Blastogenic Response Research Articles

Therapeutic efficacy and toxicity of tamoxifen loaded PLA nanoparticles for breast cancer

This study was carried out to assess the therapeutic efficacy and toxicity of tamoxifen (Tmx) loaded poly(d,l-lactic acid) (PLA) nanoparticles (Tmx-NPs) for breast cancer. An in vivo study was conducted to determine the effect of Tmx-NPs on DMBA induced mammary tumor in female Wistar rat. The experimental results showed that the mean diameter of Tmx-NPs was 224±3nm with 68±2% (w/w) of entrapment efficiency. In in vivo study, the tumor size in rat was significantly reduced (P<0.001) by treating Tmx-NPs as compared to pure Tmx and untreated group (control DMBA). Tmx-NPs showed the marked reduction in hepatotoxicity and renal toxicity when compared to pure Tmx as evidenced by histopathological examination of liver and kidney tissues as well as estimation of AST, ALT levels, and creatinine, urea, blood urea nitrogen levels. Oxidative stress and lipid peroxidation was estimated in spleen, liver and kidney and was found significantly high in pure Tmx treated group as compared to Tmx-NPs and control group. Immunological parameters like blastogenic response of splenocytes, TLC, DLC were studied and found significantly high in pure Tmx treated group but the variations were nonsignificant in Tmx-NPs group as compared to control. Thus, Tmx-NPs have significant therapeutic efficacy with reduced side effects.

Antigen-specific lymphocyte proliferation as a marker of immune response in guinea pigs with sustained Helicobacter pylori infection.

Helicobacter pylori (H. pylori) bacteria are human pathogens causing symptomatic gastritis, peptic ulcer or gastric cancer. Little is known about the kinetics of immune responses in H. pylori infected patients because the initial moment of infection has not been identified. Various animal models are used to investigate the immune processes related to H. pylori infection. In this study we checked whether H. pylori infection in guinea pigs, mimicking natural H. pylori infection in humans, resulted in the development of specific immune responses to H. pylori antigens by measuring the proliferation of lymphocytes localized in mesenteric lymph nodes, spleen and peripheral blood. The maturity of macrophages and cytokines, delivered by monocyte-macrophage lineage or lymphocytes, were considered as mediators, which might influence the lymphocyte blastogenic response. The obtained results showed the activation of T cells localized in mesenteric lymph nodes by H. pylori antigens in H. pylori infected guinea pigs four weeks postinfection. The blastogenic activity of lymphocytes was shaped by their interaction with antigen presenting cells, which were present in the cell cultures during the whole culture period. Moreover, the balance between cytokines derived from adherent leukocytes including interleukin 8--IL-8 as well as interferon gamma--IFN-γ, and transforming growth factor beta--TGF-β delivered by lymphocytes, was probably important for the successful proliferation of lymphocytes. The H. pylori specific lymphocytes were not propagated in peripheral blood and spleen of H. pylori infected animals. The modulation of immunocompetent cells by H. pylori antigens or their different distribution cannot be excluded.

English

The present study was designed to study cytokine profile and lymphocytic proliferation activity in chicken infectious anaemia virus (CIAV) infected chicks at various intervals of clinically susceptible age. Briefly, ninety specific pathogen free (SPF) chicks were divided into five experimental groups (A to E). Groups A to D were inoculated&nbsp;with 0.5 ml of 104.5TCID50&nbsp;of CIAV intramuscularly at 1, 14, 21 and 28-day-old, respectively, while&nbsp;group E served as uninfected control. Comparative cytokine gene expression levels of interleukin (IL)-1&beta;, IL-10, IL-12&beta; and granulocyte macrophage-colony stimulating factor (GM-CSF) were measured at 4 and 7 days post infection (dpi) by qRT-PCR. Also, lymphocytic proliferation activity was measured at 1, 2 and 3 week post infection (wpi) by lymphocyte transformation test (LTT).&nbsp;The down regulatory changes in cytokines and depressed&nbsp;cell mediated immune (CMI) responses were observed in chicks of all infected groups (A to D), which were significantly&nbsp;(p&lt;0.05)&nbsp;low as compared to control group. Higher intensities and more pronounced effects on cytokine downregulation and blastogenic responses of lymphocytes were seen at younger ages. Apoptosis in thymic DNA as detected by nucleosomal fragmentation was highest on 7 and 10 dpi and lowest on 14 dpi, which correlated with down regulated cytokine profile in all infected groups.&nbsp;The development of clinical signs and lesions along with PCR testing confirmed the establishment of CIAV infection in experimental chicks.&nbsp;Increased apoptosis, decreased PCV, depletion of lymphocytes, down regulated cytokine profile and reduced lymphocyte activity explain involvement of thymic and hematopoietic precursor cells leading to CMI suppression caused by CIAV. Altogether, immunopathological changes were more severe in chicks infected at younger ages as compared to later ages of the clinically susceptible period.However, further studies are suggested regarding the impact of viral load on the various cytokine profiles and revealing more about immunopathogenesis of CIAV both in young and adult birds. &nbsp; Key words:&nbsp;Chicken infectious anemia virus, cell mediated immune response, polymerase chain reaction (PCR), qRT-PCR, cytokine, apoptosis.

