Effects of vaccination on acute-phase protein response in broiler chicken.

Acute phase protein response during acute ruminal acidosis in cattle

Correlation of haemagglutinin-neuraminidase and fusion protein content with protective antibody response after immunisation with inactivated Newcastle disease vaccines

Serum Amyloid A, but Not C-Reactive Protein, Stimulates Vascular Proteoglycan Synthesis in a Pro-Atherogenic Manner

Purpose: To investigate the antibody effect of immunace on the adjuvanticity of 1,2-dioleoyl-3- trimethylammoniumpropane (DOTAP)- based liposomes.Method: The vesicles of the liposome-based ND vaccine were prepared by lipid film hydration method and physically characterized for shape, particle size and zeta potential. Forty experimental birds were divided into an unvaccinated group, a liposomal Newcastle disease (ND) vaccine group, combined liposomal ND vaccine and immunace group, and a live La Sota® vaccine group. The liposomal ND vaccine, liposomal ND vaccine and immunace and a live La Sota® vaccine groups were vaccinated orally at 3 and 6 weeks of age. Haemagglutination inhibition test was carried out after primary and booster doses.Results: The log2 mean antibody titre induced by the liposomal ND vaccine after secondary immunization of the birds was 9.60 ± 0.95 while that of the combined liposomal ND vaccine and immunace group was 7.00 ±1.71, and that of the live La Sota vaccine® was 6.00 ±0.63. There was no detectable antibody in the unvaccinated group throughout the experiment. At p < 0.05, the liposomal ND vaccine group, after secondary immunization, produced antibodies which were significantly higher than those of the combined immunace-liposomal ND vaccine group.Conclusion: There was a boost in the immune response of the birds immunized with liposomal ND vaccine and immunace after primary immunization only.Keywords: Antibody, Newcastle disease vaccine, La Sota, DOTAP, Immunace, Immunity, Adjuvant

