Effects of supplementation with green tea by-products on growth performance, meat quality, blood metabolites and immune cell proliferation in goats.

Forty-eight castrated male goats were used to determine the effects of feeding green tea by-products (GTB) on growth performance, meat quality, blood metabolites and immune cell proliferation. Experimental treatments consisted of basal diets supplemented with four levels of GTB (0%, 0.5%, 1.0% or 2.0%). Four replicate pens were assigned to each treatment with three goats per replicate. Increasing dietary GTB tended to linearly increase the overall average weight gain and feed intake (p = 0.09). Water holding capacity, pH and sensory attributes of meat were not affected by GTB supplementation, while cooking loss was reduced both linearly and quadratically (p < 0.01). The redness (linear; p = 0.02, quadratic; p < 0.01) and yellowness (quadratic; p < 0.01) values of goat meat were improved by GTB supplementation. Increasing dietary GTB quadratically increased protein and decreased crude fat (p < 0.05), while linearly decreased cholesterol (p = 0.03) content of goat meat. The proportions of monounsaturated fatty acid, polyunsaturated fatty acid (PUFA) and n-6 PUFA increased linearly (p < 0.01) and n-3 PUFA increased quadratically (p < 0.05) as GTB increased in diets. Increasing dietary GTB linearly increased the PUFA/SFA (saturated fatty acid) and tended to linearly and quadratically increase (p ≤ 0.10) the n-6/n-3 ratio. The thiobarbituric acid-reactive substances values of meat were lower in the 2.0% GTB-supplemented group in all storage periods (p < 0.05). Dietary GTB linearly decreased plasma glucose and cholesterol (p < 0.01) and quadratically decreased urea nitrogen concentrations (p = 0.001). The growth of spleen cells incubated in concanavalin A and lipopolysaccharides medium increased significantly (p < 0.05) in response to GTB supplementation. Our results suggest that GTB may positively affect the growth performance, meat quality, blood metabolites and immune cell proliferation when supplemented as a feed additive in goat diet.

Effects of fermented corni fructus and fermented kelp on growth performance, meat quality, and emission of ammonia and hydrogen sulphide from broiler chicken droppings.

1. Corni fructus is the fruit of Cornus officinalis, a dogwood species. This study was conducted to prepare fermented corni fructus preparation (FCFP) and fermented kelp (FK) from corni fructus and by-products of Laminaria japonica fermented with Bacillus subtilis and Aspergillus oryzae. 2. The effects of dietary FCFP and FK as replacer of oxytetracycline (OTC) on growth performance, meat composition, meat oxidative stability, and emissions of ammonia (NH3) and hydrogen sulphide (H2S) from broiler chicken droppings were investigated. 3. A total of 140 d-old broiler chicks were randomly allotted to 4 dietary treatments including control, OTC (0.05 g/kg), FCFP (5 g/kg), and FK (5 g/kg). 4. Overall, inclusion of FCFP resulted in lower weight gain and feed intake during the overall experimental period. Broilers fed FCFP diets tended to have lower crude fat and higher crude ash content in the carcasses. 5. In the fresh state, the malondialdehyde (MDA) value of broiler meat was lower in the FK supplemented group. At one week, meat from broilers fed antibiotic and FK diets had lower MDA values, whereas at 2 weeks broiler meat from all dietary treatment groups had lower MDA values than the control. 6. Dietary supplementation with FK significantly reduced faecal NH3 emissions throughout the experimental period, whereas dietary OTC and FCFP supplementation increased NH3 emissions at 2 and 4 weeks. There were no significant effects of dietary treatments on H2S emissions throughout the experimental period, except during week one, when FCFP supplementation reduced the emission. 7. In conclusion, dietary supplementation with 5 g/kg FK improved the oxidative stability of broiler meat and reduce faecal NH3 emissions without affecting growth performance.

Effects of Citrus junos by-products fermented with multistrain probiotics on growth performance, immunity, caecal microbiology and meat oxidative stability in broilers.

