Blood Lymphocyte Proliferative Responses Research Articles

Fever-range hyperthermia inhibits cells immune response to protein-bound polysaccharides derived from Coriolus versicolor extract.

The aim of the study was to explore whether fever-range hyperthermia (FRH) might enhance the anticancer and immunoregulatory activities of protein-bound polysaccharides (PBP), a class of fungus derived immunomodifiers used in the cancer adjuvant therapy. Blood lymphocytes and breast cancer cells (MCF-7) were cultured at 39.5°C in humidified atmosphere containing 5% CO2 for 2h. After rested at 37°C for 6h, the cells were treated with PBP extract at 100- and 300μg/ml concentration. After indicated time, the proliferative response was analyzed and cytokine mRNA expression assessment was performed by qRT-PCR. In animal model, the FRH was induced by placing rats in the Homeothermic Controller with heating blanket. Animals were heated until Tb reached 39.5°C (±0.2°C) and were maintained at this temperature for 30min. The protein-bound polysaccharides solution was injected i.p. at a dose of 100 mg/kg 6h post FRH. Twenty four hours after treatment, the blood was collected and cytokines expression analysis were performed. The results have shown that fever-range hyperthermia has an inhibitory effect on PBP extract-induced proliferative response of blood lymphocytes, as well as IL-1β and IL-6 mRNA expression. Moreover, the temperature of 39.5°C blocks PBP-induced cytotoxicity against MCF-7 cells, which correlates with significant reduction in TNF-α level. Combined treatment of rats (FRH+PBP) results in decrease of IL-1β, IL-6 and TNF-α mRNA expression in peripheral blood mononuclear cells compared to cells derived from rats treated with protein-bound polysaccharides extract alone. This study demonstrates that fever-range temperature inhibits immunostimulatory as well as anticancer effects mediated by protein-bound polysaccharides.

Polysaccharide peptides from Coriolus versicolor exert differential immunomodulatory effects on blood lymphocytes and breast cancer cell line MCF-7 in vitro

The protein-bound polysaccharides (PBP), isolated from Coriolus versicolor (CV) fungus, are considered as natural compounds with potential therapeutic applications. The immunopotentiating and antitumor activity of polysaccharopeptides has been previously examined, however similar findings could not be achieved. The source of PBP, variations in extraction process as well as environmental factors seems to affect the biological properties of these active CV components. Since further analysis are needed to draw more definite conclusion, the present study aimed to investigate the immunomodulatory properties of the PBP extract, isolated from commercially available capsules of C. versicolor. Our results revealed that the effect mediated by PBP extract depends on the target cells. We reported that the polysaccharopeptides induced a significant decrease in breast cancer MCF-7 cells growth, which was TNF-α-dependent phenomenon. Interestingly, the level of two others cytokines, IL-1β and IL-6 was not affected. On the other hand, in this study we noticed that protein-bound polysaccharides extracted from CV significantly augmented the proliferative response of blood lymphocytes in a time-dependent manner, which was associated with IL-6 and IL-1β mRNA upregulation. Moreover we found that the cells response to PBP stimuli might be inversely related to its concentration.

Reactivity of the immunological system of rats stimulated with Biolex-Beta HP after cyclophosphamide immunosuppression.

The objective of this study was to determine the stimulating effect of the Biolex-Beta HP (β-1,3/1,6-D-glucan) dietary supplement on selected parameters of specific and non-specific humoral and cellular immunity in rats immunosuppressed with cyclophosphamide. The experimental material comprised 40 Wistar rats, divided into two equal groups: control and experimental. In the course of 3 successive days, the rats from the experimental group were administered cyclophosphamide intramuscularly at a rate of 50 mg/kg BW per day. On the 8th day of the experiment, 10 control and 10 experimental rats were sacrificed, and total protein and γ-globulin levels, lysozyme and ceruloplasmin activity were determined in the blood serum. The proliferative response of blood lymphocytes after stimulation with lipopolysaccharide or concanavalin A, respiratory burst activity and the potential killing activity of phagocytes were determined in whole heparinised blood. Starting on the 8th day of the experiment, the feed of the remaining rats from the experimental and control groups was supplemented for 14 consecutive days with Biolex-Beta HP at a rate of 50 mg/kg BW per day. On day 22, arterial blood samples were collected and immune parameters were determined. The results indicate that β-1,3/1,6-D-glucan has a positive effect on the analysed parameters of non-specific cellular and humoral immunity after cyclophosphamide-induced suppression. Nevertheless, the observed effect only marked a return to the norm, as most of the analysed parameters were merely restored to their initial levels, with the exception of lysozyme activity, which considerably exceeded the level noted before immunosuppression.

