PHA Response Research Articles

Comparison of mitogen-induced proliferation in child and adult healthy groups by flow cytometry revealed similarities.

The proliferation of antigen-specific lymphocyte clones, the initial step in acquired immunity, is vital for effector functions. Proliferation tests both in immunology research and diagnosis are gaining attendance gradually, while the use of adult healthy individuals as controls of pediatric patients is a question. This study aimed to investigate and compare mitogen-stimulated proliferation responses of total lymphocytes and T- and B-lymphocyte subsets in adult and children healthy donors. Nineteen children and 20 adult healthy donors were enrolled in this study. Peripheral blood mononuclear cells (PBMCs) purified from peripheral blood samples of the donors, by Ficoll gradient centrifugation, were stained with CFSE and were cultured in a 37℃ CO2 incubator for 120h with the absence or existence of polyclonal activators: PHA and CD-Mix. After cell culture, PBMCs were stained with monoclonal antibodies against CD4 and CD19, and proliferation percentages of CD4+ T and CD19+ B cells, together with total lymphocytes were determined by flow cytometry. This study revealed similarities between children and adult age groups, concerning mitogenic stimulation of the lymphocytes. The only difference was a significantly high proliferation of pediatric CD4+ T cells in response to PHA. CD4+ T cell responses against PHA were inversely correlated with altering age. When pediatric individuals were distributed into age groups of 0-2years, 3-5years, and 6-18years, PHA responses of CD4+ cells were found to be diminished with advancing age. These findings propose the possibility of enrollment of adult healthy individuals as controls for pediatric patients.

Moonlight influence on quality of nestlings of scops owls depends on paternal coloration

Abstract Lunar phases might favor the maintenance of color polymorphism via disruptive selection if the different color variants performed differently in terms of prey capture under different moonlight levels. Moonlight, however, may affect prey capture as a side effect of its influence on prey behavior. Here we combine data of parental provisioning and quality of owlets with one ex-situ study of grasshopper activity to test whether Eurasian scops owls (Otus scops) with different plumage color and their prey are differently affected by moonlight. Food provisioning increased from new- to full-moon. However, the effect of moonlight on owlet mass gain and immune response depended on paternal coloration. On the one hand, body mass gain of nestlings of the greyest fathers increased from nights with new- to full-moon, whereas it did not change with moonlight for the brownest fathers. On the other hand, PHA response of nestlings of the brownest fathers increased with high moonlight levels during the first week of life, whereas it did not change with moonlight levels for the greyest fathers. Grasshoppers were more active at new moon than at full or waning moon. Our study provides supporting evidence that moonlight influences the behavior of both scops owls and its prey and suggests that fluctuations in environmental conditions can modulate the advantages of morphs. These results are important because they provide a general insight into a little appreciated mechanism for the maintenance of color polymorphism in natural populations based on the interactive effect of different environmental factors.

Prenatal aromatase inhibition alters postnatal immunity in domestic chickens (Gallus gallus)

In birds, exposure to exogenous testosterone during embryonic development can suppress measures of immune function; however, it is unclear whether these effects are due to direct or indirect action via aromatization. Estradiol (E2) is synthesized from testosterone by the enzyme aromatase, and this conversion is a necessary step in many signaling pathways that are ostensibly testosterone-dependent. Many lines of evidence in mammals indicate that E2 can affect immune function. We tested the hypothesis that some of the immunomodulatory effects observed in response to in ovo testosterone exposure in birds are mediated by conversion to E2 by aromatase, by using fadrozole to inhibit aromatization of endogenous testosterone during a crucial period of embryonic immune system development in domestic chickens (Gallus gallus). We then measured total IgY antibody count, response to PHA challenge, mass of thymus and bursa of Fabricius, and plasma testosterone post-hatch on days 3 and 18. Because testosterone has a reputation for immunosuppression, we predicted that if modulation of an immune measure by testosterone is dependent on aromatization, then inhibition of estrogen production by fadrozole treatment would lead to elevated measures of that parameter. Conversely, if testosterone inhibits an immune measure directly, then fadrozole treatment would likely not alter that parameter. Fadrozole treatment reduced circulating E2 in female embryos, but had no effect on males or on testosterone in either sex. Fadrozole-treated chicks had decreased day 3 plasma IgY antibody titers and a strong trend towards increased day 18 thymic mass. Furthermore, fadrozole treatment generated a positive relationship between testosterone and thymic mass in males, and tended to increase day 18 IgY levels for a given bursal mass in females. There was no effect on PHA response, bursal mass, or plasma testosterone at either age post-hatch. The alteration of several indicators of immune function in fadrozole-treated chicks implicates aromatization as a relevant pathway through which developmental exposure to testosterone can affect immunity in birds.