1. The present study was conducted to develop Citrus junos probiotics (CJP), using by-products of Citrus junos fermented with multispecies probiotic bacteria including Saccharomyces cerevisiae, Enterococcus faecium, Lactobacillus acidophilus and Bacillus subtilis. The effects of dietary CJP on the growth performance, immune status, caecal microbiology and meat oxidative stability of broiler were investigated. 2. A total of 240 one-day-old Ross broiler chicks were used in a 35-d experiment in which the chicks were randomly allotted to one of the 4 dietary treatments (0, 5, 10 and 20 g CJP/kg diet) in a completely randomised design. 3. Dietary supplementation of 5 g/kg CJP significantly increased body weight, average daily gain and average daily feed intake of broiler during the overall experimental period. 4. Serum immunoglobulin (Ig)M concentration was significantly increased by 10 and 20 g/kg CJP, whereas the IgG and IgA concentration remained unaffected. In addition, 20 g/kg CJP significantly inhibited proliferation of Escherichia coli without affecting the concentration of Lactobacillus or Bacillus spp. 5. A significant reduction in the thiobarbituric acid reactive substances values of breast and thigh meat was observed in response to increasing concentration of dietary CJP. 6. Thus, the results suggest that CJP up to a concentration of 20 g/kg can be used in the diet of broilers to improve immunity and to reduce caecal E. coli and TBARS values of breast and thigh meat without any adverse effects on growth performance.

Comparison of single and blend acidifiers as alternative to antibiotics on growth performance, fecal microflora, and humoral immunity in weaned piglets.

The banning of the use of antibiotics as feed additive has accelerated investigations of alternative feed additives in animal production. This experiment investigated the effect of pure citric acid or acidifier blend supplementation as substitute for antibiotic growth promoters on growth performance, fecal microbial count, and humoral immunity in weaned piglets challenged with Salmonella enterica serover Typhimurium and Escherichia coli KCTC 2571. A total of 60 newly weaned piglets (crossbred, 28-d-old; average 8 kg initial weight) were randomly assigned to four dietary treatments in a completely randomized design. Dietary treatments included NC (negative control; basal diet), PC (positive control; basal diet+0.002% apramycin), T1 (basal diet+0.5% pure citric acid), and T2 (basal diet+0.4% acidifier blend). All piglets were orally challenged with 5 mL of culture fluid containing 2.3×10(8) cfu/mL of E. coli KCTC 2571 and 5.9×10(8) cfu/mL of S. typhimurium at the beginning of the experiment. The PC group showed the highest ADG and ADFI, whereas gain:feed was improved in the PC and T1 group (p<0.05). All dietary treatments showed significant reduction in fecal counts of Salmonella and E. coli, compared to NC (p<0.05), with PC being better than T1 and T2. Significant elevation in fecal Lactobacillus spp. counts was shown by treatments with T1, T2, and PC, whereas Bacillus spp. counts were increased by treatment with T1 and T2 compared to NC and PC diet (p<0.05). Serum IgG concentration was increased by T1 diet (p<0.05), whereas IgM and IgA were not significantly affected by any of the dietary treatments (p>0.05). From these above results, it can be concluded that, as alternatives to antibiotics dietary acidification with pure citric acid or acidifiers blend did not fully ameliorate the negative effects of microbial challenges in respect of growth performance and microbial environment, however improved immunity suggested further research with different dose levels.

Seamustard (Undaria pinnatifida) Improves Growth, Immunity, Fatty Acid Profile and Reduces Cholesterol in Hanwoo Steers.

The study was designed to evaluate the effect of 2% seamustard (Undaria pinnatifida) by-product (SW) on growth performance, immunity, carcass characteristics, cholesterol content and fatty acid profile in Hanwoo steers. A total of 20 Hanwoo steers (ave. 22 months old; 619 kg body weight) were randomly assigned to control (basal diet) and 2% SW supplemented diet. Dietary SW supplementation significantly (p<0.05) improved average daily gain and gain:feed ratio as well as serum immunoglobulin G concentration. Chemical composition and quality grade of meat and carcass yield grades evaluated at the end of the trial were found to be unaffected by SW supplementation. Dietary SW significantly reduced meat cholesterol concentration (p<0.05). Dietary SW supplementation significantly reduced the myristic acid (C14:0) and palmitoleic acid (C16:ln-7) concentration, while SW increased the concentration of stearic acid (C18:0) and linolenic acid (C18:3n-3) compared to control (p<0.05). Dietary SW supplementation had no effect on saturated fatty acids (SFA), unsaturated fatty acids, poly unsaturated fatty acid (PUFA) or mono unsaturated fatty acid content in muscles. A reduced ratio of PUFA/SFA and n-6/n-3 were found in SW supplemented group (p<0.05). In conclusion, 2% SW supplementation was found to improve growth, immunity and fatty acid profile with significantly reduced cholesterol of beef.

