an improved method for phenotyping of canine peripheral blood mononuclear cells (PBMC) that has significant use for researchers and veterinary clinicians. The hematological changes of senescence previously identified on PBMC could be adequately reproduced on features identified by whole blood. Furthermore, this study supplies normal range references as baseline standards for clinical purposes, besides specific immunological parameters to monitor canine aging process.

Rhodococcus equi causes severe pyogranulomatous pneumonia in foals and in immunocompromised people. In mice, both CD4+ and CD8+ T lymphocytes contribute to host defense against R. equi, but CD4+ T lymphocytes are required for pulmonary clearance of the bacteria. In this prospective study of 208 foals at two equine breeding farms with endemic R. equi infections, we collected peripheral blood samples at 2 and 4 weeks of age and at the time of diagnosis of R. equi pneumonia. Samples were analyzed for concentrations of total and differential leukocytes, EqCD4+ and EqCD8+ T lymphocytes, and B lymphocytes. Thirty (14.4%) foals developed R. equi pneumonia. At the 2nd week of life, affected foals had significantly lower concentrations of white blood cells (WBC) and segmented neutrophils, significantly lower proportions of EqCD4+ T lymphocytes, and significantly higher proportions of EqCD8+ T lymphocytes. The EqCD4:EqCD8 ratio was significantly lower for affected foals. At the 4th week of life, affected foals had significantly lower concentrations of segmented neutrophils and EqCD4+ T lymphocytes than did unaffected foals. The ratio of EqCD4:EqCD8 was significantly lower for affected foals. Two- and 4-week-old foals with ratios of EqCD4:EqCD8<3 were significantly more likely to develop R. equi pneumonia. There was a significant farm effect which diluted our statistical power to detect differences; however; after adjusting for the farm effect, 2-week-old foals with ratios of EqCD4:EqCD8<3 remained significantly more likely to develop R. equi pneumonia. There were no significant differences in immunophenotypic variables between affected foals (at the time of diagnosis) and age-matched control foals. These data suggest that there are hematologic and immunophenotypic differences between affected and unaffected foals during the first 2-4 weeks of life, prior to onset of clinical signs of R. equi pneumonia. These differences may represent important immunologic mechanisms associated with increased susceptibility of individual foals to infection with R. equi. Because there was considerable overlap between values for affected and unaffected foals, we cannot yet recommend immunophenotyping of foals at endemically-infected farms as a clinically useful screening tool to identify foals at increased risk of developing R. equi pneumonia.

Unlabelled: The present study was designed to evaluate the possible effect of dietary oregano etheric oils as non-specific immunostimulating agents in growth-retarded, low-weight growing-finishing pigs. Forty-nine growth-retarded (> 10% under average weight in a group) growing-finishing pigs of the same age were assigned to two groups and treated as follows: Group 1 (n = 25): the animals weighed 58.2 +/- 2.4 kg and were fed until slaughter ad libitum with a commercial fattening diet supplemented with 3000 ppm commercial oregano feed additive (Oregpig Pecs, Hungary). Oregpig is dried leaf and flower of Origanum vulgare, enriched with 500 g/kg cold-pressed essential oils of the leaf and flower of Origanum vulgare. Analysis of Oregpig: 60 g carvacrol and 55 g thymol/kilogram. Group 2 (n = 24): the animals weighed 57.9 +/- 2.6 kg and were fed until slaughter with the same diet without Oregpig supplementation. Oregpig-receiving pigs showed a significantly (P < 0.05) better average daily gain and feed conversion rate than the non-treated animals (Oregpig group 788.1 +/- 31.3 g, control animals 709.3 +/- 42.2 g; 2.96, vs. 3.08, respectively). Mortality was significantly (P < 0.001) higher in the non-treated animals (Oregpig group, 1 animal = 4%; control, 8 animals = 33.3%). The proportion of CD4, CD8, MHC class II antigen, and non-T/non-B cells in peripheral blood lymphocytes was significantly higher in the Oregpig-receiving pigs than in the control animals. The proportion of CD4+ CD8+ double-positive T lymphocytes in peripheral blood and mesenteric lymph nodes was higher in the Oregpig-receiving pigs than in the control animals., Implication: Dietary oregano improves growth in growth-retarded growing-finishing pigs and has non-specific immunostimulatory effects on porcine immune cells.

