High Cell-mediated Immune Response Research Articles

Comparative immunogenic and immunoprotective activities of PCV2d Cap and Rep antigens delivered by an efficient eukaryotic expression system engineered into a Salmonella vaccine vector

Porcine circovirus type 2 (PCV2) stands as a predominant etiological agent in porcine circovirus-associated diseases. To manage the spread of the disease, it is necessary to develop a next-generation vaccine expressing PCV2 antigens that target the prevailing genotype such as PCV2d. A bacterial-mediated vaccine delivery by live-attenuated Salmonella has attracted interest for its low-cost production and highly effective vaccine delivery. Thus, in this study, we utilized the advantages of the Salmonella-mediated vaccine delivery by cloning PCV2d cap and rep into a eukaryotic expression plasmid pJHL204 and electroporation into an engineered live-attenuated Salmonella Typhimurium JOL2500 (Δlon, ΔcpxR, ΔsifA, Δasd). The eukaryotic antigen expression by JOL2995 (p204:cap) and JOL2996 (p204:rep) was confirmed in vitro and in vivo which showed efficient antigen delivery. Furthermore, vaccination of mice model with the vaccine candidates elicited humoral and cell-mediated immune responses as depicted by high levels of PCV2-specific antibodies, CD4+ and CD8+ T cells, and neutralizing antibodies, especially by JOL2995 (p204:cap) which correlated with the significant decrease in the viral load in PCV2d-challenged mice. Interestingly, JOL2996 (p204:rep) may not have elicited high levels of neutralizing antibodies and protective efficacy, but it elicited considerably higher cell-mediated immune responses. This study demonstrated Salmonella-mediated vaccine delivery system coupled with the eukaryotic expression vector can efficiently deliver and express the target PCV2d antigens for strong induction of immune response and protective efficacy in mice model, further supporting the potential application of the Salmonella-mediated vaccine delivery system as an effective novel approach in vaccine strategies for PCV2d.

Higher Cell-Mediated Immune Responses in Patients With Inflammatory Bowel Disease on Anti-TNF Therapy After COVID-19 Vaccination.

BackgroundSome patients with inflammatory bowel disease (IBD) on immunosuppressive therapies may have a blunted response to certain vaccines, including the messenger RNA (mRNA) coronavirus disease 2019 (COVID-19) vaccines. However, few studies have evaluated the cell-mediated immune response (CMIR), which is critical to host defense after COVID-19 infection. The aim of this study was to evaluate the humoral immune response and CMIR after mRNA COVID-19 vaccination in patients with IBD.MethodsThis prospective study (HERCULES [HumoRal and CellULar initial and Sustained immunogenicity in patients with IBD] study) evaluated humoral immune response and CMIR after completion of 2 doses of mRNA COVID-19 vaccines in 158 IBD patients and 20 healthy control (HC) subjects. The primary outcome was the CMIR to mRNA COVID-19 vaccines in patients with IBD. The secondary outcomes were a comparison of (1) the CMIR in patients with IBD and HC subjects, (2) CMIR and humoral immune response in all participants, and (3) correlation between CMIR and humoral immune response.ResultsThe majority (89%) of patients with IBD developed a CMIR, which was not different vs HC subjects (94%) (P = .6667). There was no significant difference (P = .5488) in CMIR between immunocompetent (median 255 [interquartile range, 146-958] spike T cells per million peripheral blood mononuclear cells) and immunosuppressed patients (median 377 [interquartile range, 123-1440]). There was no correlation between humoral and cell-mediated immunity after vaccination (P = .5215). In univariable analysis, anti-tumor necrosis factor therapy was associated with a higher CMIRs (P = .02) and confirmed in a multivariable model (P = .02). No other variables were associated with CMIR.ConclusionsMost patients with IBD achieved CMIR to a COVID-19 vaccine. Future studies are needed evaluating sustained CMIR and clinical outcomes.