The innate immunity of wild Eurasian beaver from Poland - present knowledge and the need for research.

For the first time we tried to examine the basic parameters of innate immunity in beavers for developing the new methods of prevention against infectious diseases in different reintroduction and translocation programmes in Poland. The aim of the present study was to determine the selected innate immunity parameters in Eurasian beavers living in natural conditions. The analyses of the results showed that the phagocytic ability (RBA) and potential killing activity (PKA) of blood phagocytes were higher in adult beavers compared to the young animals. The similar pattern was observed in proliferative response of blood lymphocytes stimulated by mitogens Concanavalin A (Con A) and lipopolysaccharides (LPS). The highest proliferative response in adult Eurasian beavers was observed. The ceruloplasmin activity in plasma was on similar levels in adult and young Eurasian beavers. The results of humoral mediated immunity showed that the lysozyme activity, total protein and gamma-globulin levels in serum were higher in adult beavers compared to the young beavers. The results of this preliminary study are important in comparative and clinical immunology.

The effect of β-hydroxy-β-methylbutyrate (HMB) on the proliferative response of blood lymphocytes and the phagocytic activity of blood monocytes and granulocytes in calves

The objective of this study was to evaluate the effect of HMB on selected indicators of immunity in calves. The experiment was performed on 14 calves aged 30 +/- 2 days, divided into two equal groups of control (group I) and experimental (group II) animals. The feed administered to experimental group calves was supplemented with HMB at 40 mg/kg BW, whereas control calves were administered standard farm-made feed without supplementation. Blood was sampled from the jugular vein immediately before the experiment (day 0) and on experimental days 15, 30 and 60 to determine the following parameters of immunity: proliferative response of LPS- and ConA-stimulated lymphocytes (MTT), respiratory burst activity (RBA) and potential killing activity (PKA) of phagocytes. The results revealed a significant increase in RBA and MTT values in calves administered HMB in comparison with the control group throughout the experiment. In the group of animals receiving HMB, an increase in PKA values was noted only on day 30.

Lymphocyte proliferative responses to chlamydial antigens in human chlamydial eye infections

In order to study the relationship between cell-mediated immune responses to Chlamydia trachomatis and the pathogenesis of human chlamydial eye disease, we have measured the peripheral blood lymphocyte proliferative responses to whole chlamydial elementary bodies in 40 subjects with oculogenital chlamydial infection of varying severity, 13 subjects with genital chlamydial infections and 12 healthy seronegative controls. The mean stimulation index was significantly higher in those with oculogenital infections than in controls. There was a strong correlation between the response to C. trachomatis serotypes B and L1. We studied the relationship between proliferative responses and four clinical parameters: follicular conjunctivitis, papillary hypertrophy, corneal pannus and epithelial punctate keratitis, but were unable to show a significant association with any of these. Nor was there any association between proliferative response and serum antibody titre to C. trachomatis (pooled serotypes D-K), duration of disease or quantitative isolation of chlamydia from the conjunctiva. The depletion of CD8+ cells had no consistent effect on proliferative responses to serotype L1 in 13 subjects.

Successful vaccination against Onchocerca ochengi infestation in cattle using live Onchocerca volvulus infective larvae

Epidemiological evidence has led to the hypothesis that the concurrent and predominant transmission of Onchocerca ochengi by Simulium damnosum s.l. in sub-Saharan Africa could lead to the protection of humans against onchocerciasis caused by Onchocerca volvulus (zooprophylaxis). To gain support for this hypothesis, we investigated whether exposure to O. volvulus could protect cattle from O. ochengi. Gudali calves were vaccinated with live O. volvulus-infective larvae and subsequently challenged with O. ochengi-infective larvae whilst raised in a fly-proof house. Post-challenge adult parasite and microfilaria development, IgG1 and lgG2 subclass antibodies response to Ov10/Ov11 recombinant Onchocerca antigens, and peripheral blood lymphocyte proliferative responses to O. ochengi crude antigens were studied over a 1-year period. The vaccinated-challenged animals had 83-87% less adult O. ochengi parasites than non-vaccinated-challenged animals. IgG1 and lgG2 antibodies to Ov10/Ov11 recombinant Onchocerca antigens were invoked by non-vaccinated-challenged animals but not by most (80%) of the vaccinated-challenged animals. These findings support the idea of cross-protection (zooprophylaxis) due to inoculation of humans with O. ochengi-infective larvae under natural transmission conditions in endemic areas.

