Proliferation Of Peripheral Lymphocytes Research Articles

Effects of selenylation modification on immune-enhancing activity of garlic polysaccharide.

The garlic polysaccharide was modified by HNO3-Na2SeO3 method according to orthogonal design L9(34) to obtain nine selenizing garlic polysaccharides, sGPS1-sGPS9. Their effects on chicken peripheral lymphocytes proliferation in vitro were compared by MTT assay. The results showed that sGPSs could significantly promote lymphocytes proliferation, sGPS3, sGPS5 and sGPS6 presented stronger efficacy. In vivo experiment, 14-day-old chickens were injected respectively with sGPS3, sGPS5 and sGPS6 when they were vaccinated with ND vaccine taking unmodified GPS as control. The results showed that three sGPSs could significantly promote lymphocyte proliferation, enhance serum antibody titer, IFN-γ and IL-2 contents. These results indicated that selenylation modification could significantly enhance the immune-enhancing activity of GPS, sGPS6 possessed the best efficacy and could be as a candidate drug of immunoenhancer. Its optimal modification conditions were 400 mg of sodium selenite for 500 mg of GPS, reaction temperature of 70°C and reaction time of 6 h.

Sulfation of Agrocybe chaxingu polysaccharides can enhance the immune response in broiler chicks

SUMMARY In this study, our objective was to compare the effects of sulfated Agrocybe chaxingu polysaccharide (saCP) and nonsulfated aCP (naCP) on lymphocyte proliferation in vitro and immune-enhancing activity in broiler chicks. In the in vitro test, 3 saCP with different degrees of sulfation—saCP0.83, saCP1.32, and saCP 1.79—were used to study the peripheral lymphocyte proliferation using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay with naCP as the control. On the basis of this test, saCP 1.32 and saCP 1.79 were selected for the in vivo test. In the in vivo test, 300 one-day-old chicks were randomly divided into 5 groups (negative control, vaccine control, naCP, saCP 1.32, and saCP 1.79). Chicks in the vaccine control and aCP groups were vaccinated with newcastle disease (nD) vaccine at 10 d of age, with a booster dose at 21 d of age. at the time of the first vaccination, the chicks in the 3 aCP groups were injected with naCP, saCP 1.32, and saCP 1.79, whereas those in the vaccine control group were injected with 0.5 ml of physiological saline once a day for 3 successive days. In comparison to naCP, saCP 1.32 and saCP 1.79 significantly increased the relative weight of the thymus and spleen at 22 d of age, enhanced serum nD antibody titers at 16, 22, 28, 34, and 42 d of age, and enhanced splenic t- and B-lymphocyte proliferation at 22 and 28 d of age. Furthermore, saCP 1.32 and saCP 1.79 could induce significant augmented levels of interferon-γ and Il-6. Sulfation could enhance both cellular and humoral immunity in broiler chicks on a degrees of sulfation basis. the saCP 1.79 exerted the best efficacy and would be expected to be immunopotentiator molecule during nD vaccination in broilers.

Adjuvanticity of compound astragalus polysaccharide and sulfated epimedium polysaccharide per os

On the basis of previous researches, compound astragalus polysaccharide (APS) and sulfated epimedium polysaccharide (sEPS) oral liquid (AEO) was prepared. Three hundred and twenty 14-day-old chickens were randomly assigned into eight groups and vaccinated with ND vaccine except for blank control (BC) group, repeated vaccination at 28 days old. At the same time of each vaccination, the chickens in three experimental groups were taken orally with AEO, respectively, at three doses, in two component control groups with APS and sEPS, once a day for three successive days; in injection control group were injected with AEI once, and in vaccination control (VC) and BC groups were not administrated. On days 7, 14, 21, 28 and 35 after the first vaccination, peripheral lymphocyte proliferation, the serum antibody titer, IFN-γ and IL-2 concentrations and on day 35 immune organ index were measured. The results showed that AEO at high and medium doses could significantly promote lymphocyte proliferation and development of immune organ, enhance antibody titer and IFN-γ and IL-2 concentration, which was stronger than actions of AEI and two components. The results confirmed that AEO possessed reliable immunoenhancement and could be exploited into an oral immunopotentiator.