Evaluation of Crossbreeding Parameters for Immunocompetence and Serum Enzyme Profile in a Partial Diallel Cross Involving Three Genetic Groups of Chicken

Background: Poultry production in rural India is mostly a non-intensive venture comprising native chicken with low production potential but higher disease resistance and adaptability. Present study is attempted for developing a suitable cross for rural poultry production as well as identifying the genetic groups that are nicking well through a partial diallel cross. Methods: A partial diallel cross using three genetic groups/ breeds of chicken viz. coloured synthetic male line (CSML), Local native chicken (Desi) and CARI-Red as the parent lines was designed. CSML was used as male and CR as female line only. The progenies were evaluated for crossbreeding parameters viz. combining abilities and heterosis for immune response and serum enzyme profile to identify the best combining parent lines.Result: Significantly higher (P less than 0.05) cell-mediated (CMI) and humoral immune response (HIR) and immune organ (spleen, bursa of fabricius and thymus) weights were recorded in CR purebred followed by Desi purebred, while CSML purebred exhibited lowest immunity. Variances for SCA differed significantly (P less than 0.001) for HIR and immune organ weights. Inconsistent but significantly higher (P less than 0.01) serum enzymes (AST, ALT and ALP) and AST/ALT ratio were recorded in triple cross and D x CR. Variances for SCA differed significantly (P less than 0.05) for serum enzymes. Results revealed that the CARI-Red and Desi were the improver parent lines for better immunocompetence and serum enzyme profile, respectively in the crosses.

Assessment of Acute-Phase Protein Response Associated with the Different Pathological Forms of Bovine Paratuberculosis.

Simple SummaryParatuberculosis (PTB) is a chronic debilitating disease caused by Mycobacterium avium subspecies paratuberculosis (Map) that affects ruminants worldwide. Many aspects related to the pathogenesis of this disease are still unknown, including the inflammatory acute-phase response developed during the course of the infection. To clarify this, serum levels of haptoglobin and serum amyloid A, two positive acute-phase proteins, were evaluated in a total of 190 cows, among which 59 were healthy control animals and 131 cows were diagnosed post-mortem with different types of lesion associated with Map-infection. The results reflect a significant increase of these proteins’ levels in the infected animals and, more specifically, in those animals with types of lesion characterized by a low bacterial load and with predominance of a cell-mediated immune response. This suggests that these molecules would play a certain role in the pathogenesis of the PTB and a possible utility as biomarkers of different stages of the disease.In this study, the concentrations of two acute-phase proteins (APPs), haptoglobin (Hp) and serum amyloid A (SAA), were quantitatively assessed in serum samples from cattle naturally infected with paratuberculosis (PTB). APP profiles were compared across 190 animals classified according to the different pathological forms associated with infection: uninfected (n = 59), with focal lesions (n = 73), multifocal lesions (n = 19), and diffuse paucibacillary (n = 11) and diffuse multibacillary lesions (n = 28). Our results showed a significant increase in both APPs in infected animals compared to the control group, with differences depending on the type of lesion. Hp and SAA levels were increased significantly in all infected animals, except in cows with diffuse multibacillary lesions that showed similar values to non-infected animals. The expression pattern of both APPs was similar and negatively correlated with the antibody levels against PTB. These results indicate that the release of Hp and SAA is related to the presence of PTB lesions associated with a high cell-mediated immune response and a lower bacterial load, suggesting that the pro-inflammatory cytokines that are associated with these forms are the main stimulus for their synthesis. These molecules could show some potential to be used as putative biomarkers of PTB infection, particularly for the identification of subclinical animals showing pathological forms related to latency or resistance to the development of advanced lesions.

114 The relationship of cell mediated immune response of Brahman cows on calving interval

Abstract The relationships between cell-mediated immune response (CMIR) of cows and calving interval and calf growth were evaluated. Calving interval was evaluated for each of the initial 7 life-time opportunities that females had to produce a calf. The number of days from when the cow had a calf until she calved again determined the cow’s calving interval for each year. The Spring breeding seasons were divided into 45-d artificial insemination and 45-d natural service periods. The Fall breeding season was natural service each year. Multiparous Brahman cows (n = 435) had records for breeding and calving dates and calf weaning weights. The CMIR (hypersensitivity response to Candida albicans measured using tail-fold thickness) was determined once for each female in the Fall and early Winter 2015. Cows were classified into response groups based on mean and SD of CMIR. High responders were those with CMIR ½ SD ≥ the mean (≥ 2.8 mm), Intermediate cows were within ½ SD of the mean (2.7-1.8 mm), and cows ½ SD ≤ the mean were Low (≤ 1.7 mm). Data were analyzed using repeated measures, mixed, and Chi Square procedures (SAS 9.4). Variables included calving interval, cow age, CMIR response group, pregnancy status, calf count, calf sex, cow sire, calf sire, and calf adjusted 180-d weaning weight. A greater proportion of High CMIR cows made it to their 6th calving interval. Chi Square analysis indicated that High CMIR cows were more likely to remain in the herd longer and produce a calf each year. Cows with High CMIR had a greater stayability (P = 0.1). Selection for High CMIR can result in selection of cows which are more likely to remain in the herd long enough to be profitable as they have a greater stayability and shorter calving intervals (about 25 days) than their Low CMIR herdmates.