Canine pyometra is a common reproductive disorder of intact diestrual bitches. The disease is associated with inflammation and infection of the uterus leading to generalized illness. The varied reactions of the host to infection, inflammation, or trauma are collectively known as the acute-phase response (APR) and encompass a wide range of pathophysiological responses. One of these systemic responses to disease is an increase in the production by the liver of a number of serum proteins, which are known collectively as the acute-phase proteins (APP). The most important APPs in dogs are C-reactive protein (CRP) and serum amyloid A (SAA). When diagnosed with pyometra, the preferred and most effective treatment is ovariohysterectomy, a procedure that in itself causes systemic inflammation due to the tissue trauma caused by surgery. The purpose of this study was to identify the effect pyometra and ovariohysterectomy have on acute phase proteins and routinely measured biochemistry variables in serum of affected bitches. We achieved this by evaluating concentrations of CRP and SAA, as well as of some biochemistry parameters in serum before and after ovariohysterectomy in bitches with pyometra. Blood samples were collected from the cephalic vein immediately prior to and 24h, 72h and 168 hours after initiation of surgery. The levels of CRP, SAA and some biochemistry parameters - total protein (TP), albumin (ALB), globulin (GLOB), as well as activity of some enzymes: alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKLP) and lactate dehydrogenase (LDH) were determined
 Aims: The aim of this study was to investigate the impact of pyometra and its surgical treatment, ovariohysterectomy, on acute phase proteins, specifically C-reactive protein (CRP) and serum amyloid A (SAA), as well as on routine serum biochemistry variables in affected bitches. By evaluating the concentrations of CRP and SAA, alongside key biochemistry parameters before and after surgery, this research seeks to elucidate the physiological and biochemical responses to both the disease and its treatment, providing insight into the systemic effects of pyometra and the surgical stress response in dogs.
 Study Design: The study design involved a cohort of twenty-seven female dogs, divided into two groups: seventeen clinically healthy intact female dogs serving as the control group, which were examined for routine health care or vaccinations, and ten female dogs diagnosed with pyometra at the Faculty of Veterinary Medicine, University of Sarajevo. The diagnosis of pyometra was made based on case history, clinical signs, ultrasonography, and confirmed by gross examination of the pus-filled uterus during and after ovariohysterectomy. The surgical procedure was performed using a standard mid-line approach under general inhalation anesthesia, with no post-operative complications reported.
 Place and Duration of Study: Faculty of Veterinary Medicine, University of Sarajevo, between June 2012 and September 2012.
 Methodology: Blood samples were collected from the distal cephalic vein into serum separating tubes (Vacutainer SST; Becton Dickinson, USA) for CRP and SAA measurement and serum biochemical evaluation just before the ovariohysterectomy (base values) as well as after 24 hours (day one), 72 hours (day three) and 168 hours (day seven). Blood samples from control dogs were collected only once. Serum samples were prepared by centrifugation (1.500 × g for 10 min) and stored in plain micro tubes (Eppendorf, Germany) at −20°C prior to analysis. Serum CRP concentrations in the dogs were measured with ELISA microplate reader (Anthos 2001 ELISA-reader, Anthos Mikrosysteme, Krefeld, Germany) using a commercial canine CRP enzyme-linked immunosorbent assay (ELISA) kit (Tridelta PhaseTM Range CRP Assay, Tridelta Development Limited, County Kildare, Ireland). For SAA, the analyses were performed using also a commercially available ELISA kit (Tridelta PhaseTM Range SAA Assay, Tridelta Development Limited, County Kildare, Ireland). Total protein (TP), albumin (ALB), globulin (GLOB) concentrations and alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALKP) and lactate dehydrogenase (LDH) activities were determined by using standard procedures and slides by Catalyst Dx Chemistry Analyzer, Idexx.
 Statistical analyses were performed using the SPSS software package (for Windows, Version 11.5, SPSS Inc, USA). All data were shown as mean ± standard deviation. The results were analyzed using the independent t-test. A P-value less than 0.05 were considered statistically significant.
 Results: Our results indicated significantly higher concentrations of CRP and SAA in dogs with pyometra throughout the study period compared to the control group, with CRP concentrations rising up to 15-fold and SAA up to 10-fold. Notably, these increases in CRP and SAA concentrations were mostly observed before surgery, with only limited additional increases post-surgery. This suggests that while ovariohysterectomy induces a proinflammatory response, it does not significantly escalate the levels of these proteins beyond the initial disease impact. The variability observed in CRP and SAA levels points to potential interindividual differences, impacting their diagnostic value. However, on the last postoperative day, the significantly elevated levels of both proteins in dogs with pyometra suggest a persistent, though less severe, inflammatory process, with a subsequent decline as homeostasis was gradually restored.
 Conclusion: The role of the acute phase response is to limit inflammation, remove the damaging factor and restore homeostasis. The systemic effects of pyometra are reflected by acute phase proteins and several biochemistry parameters.