Predicting Autoimmunity Development in 22q11.2DS

Autoimmunity is a known complication of the 22q11.2 deletion syndrome (22q11.2DS) but predictive biomarkers for risk have not been identified. In most cases, autoimmunity develops 8 years after 22q11.2DS diagnosis. We sought to determine if immune biomarkers during early childhood could predict autoimmunity later in life. Retrospective chart investigation of patients with 22q11.2 from Duke University from 2008 to 2019. Only participants who had 22q11.2 deletion by FISH or microarray were evaluated. Fisher & Mann Whitney U tests were used for statistical analysis comparing 22q11.2DS patients with and without autoimmune disease. Study was IRB approved. Seventy-one patients with 22q11.2DS were identified, and 19 of 71 (26.8%) had a physician diagnosis of autoimmune disease. The most common autoimmune diagnoses were cytopenias followed by psoriasis and juvenile arthritis. Prevalence of an autoimmune diagnosis was not significantly different for those with complete versus partial DiGeorge Syndrome. The presence of hypoparathyroidism (OR 3.5, p=0.03) and allergic rhinitis (OR 0.3, p=0.03) significantly correlated with autoimmunity. T cell lymphopenia at 22q11.2 diagnosis (median age 4 years) was correlated with autoimmunity (p=0.01). Multiple T cell subsets were independently associated with autoimmunity including CD4 (p=0.01), CD8 (p=0.01), and naïve CD4 T cells (p=0.01). NK cell numbers, B cell numbers, PHA response, vaccine responses, and size of 22q11.2 deletion were not correlative with autoimmunity. 22q11.2DS patients with hypoparathyroidism and T cell lymphopenia were associated with a higher risk of autoimmune disease. This study indicates that immune biomarkers present in early childhood may predict risk for autoimmunity later in life.

Characterizing Pneumocystis jirovecii Pneumonia in patients with Primary Immunodeficiency Disorders, Data from the US Immunodeficiency Network (USIDNET) Registry

Pneumocystis jirovecii pneumonia (PJP) is a life-threatening infection that affects immunocompromised individuals, including those with primary immunodeficiencies (PID). The US Immunodeficiency Network (USIDNET) registry was utilized to perform retrospective analysis of PID patients who developed PJP. The cohort was categorized into those who developed PJP before or at PID diagnosis versus those who developed PJP after PID diagnosis. Frequency of PJP in PID patients within USIDNET was 63/5204 (1.2%). Forty-nine patients developed PJP prior to history of transplantation and were included in final analysis. This cohort was 22% female (11/49), 78% male (38/49). The majority developed PJP before/at PID diagnosis (40/49,82%). Median age at PID diagnosis in the PJP before/ at PID diagnosis group was 0.65 years [IQR 0.40,2.08]. PID diagnoses varied, including Hyper IgM syndromes (17/49,34.7%), Severe Combined Immunodeficiency (SCID) (15/49,30.6%) and Common Variable Immunodeficiency (CVID) (8/49,16.3%), among others. Hyper IgM patients with PJP before/ at diagnosis of PID median first CD4 count was 1233/uL [IQR 929,4100]. All other patients with PJP before/at diagnosis of PID median first CD4 count was 197/uL [IQR 12,560], though for CVID patients, median first CD4 was 580/uL. Of patients with phytohemagglutinin data, 16/27 (59%) had normal PHA response. This characterization of PJP in PID from the USIDNET registry demonstrates the importance of considering the particular PID defect when contemplating PJP prophylaxis. However, PJP often precedes or is part of the initial consideration of a PID diagnosis. T cell lymphopenia and poor mitogen response may not adequately identify PID patients at risk of PJP.

