Proliferation Stimulation Index Research Articles

Immunomodulatory Activity of Robusta Green Coffee Extract on Macrophage Phagocytosis and Lymphocyte Proliferation in Vitro

Robusta green coffee contains a large quantity and variety of polyphenols and flavonoids which are known to have antioxidant and anti-inflammatory properties. This research to investigate the immunomodulatory activity of Robusta green coffee extract on macrophage activity stimulated with lipopolysaccharide (LPS) and lymphocyte proliferation. Macrophages were isolated from mice peritoneal tissue, and Lymphocytes were isolated from lymph organs. The immunomodulatory activity on macrophages was evaluated based on Active Phagocytes Cells (APC) and the proliferation of lymphocytes was evaluated by the MTT Assay which was previously given extracts with varying concentrations (20 µg/ml, 40 µg/ml, 50 µg/ml, 60 µg/ml, 70 µg/ml), then the data were analyzed by ANOVA (α=0.05). The aqueous extracts of Robusta green coffee significantly affects macrophage activity and lymphocyte proliferation in vitro. The extract was decrease the percentage of phagocytosis (PP) macrophage stimulated with LPS, however there was increase phagocytosis capacity (PC) and lymphocyte proliferation stimulation index is 3, illustrating the extract has a low immunomodulatory effect on lymphocyte proliferation. Based on these results, Robusta green coffee extract holds potential as an immunomodulatory agent, affecting against macrophage phagocytosis and lymphocyte proliferation.

Immunomodulatory and Antioxidant Activities of Select Indonesian Vegetables, Herbs, and Spices on Human Lymphocytes.

Edible plants have attracted increasing attention as functional foods as they are rich in bioactive compounds with health benefits, including antioxidant and immunomodulatory activities. However, scientific evidence of these health effects is limited. This study is aimed at determining antioxidant and immunomodulatory activities of 25 select vegetables, herbs, and spices commonly consumed in Indonesia. Phytochemical profiles were determined by measuring total flavonoid content and 1H-NMR. Human blood lymphocyte cells were used to probe the immunomodulatory potency and treated with the methanol extract of these vegetables, herbs, and spices. The results showed the enhanced propensity for all tested plant extracts to stimulate lymphocyte proliferation, except Pandanus amaryllifolius. Etlingera elatior, Ocimum xcitriodorum, Kaempferia galanga, and Apium graveolens had the highest lymphocyte cell proliferation stimulation index (SI) at concentrations of 41.67, 16.67, 4.17, and 2.5 mg/mL culture, respectively (SI 2.21 ± 0.05, 2.62 ± 0.12, 3 ± 0.05, and 2.64 ± 0.07, respectively). The NMR spectra of these four most potent plants showed low peaks in the aromatic/phenolic area and several other peaks indicating the presence of terpenoid, steroid, amino acid, and sugar compounds. The results demonstrate the immunomodulatory potential of all vegetables, herbs, and spices, except P. amaryllifolius, although this potential did not necessarily correlate with flavonoid content and antioxidant activity. Nevertheless, this research showed promising health effect, particularly immunomodulation, of the various local plants. Further elaboration on the specific immunomodulatory activity will be interesting.

Neutral polysaccharide from Panaxnotoginseng enhanced cyclophosphamide antitumor efficacy in hepatoma H22-bearing mice

BackgroundOur previous studies demonstrated that the administration of crude Polysaccharide from Panax notoginseng (CPPN) can effectively prolong the lifespan of tumor-bearing mice via boosting the host immune system as well as weak cytotoxicity against hepatocellular carcinoma (HCC). In the present study, Neutral Polysaccharide (NPPN) were further purified from crude polysaccharide isolated from panax notoginseng. The effects of NPPN on the immune function and hematopoietic function of mice with low immunity and myelosuppression induced by cyclophosphamide (CTX) were investigated. The effect of NPPN combined with CTX on the tumor inhibition rate of the H22 tumor-bearing mice and the impact of NPPN on the proliferation of H22 liver cancer cells in vitro were investigated.MethodsCPPN was obtained by water extraction and alcohol precipitation method, and further purified by DEAE Sepharose Fast Flow ion exchange resin column. NPPN was added to the immunosuppressed with myelosuppression mice induced by CTX. Thymus index, spleen index, lymphocyte proliferation stimulation index by adding of concanavalin A, determination of serum hemolysin, NK cell activity assay, mice carbon clearance experiment, blood count tests were detected. The tumor inhibition rate of the H22 tumor-bearing mice treated with NPPN combined with CTX was recorded.ResultsNPPN and 4 kinds of acid polysaccharide from Panax notoginseng (APPN) were successfully isolated from the CPPN by DEAE Sepharose Fast Flow ion exchange resin column. NPPN inhibited the growth of H22 cells and significantly increase the tumor inhibition rate of the H22 tumor-bearing mice combined with CTX. The elevation of the cellular and humoral immunity levels as well as a variety of blood count tests indicators of immunosuppressive with myelosuppression mice may contribute to the antitumor activity of NPPN.ConclusionNPPN has a potential antitumor activity for the treatment of liver cancer combined with cyclophosphamide.