Effects of composite antimicrobial peptides in weanling piglets challenged with deoxynivalenol: I. Growth performance, immune function, and antioxidation capacity1

The mycotoxin deoxynivalenol (DON) is a food contaminant that leads to reduced feed intake and reduced BW gain, as well as organ impairment. On the other hand, antimicrobial peptides have been shown to have positive effects on growth performance, nutrient digestibility, and immune function. The purpose of this study was to investigate the protective effects of composite antimicrobial peptides (CAP) on piglets challenged with DON. After a 7-d adaptation period, 28 individually housed piglets (Duroc × Landrace × Large Yorkshire) weaned at 28 d of age were randomly assigned to receive 1 of 4 treatments (7 pigs/treatment): negative control, basal diet (NC), basal diet + 0.4% CAP (CAP), basal diet + 4 mg/kg DON (DON), and basal diet + 4 ppm DON + 0.4% CAP (DON + CAP). On d 15 and 30 after the initiation of treatment, blood samples were collected for the determination of blood profile. Piglets were monitored for 30 d to assess performance and then were slaughtered to obtain organs for the determination of the relative weight of organs. The results showed that dietary supplementation with DON decreased (P < 0.05) ADFI, ADG, and G:F, whereas dietary supplementation with CAP improved ADG and G:F (P < 0.05). The relative weight of the kidney and pancreas was greater and the relative weight of the spleen was lighter in the DON treatment than in the other 3 treatments (P < 0.05). There were no effects (P > 0.05) on other relative weights of viscera, except the relative weight of the gallbladder, but the diamine oxidase activity in the liver decreased in DON-treated piglets (P < 0.05). Piglets in the DON treatment had increased serum concentrations of alkaline phosphatase, alanine transaminase, and aspartate aminotransferase and a dramatic decrease in total protein (P < 0.05), whereas there were no differences (P > 0.05) between the DON + CAP treatment and the other treatments. The DON treatment decreased the numbers of red blood cells and platelets, as well as the serum catalase concentrations, and decreased the serum concentrations of H2O2, maleic dialdehyde, and nitric oxide (P < 0.05). The numbers of platelets and thrombocytocrit, as well as the serum concentrations of catalase, were greater, whereas the maleic dialdehyde concentrations were decreased, in both the CAP and DON + CAP treatments compared with the other treatments (P < 0.05). Compared with the control treatment, DON decreased peripheral lymphocyte proliferation on d 15, whereas supplementation with CAP increased it on d 15 and 30 (P < 0.05). These findings indicate that CAP could improve feed efficiency, immune function, and antioxidation capacity and alleviate organ damage, and thus, it has a protective effect in piglets challenged with DON.

Leukocyte Proliferation and Immune Modulator Production in Patients with Chronic Kidney Disease