Background: Tarantula cubensis alcoholic extract is used to accelerate wound healing and to relieve edema in many animal species. In addition, it may be useful for many infectious diseases. Considering to these effects, it is believe that these effects may be on immune system. Cytokines (tumor necrosis factor alpha, interleukin-1 beta, interleukin-6, interleukin-10 and interferon gamma) secreted by immune cells and acute phase proteins (haptoglobin, alpha 1 acid glycoprotein, serum amyloid A) secreted by liver play role in acute phase response. The aim of the present study was to determine the effect of Tarantula cubensis alcoholic extract on cytokine and acute phase protein levels in sheep.Materials, Methods & Results: Tarantula cubensis alcoholic extract (6 mL/sheep, subcutaneously, single dose) was administered to 6 healthy sheep. Blood samples were obtained before (0 h) and after treatments at 2, 4, 8, 12, 24 and 48 h. Then, blood samples were centrifuged to obtain serum samples. Acute phase cytokines such as serum tumor necrosis factor alpha, interleukin-1 beta, interleukin-6, interleukin-10, interferon gamma and acute phase proteins such as haptoglobin, alpha 1 acid glycoprotein and serum amyloid-A concentrations were determined with commercially available kits on ELISA reader. Administration of Tarantula cubensis alcoholic extract caused fluctuations in tumor necrosis factor alpha, interleukin-1 beta, interleukin-6, interleukin-10, interferon gamma levels in sheep. In addition, levels of haptoglobin, alpha 1 acid glycoprotein, serum amyloid A showed fluctuations. But, these fluctuations in acute phase cytokines and acute phase proteins were not statistically significant (P > 0.05).Discussion: Tarantula cubensis alcoholic extract, homeopathic medicine, is used trauma, retentio secundinarium, tendinitis, bluetongue, foot and mouth, metritis and arthritis in many animal species including sheep. Cytokines, secreted against various stimulus including infectious diseases, play role in wound healing and in the regulation of the immune system. In current study, administration of Tarantula cubensis alcoholic extract lead to fluctuations in tumor necrosis factor alpha, interleukin-1 beta, interleukin-6, interleukin-10 and interferon gamma levels, but these changes were not statistically significant (P > 0.05). Non-statistical fluctuations in cytokines result from inadequate immunological response of sheep against to Tarantula cubensis alcoholic extract. Also, use of molecular analysis techniques may be changed these results. Acute phase proteins are significantly secreted from the liver during the acute phase response. In current study, administration of Tarantula cubensis alcoholic extract in sheep caused non-statistifical fluctuations on haptoglobin, alpha 1 acid glycoprotein and serum amyloid A levels (P > 0.05). Tumor necrosis factor alpha and interleukin-1 beta stimulate synthesis of interleukin-6. Interleukin-6 provides synthesis of acute phase proteins in liver. Non-statistical fluctuations in acute phase proteins result from inadequate stimulus of IL-6. In conclusion, it may be stated that administration of Tarantula cubensis alcoholic extract has no distinctive effect on the acute phase response. However, when Tarantula cubensis alcoholic extract is administered repeated times or other acute phase parameters are evaluated, different results may be observed.

In this study, 8000 Hubbard broiler chicks having maternal antibodies to Newcastle disease (ND) and infectious bronchitis disease (IB) were reared on deep litter in a poultry farm. The chicks were received Gumbopest (IBD + Newcastle Inactivated vaccines) and Avinew (ND live vaccine –VG/GA strain) with Bioral H120 strain (IB live vaccine) via spray and drinking water to investigate its immunogenicity and its effect on broiler performance. Neither IB nor ND related clinical signs or mortalities or lesions were observed after vaccination till the end of the breeding period (7 weeks). The result of serological response to ND vaccines showed that the maternal antibody titer to NDV in 1day old chicks was 10655.5 ± 3420.9 and weaned 3516 ± 3784.4 at 2 weeks. At the 3rd and 4th weeks after vaccination ND ELISA titres were increased to 1181.5 ± 1126.4 and 2326 ± 2.117 ; respectively. On theother hand the maternal antibody titer to IB Vaccine in the used day old chicks was 8271.5 ± 10642.7; this titer waned to reach 9142.5 ± 4323.2 at 1 week of age and 92 ± 130.11 at 2 weeks after vaccination. At the 3rd and 4th weeks after vaccination IB ELISA titres were reached to 0 ± 0 and 2657 ± 3757.6 respectively. This result revealed that the used live ND and IBV vaccines were able to induce antibody levels in chickens with maternal ND and IBV antibodies. The best weekly feed conversion rate (FCR) was recorded at the 4th week (1.31) and the 6th (1.71) with total FCR of 1.89. Mortality rate in the susceptible age (2-6 week) was the lowest (0.4-.73%) with total mortality of 3.27 at the end of the 7thweek. These findings pointed out that live ND Avinew vaccine and live IB Bioral H120 vaccine were safe, immunogenic to maternally immune chicks and has no dverse effect on performance