Effect of feeding different levels of Azolla pinnata on blood biochemicals, hematology and immunocompetence traits of Chabro chicken.

Aim:The present study was conducted to see the effect of feeding different levels of Azolla meal on blood biochemicals, hematology and immunocompetence traits of Chabro chicken.Materials and Methods:The study was conducted on 160 Chabro chicks, which were randomly divided into four treatment groups each with four replicates of 10 birds. The first treatment (T1) served as a control in which basal diets was offered without Azolla supplementation while in T2, T3, and T4 groups, basal diet was replaced with Azolla meal at 5%, 7.5%, and 10% levels, respectively. A feeding trial was conducted upto 8 weeks. At the last week of trial, blood samples were collected randomly from one bird of each replicate and plasma was separated to estimate certain biochemical parameters, some blood metabolites, minerals and enzymes like alanine aminotransferase and aspartate aminotransferase (AST). Hematological parameters such as hemoglobin, packed cell volume, total leukocytes count and differential leukocytes count were estimated in fresh blood just after collection. The humoral immune response was measured against sheep red blood cells,and cell-mediated immune response was measured against phyto hemagglutinin lectin from Phaseolus vulgaris (PHA-P).Results:The study showed that hematological profile of the Chabro bird was not affected by any treatment except heterophil and lymphocyte which was found higher in T2 and T3 groups and eosinophil was found higher in a T3 group than control. Blood glucose, creatinine, cholesterol, total protein, albumin, uric acid, and triglycerides were found similar in all the groups and within the normal values for broiler chicken. Liver enzymes and macro mineral content in blood were found similar in all the treatment groups and within normal physiological range. Although AST was found higher in 10% replacement group than control, the value was within normal range for broiler chicken. Although antibody titer was found similar in all the experimental groups in the present study, cell-mediate immune response (response to PHA-P) was found higher in 5%, 7.5%, and 10% replacement groups than control(p<0.05).Conclusion:Similar blood biochemical parameters and higher cell-mediated immune response in Azolla replacement group indicated immune-modulatory effect of Azolla meal without any toxicity.

A trial to improve stability and immunogenicity of inactivated NDV vaccine with paraffin oil adjuvant using aluminum stearate

Inactivated Newcastle disease virus (NDV) vaccines were prepared using paraffin oil, paraffin oil andaluminum stearate (Al.St.) and Montanide oil ISA 71VG as adjuvants. All experiments gave the priorityto the inactivated NDV vaccines with paraffin oil adjuvant and Al.St. 4- 5 mg/dose including stability(drop test, centrifugation and real time test) and viscosity (dynamic and relative) over NDV vaccineadjuvant with paraffin oil alone that showed reduced stability, however NDV vaccine with paraffin oiland Al.St. 6mg/dose and 7mg/dose showed high values of viscosity indicating hard injectability. Chicksvaccinated with inactivated NDV vaccines with paraffin oil adjuvant and Al.St. (4, 5, 6 and 7 mg/dose)gave higher cell mediated immune response (lymphocyte blastogenesis and macrophage activity),humoral immune responses (using haemagglutination inhibition test) and 100% protection uponchallenge experiment in comparison to that vaccinated with NDV vaccine with paraffin oil adjuvantalone that showed 93.3% and 86.66% protection for vaccines used immediately after preparation andthe others used after one year of storage at 4°C, respectively. Chicks vaccinated with NDV vaccineswith paraffin oil adjuvant and Al.St showed slightly lower values of both cellular and humoral immuneresponses than that of NDV vaccine adjuvant with Montanide ™ ISA 71VG. In conclusion, inactivatedNDV vaccine prepared with paraffin oil adjuvant and aluminum stearate (4 mg/dose and 5 mg/dose)were superior for that adjuvant on paraffin oil alone, as they improved stability and potency of thevaccine, and were with the same quality to that of NDV vaccine adjuvant on the imported MontanideISA 71VG.