The canine model of dietary hypersensitivity

IgE-mediated dietary hypersensitivity affects approximately 1% of the canine population. There are no breed associations and < or =50% of the patients are aged <1 year at presentation. The most common causative allergens are beef, chicken, milk, eggs, maize, wheat and soyabean. Affected dogs generally display cutaneous disease and 10-15% of the patients may have concurrent alimentary involvement. Diagnosis is currently based on dietary restriction followed by provocation. Procedures for the detection of serum allergen-specific IgE and IgG antibodies are widely available, but these tests correlate poorly with clinical presentation and dietary testing. Recent studies have demonstrated the allergen specificity of IgE antibodies by immunoblotting and have described blood lymphocyte proliferative responses to food allergens. In addition to investigations of spontaneously-arising dietary hypersensitivity, it has also proved possible to study this disorder experimentally. Small colonies of dogs sensitive to particular dietary proteins have been used to study clinical and serological responses to allergen challenge. Hypersensitivity has been experimentally induced in dogs of an atopic phenotype by repeated subcutaneous injection of alum-adjuvanted dietary allergen during neonatal life. These models have been used to trial a range of modified protein or hydrolysate diets. The dog provides a unique large-animal model for investigation of the immunopathogenesis of human dietary hypersensitivity. The dog is closely related genetically to man and shares environmental disease triggers with man. Spontaneously arising canine dietary hypersensitivity is a good clinical mimic of the human disease, and ability to therapeutically manipulate this adverse response in the dog might lead to benefits for human patients.

Severe feed restriction enhances innate immunity but suppresses cellular immunity in chicken lines divergently selected for antibody responses

The main aim of this study was to investigate the effects of different levels of feed restriction (FR) on immune responses of chicken lines divergently selected for high (H) and low (L) antibody responses to SRBC. We hypothesized that severe feed restriction suppresses immune responses and the level of immune suppression differs between birds with different genetic background. Therefore, we tested antibody responses, blood lymphocyte proliferative responses, and production of reactive oxygen intermediates (ROI) by Zymosan A-stimulated blood cells in chicken lines maintained on 3 levels of FR. The H line birds had significantly higher antibody responses, higher ROI production, and lower corticosterone (CORT) levels when compared with the L line birds. Feed restriction induced no significant effect on specific antibody responses to either a T helper 1- (Mycobacterium butyricum) or a T helper 2- (keyhole limpet hemocyanin) type antigen. Feed-restricted birds showed a marked reduction in natural antibodies binding lipoteichoic acid, in vitro lymphocyte proliferation in response to stimulation with concanavalin A, BW gain, and relative lymphoid organ weights compared with the birds fed ad libitum. However, FR birds showed a markedly enhanced ROI production, and plasma CORT levels compared with the birds fed ad libitum. The enhanced ROI production and suppressed lymphocyte proliferation coinciding with enhanced plasma CORT levels suggest stress-mediated immunomodulating effects of FR. A significant treatment by line interaction was found for ROI production; the increase of ROI production was larger in the H line than in the L line under severe FR. The increase in CORT levels was larger in the L line than in the H line under severe FR. Furthermore, the L line gained more BW than the H line under ad libitum conditions. Finally, under severe FR, relative spleen weight was lower in the L line than in the H line. The present findings suggest genetic differences affecting physiological and immunological responses under FR conditions.