Acute phase protein (APP) concentrations can change due to inflammation and be used to monitor disease in the Iberian ibex (Capra pyrenaica). This study aimed to validate Haptoglobin (Hp) and serum amyloid A (SAA) analytes, establish reference values, and characterize Hp and SAA responses in the Iberian ibex after experimentally induced inflammation and experimental bluetongue virus (BTV) infection. Sera from 40 free-ranging box-trapped ibexes were used to establish Hp and SAA reference values. Six healthy ibexes were subcutaneously injected with 5mL of turpentine, then, blood samples were taken, and clinical evaluations were performed on days 0, 1, 2, 3, 4, 7, and 14 postinjection. Anotherseven ibexes were challenged with BTV. Serum Hp and SAA concentrations were quantified using commercial assays following the manufacturer's instructions. Intra-assay precision and linearity were acceptable for both Hp and SAA. Intra-assay variation for high and low concentration of Hp and SAA were 9.74% and 17.31% and 16.49% and 12.89%, respectively. Inter-assay variation was higher for the low APP concentrations. Reference values for the healthy Iberian ibexes were (median, minimum, and maximum values) 0.2 (0.12-0.64) g/L for Hp and 4.74 (0.05-29.54) mg/L for SAA. Both Hp and SAA acted as a moderate and a major APP, respectively, and each could distinguish animals with turpentine-induced inflammation from those without. Hp and SAA did not change in asymptomatic BTV-infected animals. This study validated Hp and SAA analytes and provided basal reference values for these analytes in the Iberian ibex. Both APPs were able to discriminate between healthy and diseased Iberian ibexes animals during turpentine-induced inflammatory processes.

In addition to the well-recognized transport function of lipoproteins, a large body of evidence has demonstrated that lipoproteins also play an important role in host defense as part of the innate immune system (for review, see Ref. 1). One of the key defensive functions is the ability of HDL and other lipoproteins to bind endotoxin (lipopolysaccharide, LPS) and other bacterial products and neutralize their toxic effects. In this issue of the Journal of Lipid Research, Wang et al. (2) provide insights into the structural requirements for LPS neutralization by apolipoprotein A-I. The helical structure of apolipoprotein A-I is based on eight similar 22 amino acid and two 11 amino acid tandem repeats, but the areas required for HDL formation and function can now be attributed to specific regions based on studies of specific mutations (3). The central region (amino acids 144–186) activates LCAT and contributes to HDL maturation and stability. The N(44–65) and C(220–241) terminal repeats are necessary to initiate lipid binding, form nascent HDL, and remove cholesterol from macrophages. A larger portion of the C-terminal region (190–243) is critical for phospholipid binding and promoting cholesterol efflux. Naturally occurring mutations of cysteines in apolipoprotein A-I, such as A-IMilano and A-IParis, are associated with protection against atherosclerosis even when HDL cholesterol levels are decreased (4). The paper by Wang et al. (2) addresses what regions of apolipoprotein A-I are required for neutralization of LPS by substituting other amino acids for specific cysteine residues. Serine substitution of one cysteine (228) in the C-terminal domain dramatically reduced the ability of HDL to neutralize LPS, while another C-terminal substitution (cysteine 195), proximal to the last 22 residue repeat, had little effect. Midregion substitutions (cysteines 107, 129, and 173) also had little effect. On the other hand, substitution in the first N-terminal repeat (cysteine 52) and especially the next region (cysteine 74) formed HDL that was more effective at neutralizing LPS and protecting from LPS induced lung injury. Thus, Wang et al. (2) have shown that, as with cholesterol and phospholipid metabolism, specific regions of apolipoprotein A-I are essential for LPS neutralization. Furthermore, the regions involved in LPS neutralization are different than those involved in cholesterol and phospholipid metabolism. For example, these authors have previously shown (5) that substitution for cysteine residues 129 and 195 impaired lipid binding, while substitutions at 173 and 195 impaired the ability of HDL to promote cholesterol efflux. In contrast, a substitution at 107 had an increased capacity to promote cholesterol efflux. As noted above, the substitutions at 52 and 74 enhanced the ability of HDL to neutralize LPS, yet these substitutions had no effect on HDL structure or the ability of HDL to remove cholesterol from macrophages. Consequently, the increase in protection from LPS makes the 52 and 74 substitutions “super” apolipoproteins A-Is for host defense, with little downside in adversely affecting reverse cholesterol transport and increasing the risk of atherosclerosis. It remains to be seen whether similar naturally occurring mutations occur in humans and whether such mutations in apolipoprotein A-I will have a beneficial effect during gram-negative infections. Additionally, studies should examine whether similar modifications of apolipoprotein A-I will also enhance the neutralization of other toxic bacterial products, such as lipoteichoic acid. Infections activate Toll-like receptors stimulating the secretion of cytokines, which have profound effects on lipid and lipoprotein metabolism (for review, see Ref. 1). The changes in lipid and lipoprotein metabolism are part of the acute phase response (APR), a pattern best known for increases in serum proteins (6). Positive APR proteins are those whose circulating levels increase during the APR while negative APR proteins decrease. Two positive APR proteins, C-reactive protein and serum amyloid A, bind to lipoproteins and hence can be considered to be apolipoproteins. The increases in serum proteins during the APR are transcriptionally mediated and usually driven by activation of transcription at NF-kB and NF-IL-6 response elements (6). However, many of the changes in lipid and lipoprotein metabolism are part of the negative