Effect of Sea Tangle (Laminaria japonica) and Charcoal Supplementation as Alternatives to Antibiotics on Growth Performance and Meat Quality of Ducks.

A total of 150 growing ducks were assigned to five dietary treatments to study the effect of sea tangle and charcoal (STC) supplementation on growth performance and meat characteristics in a completely randomized design. There were six replicates and five ducklings in each replication. The five dietary treatments were control, antibiotic, and 0.1%, 0.5%, and 1% STC supplemented diets. No significant differences were found on ADG, ADFI, and gain:feed among treatments in different weeks. The overall (0 to 3 weeks) ADFI decreased in antibiotic treatment (p<0.05) whereas the gain:feed increased significantly upon 1.0% STC supplementation compared to control (p<0.05). No significant variation was found in meat chemical composition except crude fat content which was high in 1.0% STC dietary group (p<0.05). Meat cholesterol was reduced in 0.1% STC group (p<0.05) compared to other dose levels while serum cholesterol was unaffected. High density lipoprotein (HDL) content was high in 1.0% STC (p<0.05) and low density lipoprotein (LDL) was low in 0.1% and 1.0% STC dietary groups (p = 0.06). No significant effect was found on the thiobarbituric acid reactive substances (TBARS) of fresh meat, whereas the TBARS value of meat preserved for 1 week was reduced significantly in STC dietary groups (p<0.05). The 0.1% STC dietary group showed an increased myristic acid (p = 0.07) content whereas, the content of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids increased in STC supplementation than antibiotic group (p<0.05). An increased concentration of omega-3 fatty acids and a reduced ratio of n-6/n-3 PUFA ratio was found upon 1.0% STC supplementation compared to antibiotic dietary group (p<0.05). Therefore, 1.0% STC dietary supplementation can be used as alternatives to antibiotics in duck production.

Screening-detected calcified and non-calcified ductal carcinoma in situ: differences in the imaging and histopathological features.

AIM: To compare the imaging and histopathological features of screening-detected calcified and non-calcified ductal carcinoma in situ (DCIS). MATERIALS AND METHODS: The study involved 217 DCIS cases in 212 asymptomatic patients admitted between May 2007 and December 2010. All lesions were divided into calcified and non-calcified DCIS according to the presence of calcifications on mammography. Two radiologists reviewed the findings from mammography, ultrasound, and magnetic resonance imaging (MRI) in consensus according to the Breast Imaging Reporting and Data System. The histopathological features of the lesions were obtained from medical records. Statistical comparisons were performed using the chi-square, Fisher's exact test, and intraclass correlation coefficient (ICC) analyses. RESULTS: On mammography, most non-calcified DCIS presented as either a false-negative finding (49%) or mass lesion (30%), whereas most calcified DCIS (68.5%) presented as calcification alone (p < 0.001). At ultrasound, all of the non-calcified DCIS appeared as a mass, whereas 62% of the calcified DCIS appeared as a mass (p < 0.001). At histopathology, high nuclear grade (p = 0.017), necrosis (p < 0.001), positive progesterone receptor (p = 0.027), and presence of the HER-2/neu oncogene (p < 0.001) were more common in the calcified DCIS than in the non-calcified DCIS. There were no significant differences in the MRI features between the two groups. The ICC values of the non-calcified and calcified DCIS between predicted tumour size and pathologic size were 0.625 versus 0.705 for mammography, 0.801 versus 0.552 for ultrasound, and 0.760 versus 0.767 for MRI. CONCLUSIONS: Screening-detected calcified and non-calcified DCIS have different mammographic and sonographic features. Ultrasound could be helpful to predict the pathological size of the non-calcified DCIS.