Increased protection against bovine tuberculosis in the brushtail possum (Trichosurus vulpecula) when BCG is administered with killed Mycobacterium vaccae

Setting: The Australian brushtail possum is the major wildlife reservoir for Mycobacterium bovis infection in New Zealand. Development of an effective tuberculosis vaccine for possums will reduce the spread of infection to cattle and farmed deer.Objectives: To determine whether killed M. vaccae can improve the efficacy of vaccination with M. bovis bacillus Calmette Guerin (BCG) against bovine tuberculosis in the possum.Design: Groups of possums (n=6–8) were vaccinated via intranasal and intraconjunctival routes with BCG alone or BCG in combination with heat-killed M. vaccae. Controls were non-vaccinated or vaccinated with heat-killed M. vaccae alone. After challenge with virulent M. bovis, protection was assessed by a reduction in loss of body weight and bacterial counts in lungs and spleens. Blood lymphocyte proliferative responses to M. bovis purified protein derivative were monitored throughout.Results: The earliest lymphocyte responses following vaccination were from animals inoculated with BCG plus 100 μg heat-killed M. vaccae. Loss of body weight was significantly reduced in all BCG-vaccinated groups compared control groups. Spleen bacterial counts were significantly lower in animals vaccinated with M. vaccae plus BCG compared to the non-vaccinated group. Furthermore, vaccination with 100 μgM. vaccae plus BCG significantly reduced spleen bacterial counts compared to vaccination with BCG alone.Conclusion: The possum infection model is one of the first to show that novel vaccine strategies may offer better protection against tuberculosis than BCG alone.

Effect of Subcutaneous Injection of Ginseng on Cows with Subclinical Staphylococcus aureus Mastitis

Cows with subclinical mastitis caused by Staphylococcus aureus were subjected to subcutaneous injections with either an extract from the root of Panax ginseng CA Meyer at a dose of 8 mg/kg body weight per day for 6 days, or with saline as a control. The injection areas were checked for adverse reactions. The daily milk production was measured before and after treatment. Blood was collected for total and differential leucocyte counts, identification of lymphocyte subpopulations using flow cytometry, lymphocyte proliferation test, and neutrophil phagocytosis and oxidative burst assay. Quarter milk samples were collected for bacteriological analysis and somatic cell counts (SCC). After the end of treatment, the numbers of S. aureus‐infected quarters and milk SCC tended to decrease in ginseng‐treated cows. Phagocytosis and oxidative burst activity of blood neutrophils were significantly increased 1 week after ginseng treatment, but the proliferative response of blood lymphocytes did not change significantly. The number of monocytes in ginseng‐injected cows was significantly higher 1 week post‐treatment than pre‐treatment, and the number of lymphocytes was significantly higher than pre‐infusion at 2 and 3 weeks after ginseng treatment. Similar changes were not observed in the control group. The present findings indicate that ginseng treatment can activate the innate immunity of cows and may contribute to the cow’s recovery from mastitis. It is therefore suggested that ginseng has a potential as a stimulator of the immune system of dairy cows.

Role of central opioid receptor subtypes in morphine-induced alterations in peripheral lymphocyte activity

Morphine has been shown to decrease proliferative responses of rat T-lymphocytes via central opioid receptors, however, the specific receptor subtype(s) mediating this effect have not been established. To determine the potential role of central μ opioid receptors in morphine-mediated suppression of T-lymphocyte proliferation, 20 nmol/2 μl of either morphine sulfate or DAMGO ( μ-selective agonist) were administered into the lateral ventricle of freely moving Sprague–Dawley rats. Lymphocyte proliferative response to the polyclonal T cell mitogen concanavalin A (ConA), changes in splenic natural killer cell (NK) cytolytic activity, activation of the hypothalamic–pituitary–adrenal (HPA) axis and antinociception (tail-flick latency) were examined. Results indicated that like morphine, DAMGO decreased blood lymphocyte proliferative responses by 80% and elevated both tail-flick latency and plasma corticosterone when compared to saline-treated animals. The proliferation response of lymphocytes from the spleen or thymus and splenic NK cell activity were not significantly altered by either morphine or DAMGO treatment. The effects of DAMGO were determined to be dose-dependent and completely antagonized by naltrexone pretreatment. Central administration of DPDPE ( δ-selective agonist) and U-50488 ( κ-selective agonist) produced between 40–50% suppression of blood lymphocyte proliferation responses only at a dose five times greater (100 nmol) than DAMGO treatment, without altering antinociception or activation of the HPA axis. To determine the central opioid receptor subtype(s) involved in the effects of morphine, selective opioid antagonists were microinjected into the lateral ventricle prior to morphine treatment (6 mg/kg, s.c.). CTOP ( μ-selective antagonist, 5 μg/2 μl) completely blocked the effects of morphine on all parameters measured, however, naltrindole ( δ-selective antagonist, 2 μg/2 μl) or nor-binaltorphimine ( κ-selective antagonist, 73.5 μg/2 μl) failed to block the effects of morphine. Collectively, these results provide evidence that morphine acts primarily through central μ receptors to modulate peripheral blood lymphocyte proliferation responses. Further, the antinociception and blood lymphocyte effects show greater sensitivity to opioids than either natural killer cell cytolytic activity or activation of the HPA axis.