Immune response of the mammary gland during different stages of lactation cycle in high versus low yielding Karan Fries crossbred cows

To investigate the magnitude of innate immunity of mammary glands during different stages of the lactation cycle, milk samples were collected from both high (n=10) and low (n=10) yielding Karan Fries (KF) crossbred cows during early, mid and late lactation stages. Milk somatic cells released during different stages of the lactation cycle were measured microscopically. In vitro immune activity of each milk leukocytes (viz. neutrophils, lymphocytes and macrophages) were also evaluated after isolating each of the cells from the total milk somatic cell pellet. Relative mRNA expression profiles of TNF-α, IL-8 and TLR-4 by RT polymerase chain reaction were also studied in total milk somatic cells during various stages of the lactation cycle in high yielding cows, whereas low yielding KF cows were taken as control. The results were analyzed and significance was tested by employing two way ANOVA, and the relative expression ratio of the target genes of high yielding cows was tested and analyzed for significance by Relative Expression Software Tool REST version 2009. Irrespective of the stage of lactation, high yielding animals were having higher (P<0.01) somatic cell counts (SCC) compared to low yielders. Milk SCC was found to be significantly higher (P<0.01) during mid-lactation compared to other stages in high yielding cows. In vitro immune response of milk leukocytes were lower in high yielders (P<0.01) compared to low yielders, irrespective of stage of lactation. In vitro immune response of all the milk leukocytes was found to be lower (P<0.01) in high yielding cows compared to low yielding cows. In vitro phagocytic index (PI) of milk neutrophils was higher (P<0.01) during mid lactation periods in both the groups of cows. PI of milk macrophages was found to be higher (P<0.01) during early lactation period in both high and low yielding cows. LPS induced milk lymphocyte blastogenic response was found to be higher (P<0.01) in early lactation period of low yielders. Abundance of IL-8, TNF-α and TLR-4 transcripts were up-regulated (P<0.01) during mid lactation of high yielders as compared to control cows (low yielding cows). Abundance of TNF-α and TLR-4 transcripts were down-regulated (P<0.01) during late lactation in high yielding cows. This study indicated that innate immune responses of milk somatic cells in KF cows are modulated by both lactation stage and milk yield. It was also found that mammary glands of high yielding cows have better magnitude of innate immune response during mid lactation stage compared to early and late lactation in terms of milk SCC and in vitro immune response of isolated milk leukocytes.

Assessment of cell mediated immune response in rabbits immunized with affinity purified 35kDa midgut antigen of Rhipicephalus haemaphysaloides ticks.

Cell mediated immune response to immunoaffinity chromatography purified midgut antigen of Rhipicephalus haemaphysaloides ticks in rabbits was studied by using lymphocyte transformation test. This test was carried out by using 5-bromo-2'-deoxy-uridine kit method. The blastogenic response of peripheral blood lymphocytes of normal rabbit to different concentrations of antigen and mitogen (Con A) showed that 2μg of antigen and 2μg of mitogen gave maximum stimulation index. The antigen specific responsiveness of immunized rabbits with affinity purified 35kDa midgut antigen was highly significant (P≤0.01) compared to mitogen. The maximum lymphocyte stimulation index (LSI) of 2.47 was observed on 49 day post immunization in immunized group. The lymphocytes separated from control animal cultured in RPMI1640 medium with 2μg of antigen and 2μg of mitogen (Con A) were never stimulated and their LSI values were below 2.0.