IntroductionIn Chronic Kidney Disease (CKD), immune cells are affected by uremic retention toxins. Given this effect, we analyzed lymphocyte proliferative response and immune modulators production following in vitro stimulation.MethodsWhole blood was drawn from healthy controls, patients with eGFR <20 ml/min/1.73 m2 (Pre-dialysis, CKD stages 4 and 5) and hemodialysis patients (stage 5D). Peripheral cells were incubated for six days with pokeweed mitogen, concanavalin A, Staphylococcus enterotoxin A or influenza A vaccine. Peripheral lymphocyte proliferation was then analyzed by the “Flow-cytometric Assay of Specific Cell-mediated Immune response in Activated whole blood” (FASCIA) method, and cytokine profile in the cell supernatants was analyzed by the Milliplex multi-array method.ResultsThe absolute number of lymphoblasts in response to mitogenic stimulation and the number of cells in each CD4+ and CD8+ subpopulation were similar comparing the three groups, except for a single decline in number of lymphoblasts after stimulation with Staphylococcus enterotoxin A, comparing dialysis patients with healthy controls. Levels of interleukin (IL)-2 (p=0.026), -10 (p=0.019) and -15 (p=0.027) in the Staphylococcus enterotoxin A-stimulated supernatant were lower in hemodialysis patients compared to healthy controls. Levels of IL-15 (p=0.017) from pre-dialysis patients and levels of IL-5 (p=0.019) from hemodialysis patients in influenza A vaccine-stimulated supernatants were also lower compared to controls. In pokeweed mitogen–stimulated supernatant, IL-2 levels (p=0.013) were lower in hemodialysis patients compared to pre-dialysis patients. TNF-α, IL-10, IL-12, IL-15, IL-8, MCP-1, IP-10, IFN-α2, IL-1α and eotaxin levels were all significantly higher in plasma obtained from CKD patients.ConclusionOur results suggest that T-cells from CKD patients have similar proliferative response to stimulation compared with healthy individuals. Moreover, however the immune cells show inability to produce selected cytokines, most likely due to the uremic milieu or dialysis procedure.

The comparison of immune-enhancing activity of sulfated polysaccharidses from Tremella and Condonpsis pilosula

Based on our previous research, four sulfated polysaccharide (sPSs) from Tremella and Condonpsis pilosula, sTPStp, sTPS70c, sCPPStp and sCPPS50c, were prepared and their effects on splenic lymphocytes proliferation in vitro and the immune response of ND vaccine in chicken were compared taking the unmodified polysaccharide (uPS) TPStp as control. The results showed that four sPSs could significantly or numerically stimulate splenic lymphocyte proliferation singly or synergistically with LPS in vitro, sTPS70c and sCPPStp demonstrated better effect; promote peripheral lymphocytes proliferation and enhance serum HI antibody titer in chickens vaccinated with ND vaccine, the actions of sPSs were stronger than that of uPS, and sTPS70c at medium dosage presented the best efficacy. These indicated that sulfation modification could improve the immune-enhancing activity of TPS and CPPS, sTPS70c possessed the strongest activity and would be expected as a component of new-type immunopotentiator.

Cordyceps militaris polysaccharides can improve the immune efficacy of Newcastle disease vaccine in chicken

Cordyceps militaris polysaccharide (CMP) was prepared by water decoction and ethanol precipitation. The fractional CMP40 and CMP50 were extracted from the CMP solution by stepwise precipitation with ethanol at 40% and 50% of working concentration, respectively. The immune-enhancing activities of two polysaccharides were researched. In vitro experiment, the effects of two polysaccharides on chicken peripheral lymphocyte proliferation were determined by MTT assay. The result displayed that two polysaccharides could significantly stimulate lymphocyte proliferation, the action of CMP40 was significantly or numerically stronger than those of CMP50. In vivo experiment, 320 14-day-old chickens were averagely divided into eight groups. The chickens, except blank control (BC) group, were vaccinated with Newcastle disease vaccine, repeated vaccination at 28 days old. At the same time of the first vaccination, the chickens in three CMP40 fraction groups and three CMP50 fraction groups were injected respectively with the polysaccharide at low, medium and high concentrations, in vaccination control (VC) and BC group, with equal volume of physiological saline, once a day for three successive days. On days 7, 14, 21, 28, 35 and 42 after the first vaccination, the lymphocyte proliferation, serum antibody titre and interferon-gamma and interleukin-4 were measured. The results showed that CMP40 and CMP50 at suitable dose could significantly promote lymphocyte proliferation, enhance serum antibody titre, and improve serum interferon-gamma and interleukin-4 concentrations. It indicated that CMP40 fraction and CMP50 fraction could significantly improve the immune efficacy of Newcastle disease vaccine, and would be as the candidate of a new-type immune adjuvant.