16alpha-Bromo-epiandrosterone (epiBr), a synthetic derivative of the natural hormone dehyroepiandrosterone (DHEA), was evaluated for its effects on feline immunodeficiency virus (FIV) infection in experimental cats. The rationale for this study was based on the ability of DHEA to significantly reduce the mortality to viral infections in mice. DHEA and epiBr also have demonstrable in vitro anti-viral activity for both HIV-1 and FIV. Preliminary pharmacokinetic studies in cats demonstrated that subcutaneously injected epiBr was rapidly absorbed, completely metabolized, and nontoxic. Metabolites were excreted in both urine and feces, with the latter having the most complex pattern of breakdown products. Cats were then divided into four groups; two groups were infected with FIV and two uninfected. Two groups, one infected and one uninfected were treated on 5 consecutive days of weeks 0, 4, 8, 12 and 16 with epiBr. The remaining two groups were mock treated with the drug vehicle alone. Treatment started 1 week prior to infection and extended for 4 weeks after infection. Cats were observed for 20 weeks post-FIV infection. Infected cats had identical decreases in blood neutrophil and lymphocyte counts following, regardless of whether they were treated with epiBr or vehicle alone. The CD4/CD8 T-cell ratio was decreased following FIV exposure, but was significantly more decreased for the epiBr treated animals from week 2 post-infection onward. CD4+ T cells were decreased in FIV-infected cats treated with epiBr compared to their untreated cohort, while CD8+ T cells tended to be higher in treated animals. FIV infected cats that were treated with epiBr had over one-log higher virus loads at week 2 post-infection than non-epiBr treated cohorts. In spite of this enhanced initial viremia, the subsequent levels of virus in the blood were significantly lower in epiBr treated versus untreated animals. EpiBr treated cats had significantly higher FIV-p24 antibody responses than control cats receiving vehicle alone, although primary and secondary antibody responses to a T-cell dependent non-FIV antigen, keyhole limpet hemocyanin (KLH), were unaffected. EpiBr treatment significantly decreased the expected FIV-induced suppression of IL-12 p40 mRNA levels in peripheral blood mononuclear cells (PBMCs) observed at weeks 4, 5, 8, 9 and 16 post-infection, but had no influence on FIV-induced changes in IL-4, IL-6, IL-10, IFN-gamma, MIP-1alpha and RANTES.

1. An experiment was carried out to evaluate the effect of early feed restriction (FR) on immunocompetence of Ross and Arian chickens with separated sexes under heat stress (HS) conditions. 2. Chickens consumed feed ad libitum (AL) or were restricted on alternate days from 11 to 20 d of age. From 35 to 41 d of age, the HS groups were exposed to a high ambient temperature of 39 +/- 1 degreesC for 7 h each day, while the thermoneutral groups (TN) were at 33 degrees C. 3. At 21 and 42d of age, the percentage of CD4+ (helper T cells) and CD8+ (cytotoxic T cells) were determined by flow cytometry. Antibody response to sheep red blood cells (SRBC) and heterophil/lymphocyte (H/L) ratio were determined on d 21 and 42. 4. On d 21, FR elevated the CD4+, antibody titre and H/L ratio, but it decreased the CD8+ T cells. On d 42, HS decreased CD4+, CD8+, and antibody titre, but it increased H/L ratio. Under TN conditions, FR chickens had higher CD4+ than AL chickens. On d 42, FR/HS chickens had higher CD4+ and antibody titre, but they had lower CD8+ and H/L ratio than AL/HS chickens. 5. On d 42, the TN-Ross strain had lower CD4+, but they had higher CD8+ and antibody titres than the TN-Arian strain. On d 42, the HS-Arian strain had higher antibody titres and a lower H/L ratio than the HS-Ross strain. 6. Male chickens had higher CD4+, CD8+, antibody titres and H/L ratios 25 in all treatment groups. 7. In conclusion, FR early in life reduced some of the negative effects of the heat stress on the immune system of broiler chickens when exposed to high environmental temperatures later in life.

Lesional skin of cats with allergic dermatitis has a cellular infiltrate and a CD4/CD8 ratio comparable to that in humans with atopic dermatitis. CD4+ helper T cells and in particular cells belonging to the Th2 subset play an important role in disease pathogenesis in humans. We investigated the cytokine pattern of CD4+ T cells in situ, with special emphasis on the putative presence of cells producing interleukin 4 (IL4), in cats with allergic dermatitis. Immunohistochemical procedures were used to determine that CD4+ T cells in lesional and nonlesional skin of cats with allergic dermatitis can produce IL4, as occurs in humans. Lesional and nonlesional skin of cats with allergic dermatitis had significantly more IL4+ T cells (P = 0.001) than did skin of healthy control cats. Double staining indicated that all IL4+ cells were positive for pan-T or CD4 markers. Double labeling for mast cell chymase and IL4 stained primarily different cells. Western blotting demonstrated cross-reactivity between the antibody against human IL4 and a feline recombinant IL4. These results indicate that IL4 is primarily produced by CD4+ T cells and is also present in clinically uninvolved skin, indicating a role in the pathogenesis of allergic dermatitis in cats.