Relationship between neural activity and immunity in patients with undifferentiated somatoform disorder

It has been suggested that somatoform disorders are related to both the brain and the immune system, and that immune functions may be influenced by cerebral asymmetry. However, few studies have examined the relationship between brain activity and immune function in somatoform disorders. Thirty-two patients with non-medicated undifferentiated somatoform disorder were enrolled in this study. Blastogenic responses to phytohemagglutinin (PHA) were used to measure immunity. Regional cerebral perfusion was measured by 99m-Tc-ethyl cysteinate dimer single photon emission computed tomography (SPECT). Significant hypoperfusion was found at the left inferior parietal lobule and the left supramarginal gyrus in the more immune-suppressed (MIS) subgroup compared with the less immune-suppressed (LIS) subgroup. However, no regions of significant hyperperfusion were found in the MIS subgroup compared with the LIS subgroup. Decreased cerebral blood flow in the left inferior parietal lobule and the left supramarginal gyrus in the patient group was also significantly associated with reduced blastogenic responses to PHA regardless of sex and age. These results suggest that the left inferior parietal lobule and the left supramarginal gyrus might play an immunomodulating role in patients with undifferentiated somatoform disorder. In addition, these results suggest the role of cerebral asymmetry in altered immunity in the patients.

Correlation between peripheral melatonin and general immune status of domestic goat, Capra hircus: A seasonal and sex dependent variation

Caprine immune system has largely been neglected by ruminant researchers despite of its high economic importance in bioindustry. We made an attempt to note the annual/seasonal and gender dependent variation in immune status of the domestic goat, Capra hircus, Jamunapari breed and correlated with the concentration of peripheral gonadal hormones (testosterone, 17β-estradiol) and melatonin. Based on the data of general immune parameters such as total leukocyte count (TLC) and percent lymphocyte count (%LC) along with blastogenic response of splenocytes in terms of percent stimulation ratio (%SR), we found an annual variation in immune status of male, female and castrated male goats in a gender dependent manner. Female goats showed higher immune parameters than that of castrated as well as intact male goats. Testosterone in intact male goats was high throughout the year, which might be responsible for low immune status. However, in castrated male goats, plasma testosterone was undetectable and presented an immune status equivalent to that of female goats. We observed a high immune status during the winter season in all the groups of goats, which might be due to high circulatory concentration and longer duration of melatonin induced by short days. Being released by circulating lymphocytes, the concentration of interleukin-2 (IL-2) in blood plasma, showed an annual and seasonal variation being higher in female than castrated and intact male goats throughout the year, having a similar trend in plasma melatonin concentration. The higher immune status in female goats during the winter months might have helped them to overcome the winter bound stress, i.e. low temperature. Therefore, we may propose that testosterone acts as an immunosupressor while melatonin as an immunostimulator for goats as it showed a positive correlation with immune parameters noted.

Deactivation of T-2 toxin in broiler ducks by biotransformation

SUMMARY Effects of 2 dietary levels (0.6 and 1.0 ppm) of T-2 toxin, and the possible protective capacity of a mycotoxin-inactivating feed additive (Detoxa Plus) were investigated in growing White Pekin ducklings in a 49-d trial comprising 6 treatment groups of 10 ducks/group. The experimental design consisted of 1 negative and 1 positive control and 4 test groups, as follows: group 1, negative control (no T-2 toxin and no feed additive added to the feeds); group 2, positive control (no T-2 toxin added, but the feeds were supplemented with feed additive at 2 kg/t); group 3, feeds were complemented with 0.6 ppm of purified T-2 toxin (no feed additive added); group 4, feeds were complemented with 0.6 ppm of T-2 toxin and 2 kg/t of the feed additive was provided; group 5, feeds were complemented with 1.0 ppm of T-2 toxin (no feed additive added); group 6, feeds were complemented with 1.0 ppm of T-2 toxin and 2 kg/t of the feed additive was provided. From wk 4 until the end of the trial, the daily BW gain of group 3 ducks was inferior to that of control ducks, and the differences in means were statistically significant (P ≤ 0.05) at wk 4 and 7. The final BW of this group was also significantly lower than that of the control (P ≤ 0.001). The BW gain of ducks in group 5 was also depressed by the toxin treatment. The adverse effect of 0.6 ppm of T-2 toxin was fully counteracted by the feed additive in terms of daily BW gain, cumulative daily BW gain, and BW at exsanguination. The T-2 toxin at both treatment levels depressed the blastogenic response of lymphocytes to nonspecific mitogens (concanavalin A and phytohaemagglutinin), which was counteracted by the feed additive at the lower dietary concentration of T-2 toxin. No such effect was observed with 1.0 ppm of T-2 toxin. Metabolic blood parameters and hematological data showed no consistent treatment effects. We conclude that this feed additive is able to counteract the adverse effects of T-2 toxin at dietary concentrations that might be encountered under field conditions.