Reduced Toxicity Conditioning for Non-Malignant Hematopoietic Cell Transplants

Background. Reduced-intensity conditioning (RIC) regimens are commonly used for hematopoietic cell transplantation (HCT) in non-malignant disorders to minimize potential drug-related toxicity from myeloablative conditioning (MAC). However, melphalan-based RIC regimens are associated with poor engraftment and need for second HCT or donor lymphocyte infusions (DLIs). This study expands on our previous experience with targeted busulfan, fludarabine, and intravenous (IV) alemtuzumab as a low-toxicity but myeloablative regimen to achieve sustained donor engraftment. Methods. We conducted a retrospective analysis of patients ≤25 years of age with non-malignant disorders who received targeted busulfan, fludarabine and IV alemtuzumab as conditioning for their first HCT between 2004 and 2018. Graft-versus-host disease (GVHD) prophylaxis included calcineurin inhibitors in all patients, plus methotrexate (n=59), methylprednisolone (n=2), or mycophenolate (n=1). The primary end point was 3-year overall survival (OS). Secondary end points included treatment-related mortality (TRM), event-free survival (EFS) defined as alive without second HCT or disease progression, and incidence of acute and chronic GVHD (aGVHD and cGVHD). Other end points included incidence of graft failure (GF) defined as <5% donor chimerism in whole blood, sinusoidal obstruction syndrome (SOS) and CMV viremia. Analysis was stratified by era of HCT (2004-2011 vs. 2012-2018). Results. We identified 62 patients with a median age of 5.5 years and male:female of 46:16. Diagnoses included primary immune deficiencies (60%), hemoglobinopathies (22%), bone marrow failure disorders (8%) and inherited metabolic disorders (10%). The median busulfan cumulative area-under-the-curve (cAUC) was 70 mg*h/L (range, 57-108 mg*h/L). Donors were matched sibling (MSD) in 27%, 8/8 allele-matched unrelated (MUD) in 50% and Class I allele-mismatched (MM) in 23%. All but 2 of the Class I allele-MM donors were unrelated. The source was bone marrow (BM) in 74% and peripheral blood stem cells (PBSC) in 26%. The cumulative incidence (CI) of GF was 8% (95% CI, 1-16%). In engrafted patients, the median donor chimerism in whole blood, CD3, CD14/15, and CD19 at 1-year was 96%, 90%, 99%, and 99%, respectively. Only 1 patient required DLIs for poor chimerism. T-cell reconstitution (CD4 >200 x 109/L and PHA response >50% control) was achieved at a median of 182 days (range, 76-585 days). Two patients died following disease progression despite 100% donor chimerism. The CI of TRM was 10% (95% CI, 2-17%). Causes of TRM included infectious complications (n=2), hemorrhage (n=1), respiratory failure (n=3 with 2 following subsequent transplants after graft rejection). TRM incidence was significantly higher in patients with Class I MM compared to those with MSD or 8/8 allele-MUD (29% vs 6% vs 3%; p=0.024; Figure 1A). SOS occurred in 5% of patients and all resolved with supportive therapy (n=1) or defibrotide (n=2). Post-transplant CMV viremia occurred in 26% of all patients and in 71% of CMV seropositive patients. None developed CMV disease and all infections resolved with anti-viral therapy. OS and EFS at 3-years were 87% (95% CI, 78-95%) and 80% (95% CI, 70-90%), respectively. HCTs performed in the more recent era showed an OS of 97% compared to 79% in HCTs performed earlier (p=0.057; Figure 1B). OS was 97% in patients with 8/8 allele-MUD compared to 79% in those with MSD and 71% in those with Class I allele-MM (p=0.051). The 6-month CI of grade II-IV aGVHD was 7% (95% CI, 3-13%), while the 3-year CI of cGVHD was 5% (95% CI, 0-11%). GVHD incidence was higher in patients receiving PBSC compared to BM (25% vs 0%; p<0.001) and in patients ≥5 years of age compared to those <5 years of age (13% vs 0%; p=0.043). Patients with inherited metabolic disorders showed significantly worse OS (44%, p=0.016), EFS (33%, p=0.001) and aGVHD (33%, p=0.008) than those in other disease groups. Univariate analysis did not show any significant differences in survival or GVHD incidence by race or sex. Conclusions. Use of targeted busulfan, fludarabine and IV alemtuzumab as myeloablative conditioning in pediatric patients with primary immunodeficiencies, hemoglobinopathies, and bone marrow failure syndromes undergoing matched sibling or well-matched unrelated donor HCT provides a well-tolerated option to achieve sustained engraftment with low incidence of GVHD and toxicities such as SOS. Figure 1 Disclosures Shimano: Pfizer: Research Funding; Daiichi Sankyo: Research Funding; Novartis: Research Funding. Huang:Novartis: Research Funding. Cowan:Homology Medicine: Equity Ownership, Membership on an entity's Board of Directors or advisory committees; Leadiant: Consultancy; California Institute Of Regenerative Medicine: Research Funding; NIH NIAD: Research Funding; Rocket Pharma: Consultancy; UpToDate: Honoraria; bluebird bio: Consultancy. Dvorak:Alexion Inc: Consultancy; Jazz Pharmaceuticals: Consultancy.