Tumor necrosis factor-α and acute-phase proteins in early pregnant ewes after challenge with peptidoglycan-polysaccharide

Due to the weak immunogenicity, traditional inactivated vaccines need to use adjuvants and multiple doses to generate effective immune protection. Therefore, in theory and practice, it is valuable to develop a vaccine delivery system with an adjuvant effect, which can be carried out in a single dose. In view of this, inactivated Newcastle disease virus (NDV)-loaded poly (lactic-co-glycolic acid) (PLGA) microspheres with different release characteristics were prepared. Two release peaks of four groups of microspheres were observed during in vitro release test. The first release peak came out on the first day. The second release peak of A microspheres appeared at the 7th week. The second release peak of B microspheres appeared at the 5th week. The second release peaks of C and D microspheres all appeared at the 11th week. The release of C microspheres contrast to D microspheres was more smooth at release peak, so A, B, C microspheres were selected to construct the pulsatile system. Thus inactivated Newcastle disease(ND) vaccine pulsatile release systems using an appropriate combination of vaccine microspheres with a suitable release characteristics was constituted . Then their immune effects on chickens were evaluated. Chickens without maternal antibodies against NDV were grouped randomly and then immunized with NDV-PLGA microsphere vaccine and inactivated ND vaccine respectively. The results show that inactivated NDV-PLGA microsphere vaccine had the effect of pulsatile release in chickens and were able to maintain high and constant antibody level. And the pulsatile release system composed of mixed microspheres had the best immune effect. This study demonstrated that inactivated NDV-PLGA microsphere pulsatile release system with reasonable release interval could realize effective immune protection in a long time by a single injection. This was the first ever report on PLGA microspheres-based pulsatile release system for inactivated NDV vaccine.

Interleukin-18 (IL-18) has been known to induce interferon-γ (IFN-γ) production and promote Th1 immunity. Although mammalian IL-18 has been characterized in great detail, the properties and application of chicken IL-18 remain largely uninvestigated as of yet. In this study, we evaluated the immunomodulatory properties of Salmonella enterica serovar Typhimurium expressing chicken interleukin-18 (chIL-18) on immune responses induced by avian influenza (AI) and Newcastle disease (ND) vaccines. After oral administration of S. enterica serovar Typhimurium expressing chIL-18, chickens were vaccinated intramuscularly with the recommended dose of either inactivated AI H9N2 vaccine or ND (B1 strain) vaccine. Chickens receiving a primary vaccination were boosted using the same protocol 7 days later. Humoral and cell-mediated immune responses were evaluated in terms of HI antibody titers and proliferation and mRNA expression of IFN-γ and IL-4 of peripheral blood mononuclear cells (PBMC) in response to specific antigen stimulation. According to our results, oral administration of S. enterica serovar Typhimurium expressing chIL-18 induced enhanced humoral and Th1-biased cell-mediated immunity against AI and ND vaccines, compared to that of chickens received S. enterica serovar Typhimurium harboring empty vector. Therefore, we conclude that our proposed vaccination regimen using inactivated AI and ND viruses along with oral administration of S. enterica serovar Typhimurium expressing chIL-18 may provide a novel approach in protecting chicken from currently circulating AI and ND virus strains.