Novel immunogenic baculovirus expressed virus-like particles of foot-and-mouth disease (FMD) virus protect guinea pigs against challenge

Vaccination is a well accepted strategy for control of foot-and-mouth disease (FMD) in endemic countries. Currently, chemically inactivated virus antigens are used for preparation of FMD vaccine. To develop a non-infectious and safe recombinant vaccine, we expressed structural polypeptide of FMDV (O/IND/R2/75) using baculovirus expression system. We show that inclusion of mutated viral 3C protease in frame with the polypeptide (P1-2A), enhanced the yield of structural proteins. The structural proteins retained antigenicity and assembled into empty virus-like particles (VLPs). Immunization of guinea pigs with purified fractions of the VLPs resulted in humoral and cell mediated immune response by 4weeks. The VLPs elicited comparable humoral immune response and relatively higher cell mediated immune response, when compared to conventional vaccine in guinea pigs. Further, up to 70% of the VLP immunized guinea pigs were protected against challenge with homologous guinea pig adapted virus. Our results highlight the application of recombinant FMDV VLPs in FMD vaccination.

Carotenoid supplementation and GnRH challenges influence female endocrine physiology, immune function, and egg-yolk characteristics in Japanese quail (Coturnix japonica)

Androgens and carotenoids circulating in plasma affect the physiology and behavior of vertebrates. Much is known about control mechanisms and functions of each of these substances, yet their interactive effects are not well understood. Here we examine possible additive, multiplicative, and interactive effects of testosterone and carotenoids on female endocrine physiology, immunocompetence, and investment in eggs by simultaneously manipulating levels of testosterone [via gonadotropin releasing hormone (GnRH) challenges] and carotenoids (via diet supplementation) in captive female Japanese quail (Coturnix japonica). Females were randomly assigned to one of four treatments: carotenoid supplementation, GnRH challenge, GnRH challenge+carotenoid supplementation, or control. Carotenoid supplementation significantly increased circulating plasma carotenoid levels and acquired immune system performance, but not innate immunity. GnRH challenges elevated circulating testosterone and carotenoid levels, and induced immunosuppression in females. However, females in the GnRH challenge+carotenoid supplementation treatment had higher cell-mediated immune responses than control females and similar responses to those of carotenoid-supplemented females. Hence, availability of carotenoids in female quail seemed to counteract immunosuppressive effects of GnRH challenges. Our results provide further evidence for synergistic effects of carotenoids and testosterone on endocrine physiology and immune function in female birds. Elevated plasma testosterone or carotenoids levels resulted in increased deposition of those compounds to eggs, respectively. Furthermore, because we found that concentrations of testosterone and carotenoids in yolks were correlated within each treatment group, differential deposition of hormones and carotenoids in eggs may not only respond to surrounding social and environmental conditions, but also to other components of the egg.

Immunogenic properties of a recombinant fusion protein containing the C-terminal 19 kDa of Plasmodium falciparum merozoite surface protein-1 and the innate immunity agonist FliC flagellin of Salmonella Typhimurium