Assessment of extent of breast cancer: comparison between digital breast tomosynthesis and full-field digital mammography.

AIM: To compare the accuracy of digital breast tomosynthesis (DBT) and full-field digital mammography (FFDM) in preoperative assessment of local extent of breast cancer. MATERIALS AND METHODS: Lesion sizes of breast cancers on DBT and FFDM images were independently evaluated by breast radiologists. Each lesion was flagged as either mis-sized or not depending on whether the assessment of size at imaging was within 1 cm of the lesion size at surgery. Additional analyses were made by mammographic parenchymal density and by lesion size, using 2 cm as the boundary to separate the two subgroups. Statistical comparisons were performed using a repeated measures linear model on the percent mis-sized. P-values < 0.05 were considered statistically significant. RESULTS: The dataset included 173 malignant breast lesions (mean size 23.8 mm, 43% of lesions were ≤2 cm in size) in 169 patients, two-thirds of which had heterogeneously or extremely dense breasts. Overall, the percentage of lesions mis-sized at DBT was significantly lower than at FFDM (19% versus 29%, p = 0.003). There was significantly less mis-sizing at DBT in both heterogeneously dense breasts (11.1% difference between DBT and FFDM, p = 0.016) and extremely dense breasts (15.8% difference, p = 0.024). DBT also had significantly less mis-sizing than FFDM in the subgroup of lesions that were ≤2 cm in size (14.7% difference, p = 0.005). CONCLUSION: DBT was significantly superior to FFDM for the evaluation of lesion size overall, and specifically for small lesions and for lesions in dense breasts. The superiority of DBT versus FFDM increased with parenchymal density.

Acute myocardial infarction due to sinus of Valsalva aneurysm in a patient with Behçet's disease.

Cardiovascular manifestations have been reported in 7-38% of patients with Behçet's disease (BD), and mortality occurs in up to 20% of those with marked vascular involvement. Sporadic cases of endocarditis, myocarditis, pericarditis, acute myocardial infarction, aortic aneurysm, ventricular thrombosis, congestive cardiomyopathy, and valvular dysfunction have been reported. Here we report a case of acute myocardial infarction that resulted from the compression of coronary arteries by a sinus of Valsalva aneurysm in a patient with BD.

Heat stress-induced modulation of host defense against Toxoplasma gondii infection in mice.

This study examined the effects of burn injury on murine immune response against Toxoplasma gondii infection. Male C57BL/6 mice were divided into 3 groups: T. gondii infection (group T), burn injury (group B), and burn injury followed by T. gondii infection (group BT). The survival of group BT was significantly lower than those of group B and group T. Parasite abundance in the tissues was determined by quantitative competitive-polymerase chain reaction. Group BT exhibited significantly higher numbers of T. gondii than group T. Antibody production against T.g.HSP30 in group BT was significantly lower than that in group T, whereas no significant difference was observed in SAG1-specific antibody production. Delayed-type hypersensitivity (DTH) specific for 2,4-dinitrofluorobenzene (DNFB) of both group B and group BT was significantly lower than that of group T. One week after infection, serum interferon-gamma (IFN-gamma) and interleukin (IL)-10 levels in group BT were significantly lower, whereas serum IL-6 levels were significantly higher than in group T Serum TNF-alpha levels in both group T and group BT were elevated at 1 wk after infection, although there was no significant difference between them. Serum IFN-gamma, IL-10, and TNF-alpha levels in group B were not elevated during the experimental term. In conclusion, the impaired antigen-specific antibody production and DTH response, together with the modulated patterns of cytokine responses, seemed to be strongly involved in the development of burn-induced immunosuppression and the consequent increased susceptibility to T. gondii infection in mice.

Monitoring of behavior using a video-recording system for recognition of Salmonella infection in experimentally infected growing pigs.