Lymphocyte Blastogenic Responses to Food Antigens in Cats Showing Clinical Symptoms of Food Hypersensitivity

Three cats were diagnosed as having food hypersensitivity by food elimination and oral food provocation tests. Twelve allergenic food ingredients were identified by oral food provocation test in the 3 cats. Of the 12 food ingredients, 9 offending food antigens were shown to be positive in a lymphocyte stimulation test; however, none of them were positive in antigen-specific IgE testing, and only four food antigens were positive in intradermal testing. The stimulation indices in the lymphocyte stimulation tests for the 9 food ingredients were found to be decreased after the cats were fed elimination diets. The present study demonstrates that the lymphocyte stimulation test reflects an immunologic reaction involved in food hypersensitivity and can help identify allergenic food ingredients in feline food hypersensitivity.

Melatonin ameliorates oxidative stress and induces cellular proliferation of lymphoid tissues of a tropical rodent, Funambulus pennanti, during reproductively active phase

Effect of melatonin treatment on free radical production was assessed with simultaneous investigation of hormonal level (melatonin and testosterone), blastogenic response, stimulation index, and histological observation of lymphoid organs (spleen, thymus, and bone marrow) in male Indian palm squirrel (Funambulus pennanti) during reproductively active phase (RAP). Low endogenous melatonin and high testosterone level were noted during RAP. Daily subcutaneous injection of melatonin (25 μg/100 g B wt.) at 17.30-18.00 h to squirrels for 60 consecutive days during May-June significantly decreased the thiobarbituric acid reactive substances (TBARS) level compared to control squirrels. Melatonin treatment significantly increased % stimulation ratio (%SR) of splenocytes and thymocytes against T cell mitogen concanavalin A. Pinealectomy (Px) led to a significant increase in TBARS level whereas a significant decrease was observed in blastogenic response and stimulation index was noted. Melatonin injection to Px squirrels showed restoration in %SR of thymocytes and splenocytes with a significant decrease in the TBARS level of the lymphoid tissues. Further, free radical load was induced by lipopolysaccharide (LPS; 400 μg/ml) in lymphatic tissue homogenates and noted that melatonin supplementation (2 mM/ml) led to a significant decrease in TBARS level compared to the control and LPS-supplemented groups. Histological observation showed dense cellularity of thymocytes and splenocytes. Acridine orange staining technique shows a significant increase in thymocyte apoptosis Px squirrels when compared with melatonin-treated squirrels. These findings suggest that endogenous and exogenous melatonin might be responsible for the maintenance of immune system to adapt this seasonal breeder for the rigors of the environmental changes.

Effects of yeast cell walls on performance and immune responses of cyclosporine A-treated, immunosuppressed broiler chickens

The present study was conducted to investigate the effects of dietary yeast (Saccharomyces cerevisiae) cell walls (YCW) from the yeast extract industry on performance and immune function of cyclosporine A (CSA)-treated, immunosuppressed broiler chickens. A total of 240 day-old male broilers were allocated randomly into four treatments: (1) non-challenged control; (2) non-challenged control+0·3 % YCW; (3) CSA-challenged group; (4) CSA-challenged+0·3 % YCW. On days 1-4 and 22-25 of age, broilers were subcutaneously injected with CSA or sterile saline. The results showed that supplementation of YCW significantly improved daily weight gain (DWG) during the starter (days 1-21, P < 0·01), finisher (days 22-42, P < 0·01) and overall (days 1-42, P < 0·05) periods compared with the control birds, but had no effect on feed conversion ratio (FCR, P>0·05). Compared with the CSA-treated birds, YCW alleviated the decrease of DWG (P < 0·01) and increase of FCR (P < 0·05) caused by CSA challenge at different periods and cumulatively. On days 21 and 42, YCW mitigated the CSA-induced decrease of peripheral blood lymphocyte blastogenic response (P < 0·01). In addition, YCW improved the relative weights of the bursa of Fabricius (P < 0·01) and thymus (P < 0·01) and up-regulated the splenic expression of pro-inflammatory cytokines IL-1β (P < 0·01) and IL-6 (P < 0·01) on day 42 compared with the CSA-treated birds. These results indicate that YCW supplementation has beneficial effects in attenuating the immunosuppressive effects of CSA challenge, therefore improving growth performance of broiler chickens.