In a recent study, we demonstrated the immunogenic properties of a new malaria vaccine polypeptide based on a 19 kDa C-terminal fragment of the merozoite surface protein-1 (MSP1 19) from Plasmodium vivax and an innate immunity agonist, the Salmonella enterica serovar Typhimurium flagellin (FliC). Herein, we tested whether the same strategy, based on the MSP1 19 component of the deadly malaria parasite Plasmodium falciparum, could also generate a fusion polypeptide with enhanced immunogenicity. The His 6FliC-MSP1 19 fusion protein was expressed from a recombinant Escherichia coli and showed preserved in vitro TLR5-binding activity. In contrast to animals injected with His 6MSP1 19, mice subcutaneously immunised with the recombinant His 6FliC-MSP1 19 developed strong MSP1 19-specific systemic antibody responses with a prevailing IgG1 subclass. Incorporation of other adjuvants, such as CpG ODN 1826, complete and incomplete Freund's adjuvants or Quil-A, improved the IgG responses after the second, but not the third, immunising dose. It also resulted in a more balanced IgG subclass response, as evaluated by the IgG1/IgG2c ratio, and higher cell-mediated immune response, as determined by the detection of antigen-specific interferon-γ secretion by immune spleen cells. MSP1 19-specific antibodies recognised not only the recombinant protein, but also the native protein expressed on the surface of P. falciparum parasites. Finally, sera from rabbits immunised with the fusion protein alone inhibited the in vitro growth of three different P. falciparum strains. In summary, these results extend our previous observations and further demonstrate that fusion of the innate immunity agonist FliC to Plasmodium antigens is a promising alternative to improve their immunogenicity.

Differences in udder health and immune response traits of Holstein-Friesians, Norwegian Reds, and their crosses in second lactation

The objective of this study was to investigate potential differences in udder health and immune response traits among Holstein-Friesian (HF), Norwegian Red (NR), and NR×HF (NRX) cows on 30 commercial Irish dairy farms. A total of 648 second-lactation cows (HF n=274, NR n=207, and NRX n=167) were immunized with hen egg white lysozyme (HEWL) to induce antibody-mediated immune response (AMIR). Candida albicans was used to induce a cell-mediated immune response, with in vivo delayed-type hypersensitivity used as the indicator. Antibody response to HEWL was measured by ELISA. Udder health defined as mean somatic cell score (SCS) over the lactation, mean SCS within 30 d of the beginning of the immunization, peak SCS for each individual cow during lactation, and incidence of mastitis were statistically superior for the NR. The NR had a greater primary AMIR, producing greater concentrations of anti-HEWL immunoglobulin G on d 14 compared with HF and NRX. No difference was observed among the breed groups in the magnitude of secondary AMIR (d 21 post-immunization) or cell-mediated immune response. The proportion of high and low responders was similar across breed groups. Cows with high AMIR and high cell-mediated immune response had significantly lower mean SCS within 30 d of the start of the immunization, but greater occurrence of clinical mastitis, recorded as a binary trait over the course of the lactation. Otherwise, no significant difference in udder health was evident between cows designated as high and low responders. Although differences in mean breed group SCS values were in line with group mean AMIR values, no association was found among the traits when correlated on an individual cow basis. Results highlight the superiority of the NR with regard to udder health and suggest that improvements to udder health may result from crossbreeding with the NR. However, the immune response traits investigated proved to be inconsistent indicators of udder health.

Circulating Corticosterone Reaction to Restraint and Adrenocorticotropin Hormone Administration in White Leghorns Selected for Immune Response Traits

Corticosterone plasma concentration was measured in a random-bred control line and in 3 White Leghorn chicken lines previously selected over 9 generations for 3 different in vivo immune responses: high antibody response to Newcastle disease virus vaccine 3 wk after vaccination (ND3), high cell-mediated immune response (response to phytohemagglutinin, PHA), and high phagocytic activity measured as carbon clearance (CC). The objective of the study was to estimate if selection on immune response had an effect on the response to stress assessed by measures of corticosterone concentration before and after physical stress or adrenocorticotropin hormone injection and if the effect was dependent on the immune response trait that had been selected for, by joint analyses of immune responses and concentrations. The mean values of plasma corticosterone measures did not differ between lines, indicating that selection for different high immune responses had little effect on response to stress. Within line, however, significant negative correlations (−0.46 ≤ r ≤ −0.39) were found between cell-mediated immunity and corticosterone plasma concentrations in 3 of the 4 lines. Moreover, in the line selected for high antibody titers (ND3-L), corticosterone levels were positively correlated to ND3 (r = 0.41 and 0.47) and negatively correlated to CC (r = −0.48).