Sense of coherence in lung cancer patients: Association with coping, immune function, and fatigue

Sense of coherence (SOC) reflects the degree to which an individual perceives their world as comprehensible, manageable, and meaningful. Among cancer populations, SOC has been linked with greater quality of life and reduced mortality, suggesting that SOC may serve as a marker of resilience in the context of chronic disease. Utilizing a published adaptation of the transactional model of stress and coping, we hypothesized greater SOC would be associated with adaptive coping, more regulated cytokine response, and lower fatigue. Sixty-two lung cancer patients provided self-report data on SOC, coping, and fatigue. PBMC were incubated with stimulators of acquired and innate immunity (PHA, LPS), and cytokines were measured (pro-inflammatory: IL-1beta IL-6, TNF-alpha; Type 1/TH1: IFN-gamma; Type 2/TH2: IL-5). Linear regressions tested hypotheses. Greater SOC was associated with emotionally expressive coping (p = 0.005), an attenuated IL-5 response to PHA (p = 0.038) and IFN-gamma response to LPS (p=.001); and lower fatigue (p

Studies on the mechanisms of nickel induced immunosuppression

Abstract Nickel (Ni) inhibits lymphoproliferative responses. Proliferation was performed on PBMC of 20 controls and 10 patients with Ni allergy. NiSO4 was used at a concentration of 0.1mM. PHA was added at a concentration of 5 μg/ml. Cells were incubated for 5 days and proliferation assessed by CFSE staining. At 0.1mM Ni strongly reduced (>40%) PHA-induced proliferation. The inhibition decreases in a dose-dependent manner and response to PHA is restored at 20 mM Ni. Ni alone induces small (<5%) proliferative response in both allergic and non allergic donors. Nickel inhibition requires the presence of Ni in the supernatant and is abrogated when Ni is incubated with the cells for up to 4 hours and the supernatant is washed. Strong inhibition was also observed in preliminary experiments with tuberculin purified protein derivative (PPD) stimulated PBMC. Staining for CD8 on CFSE cultures showed that PHA response was due to both CD8 and CD4 cells. Ni increased % of CD4 cells and decreased the % of CD8 cells. PPD response was predominantly a CD4 response, with Nickel reducing the % of CD4 cells at higher concentations, while unaffecting the CD8. Ni inhibits IL-6 production essential to normal PHA response. The addition of 10–100 IU of IL-6 and/or IL-2 to the cultures did not affect Ni inhibition of PHA proliferation. Addition of IL-6 increased CD25+ cells. Our data suggest that Ni induced inhibition of PHA response is due to steric interference between PHA and its cellular ligand. Further studies will clarify the precise mechanism of Ni mediated inhibition of immune responses and its the role in the pathogenesis of Ni allergy.