Impact of breast cancer recurrence and cancer-specific stress on spouse health and immune function

Spouses of cancer patients are at-risk for poor psychological and physical health as they cope with the complex nature of the disease and fears of losing their partner. Moreover, spouses often serve as patients' primary informal caregivers, a group that evidences poor outcomes across a variety of domains. The present study examines the relative contributions of cancer recurrence - a cancer-specific stressful event - and the subjective experience of cancer-specific stress (IES) in a sample of male spouses of breast cancer survivors. We hypothesized that stress would contribute to poorer physical health and compromised immune function. Spouses (recurrence; n=16) of patients who were coping with their first recurrence were matched to spouses of patients with no evidence of disease (disease-free; n=16). Self-reported physical health (physical symptoms and fatigue) and immune function [T-cell blastogenic response to the mitogens Concanavalin A (ConA) and phytohemagglutanin (PHA) and T3 monoclonal antibody (T3 Mab)] were included as outcomes. Results indicated that patient recurrence status was not a significant unique predictor of physical health or immune function; rather, among all spouses, cancer-specific stress symptoms were associated with increased physical symptoms and altered T-cell blastogenesis. These data suggest that the health implications of caregiving for spouses of cancer survivors is more strongly linked to their subjective experience of cancer as stressful, rather than simply the patients' disease status.

Treatment of rats with heat killed cells (HKC) of Cryptococcus neoformans var. grubii induces cellular activation in spleen and lymphatic nodes

Our previous studies showed that the subcutaneous pretreatment of rats with heat killed cells of Cryptococcus neoformans (HKC) emulsified in complete Freund adjuvant (CFA) promotes protection against an intraperitoneal challenge with viable C. neoformans. In this model, an appropriate activation of adherent peritoneal cells after antigenic treatment is very important for the control of the infection. Here, we investigated the immune response developed in spleen and lymphatic nodes as a result of treatment with HKC-CFA, which might also contribute in the protective phenomenon of this treatment against cryptococcal infection. The results show that, compared with adjuvant alone, rats which received treatment with HKC-CFA presented a greater activation of adherent splenic cells, with up-regulation of major histocompatibility complex class II (MHC II) and CD86 expression and secretion of anticryptococcal metabolites. Furthermore, this treatment also induced an increase in the blastogenic response and the secretion of Th1 and Th2 cytokines by spleen cells in comparison with cells from CFA-phosphate-buffered saline (PBS) treated rats. On the other hand, lymph node cells from animals treated with HKC-CFA presented a rise in the expression of MHCII but not of CD86 with respect to control cells from rats treated with CFA-PBS. These cells also showed a high proliferative response and secretion of Th1-related cytokines, interleukin (IL)-12 and tumor necrosis factor (TNF). These results show that treatment of rats with HKC-CFA is able to induce an early immune response in secondary lymphoid organs, which may contribute to the protective effect induced by this treatment.

The effect of fermented wheat germ extract on production parameters and immune status of growing pigs

Model experiment was conducted with growing pigs fed diets containing zero or two levels (1 or 2 g/kg) of fermented wheat germ extract (FWGE). In the first part of the experiment (19 days between 49 and 68 days of age) weight gain and feed conversion were tested. The 2 nd part of the experiment (21 days between 68 and 89 days of age) served to measure the effect of FWGE on some compartments of the immune system. In the first part of the experiment supplementation of the starters’ diet with FWGE yielded 7.8 and 14.2% additional daily weight gain in the 1 st and 2 nd experimental groups, respectively (P<0.05). Humoral immune response to purified horse globulin (PHG) antigen was unaffected by the treatment. The blastogenic response of lymphocytes to non specific mitogens (PHA, Con-A, PWM), phagocytic activity and phagocytic index as well as the intradermal PHA test was consistently and significantly enhanced by the treatments.