Field evaluation of Eimeria tenella (local isolates) gametocytes vaccine and its comparative efficacy with imported live vaccine, LivaCox®

The present paper describes the field evaluation of local gametocyte vaccine and its comparative efficacy with commercial anticoccidial vaccine, LivaCox, used in breeder and broiler flocks in Pakistan. Humoral immune response in vaccinated and control chickens was monitored by enzyme-linked immunosorbent assay. Results demonstrated significantly elevated antibody titres in vaccinated groups as compared to control groups conducted both in Laboratory and field experiments. Significantly (P < 0.01) higher antibody titres in local gametocyte-vaccinated group as compared to LivaCox-vaccinated chickens were recorded. Splenic cell migration inhibition assay was used to detect the cell-mediated immune (CMI) response, and results were expressed in terms of per cent migration index. Lower per cent migration index in LivaCox-vaccinated chickens indicated the higher CMI response, as compared to local gametocyte-vaccinated chickens, although the difference was statistically non-significant (P > 0.05). Results of the challenge studies in laboratory experiments revealed significantly higher (P < 0.05) oocyst count in LivaCox-vaccinated group as compared to local gametocyte-vaccinated chickens.Maximum protection (75%) against mixed species of genus Eimeria was recorded in chickens vaccinated with gametocyte vaccines as compared to LivaCox-vaccinated group. The mean body weight gains in chickens vaccinated with local gametocyte vaccine were significantly better (P < 0.05) than in chickens vaccinated with LivaCox vaccine, both in laboratory and field experiments. Majority of the chickens (70-72%) in control group demonstrated severe lesions (3.0-4.0), while 20-26% chickens showed moderate lesions (2.0). On the other hand, local gametocyte- and LivaCox-immunized chickens developed 78% and 85% mild to moderated lesions (1.0-2.0), respectively. Results of the present study provide a probable explanation for cross-protection induced by Eimeria tenella gametocyte vaccines against other species of genus Eimeria.

Correlated effects of selection for immunity in White Leghorn chicken lines on natural antibodies and specific antibody responses to KLH and M. butyricum

BackgroundThe effect of selection for three general immune response traits on primary antibody responses (Ab) to Mycobacterium butyricum or keyhole limpet hemocyanin (KLH) was studied in four experimental lines of White Leghorn chicken. Birds underwent 12 generations of selection for one of three different general immune criteria; high antibody response to Newcastle disease virus 3 weeks after vaccination (ND3), high cell-mediated immune response, using the wing web response to phytohemglutinin (PHA) and high phagocytic activity, measured as carbone clearance (CC). Line ND3-L was selected on ND3, line PHA-L was selected on PHA, and line CC-L on CC, but all lines were measured for all three traits. The fourth line was a contemporary random bred control maintained throughout the selection experiment. Principal component analysis was used to distinguish clusters based on the overall set of immune measures.ResultsIn the KLH immunised group, no differences were present between lines for natural antibodies binding to KLH and LPS, and, lines ND3-L and PHA-L had higher titers to LTA and anti-Gal titers measured before the immunisation protocol. The measure of ND3 was correlated positively with LPS titers measured post KLH immunisation and with the difference between LPS titers measured at day 0 and 7 post immunisation. In the M. butyricum immunised group, Line ND3-L showed significantly higher specific antibody response to M. butyricum, and this result agrees well with the hypothesis that the Th-1 pathway was expected to be selected for in this line.ConclusionThis study has shown that the two different antigens KLH and M. butyricum gave rise to different responses in the set of selected lines, and that the response was only enhanced for the antigen associated with the same response mechanism as that for the trait (ND3, PHA or CC) for which the line was selected. Interactions between innate and acquired immunity have been observed mainly for the high antibody selected trait, indicating there was a specific interaction due to the selection criterion. Furthermore, the results confirmed the independence between the three selected traits. Finally, principal component analysis contributed to visually discriminate high and low responders to the two new antigens in the four lines.

Antibody Responses to Keyhole Limpet Hemocyanin, Lipopolysaccharide, and Newcastle Disease Virus Vaccine in F2 and Backcrosses of White Leghorn Lines Selected for Two Different Immune Response Traits