Cord Blood Lymphocyte Responses to Antigens for the Prediction of Allergy

Background It is not sufficient to predict a high allergic risk for newborns on the basis of an ncreased cord blood immunoglobulin E (IgE) concentration alone, because an increased cord blood IgE concentration is specific but not sensitive in the prediction of the developn,ent of allergic disorders. Objective We describe two cases in which cord blood lymphocyte proliferative responses to food antigens were useful for prediction of the development of allergic disorders. Methods and results The cases of two full-term newborn infants who developed food-sensitive atopic dermatitis after the age of 10 months were investigated. The cord blood IgE concentrations and the proliferative responses of cord blood lymphocytes to food antigens were measured. In both cases, the clinical features from 10 months after birth were closely related to the proliferative responses of the cord blood lymphocytes to each of the food antigens but not to the cord blood IgE concentration. Conclusion Analysis of the proliferative res...

Induction of a T- and B-cell response against a unique amino acid sequence of the mouse IgG2A hinge region in a MAb-treated patient

A patient with malignant glioblastoma was treated with intratumoral infusions of the murine MAb425 (IgG2A) directed against the epidermal growth factor receptor. At the 10th infusion, the patient developed somnolence, fever and headache. The symptoms increased during the subsequent 48 hr but then gradually disappeared within a week. The cerebrospinal fluid (CSF) contained increased concentrations of interleukin-2. The main CSF cell subset was CD4 T-cells. A marked blood lymphocyte proliferative response against mouse IgG2A was noted. The reactive T-cell epitope(s) could be localized to a 14 amino acid (RGPTIKPCPPCKCP) long peptide of the hinge region. A B-cell response (IgG antibodies) against this peptide was also induced.

Characterization of immune response to Eimeria tenella antigens in a natural immunity model with hosts which differ serologically at the B locus of the major histocompatibility complex.

A model to simulate natural immunity to Eimeria tenella was developed in three chicken lines which differ at the B locus of the major histocompatibility complex. Homozygous, 1-day-old chicks of the B19B19, B24B24, or B30B30 genotype were trickle immunized by being orally fed a small infectious dose of E. tenella oocysts for 5 consecutive days. These naturally exposed birds were then challenged at different times between 5 and 24 days after the final dose, and the level of protection was assessed 6 days after challenge, using body weight gain and intestinal lesion scores. The duration of immunity in naturally exposed birds differed among the major histocompatibility complex lines. Trickle immunization of the B19B19 haplotype afforded the longest and strongest level of protection compared to the other two haplotypes tested. In addition, in vitro splenic and peripheral blood lymphocyte proliferative responses in trickle-immunized birds were measured against sporozoite, merozoite, and tissue culture-derived E. tenella parasite antigens isolated from the recently described SB-CEV-1/F7 established cell line. The lymphocytes obtained from B19B19 birds trickle immunized responded in vitro to the E. tenella-infected SB-CEV-1/F7 tissue culture-derived parasite antigen. Furthermore, antigen-specific immune responses appeared earlier in immune, challenged B19B19 birds than in their naive, challenged counterparts. The development of a model simulating natural immunization will serve as a foundation to further characterize both humoral and cell-mediated responses to E. tenella tissue culture-derived parasite antigens and to better understand host protective immune responses to avian coccidiosis.

Absence of involvement of nitric oxide in LP-BM5-induced immunodeficiency syndrome

To examine the role of nitric oxide (NO) in murine AIDS (MAIDS) pathogenesis, we determined NO production and inducible NOS (iNOS) mRNA expression in the macrophages of LP-BM5-infected mice, together with the in vivo effects of L-NAME, a competitive inhibitor of NO synthase. LP-BM5 infection induced neither spontaneous nitrite production nor iNOS mRNA expression. No differences in IFN gamma + LPS-induced nitrite production or iNOS mRNA expression were observed in macrophages, from non-infected or infected mice. Spleen weight, ecotropic MuLV replication, the blood lymphocyte phenotype and proliferative response of splenocytes were not modified by L-NAME. LP-BM5 infection did not increase macrophage NO production and NO production did not appear to protect against LP-BM5-induced immunodeficiency.