Perthamide C Inhibits eNOS and iNOS Expression and Has Immunomodulating Activity In Vivo

Here we have characterized perthamide C, a cyclopeptide from a Solomon Lithistid sponge Theonella swinhoei, which displays an anti-inflammatory/immunomodulatory activity. The study has been performed using the carragenan-induced mouse paw edema that displays an early (0–6 h) and a late phase (24–96 h). Perthamide C significantly inhibits neutrophils infiltration in tissue both in the early and late phases. This effect was coupled to a reduced expression of the endothelial nitric oxide synthase (eNOS) in the early phase while cyclooxygenase-1 and 2 (COX-1, COX-2), and inducible NOS (iNOS) expression were unaffected. In the late phase perthamide C reduced expression of both NOS isoforms without affecting COXs expression. This peculiar selectivity toward the two enzymes deputed to produce NO lead us to investigate on a possible action of perthamide C on lymphocytes infiltration and activation. We found that perthamide C inhibited the proliferation of peripheral lymphocytes, and that this effect was secondary to its metabolic activation in vivo. Indeed, in vitro perthamide C did not inhibit proliferation as opposite to its metabolite perthamide H.In conclusion, perthamide C selectively interferes with NO generation triggered by either eNOS or iNOS without affecting either COX-1 or COX-2. This in turn leads to modulation of the inflammatory response through a reduction of vascular permeability, neutrophil infiltration as well as lymphocyte proliferation.

Selenylation modification can enhance immune-enhancing activity of Chinese angelica polysaccharide

On the basis of previous test that selenizing Chinese angelica polysaccharides (sCAPs) with stronger immune-enhancing activity in vitro were picked out, the immune-enhancing activity in vivo of three sCAPs, sCAP2, sCAP6 and sCAP8, at high and low dosage were compared taking the unmodified Chinese angelica polysaccharide (CAP) as control by determination of peripheral lymphocyte proliferation, serum antibody titer, IFN-γ and IL-6 contents in chicken vaccinated with Newcastle Disease vaccine. The results showed that three sCAPs at suitable dosage could significantly promote lymphocyte proliferation, enhance serum antibody titer, IFN-γ and IL-6 contents as compared with unmodified CAP, sCAP2 at low dosage possessed the strongest action. These results indicated that selenylation modification could significantly enhance the immune-enhancing activity of CAP, sCAP2 possessed the best efficacy and would be as a component drug of new-type immunoenhancer.

NFATc1 Induction in Peripheral T and B Lymphocytes

NFAT transcription factors control the proliferation and survival of peripheral lymphocytes. We have reported previously that the short isoform NFATc1/αA whose generation is induced by immune receptor stimulation supports the proliferation and inhibits the activation-induced cell death of peripheral T and B cells. We will show in this study that in novel bacterial artificial chromosome transgenic mice that express EGFP under the control of entire Nfatc1 locus the Nfatc1/Egfp transgene is expressed as early as in double-negative thymocytes and in nonstimulated peripheral T and B cells. Upon immune receptor stimulation, Nfatc1/Egfp expression is elevated in B, Th1, and Th2 cells, but only weakly in T regulatory, Th9, and Th17 cells in vitro whose generation is affected by TGFβ. In naive lymphocytes, persistent immune receptor signals led to a 3-5 increase in NFATc1/αA RNA levels during primary and secondary stimulation, but a much stronger induction was observed at the protein level. Whereas anti-CD3(+)CD28 stimulation of primary T cells induces both NFATc1/αA and their proliferation and survival, anti-IgM stimulation of B cells induces NFATc1/αA and proliferation, but activation-induced cell death after 3-d incubation in vitro. The anti-IgM-mediated activation-induced cell death induction of B cells in vitro is suppressed by anti-CD40-, LPS-, and CpG-mediated signals. In addition to inducing NF-κB factors, together with anti-IgM, these signals also support the generation of NFATc1/αA. According to these data and the architecture of its promoter region, the Nfatc1 gene resembles a primary response gene whose induction is affected at the posttranscriptional level.