Behavior is one of the most commonly used indicators of illness; however, few studies have investigated how different common diseases affect animal behavior. This experiment was conducted to investigate behavioral and clinical alterations in growing pigs experimentally infected with Salmonella spp. during a 4-week post-infection period. A total of 48 growing pigs were divided into one of the three treatment groups (1) control, (2) infection with Salmonella Typhimurium or (3) infection with Salmonella Enteritidis. Individual pigs' behavior was recorded daily (0900 to 1100 and 1600 to 1800 h) using a video-recording system. Pigs in both infected groups had lower weight gain and feed intake during week 0 to 2 and 0 to 4 experimental period. Bacteriological data revealed that pigs in both infected groups persistently shed bacteria throughout the period of study. Oral infection of growing pigs with S. Typhimurium and S. Enteritidis significantly reduced the frequency of morning large (except week 1) and small movement throughout the study period. In the evening, significantly lowest frequency of movements were observed in the S. Enteritidis-infected group compared with the control. The standing and sitting frequency were significantly lower in both infected groups only at the morning of week 4. Infection with Salmonella spp. led to a significant reduction in the frequency and duration of morning eating and drinking throughout the experimental period, with the exception of 4th week drinking duration. The lowest frequency of evening eating during week 1 and 4 was recorded in both infected groups; whereas, the duration differed only at week 1. The evening drinking frequency only tended to decrease in response to S. Typhimurium infection at week 1. This study shows that, pigs infected with Salmonella spp. had poor performance, shedding high levels of Salmonella with their feces and reduced feeding and drinking activity, which are adaptive responses to infection and may help caretakers to detect ill health.

Heparin bridging in warfarin anticoagulation therapy initiation could increase bleeding in non-valvular atrial fibrillation patients: a multicenter propensity-matched analysis.

BACKGROUND: The efficacy of heparin-bridging therapy during the initiation of oral anticoagulation therapy (OAC) in non-valvular atrial fibrillation (NVAF) is unclear. OBJECTIVES: To evaluate the efficacy and the safety of heparin-bridging therapy during OAC initiation in NVAF patients. PATIENTS/METHODS: This study included 5327 consecutive warfarin-naïve NVAF patients who received OAC that was initiated with (n = 1053) or without (n = 4274) heparin bridging at four tertiary hospitals. Stroke and bleeding events within 30 days of OAC were evaluated. RESULTS: While there was no difference in the incidence of stroke (0.5% vs. 0.3%, P = 0.381), major bleeding rate (0.9% vs. 0.3%, P = 0.004) was higher in heparin-bridged than in non-bridged patients. This trend remained in the propensity score-matched population (stroke 0.5% vs. 0.6%, P = 0.762; major bleeding 0.8% vs. 0.1%, P = 0.019). A high CHA2 DS2 -VASc score was an independent predictor for stroke, whereas bridging therapy had no beneficial effect in preventing stroke regardless of CHADS2 or CHA2 DS2 -VASc scores. The HAS-BLED score had a predictive value for major bleeding (odds ratio 1.80, 95% confidence interval 1.11-2.92, P = 0.018), and heparin-bridging therapy was associated with a higher major bleeding rate (odds ratio 4.44, 95% confidence interval 1.68-11.72, P = 0.003), especially in patients with a HAS-BLED score of ≥ 1. CONCLUSIONS: The heparin-bridging therapy increased bleeding without the benefit of preventing stroke at the initiation of OAC in NVAF. Our data suggest that heparin bridging should not be considered at the initiation of OAC in NVAF patients.

Toxoplasma gondii Hsp70 as a danger signal in toxoplasma gondii-infected mice.

Toxoplasma gondii Hsp70, T gondii Hsp30/bag1, and surface antigen 1 messenger RNAs were shown to be useful in analyzing stage conversion of T gondii between bradyzoites and tachyzoites. The high-level expression of T gondii Hsp70 was correlated with mortality in interferon-gamma knockout mice infected with T gondii. Tgondii Hsp70 inhibited the induction of nitric oxide release by peritoneal macrophages of T gondii-infected mice. These findings identify T gondii Hsp70 as a danger signal during lethal, acute T gondii infection.

Toxoplasma gondii: difference of invasion into tissue of digestive organs between susceptible and resistant strain and influence of IFN-gamma in mice inoculated with the cysts perorally.