Transgenerational effects enhance specific immune response in a wild passerine.

Vertebrate mothers transfer diverse compounds to developing embryos that can affect their development and final phenotype (i.e., maternal effects). However, the way such effects modulate offspring phenotype, in particular their immunity, remains unclear. To test the impact of maternal effects on offspring development, we treated wild breeding house sparrows (Passer domesticus) in Sevilla, SE Spain with Newcastle disease virus (NDV) vaccine. Female parents were vaccinated when caring for first broods, eliciting a specific immune response to NDV. The immune response to the same vaccine, and to the PHA inflammatory test were measured in 11-day-old chicks from their following brood. Vaccinated chicks from vaccinated mothers developed a stronger specific response that was related to maternal NDV antibody concentration while rearing their chicks. The chicks’ carotenoid concentration and total antioxidant capacity in blood were negatively related to NDV antibody concentration, whereas no relation with PHA response was found. Specific NDV antibodies could not be detected in 11-day-old control chicks from vaccinated mothers, implying that maternally transmitted antibodies are not directly involved but may promote offspring specific immunity through a priming effect, while other immunity components remain unaffected. Maternally transmitted antibodies in the house sparrow are short-lived, depend on maternal circulation levels and enhance pre-fledging chick specific immunity when exposed to the same pathogens as the mothers.

A homozygous STIM1 mutation impairs store-operated calcium entry and natural killer cell effector function without clinical immunodeficiency

A homozygous STIM1 mutation impairs store-operated calcium entry and natural killer cell effector function without clinical immunodeficiency

Unrelated donor hematopoietic stem cell transplantation for the treatment of non-malignant genetic diseases: An alemtuzumab based regimen is associated with cure of clinical disease; earlier clearance of alemtuzumab may be associated with graft rejection.

Hematopoietic stem cell transplantation (HSCT) with matched unrelated donors (MUD), offers potentially curative therapy for patients with non-malignant genetic diseases. In this pilot study conducted from 2006 to 2014, we report the outcomes of 15 patients with non-malignant genetic diseases who received a myeloablative regimen with a reduced cyclophosphamide dose, adjunctive serotherapy and MUD HSCT [intravenous alemtuzumab (52 mg/m(2) ), busulfan (16 mg/kg), fludarabine (140mg/m(2) ), and cyclophosphamide (105 mg/kg)]. Graft-versus-host-disease (GVHD) prophylaxis consisted of tacrolimus/cyclosporine and methylprednisolone. Median (range) time to neutrophil engraftment (>500 cells/µL) and platelet engraftment (>20,000/mm(3) ) were 15 (12-28) and 25 (17-30) days, respectively. At a median follow-up of 2 (0.2-5.4) years, the overall survival (OS) was 93.3% (95% CI: 0.61-0.99) and disease-free survival (DFS) was 73.3% (95% CI: 0.44-0.89). Among this small sample, earlier alemtuzumab clearance was significantly associated with graft rejection (P = 0.047), earlier PHA response (P = 0.009) and a trend toward earlier recovery of recent thymic emigrants (RTE) (P = 0.06). This regimen was associated with durable donor engraftment and relatively low rates of regimen related toxicity (RRT); future alemtuzumab pharmacokinetic studies may improve outcomes, by allowing targeted alemtuzumab clearance to reduce graft rejection and promote more rapid immune reconstitution.