Clindamycin is a lincosamide antibiotic that binds primarily to alpha 1-acid glycoprotein (AAG), an acute-phase serum protein. Many studies have shown that AAG concentrations increase in response to stress, including infection, myocardial infarction, and trauma. The objectives of this study were to determine the serum protein binding of various clindamycin concentrations in sera with normal and elevated AAG concentrations. Serum was obtained from 4 healthy volunteers and 12 patients with pathophysiologic conditions known to elevate serum AAG concentrations. Timing for collection was determined from the literature, corresponding with the expected peak concentration for each disease state. Samples were assayed for AAG by radial immunodiffusion and were spiked with clindamycin to achieve total concentrations of 10 micrograms/ml (n = 18), 4 micrograms/ml (n = 10), and 2 micrograms/ml (n = 7). Protein binding was determined by ultrafiltration and subsequent high-performance liquid or gas chromatography. Protein binding was dependent on the serum concentrations of both AAG and clindamycin. When AAG concentrations increased from 101-150 mg/dl to 201 mg/dl or greater, mean protein binding increased from 81.2% to 92.4% (p = 0.1265) and from 61.3% to 88.6% (p less than 0.05) at clindamycin concentrations of 2 and 4 micrograms/ml, respectively. With AAG concentrations between 101 and 150 mg/dl, mean protein binding increased from 62.4% at 10 micrograms/ml to 81.2% at 2 micrograms/ml (p = 0.1514). Since AAG concentrations may increase in certain patients, the concentration of free (pharmacologically active) drug may fall below the minimum inhibitory concentration for several pathogens earlier in a dosing interval.

To investigate the correlation of serum long noncoding RNA-metastasis associated lung adenocarcinoma transcript 1(LncRNA MALAT1) and serum amyloid A(SAA) with diabetic kidney disease. Retrospective research was used, and 40 patients with type 2 diabetes and 80 patients with type 2 diabetic kidney disease patients who were treated in Tianjin Medical University Chu Hsien-I Memorial Hospital from August 2021 to February 2022 were selected, and 40 healthy subjects were selected during the same period. Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect serum LncRNA MALAT1. SAA were detected with enzyme linked immunosorbent assay (ELISA). Automatic biochemistry analyzer was used to detect serum creatinine (CREA) and low-density lipoprotein cholesterol(LDL-C),automatic blood glucose analyzer to detect serum fasting plasma glucose (FPG), automatic glycated hemoglobin analyzer to detect hemoglobin A1C (HbA1c), and automatic immunoassay analyzer to detect urinary albumin to creatinine ratio(UACR). Differences between groups were compared by t test and analysis of variance. Pearson analysis was used to analyze the correlation between MALAT1, SAA and other indicators. Receiver operating characteristic curve(ROC) was used to evaluate the auxiliary diagnostic value of MALAT1 and SAA for diabetic kidney disease. The results showed that MALAT1 and SAA in the diabetic kidney disease with mass albuminuria group were higher than those in the type 2 diabetes mellitus group (q=8.57, P<0.01; q=11.09, P<0.01) and the diabetic kidney disease with microalbuminuria group (q=3.96, P<0.05; q=7.85, P<0.01). MALAT1 had a high correlation with UACR, CREA, SAA, HbA1c and FPG (r value was 0.706, 0.643, 0.578, 0.553, and 0.524, all P<0.01), and SAA had a high correlation with UACR, HbA1c and FPG (r value was 0.664, 0.617, and 0.595, all P<0.01). ROC curve analysis of the diagnostic value of LncRNA MALAT1 and protein SAA for diabetic kidney disease showed that the areas under curve (AUC) were 0.741 and 0.744, respectively. The combined diagnostic value of the two was the greatest (AUC=0.801). In summary, MALAT1 and SAA were elevated in the serum of patients with type 2 diabetes. Their concentrations in the serum of group with diabetic kidney disease were higher than that in the type 2 diabetes group, and the serum concentrations of MALAT1 and SAA in group with mass albuminuria are higher than the group with microalbuminuria. MALAT1 and SAA were both closely related to UACR and HbA1c, and there is a correlation between them. Both of them may have ancillary diagnostic value for diabetic kidney disease.

SAA, HDL biogenesis, and inflammation