Because it is widely accepted that there is a significant difference in susceptibility to chronic infection by Toxoplasma gondii among inbred mouse strains with different genetic backgrounds, we compared the distribution of the protozoa in digestive organs at early stages of infection between resistant (BALB/c) and susceptible (C57BL/ 6) mice after peroral infection with Fukaya strain cysts. Furthermore, to determine the influence of interferon gamma (IFN-gamma) on the infectivity of the cysts to the digestive tract, homozygous IFN-gamma knockout mice were utilized. Quantitative competitive polymerase chain reaction (QC-PCR) was employed to assess the distribution of T. gondii in different organs at various times after ingestion of cysts. SAG1, a T. gondii-specific gene, was detected in the small intestine and the caecum in wild-type C57BL/6 mice and in the whole digestive tract in IFN-gamma knockout C57BL/6 at 24 hr after infection. No detectable reaction in QC-PCR was observed in BALB/c mice at 24 hr after ingestion of the cysts. Destruction of the IFN-gamma gene showed less effect on the resistance to infection in BALB/c mice, but remarkable augmentation of infectivity of T. gondii to the rectum and peripheral blood was observed in C57BL/6 mice.

Involvement of MyD88 in host defense and the down-regulation of anti-heat shock protein 70 autoantibody formation by MyD88 in Toxoplasma gondii-infected mice.

This study investigated the influence of TLR (toll-like receptor)4, TLR2, and MyD88 in Toxoplasma gondii-infected wild-type (WT) mice and TLR4-, TLR2-, and MyD88-deficient mice. Ninety-five percent of MyD88-deficient mice died 10-16 days after intraperitoneal infection with 100 cysts of T. gondii Fukaya strain, whereas 95-100% of TLR4- and TLR2-deficient mice and WT C57BL/6 (B6) mice survived for more than 7 wk after T. gondii infection. The distribution of T. gondii in various organs of TLR4-, TLR2-, and MyD88-deficient mice and WT B6 mice was assessed 2 wk after T. gondii intraperitoneal infection using quantitative competitive polymerase chain reaction. In MyD88-deficient mice, high levels of T. gondii load were observed in the brain, tongue, heart, lungs, spleen, liver, mesenteric lymph node, and kidneys after infection. The T. gondii load was significantly increased in the lungs in both TLR4- and TLR2-deficient mice compared with WT B6 mice. High levels of anti-mouse heat shock protein (mHSP)70 autoantibody and anti-T. gondii HSP70 antibody production were detected in the sera from MyD88-deficient mice.

Organ infectivity of Toxoplasma gondii in interferon-gamma knockout mice.

To determine the influence of interferon (IFN)-gamma on the organ infectivity and on the genetic susceptibility of susceptible (C57BL/6) and resistant (BALB/c) strains after peroral infection with cysts of Toxoplasma gondii. IFN-gamma knockout (KO) mice in C57BL/6 and BALB/c backgrounds were utilized. The kinetics of the changes in T. gondii abundance were evaluated with a quantitative competitive polymerase chain reaction assay in various organs at different times after peroral infection. In IFN-gamma KO mice, a T. gondii-specific gene, SAG1, was detected in all organs examined, and the protozoan proliferated much more actively than in wild-type mice. The abundance of T. gondii was much higher in mesenteric lymph nodes and the heart than in other organs. In contrast, in the nervous system organs and kidneys, only a weakly detectable reaction was observed. Toxoplasma gondii grew at a more rapid rate in the organs of IFN-gamma KO C57BL/6 mice than in the organs of IFN-gamma KO BALB/c mice during the course of infection. Destruction of the IFN-gamma gene showed remarkable effects on the infectivity in both susceptible and resistant mice.

Role of Toxoplasma gondii HSP70 and Toxoplasma gondii HSP30/bag1 in antibody formation and prophylactic immunity in mice experimentally infected with Toxoplasma gondii.