Transplantation-potential-related biological properties of decidua basalis mesenchymal stem cells from maternal human term placenta

Human placental decidua basalis originates from the maternal side of the placenta and has been described as a source of mesenchymal stem cells (MSCs). However, for its application in tissue regeneration and repair, the transplantation-potential-related biological properties of decidua-basalis-derived mesenchymal stem cells (DBMSCs) remain to be elucidated. We obtained DBMSCs through enzymatic digestion and density gradient centrifugation and confirmed their capacity to differentiate into cell types of the mesodermal lineage, such as osteoblasts, adipocytes and chondroblasts. Karyotype analysis showed that the isolated DBMSCs maintained chromosomal stability after long-term culture in vitro. Growth kinetics and ultrastructural observation revealed a high level of DBMSC proliferative activity. In addition, DBMSCs showed immunosuppressive properties by suppressing both mitogen- and alloantigen-induced peripheral lymphocyte proliferation. All of these properties suggest that DBMSCs, which are abundant and easily accessible, are a novel potential source of seed cells for cell transplantation treatments.

Effect of epimedium polysaccharide-propolis flavone immunopotentiator on immunosuppression induced by cyclophosphamide in chickens

Two hundred and fifty 11-day-old chickens were randomly assigned into 5 groups and except normal control group injected with cyclophosphamide once a day for 3 successive days. At day-14-old, all chickens were vaccinated with Newcastle disease vaccine. At the same time of the first vaccination, the chickens in three experimental groups were injected respectively with epimedium polysaccharide-propolis flavone immunopotentiator (EPI) at three dosages, once a day for 3 successive days. On days 7, 14, 21 and 28 after the first vaccination, the serum antibody titer and IgG, IgM, IFN-γ and IL-6 concentrations, peripheral lymphocyte proliferation, including immune organ index on day 28, were measured. The results demonstrated that EPI at high and medium doses could significantly enhance antibody titer and IgG, IgM, IFN-γ and IL-6 concentrations, promote lymphocyte proliferation and enlarge immune organ index as compared with model control group. This indicated that EPI could effectively resist the immunosuppression.

Higher Sensitivity of Peripheral Blood Lymphocytes to Endogenous Glucocorticoid in Renal Transplant Recipients Treated with Tacrolimus, as Compared to Those Treated with Cyclosporine