Immunomodulatory effect of glucan on specific and nonspecific immunity after vaccination in puppies

The objective of the study was to determine the immunostimulatory effect of β-(1,3/1,6)-D-glucan in puppies. The effect exerted on the efficacy of vaccination, especially against canine parvovirus and rabies infection, was studied. The application of vaccine and glucan leads to significant increases in the nonspecific immunological parameters (phagocytic ability of leukocytes, blastogenic response of lymphocytes, metabolic and chemotactic activity of polymorphonuclear cells). The level of antibodies against canine parvovirus (Ab CPV) and rabies infection reached the most statistically significant values on the 28th day after the application of vaccine and a syrup containing β-(1,3/1,6)-D-glucan (Group GV) as compared to the control group (Group V, puppies receiving only vaccine). Dogs without glucan supplementation did not produce such significant levels of antibodies. We can conclude that glucan has relevant immunostimulatory effects in dogs with altered immunity. The glucan product tested in this study (PleraSAN V, PLEURAN, Bratislava, Slovakia) could be used in the small animal clinical practice.

Effect of L‐arginine supplementation on immune responsiveness in patients with sickle cell disease

L-arginine (L-Arg) is deficient in sickle cell disease (SSD) during vasoocclusion. We investigated possible causal relationship between L-Arg deficiency and immune dysfunction in SSD in steady-state. Fifteen patients with SSD in steady-state and 13 controls were studied. Plasma L-Arg levels were measured using liquid chromatography. T cell subsets and CD3zeta (CD3zeta) chain expression were analyzed using flow cytometry. Lymphocyte proliferative response to phytohemagglutinin (PHA) and production of IL-6 and interferon-gamma (IFN-gamma) were evaluated with and without L-Arg. SSD patients had significantly lower L-Arg levels than controls. CD3 and CD19 cell populations were comparable for both groups, but SSD patients had above normal numbers of natural killer cells (P = 0.06). Patients and controls exhibited significantly increased lymphocyte blastogenesis to PHA after introduction of L-Arg to cultures; response of patients was significantly greater than values for control individuals. Proliferative response to candida in SSD patients was significantly lower than in controls; L-Arg supplementation did not increase this response. L-Arg had no effect on blastogenic response to PPD and candida albicans. No effect was likewise seen in production of IL-6 and IFN-gamma after addition of L-Arg. CD3zeta chain expression increased after addition of L-Arg in both groups; differences were insignificant. L-Arg levels in steady-state SSD are significantly lower than in controls. L-Arg supplementation enhanced lymphocyte blastogenesis to PHA for both controls and patients, but not in response to antigen. There were no significant differences in CD3zeta chain expression although upregulation of expression occurred after L-Arg supplementation for both groups.

Effect of Glucocorticoid and Melatonin on Immune Function of an Indian Tropical Bird, Perdicula Asiatica: An in Vivo and in Vitro Study

Inverse relationship between circulatory levels of glucocorticoids and melatonin has been reported for most of the vertebrates including human beings. We report the importance of glucocorticoid along with melatonin in avian immune regulation in an Indian tropical bird, Perdicula asiatica with in vivo and in vitro approach. Dexamethasone (30μg/bird/day) treatment of the male birds suppressed the immune activity as judged by the spleen activity (mass, anatomy), circulating total leukocyte, and lymphocyte count, blastogenic response, increased % apoptosis and cytokine (IL-2) production by splenocytes. Melatonin (25μg/100g B.wt./day) treatment increased the above-mentioned immune parameters whereas melatonin together with dexamethasone restored the suppressed immune parameters by dexamethasone to control level. In vitro melatonin (2.5 pM) supplementation to splenocyte cultures restored the dexamethasone (2 μM) suppressed splenocyte proliferation, % apoptosis and IL-2 production. Therefore, melatonin antagonized the suppressive effect of synthetic glucocorticoid on all immune parameters studied in vivo as well as in vitro. Furthermore, exogenous administration of dexamethasone and melatonin treatment altered the circulatory level of corticosterone and melatonin in an inverse manner. It is therefore, suggested that a hormonal trade-off between glucocorticoid and melatonin exists under in vivo and in vitro conditions, being involved in maintenance of the immune function of P. asiatica probably by involving cytokines i.e. IL-2 mediated pathway.