A mathematical model approach quantifying patients' response to changes in mechanical ventilation: Evaluation in pressure support

PurposeThis article evaluates how mathematical models of gas exchange, blood acid-base status, chemical respiratory drive, and muscle function can describe the respiratory response of spontaneously breathing patients to different levels of pressure support. MethodsThe models were evaluated with data from 12 patients ventilated in pressure support ventilation. Models were tuned with clinical data (arterial blood gas measurement, ventilation, and respiratory gas fractions of O2 and CO2) to describe each patient at the clinical level of pressure support. Patients were ventilated up to 5 different pressure support levels, for 15 minutes at each level to achieve steady-state conditions. Model-simulated values of respiratory frequency (fR), arterial pH (pHa), and end-tidal CO2 (FeCO2) were compared to measured values at each pressure support level. ResultsModel simulations compared well to measured data with Bland-Altman bias and limits of agreement of fR of 0.7 ± 2.2 per minute, pHa of −0.0007 ± 0.019, and FeCO2 of −0.001 ± 0.003. ConclusionThe models describe patients' fR, pHa, and FeCO2 response to changes in pressure support with low bias and narrow limits of agreement.

Sublethal Pb exposure produces season-dependent effects on immune response, oxidative balance and investment in carotenoid-based coloration in red-legged partridges.

Ingestion of lead (Pb) shot pellets constitutes the main cause of Pb poisoning in avifauna. We studied the effects of sublethal Pb exposure on immunity, carotenoid-based coloration, oxidative stress and trade-offs among these types of responses during spring and autumn in red-legged partridges (Alectoris rufa). We evaluated constitutive immunity testing lysozyme and natural antibody levels, and blood bactericidal and phagocytic activities. We studied induced immunity by testing PHA and humoral responses. We analyzed fecal parasite and bacterial abundance and oxidative stress biomarkers. Pb exposure in spring reduced natural antibody levels, whereas in autumn, it reduced lysozyme levels and increased phagocytic activity. Pb exposure increased PHA response in both seasons, and decreased T-independent humoral response in autumn. Pb exposure also increased noncoliform and decreased coliform Gram-negative gut bacteria. In spring, Pb exposure decreased antioxidant levels and increased coloration in males, whereas in autumn, it increased retinol levels but reduced coloration in both genders. Our results suggest that in spring, Pb-exposed females used antioxidants to cope with oxidative stress at the expense of coloration, whereas Pb-exposed males increased coloration, which may reflect an increased breeding investment. In autumn, both genders prioritized oxidative balance maintenance at the expense of coloration.

Changes in phytohaemagglutinin skin-swelling responses during the breeding season in a multi-brooded species, the Eurasian Tree Sparrow: do males with higher testosterone levels show stronger immune responses?

Life-history theory assumes that the fitness costs of immunity may have negative effects on reproductive success. Similarly, the immunocompetence handicap hypothesis is based on findings that testosterone (T) has immunosuppressive effects, although the basis of this hypothesis has recently been challenged. As much of the work examining the relationship between T levels and immune function has been carried out in captive-housed species, these results may not accurately reflect the situation of animals living in natural environments. To better understand the relationship between plasma T levels and immune function, studies focusing on free-living animals are needed. A previous study by our group determined the changes in both baseline and stress-induced T levels in free-living Eurasian Tree Sparrows (Passer montanus) across different annual cycle stages. In this study, we further report the phytohaemagglutinin skin-swelling (PHA) immune response in this multi-brooded species during different breeding sub-stages, and then determine the relationships between the PHA response and both baseline and stress-induced T levels. Our results show that the PHA response varied across the different sub-stages and differed significantly between the first and second brood stage. Furthermore, T levels in male Tree Sparrows are positively correlated with the PHA response during the breeding season, whereas this relationship is negative in females, suggesting that the biological function of T differs between the sexes. Therefore, our results suggest that free-living animals have evolved the ability to orchestrate trade-offs between reproduction and immune functions based on changes in physiology and the environment, which should provide further opportunities to study the flexibility and plasticity of physiological and ecological adaptations in natural environments.

Genetic and environmental variation in condition, cutaneous immunity, and haematocrit in house wrens.