Background: Steroids in maintenance immunosuppressive drugs produce severe adverse events, and therefore, the use of steroids is withdrawn for many renal transplant recipients. However, steroids need to be restarted to prevent the worsening of renal function because of rejection episodes in some patients or in patients who experience steroid withdrawal syndrome. Currently, there are no indicators to predict which patients can sustain steroid withdrawal. Lymphocyte sensitivity to endogenous glucocorticoid cortisol could be a biological marker for safety reduction and steroid withdrawal. We examined relationship between lymphocyte sensitivity to cortisol in vitro and clinical outcome after steroid reduction or withdrawal in the renal transplant recipients. We also compared peripheral lymphocyte sensitivity to cortisol between transplant recipients treated with tacrolimus (Tac) and those treated with cyclosporine. Methods: The suppressive efficacies of cortisol against T-cell mitogen stimulated proliferation of peripheral lymphocytes was investigated in totally 41 patients with renal tramsplant patients, who were reducedor withdrawnsteroid. Nineteen out of the 41 patients were treated with Tac and the other 22 patients with cyclosporine A (CyA). Relationship between lymphocyte sensitivity to cortisol invitro and clinical outcome after steroid reduction or withdrawal was evaluated in 30 out of 41 renal transplant recipients. The lymphocyte sensitivity to cortisol was compared between 19 patients treated with Tac and 22 patients treated with CyA. Results: The cortisol IC50 values in the Tac and CyA groups were 0.09 ± 0.12 and 14.2 ± 12.7ng/ml, respectively. Lymphocyte sensitivity to cortisol in the Tac-treated group was significantly higher than that in the CyA treated group (p =0.0283). On the other hand, incidences of steroid withdrawal syndrome and increase in serum creatinine concentration were not significantly different between the Tac and CyA groups. Lymphocyte sensitivity to cortisol and its relationship with the clinical outcome after steroid reduction and withdrawal was investigated in 30 long stable renal transplant recipients. Serum creatinine levels increased in a significantly higher number of patients with lymphocytes hyposensitive to cortisol (IC50 ≥ 10000 ng/mL) than in normally sensitive patients (IC50 < 10000 ng/mL) (P < 0.005). The incidences of steroid withdrawal syndrome (P< 0.05) and necessity for increasing steroid dose or restarting steroid administration (P< 0.005) were also higher in the patients hyposensitive to cortisol than in normally sensitive patients. The patients in whom the lymphocyte proliferation rate was less than 60% did not show increase in S-Cr levels, experience steroid withdrawal symptoms, or require an increase in the steroid dose or restart of steroid administration. Conclusion: We considered from these observations that the lymphocyte sensitivity to cortisol would be useful for selecting patients for whom steroids should be withdrawn, and as a biomarker to safely reduce and withdraw steroids. Lymphocyte sensitivity to cortisol was higher in the Tac treated patients than that in the CyA treated ones. Since the cortisol sensitivity of peripheral lymphocytes is suggested to be a predictive marker for safe steroid withdrawal as described above, Tac administration shows promise in aiding successful withdrawal of steroid in long-term renal transplant recipients.

Immune-enhancing activity comparison of sulfated ophiopogonpolysaccharide and sulfated jujube polysaccharide

The immune-enhancing activities of four sulfated polysaccharides, sOPSt, sOPS80, sJPSt and sJPS50 picked out in our previous researches, were compared taking four corresponding unmodified polysaccharides as control. In vitro experiment, the effects of eight polysaccharides on chicken peripheral lymphocyte proliferation were determined by MTT assay. The result displayed that four sulfated polysaccharides could significantly stimulate lymphocyte proliferation, their actions were significantly or numerically stronger than those of corresponding unmodified polysaccharides, sOPS80 presented better efficacy. In vivo experiment, 300 14-day-old chickens were averagely divided into ten groups. The chickens except blank control (BC) group were vaccinated with Newcastle disease vaccine, repeated vaccination at 28 days old. At the same time of the first vaccination, the chickens in eight polysaccharides groups were injected respectively with 0.5ml (1mg) of eight polysaccharides, in vaccination control (VC) and BC group, with 0.5mL of physiological saline, once a day for three successive days. On days 7, 14, 21 and 28 after the first vaccination, the peripheral lymphocyte proliferation and serum ND antibody titer were determined. The result showed that four sulfated polysaccharides could significantly promote lymphocyte proliferation and enhance serum antibody titer. Their actions were significantly or numerically stronger than those of corresponding unmodified polysaccharides, sOPS80 possessed the best efficacy. These results indicated that sulfation modification could enhance the immune-enhancing activity of OPS and JPS, sOPS80 possessed the best efficacy and would be expected as a component drug of new-type immunopotentiator.

Optimization of selenylation conditions for Chinese angelica polysaccharide based on immune-enhancing activity

Chinese angelica polysaccharide (CAP) was extracted by water decoction and ethanol precipitation, purified through eliminating protein by Sevage method and column chromatography of Sephadex G-200, then selenizingly modified by nitric acid–sodium selenite method according to L9(34) orthogonal design of three-factors, the usage amount of sodium selenite, reaction temperature and reaction time, at three level to obtain nine selenizing CAPs, sCAP1–sCAP9. Their effects on chicken peripheral lymphocytes proliferation in vitro were compared by MTT assay taking the non-modified CAP as control. The results showed that selenylation modification could significantly enhance the immune-enhancing activity of CAP, sCAP2 presented best effect and the optimal modification conditions were 200mg of sodium selenite for 500mg of CAP, the reaction temperature of 70°C and the reaction time of 6h.