Immunogenicity of recombinant GP5 protein of porcine reproductive and respiratory syndrome virus expressed in tobacco plant

The aim of the study was to evaluate the immunogenicity of the ORF5-encoded major envelop glycoprotein 5 (GP5) of porcine reproductive and respiratory syndrome virus (PRRSV) expressed in tobacco plant as a potential pig oral vaccine in protection against PRRSV infection. Six-week-old PRRSV-free pigs were fed four times orally with 50 g of chopped fresh GP5 transgenic tobacco leaves (GP5-T) (GP5 reaching 0.011% of total soluble protein) or wild-type tobacco leaves (W-T) each on days 0, 14, 28, and 42. Samples of serum, saliva, and peripheral blood mononuclear cells (PBMCs) were collected on days −1, 6, 13, 20, 27, 34, 41, and 48 after the initial oral vaccination. A similar vaccination-dependent gradual increase in the responses of serum and saliva anti-PRRSV total IgG and IgA, respectively, and in the levels of PRRSV-specific blastogenic response of PBMCs was seen in GP5-T-treated pigs; all statistically significant elevations occurred after the 2nd vaccination and were revealed after 20 days post-initial oral vaccination (DPIOV). Pigs fed on GP5-T also developed serum neutralizing antibodies to PRRSV at a titer of 1:4–1:8 after the 4th vaccination by 48 DPIOV. No detectable anti-PRRSV antibody responses and PRRSV-specific blastogenic response were seen in W-T-treated pigs. The present study has demonstrated that pigs fed on GP5-T could develop specific mucosal as well as systemic humoral and cellular immune responses against PRRSV. The results also support that transgenic plant as GP5-T can be an effective system for oral delivery of recombinant subunit vaccines in pigs.

IGF-I Stimulates In Vivo Thymopoiesis After Stem Cell Transplantation in a Child with Omenn Syndrome

Children with severe combined immunodeficiency who receive a T cell-depleted hematopoietic stem cell transplantation (HSCT) have delayed immune reconstitution. To examine the use of recombinant human insulin-like growth factor-I (rhIGF-I) therapy to stimulate thymopoiesis after HSCT. A child with Omenn syndrome failed T cell reconstitution 6 months after HSCT. She started rhIGF-I therapy just before age 18 months. The initial dose (40 microg/kg twice daily) was increased every 2 weeks to a maximum dose of 120 microg/kg twice daily. The patient's absolute T cells increased from 7 to 132/mm(3) and 662/mm(3) after 3 and 5 months of rhIGF-I therapy, respectively, and her blastogenic response to phytohemagglutinin normalized. Three months after discontinuation of rhIGF-I therapy, the T cells continued to increase (to 2,427/mm(3)) although the blastogenic response to phytohemagglutinin decreased. This is the first known use of rhIGF-I therapy to help restore thymopoiesis in a child.

Photoperiod–Testicular–Immune Interaction in a Seasonal Breeder Indian Palm Squirrel Funambulus pennanti During the Reproductively Inactive and Active Phases

The differential effect of long (LD; 16 : 8 h light/dark), short (SD; 10 : 14 h light/dark) and natural day length (NDL; 12 : 12 h light/dark) during the reproductively inactive (RIP) and active (RAP) phases was assessed in relation to immunity and reproductive function of a tropical rodent Funambulus pennanti. They presented high immunity and low testicular activity during RIP and an opposite during RAP. SD increased spleen and thymus weight, leukocyte and lymphocyte counts, cell mediated immunity [i.e. blastogenic response in terms of percentage stimulation ratio of splenocytes and thymocytes (when challenged with concanavalin A)] and delayed type hypersensitivity to oxazolone. SD during RIP increased the above mentioned parameters and reduced testes weight compared to NDL groups. During RAP, LD reduced all the immunological parameters when compared with NDL and SD experiencing groups of RIP and RAP phases. The LD group reduced the immunological parameters compared to RAP, suggesting that LD had always an inhibitory effect on immune status being independent of reproductive phases. The intensity of the stimulatory effects of SD and inhibitory effects of LD during both reproductive phases was significantly different. We exposed another set of squirrels to the above photoperiodic schedule for prolonged period (30 weeks) during RAP. A clear testicular refractoriness followed by immunorefractoriness was observed in the group experiencing SD and LD for 30 weeks. The photorefractoriness presented by the testes was inversely related to the immunorefractoriness. The peripheral melatonin level of those squirrels reflected the photoperiodic signal perceived by squirrels for immunomodulation and gonadal function, suggesting that immune system and gonadal function might have coevolved.