BackgroundLife-history studies of wild bird populations often focus on the relationship between an individual’s condition and its capacity to mount an immune response, as measured by a commonly-employed assay of cutaneous immunity, the PHA skin test. In addition, haematocrit, the packed cell volume in relation to total blood volume, is often measured as an indicator of physiological performance. A multi-year study of a wild population of house wrens has recently revealed that those exhibiting the highest condition and strongest PHA responses as nestlings are most likely to be recruited to the breeding population and to breed through two years of age; in contrast, intermediate haematocrit values result in the highest recruitment to the population. Selection theory would predict, therefore, that most of the underlying genetic variation in these traits should be exhausted resulting in low heritability, although such traits may also exhibit low heritability because of increased residual variance. Here, we examine the genetic and environmental variation in condition, cutaneous immunity, and haematocrit using an animal model based on a pedigree of approximately 2,800 house wrens.ResultsEnvironmental effects played a paramount role in shaping the expression of the fitness-related traits measured in this wild population, but two of them, condition and haematocrit, retained significant heritable variation. Condition was also positively correlated with both the PHA response and haematocrit, but in the absence of any significant genetic correlations, it appears that this covariance arises through parallel effects of the environment acting on this suite of traits.ConclusionsThe maintenance of genetic variation in different measures of condition appears to be a pervasive feature of wild bird populations, in contradiction of conventional selection theory. A major challenge in future studies will be to explain how such variation persists in the face of the directional selection acting on condition in house wrens and other species.

Stochastic Food Deprivation Increases Cellular Immunity in Kunming Mice

Animals’ immunity is an important factor to determine their survival and fitness. Unpredictable food shortage is common to small mammals due to fluctuation of food resources throughout a year. In general, acute food deprivation (FD) inhibited immune function in rodents. In the present study we tested the hypothesis that stochastic FD would also suppress T cell-mediated immunity in mice. Fifteen adult male Kunming mice were randomly assigned into the Fed (n = 7) and FD (n = 8) groups, in which the latter were subjected to stochastic FD regime. Unexpectedly, T cell-mediated immunity assessed by PHA response was increased in the FD mice compared with the fed controls. However, body fat mass, wet thymus and spleen mass, white blood cells, serum leptin and corticosterone concentrations did not differ between the Fed and FD groups. Taken together, stochastic food deprivation can enhance cellular immunity in Kunming mice.

Age at weaning, immunocompetence and ectoparasite performance in a precocial desert rodent.

We studied the effects of early weaning on immunocompetence and parasite resistance in a precocial rodent Acomys cahirinus. We hypothesized that if parasite resistance is energetically expensive and nutritional and immunological support from mothers are necessary for the long-term health of offspring, then early weaned animals would be immunologically weaker and less able to defend themselves against parasites than later weaned animals. We weaned pups at 14, 21 or 28 days after birth and assessed their immunocompetence and resistance against fleas Parapulex chephrenis when they attained adulthood. Immunocompetence was assessed using leukocyte concentration (LC) and a phytohaemagglutinin injection assay (PHA test). To estimate resistance against fleas, we measured performance of fleas via the number of produced eggs and duration of development and resistance to starvation of the flea offspring. We found a significant positive effect of weaning age on the PHA response but not on LC. The effect of age at weaning on flea egg production was manifested in male but not female hosts, with egg production being higher if a host was weaned at 14 than at 28 days. Weaning age of the host did not affect either duration of development or resistance to starvation of fleas produced by mothers fed on these hosts. We conclude that even in relatively precocial mammals, weaning age is an important indicator of future immunological responses and the ability of an animal to resist parasite infestations. Hosts weaned at an earlier age make easier, less-resistant targets for parasite infestations than hosts weaned later in life.

Effects of reproduction on immuno-suppression and oxidative damage, and hence support or otherwise for their roles as mechanisms underpinning life history trade-offs, are tissue and assay dependent