Immuno-enhancing activity of sulfated Auricularia auricula polysaccharides

The crude total Auricularia auricula polysaccharide (AAPct) was extracted by water decoction and ethanol precipitation, protein was removed to obtain total A. auricula polysaccharide (AAPt), then was graded into AAP1 and AAP2 through column chromatography. sAAPt, sAAP1 and sAAP2 were prepared by chlorosulfonic acid-pyridine method. In vitro test, the effects of sAAPt, sAAP1, sAAP2, AAPt, AAP1 and AAP2, on chicken peripheral lymphocytes proliferation were compared. The results showed that sAAPt and sAAP1 demonstrated better effect. In vivo test, 14-day-old chickens were injected respectively with sAAPt, sAAP1, AAPt and AAP1 at the first vaccination of ND vaccine, once a day for three days. On days 7, 14, 21 and 28 after the first vaccination, the peripheral lymphocytes proliferation and antibody titer were determined. The results indicated that sAAPt possessed the best efficacy and would be expected to be used as a component of a new-type immunopotentiator.

Higher Sensitivity of Peripheral Blood Lymphocytes to Endogenous Glucocorticoid in Renal Transplant Recipients Treated With Tacrolimus, as Compared to Those Treated With Cyclosporine.

Lymphocyte sensitivity to endogenous glucocorticoid cortisol could be a biological marker for safe reduction and withdrawal of steroids in renal transplant recipients. We compared peripheral lymphocyte sensitivity with cortisol between transplant recipients treated with tacrolimus (Tac) and those treated with cyclosporine. The suppressive efficacies of cortisol against T-cell mitogen-stimulated proliferation of peripheral lymphocytes were investigated in 44 renal transplant patients, who either had reduced or been withdrawn from steroid treatment. Twenty of the 44 patients were treated with Tac, and the other 24 patients were treated with cyclosporine A (CyA). The lymphocyte sensitivity to cortisol was compared between these two patient groups. The cortisol IC50 values in the Tac and CyA groups were 0.09 ± 0.12 and 14.2 ± 12.7 ng/ml, respectively. Lymphocyte sensitivity to cortisol in the Tac-treated group was significantly higher than that in the CyA-treated group (p = 0.0283). On the other hand, incidences of steroid withdrawal syndrome and increases in serum creatinine concentration were not significantly different between the Tac and CyA groups. Lymphocyte sensitivity to cortisol was higher in the Tac-treated patients than that in the CyA-treated ones. Since the cortisol sensitivity of peripheral lymphocytes is suggested to be a predictive marker for safe steroid withdrawal, Tac administration shows promise in aiding successful withdrawal of steroid treatment in long-term renal transplant recipients.

Comparison of the Safety and Efficacy of a New Live Salmonella Gallinarum Vaccine Candidate, JOL916, with the SG9R Vaccine in Chickens

We evaluated a recently developed live vaccine candidate for fowl typhoid (FT)-JOL916, a lon/cpxR mutant of Salmonella Gallinarum (SG)-by comparing its safety and efficacy with that of the well-known rough mutant strain SG9R vaccine in 6-wk-old Hy-Line hens. Forty-five chickens were divided into three groups of 15 chickens each. The chickens were then intramuscularly inoculated with 2 x 10(7) colony-forming units (CFUs) of JOL916 (JOL916 group), 2 x 10(7) CFUs of SG9R (SG9R group), or phosphate-buffered saline (control group). After vaccination, no clinical symptoms were observed in any of the groups. No differences in body weight increase were detected among the three groups postvaccination. A cellular immune response was observed at 2 wk postvaccination (wpv) in the JOL916 group with the peripheral lymphocyte proliferation assay, whereas no response was detected in the SG9R group. Elevation of SG antigen-specific plasma immunoglobulin was observed 2 and 3 wpv in the JOL916 and SG9R vaccine groups, respectively. After virulent challenge on day 25 postvaccination, 0, 1, and 15 chickens in the JOL916 group, SG9R group, and control group, respectively, died by 12 days postchallenge; the death rate of the SG9R vaccine group was statistically similar to that of the JOL916 group. Postmortem examination revealed that the JOL916 vaccine offered more efficient protection than the SG9R vaccine, with significantly decreased hepatic necrotic foci scores, splenic enlargement scores, necrotic foci scores, and recovery of the challenge strain from the spleen. Vaccination with JOL916 appears to be safe and offers better protection than SG9R against FT in chickens.