Absence of involvement of nitric oxide in LP-BM5-induced immunodeficiency syndrome

To examine the role of nitric oxide (NO) in murine AIDS (MAIDS) pathogenesis, we determined NO production and inducible NOS (iNOS) mRNA expression in the macrophages of LP-BM5-infected mice, together with the in vivo effects of l-NAME, a competitive inhibitor of NO synthase. LP-BM5 infection induced neither spontaneous nitrite production nor iNOS mRNA expression. No differences in IFNγ + LPS-induced nitrite production or iNOS mRNA expression were observed in macrophages from non-infected or infected mice. Spleen weight, ecotropic MuLV replication, the blood lymphocyte phenotype and proliferative response of splenocytes were not modified by l-NAME. LP-BM5 infection did not increase macrophage NO production and NO production did not appear to protect against LP-BM5-induced immunodeficiency.

Peripheral blood lymphocyte proliferative responses in cattle infected with or vaccinated against Anaplasma marginale.

An assay was developed for measurement of the peripheral blood lymphocyte proliferative response (PBLPR) in cattle infected with or immunised against Anaplasma marginale. PBLPR was not evident in all cattle that had recovered from A. marginale infection. However, A. marginale-sensitised lymphocytes were detected in the spleens of all immune cattle tested in the absence of detectable PBLPR. During the course of initial infection, cattle exhibited detectable PBLPR for a period corresponding with and up to 2 weeks after patent parasitaemia, followed by a second, usually larger peak in PBLPR corresponding to the time of sub-clinical relapse of cattle. Analysis of the PBLPR of A. marginale chronically infected cattle demonstrated highly variable PBLPR between individuals and over time. A positive PBLPR was induced in cattle by vaccination using a crude A. marginale antigen preparation. The PBLPR of vaccinated cattle subsequently infected with A. marginale was markedly different from that of naive cattle, with reduced PBLPR being associated with the onset of parasitaemia. The antigen used in the PBLPR assay was inactivated by proteolysis. Proteolysis also abolished immunity that had been induced in cattle vaccinated using the antigen preparation. A marginale-sensitised PBL did not proliferate in response to antigen from the heterologous species A. centrale. A. centrale-sensitised PBL, however, responded to A. marginale antigen. Interferon-gamma (IFN-gamma) was detected in PBLPR-assay supernatants and was associated with a strong PBLPR.

Study on immune response of goats vaccinated with a live strain of Mycobacterium paratuberculosis

A study of the immune response of goats vaccinated with a live strain (316-F) of Mycobacterium paratuberculosis has been performed, taking into consideration both cellular and serological responses using a lymphocyte transformation assay (LTA) and a counterimmuno-electrophoresis test (CIET). The analyses made at three monthly intervals reveal an in vitro proliferative response of blood lymphocytes to specific antigen and lymphocytes T mitogen over a year of post-vaccination, but no humoral response was observed when CIET was used.

Tolerance and crosstolerance to the suppressive effects of cocaine and morphine on lymphocyte proliferation

The effects of acute or daily exposure to either cocaine or morphine on lymphocyte proliferative responses and NK cytolytic activity were determined. Two hours following the IV infusion of cocaine (5 mg/kg), blood lymphocyte proliferative responses were found to be suppressed by 75%. Cocaine had no effect on proliferative responses of thymic or splenic lymphocytes or cytolytic activity of splenic NK cells following acute or 5-day repetitive dosing. Similar to the effects of cocaine, morphine (10 mg/kg) administration was also accompanied by a suppressed blood lymphocyte response, which was no longer apparent 8 days following repeated morphine injections. Animals that had received daily injections of either morphine of cocaine were also found to be resistant to the inhibitory effects of a single dose of morphine or cocaine, respectively. These data suggest repeated exposure to either morphine or cocaine results in the development of an apparent crosstolerant state to further suppression of blood lymphocyte proliferative responses by either drug.