Life history parameters appear to be traded off against each other, but the physiological mechanisms involved remain unclear. One hypothesis is that potentially energetically costly processes such as immune function and protection from oxidative stress may be compromised during reproductive attempts because of selective resource allocation. Lower temperatures also impose energy costs, and hence allocation decisions might be more pronounced when animals are forced to reproduce in the cold. Here, we experimentally tested whether reproduction at different ambient temperatures was associated with elevated oxidative stress and suppressed immune function in Mongolian gerbils (Meriones unguiculatus). Using a variety of different markers for both immune function and oxidative stress, we found that some measures of immune function (serum bactericidal capacity and size of the thymus) were significantly suppressed, while some measures of oxidative protection [serum superoxide dismutase (SOD) activity and glutathione peroxidase (GPx) activity] were also reduced, and a marker of oxidative damage (protein carbonyls in serum) was increased in lactating compared with non-reproductive gerbils. These changes were in line with the selective resource allocation predictions. However, the phytohaemagglutinin response and serum total immunoglobulin (IgG) were not suppressed, and other markers of oxidative damage [malondialdehyde (MDA) (TBARS) and protein carbonyls in the liver] were actually lower in lactating compared with non-reproductive gerbils, consistent with increased levels of SOD activity and total antioxidant capacity in the liver. These latter changes were opposite of the expectations based on resource allocation. Furthermore, other measures of protection (GPx levels in the liver and protein thiols in both serum and liver) and damage [MDA (TBARS) in serum] were unrelated to reproductive status. Ambient temperature differences did not impact on these patterns. Collectively, our results indicated that the inferred effects of reproduction on immunosuppression and oxidative damage, and hence support or otherwise for particular physiological mechanisms that underpin life history trade-offs, are critically dependent on the exact markers and tissues used. This may be because during reproduction individuals selectively allocate protection to some key tissues, but sacrifice protection of others.

What does carotenoid-dependent coloration tell? Plasma carotenoid level signals immunocompetence and oxidative stress state in birds-A meta-analysis.

Mechanisms maintaining honesty of sexual signals are far from resolved, limiting our understanding of sexual selection and potential important parts of physiology. Carotenoid pigmented visual signals are among the most extensively studied sexual displays, but evidence regarding hypotheses on how carotenoids ensure signal honesty is mixed. Using a phylogenetically controlled meta-analysis of 357 effect sizes across 88 different species of birds, we tested two prominent hypotheses in the field: that carotenoid-dependent coloration signals i) immunocompetence and/or ii) oxidative stress state. Separate meta-analyses were performed for the relationships of trait coloration and circulating carotenoid level with different measures of immunocompetence and oxidative stress state. For immunocompetence we find that carotenoid levels (r = 0.20) and trait color intensity (r = 0.17) are significantly positively related to PHA response. Additionally we find that carotenoids are significantly positively related to antioxidant capacity (r = 0.10), but not significantly related to oxidative damage (r = −0.02). Thus our analyses provide support for both hypotheses, in that at least for some aspects of immunity and oxidative stress state the predicted correlations were found. Furthermore, we tested for differences in effect size between experimental and observational studies; a larger effect in observational studies would indicate that co-variation might not be causal. However, we detected no significant difference, suggesting that the relationships we found are causal. The overall effect sizes we report are modest and we discuss potential factors contributing to this, including differences between species. We suggest complementary mechanisms maintaining honesty rather than the involvement of carotenoids in immune function and oxidative stress and suggest experiments on how to test these.

The number and coloration of white flank spots predict the strength of a cutaneous immune response in female Diamond Firetails, Stagonopleura guttata

Colour variation in birds is often used to signal functional differences between individuals and sexes, though white plumage has usually been disregarded because white feathers were thought to be cheap to produce and hence unreliable signals. Here, we provide evidence for sex-specific morphological and colour differences in the strikingly patterned but apparently monomorphic Dia- mond Firetail (Stagonopleura guttata). We found that males had longer and darker tails, wider lores and a darker bill, while females had significantly more white flank spots. These gender-specific trait differences could be used to signal individual quality in sexual or social interactions, and hence we examined the correlation between these traits and the cell-mediated immune response (phytohaemag- glutinin, PHA test). During both the breeding and non- breeding seasons, we found a positive correlation between PHA response and white flank spot number as well as their ultra-violet reflectance in females, and a negative correla- tion with white chroma and hue. In males, there was a positive correlation between PHA response and red rump reflectance (hue, red-chroma). There was no association between PHA response and either tail length or lore depth in either sex. These results add to a growing body of evidence that female spot number and white plumage reflectance signal quality.