Production of antibodies against Toxoplasma gondii (T. gondii)-derived stress proteins, T. gondii HSP70 (T.g.HSP70) and T.g.HSP30/bagl, in C57BL/6 and BALB/c mice perorally infected with cysts of the avirulent Fukaya strain of T. gondii was analyzed. Production of anti-T.g.HSP70 IgG antibodies was transient, whereas production of anti-T.g.HSP30/bag1 IgG antibodies persisted after infection in both C57BL/6 and BALB/c mice. C57BL/6 mice, a susceptible strain, predominantly produced IgG antibodies specific for T.g.HSP70, whereas BALB/c mice, a resistant strain, predominantly produced IgG antibodies specific for T.g.HSP30/bag1, after T. gondii infection. Immunization with rT.g.HSP30/bag1 enhanced, whereas immunization with rT.g.HSP70 reduced host protective immunity against T. gondii infection with a cyst-forming avirulent strain, Fukaya, and a virulent strain, RH.

A role of carboxy-terminal region of Toxoplasma gondii-heat shock protein 70 in enhancement of T. gondii infection in mice.

We investigated the role of recombinant Toxoplasma gondii heat shock protein (rT.g.HSP) 70-full length, rT.g.HSP70-NH2-terminal region, or rT.g.HSP70-carboxy-terminal region in prophylactic immunity in C57BL/6 mice perorally infected with Fukaya cysts of T. gondii. At 3, 4, 5, and 6 weeks after infection, the number of T. gondii in the brain tissue of each mouse was measured by quantitative competitive-polymerase chain reaction (QC-PCR) targeting the surface antigen (SAG) 1 gene. Immunization with rT.g.HSP70-full length or r.T.g.HSP70-carboxy-terminal region increased the number of T. gondii in the brain tissue after T. gondii infection, whereas immunization with rT.g.HSP70-NH2-terminal region did not. These results suggest that T.g.HSP70-carboxy-terminal region as well as T.g.HSP70-full length may induce deleterious effects on the protective immunity of mice infected with a cyst-forming T. gondii strain, Fukaya.

Anti-HSP70 autoantibody formation by B-1 cells in Toxoplasma gondii-infected mice.

Formation of anti-Toxoplasma gondii HSP70 (TgHSP70) antibody cross-reactive to mouse HSP70 (mHSP70) was observed in the sera of BALB/c (a resistant strain) and C57BL/6 (B6; a susceptible strain) mice after peroral infection with T. gondii cysts of the Fukaya strain. The levels of anti-mHSP70 immunoglobulin G (IgG) autoantibody production in B6 mice were higher than those in BALB/c mice. The isotype and subclass of IgG of anti-TgHSP70 monoclonal antibodies cross-reactive to mHSP70 were mu and gamma3. Anti-mHSP70 autoantibody in T. gondii-infected BALB/c and B6 mice was shown to be produced by the CD5(+) subset of B cells (B-1a cells) but not by conventional B cells (B-2 cells). The epitopes recognized by anti-mHSP70 autoantibody were located primarily in the C-terminal fragment of mHSP70.

Heat shock cognate protein 71-associated peptides function as an epitope for Toxoplasma gondii-specific CD4+ CTL.

HLA-DR-restricted CD4+ cytotoxic T-lymphocyte (CTL) lines specific for Toxoplasma gondii (T. gondii)-infected melanoma cells have been established from peripheral blood lymphocytes (PBLs) of a patient with chronic toxoplasmosis. The role of heat shock cognate protein (HSC) 71 in antigen (Ag) processing and presentation of T. gondii-infected melanoma cells to these CD4+ CTL lines was investigated. A human melanoma cell line (P36) pulsed with T. gondii-infected P36 cell-derived HSC71 was lysed by a T. gondii-specific CD4+ CTL line (Tx-HSC-1). The Tx-HSC-1 also killed T. gondii-infected P36 cells. The lytic activity of Tx-HSC-1 against P36 cells pulsed with T. gondii-infected P36 cell-derived HSC71 was inhibited by monoclonal antibodies (mAbs) against HSC71. Anti-human leukocyte antigen (HLA)-DR mAb also partially blocked the lytic activity, whereas anti-HLA-A,B,C mAb did not block the lytic activity. In addition, a flow cytometric analysis with these specific mAbs against HSC71 showed HSC71 to be expressed on the cell surface of T. gondii-infected P36 cells as well as uninfected P36 cells. These data indicate that HSC71 molecules are expressed on human melanoma cell line P36, and that HSC71 may play a potential role in Ag presentation and processing of T. gondii-infected P36 cells to CD4+ CTL.