Planned crosses were designed to produce an F(2) and 2 backcross populations from 2 lines of White Leghorn chickens previously selected over 10 generations for 2 different in vivo immune responses. The selection criteria applied on the 2 grandparental lines were as follows: high antibody response to Newcastle disease virus vaccine 3 wk after vaccination (ND3) and high cell-mediated immune response [response to phytohemagglutinin]. Furthermore a control line was kept by random breeding. The objective of the study was to estimate if the 2 selection criteria applied on the pure lines had changed the level of and type of immune (humoral) response to a new antigen, keyhole limpet hemocyanin (KLH), in the various second-generation progeny groups. In addition, correlations between parameters of acquired and innate immunity were tested. Primary total (IgT) and isotype-specific (IgG and IgM) antibody response to KLH 1 wk after immunization and levels of natural antibodies (NAB) binding to Salmonella enteriditis-derived lipopolysaccharide (LPS) were measured. Although no differences were present between IgM and IgG antibodies to KLH and the phytohemagglutinin skin-swelling response, significant differences were present between all the progeny groups for IgT to KLH and ND3 and NAB binding to LPS. The mean values for IgT to ND3 and KLH were significantly different between the crosses using the selected lines compared with the control line, indicating a contribution of the previous selection. In addition, a sex effect was found for IgM to KLH and NAB to LPS, for which females had a higher response than males in both cases. No interaction between progeny type and sex was found. Furthermore, significant positive correlations were found between NAB to LPS and specific antibody titers to KLH. Finally, the results of the present study demonstrated an interaction between innate and acquired immunity under this strategy of selection and crossbreeding and confirmed the effect of selection on general immune response to a new antigen in second-generation crosses.

P2010 Evaluation of cellular immune response against purified antigen 85 in patients with tuberculosis

Background: Tuberculosis (TB) remains an important health problem throughout the world. Despite its significance in public health, mechanisms of protective immunity against Mycobacyerium Tuberculosis in humans have not yet been understood. Objective: To evaluate cell mediated immune response against purified Ag 85, PPD and Phytohemagglutinin (PHA) in patients with tuberculosis and healthy tuberculin positive and negative individuals. Methods: Thirty patients with tuberculosis and 60 healthy tuberculin skin test positive and negative volunteers were participated in this study. Cell mediated immunity was assessed by measuring [³H]-thymidine uptake and detection of IFN- γ in the culture supernatant using commercial ELISA test. Results: In the present study, we showed that IFN- γ production and cell proliferation response to Ag 85 were significantly higher in tuberculin positive than tuberculin negative individuals (P<0.01). Among tuberculous patients, IFN-γ production and cell proliferative responses to Ag 85 was significantly lower in contrast to healthy tuberculin positive individuals (P<0.01). In addition, IFN- γ response in patients with cavitary tuberculosis was lower than patients without cavitation (P<0.05). Conclusion: Based on the higher cell mediated immune responses to Ag 85 in healthy tuberculin positive volunteers compared to patients (especially with advanced disease), purified Ag 85 can be used as a sensitive marker for analysis of immune responses in tuberculosis.

Association of Bovine Leukocyte Antigen (BoLA) DRB3.2 with Immune Response, Mastitis, and Production and Type Traits in Canadian Holsteins

Data collected from 328 Canadian Holsteins in a research herd at the University of Guelph were used to study associations among expression of bovine leukocyte antigen (BoLA) DRB3.2 alleles, immune response, mastitis resistance via somatic cell counts (SCC), and clinical mastitis, as well as to extend these results to production and type traits. Accordingly, groups of cows were evaluated in vivo for both the antibody-mediated immune response (AMIR) and the cell-mediated immune response (CMIR), which generally predominate in responses to extracellular and intracellular pathogens, respectively. Of note was that associations between BoLA DRB3.2 alleles and immune responses tended to be in the opposite sign for the 2 AMIR and CMIR traits examined. For example, alleles DRB3.2*3 and *24 were associated with higher AMIR but lower CMIR, whereas allele *22 was associated with lower AMIR but higher CMIR. This finding is in agreement with the hypothesis that both traits are genetically independent and represent opposing type 1 and type 2 immune responses. Additionally, BoLA DRB3.2*3 and *11 were associated with lower SCC, whereas alleles *22 and *23 were associated with higher SCC. Finally, allele DRB3.2*3 was also associated with less clinical mastitis, whereas allele *8 was associated with higher mastitis risk. Allele *3 was of particular relevance because it was associated with increased antibodies, as well as reduced mastitis and SCC. This could be due to an indirect relationship between the ability to produce a high antibody response and enhanced defense against intrammamary infections caused by extracellular pathogens. Consequently, the BoLA DRB3.2*3 allele should be investigated further as a candidate for resistance to some types of intramammary infections, the important caveat being its association with lower CMIR, particularly with one of the test antigens used to evaluate delayed-type hypersensitivity. The results of associations between BoLA DRB3.2 and production traits were, in some cases, antagonistic in that BoLA DRB3.2 alleles *11 and *23, which are associated with increased production traits, were associated with lower and higher SCC, respectively. Collectively, these findings advocate the use of alleles *3, *23, and *22 as reference points for more detailed mechanistic studies. This does not imply that genetic selection for mastitis resistance should be based on BoLA alleles, but that information on a variety of genes may aid in identification and selection for improved health.