To determine the immunological response in lactating dairy cows infected with Salmonella (S.) Takoradi, the relationships among distributions of peripheral blood mononuclear cell (PBMC) subpopulations, endotoxin concentrations and dynamics of inflammatory cytokines in blood were investigated. The ratio of CD4+ T cells to CD8+ T cells was significantly lower in the affected cattle than in the control cattle (p<0.05) to decrease in the number of CD4+ T cells in the infected cattle. In contrast, the numbers of gammadeltaT cells, MHC class II-positive cells were significantly higher in the affected cattle than in the control cattle (p<0.01 respectively). Endotoxemia was found in all but one of the affected cattle. Serum IL-1 and IL-6 bioactivities were significantly higher in the affected cattle than in the control cattle (IL-1, p<0.05; IL-6. p<0.01). Serum TNF-alpha activities and levels were not detected in the control and affected cattle. The activities of proinflammatory cytokines determined by the bioassay are important to the relationships between concentration of endotoxin, cytokines and clinical signs. such as leukocytosis, leukopenia, fever or bacterial shedding. Serum IL-2 levels were lower in the affected cattle than in the control cattle. Serum IFN-y was not detected in the affected cattle except one. These results by the ELISA seemed to reflect the condition of subpopulation in the PBMCs from the shedding cattle. The present results suggest that cellular immunity is suppressed while the humoral immunity is activated in acute bovine salmonellosis.

A line of White Leghorn chickens was selected for high group productivity and longevity resulting in improved survival and feather score as well as reduced cannibalism and flightiness. Improvements in survival might have also been due to improved immunity. The present study was designed to test the hypothesis that selection for high (HGPS) and low (LGPS) group productivity and survivability also altered immune and hematological parameters. The LGPS line was an intense reverse selected line of the HGPS line at the eighth generation of development. Hens were randomly assigned to individual cages at 17 wk of age. Blood samples were collected from the hens at 21 wk of age. Subsets of T lymphocytes (CD4+, CD8+, and gammadelta cells) were measured using flow cytometry. Concentrations of plasma IgG were quantified with western blot analysis and immunoprecipitation assay. Hematological parameters were collected from blood smears. The HGPS hens had significantly higher percentages of blood lymphocytes and CD4+:CD8+ ratios of circulating T cells (P < 0.01) and tended to have more, but not significantly, gammadelta T cells (P = 0.07) than the LGPS hens. In contrast, the LGPS hens exhibited eosinophilia and heterophilia and greater heterophil:lymphocyte ratios (P < 0.01). The concentrations of plasma IgG were also significantly higher in the LGPS hens (P < 0.01). These results suggest that genetic selection for group productivity and longevity also alters the immunological and hematological systems of hens. The line difference in regulation of T cells, leukocytes, and production of IgG may suggest that different genes or modes of gene action are involved.

Objectives: To compare results of a nonradioactive colorimetric microplate assay with results of a traditional radioactive proliferation assay for determination of its use as a reliable and accurate alternative method for determination of proliferative activity of feline lymphocytes., Sample Population: Blood samples from 10 clinically normal domestic shorthair cats., Procedure: Double-density gradient separation was used to isolate mononuclear cells. Isolated cells were stimulated with various concentrations of concanavalin A (Con-A) and cultured for 72 hours. Lymphocyte proliferation was measured by radioactive ([3H]thymidine) and nonradioactive (colorimetric) techniques. Immunophenotypic analysis with feline-specific CD4+ and CD8+ monoclonal antibody was performed, using flow cytometry., Results: Mononuclear cells were successfully isolated (97 to 99% purity and viability) from blood samples. A similar dose-dependent proliferative response to Con-A stimulation was measured with [3H]thymidine incorporation and the colorimetric assay. For both techniques, concentrations of 0.1 and 1.0 microg of Con-A/ml were submitogenic, and 100 microg/ml was toxic to cultured cells. For both techniques, maximal proliferation was observed with 5 microg of Con-A/ml., Conclusions and Clinical Relevance: These results indicate that the nonradioactive colorimetric technique is a reliable and accurate method for measuring proliferative activity of feline lymphocytes. Clinically, this assay can be used as part of a screening process to determine immunocompetence of at-risk cats and to evaluate treatments for cats with immune-mediated or T-cell-dependent diseases.

A system for transplanting invasive equine trophoblast (i.e., chorionic girdle) to ectopic sites has been developed as a means to study the differentiation of this tissue and to assess maternal immune responses to the conceptus tissue in a site outside the uterus. Chorionic girdle was isolated from Day 33 to 34 conceptuses and surgically placed into the vulvar mucosa or subdermal skin of recipient mares. Biopsy specimens of the graft sites for immunohistochemical staining were taken at weekly or biweekly intervals after grafting. Serum samples were collected from each recipient and tested for antibody to donor major histocompatibility complex (MHC) class I antigens using the lymphocyte microcytotoxicity assay. Transplanted trophoblast cells expressed differentiation markers associated with invading chorionic girdle and endometrial cup cells. The transplanted trophoblast cells were also labeled by an antibody to eCG. Strong cellular and humoral immune responses to the transplanted tissue were mounted by the recipients, similar to those occurring during normal equine pregnancy. Despite these responses, the invasive trophoblast transplants survived for at least 28 days after grafting and downregulated MHC class I antigens, as do the mature endometrial cup cells in equine pregnancy. These findings suggest that invasive equine trophoblast has the capacity to differentiate fully in equine nonuterine tissues, and that it can evade maternal immune responses independent of the physiological state of pregnancy and in sites other than the uterus.