Safety Evaluation and Immunogenicity of Arabinose-Based Conditional Lethal Salmonella Gallinarum Mutant Unable to SurviveEx Vivoas a Vaccine Candidate for Protection Against Fowl Typhoid

In seeking to develop a safe fowl typhoid (FT) vaccine, a novel candidate lacking cpxR, lon, and asd Salmonella Gallinarum (SG) genes was constructed with the plasmid-containing araC::P(araBAD)::asd system. A balanced-lethal host-vector system based on the essential bacterial gene for aspartate beta-semialdehyde dehydrogenase (asd) was used to construct the SG mutant strain. A plasmid (p15A ori) with an araC::P(araBAD)::asd cassette was introduced into an auxotrophic mutant to prevent ex vivo survival. The safety, immunity, and protective properties of the SG mutant were evaluated. Inoculation of the mutant at 10(6) colony-forming units (CFU) did not result in recovery in feces and internal organs, whereas inoculation at 10(8) and 10(10) CFU resulted in moderate bacterial recovery from feces and organs. Birds immunized with the mutant were challenged with a virulent SG strain at day 14 postimmunization; significantly reduced mortality and induced plasma immunoglobulin (Ig)G and mucosal IgA responses were noted. Cellular immune responses as evaluated by a peripheral lymphocyte proliferation assay were also significantly induced. The balanced-lethal host-vector system for construction of SG mutants is an effective and improved approach for safe vaccine construction against FT.

Safety and efficacy of a virulence gene-deleted live vaccine candidate for fowl typhoid in young chickens

The safety and efficacy of a live lon-and-cpxR-deleted Salmonella enterica serovar Gallinarum (SG) vaccine candidate (JOL916) was evaluated in young layer chickens. Vaccinated (n=25) and unvaccinated (n=25) groups were organized, respectively, at 1, 2, 3, and 4 weeks of age. One-week-old and 2-week-old chickens were orally inoculated with 2×107 colony-forming units of JOL916, and orally challenged with 2 x 106 colony-forming units of a wild-type SG strain at the third week post vaccination (w.p.v.). Doses of vaccination and challenge were increased 10-fold for 3-week-old and 4-week-old chickens. SG-antigen-specific peripheral lymphocyte proliferation response and concentrations of plasma IgG and secretary IgA in the intestine were examined at the second w.p.v. Gross lesions of the liver and spleen and recovery of the vaccine strain from the spleen were also examined at the second w.p.v. No evidence of side effects was detected by observation of general condition and body weight gain in all vaccinated groups. No, or very mild, gross lesions in the chickens were observed in the liver and/or spleen after vaccination. Significant cellular immune responses and systemic IgG responses were induced after vaccination in all age groups. Elevation of secretary IgA concentration was significant in the group, vaccinated at the age of 1 week. Depression scores after challenge were significantly lower in the vaccinated groups, as compared with the corresponding control groups. Significant reductions of death rates were observed in all vaccinated groups, as compared with the equivalent unvaccinated groups. Thus, the oral vaccination of young chickens with JOL916 was demonstrated to be safe. Moreover, it offered efficient protection against fowl typhoid.