Robust HIV-specific immune responses were induced by DNA vaccine prime followed by attenuated recombinant vaccinia virus (LC16m8 strain) boost

Recombinant vaccinia virus-based vaccine combined with DNA vaccine has produced a protective immune response against HIV infection in non-human primates. In this study, we explored the immunogenicity of a recombinant vaccinia virus (LC16m8 strain), which has been used in children without severe side effects. The vaccinia virus expressing an HIV 89.6 env gene (vLC-Env) alone or combined with a DNA vaccine expressing the HIV 89.6 env gene (pCAG-Env) was characterized in BALB/c mice. Vaccination of vLC-Env induced much higher HIV-specific humoral and cell-mediated immune responses than that of pCAG-Env. Priming with pCAG-Env further enhanced vLC-Env induced immune responses, especially cell-mediated immune response. Moreover, efficient expression of Env protein was achieved following infection of bone marrow dendritic cells by vLC-Env in vitro. Administration of vLC-Env-infected dendritic cells to mice generated a high cell-mediated immune response. These results demonstrate that priming with pCAG-Env and boosting with vLC-Env represents a logical candidate for vaccination against HIV infection.

Bill colour and immunocompetence in the European blackbird

The level of expression of secondary sexual characters has been suggested to signal male ability to resist parasitic infestations. To test this idea, several studies have examined the link between sexual signals and immunocompetence in birds. However, most of them have used only a single aspect of immune response to evaluate immunocompetence. We investigated the relation between bill colour and immunocompetence in captive male European blackbirds, Turdus merula, during the breeding season by assessing both cell-mediated and humoral components of the immune system. The blackbird is a sexually dimorphic species with bill colour varying from yellow to orange in males. Humoral immunity was assessed by measuring both primary and secondary responses to sheep red blood cell inoculation. Cell-mediated immunity was estimated with a delayed cutaneous hypersensitivity response to an injection of a mitogen (phytohaemagglutinin). No relation was found between male bill colour and the primary humoral response. However, males with orange bills showed a lower secondary humoral response but a higher cell-mediated immune response than males with yellow bills. Thus, the relation between immunocompetence and a secondary sexual trait may differ markedly depending on which component of the immune system is under consideration. We discuss our results in relation to mechanisms involved in sexual selection. Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour

Evaluation of immune responses of cattle as a means to identify high or low responders and use of a human microarray to differentiate gene expression.

An immune response (IR) index to identify cows with high (H) and low (L) antibody-mediated immune responses (AMIR) had been previously devised. High AMIR associated with decreased mastitis and improved response to vaccination. Measurement of cell-mediated immune response (CMIR) was not included in the index; therefore various antigen/adjuvant combinations were evaluated as inducers of DTH to be added to the IR-index. The Bacillus Calmette Guérin (BCG)-induced/purified protein derivative (PPD)-elicited tuberculin skin test is a reliable measure of DTH; however, its use to identify livestock with high CMIR may be confounded due to previous exposure to Mycobacteria tuberculosis. DTH to BCG/PPD was therefore compared with that induced by Mycobacteria phlei (saprophyte) and its derivative phlein as the test antigen. Antibody to OVA was also evaluated. The results indicated that BCG/PPD and M. phlei/phlein induced similar DTH, but cross reaction to PPD was evident following induction of DTH using M. phlei making it a less than ideal alternative for testing livestock. Nonetheless, cows could be ranked for both AMIR and CMIR. RNA from two cows with the highest and lowest IR ranks was then used to probe a human 1.7 kD microarray to determine the ability of a human array to provide information on bovine genes associated with H and L.