MiR-669b-3p regulates CD4+ T cell function by down-regulating indoleamine-2, 3-dioxygenase

ObjectiveAcute rejection is a major cause of morbidity and mortality after solid organ transplantation. Therefore, optimizing treatment strategies and improving curative effect is urgent and necessary. Reliable biomarkers for acute rejection and the underlying molecular mechanisms remain to be determined. MethodsIn this study, we established a mouse-to-mouse cardiac transplantation model and identified miR-669b-3p as a potential biomarker of acute rejection using a microRNA polymerase chain reaction (PCR)-based chip assay. ResultsFurther analyses showed that miR-669b-3p negatively regulated indoleamine-2,3-dioxygenase (IDO), a rate-limiting enzyme of tryptophan catabolism inhibiting T cell function. Using mixed lymphocyte reaction assay, we showed that miR-669b-3p increased proliferation stimulation index and inhibited apoptosis in CD4+ T cells. Moreover, miR-669b-3p regulated the expression of inflammatory cytokines such as tumor necrosis factor alpha (TNF-α) and Interleukin 10 (IL-10) and contributed to cytokine shift towards a Th2-dominant response. ConclusionOur results advance the current understanding of the immune regulatory function of miRNA and shed light on the role of miR-669b-3p in CD4+ T cells, suggesting that miR-669b-3p is a potential target for acute allograft rejection.

The Effect of Black Soybean Tempe Extract on the Increase of Proliferation Stimulation Index (PTI), Protein Tyrosine Kinase Enzyme Activity and Proliferating Cell Nuclear Antigen of Human Lymphocytes

Phytochemical components of tempe could interact with receptors on the surface of lymphocytes, which then increase the activity of the enzyme of protein tyrosine kinase and DNA polymerase. The aim of this study was to determine the effectiveness of black soybean tempe extract on the increase of proliferation stimulation index, protein tyrosine kinase enzyme activity, and proliferating cell nuclear antigen of human lymphocytes, in vitro. A number of 15 ml blood samples were drawn from a total of five respondents. The lymphocytes were then separated from these blood samples and later cultured. The black soybean tempe extract was added in varied concentrations of 0, 25, 50, 100, and 200 µg/ml in the culturing process and used as a positive control in cultures with added PHA. Lymphocyte cultures were incubated for 72 hours and after that their PSI, PTK enzyme activity and PCNA were analyzed. The results showed that the extract of black soybean tempe added to cultured human lymphocytes significantly affected the lymphocytes PSI, PTK enzyme activity and PCNA, where the increased level of black soybean tempe extract added into the lymphocyte culture tended to raise the values of PSI, PTK enzyme activity and PCNA. The addition of black soybean tempe extract to cultured human lymphocytes affected the lymphocytes PSI, PTK, and PCNA.

F1 Domain of the Leishmania (Leishmania) donovani Nucleoside Hydrolase Promotes a Th1 Response in Leishmania (Leishmania) infantum Cured Patients and in Asymptomatic Individuals Living in an Endemic Area of Leishmaniasis.

The Leishmania (Leishmania) donovani nucleoside hydrolase NH36 is the main antigen of the Leishmune® vaccine and one of the promising candidates for vaccination against visceral leishmaniasis. The antigenicity of the N-terminal (F1), the central (F2), or the C-terminal recombinant domain (F3) of NH36 was evaluated using peripheral blood mononuclear cells (PBMC) from individuals infected with L. (L.) infantum from an endemic area of visceral leishmaniasis of Spain. Both NH36 and F1 domains significantly increased the PBMC proliferation stimulation index of cured patients and infected asymptomatic individuals compared to healthy controls. Moreover, F1 induced a 19% higher proliferative response than NH36 in asymptomatic exposed subjects. In addition, in patients cured from visceral leishmaniasis, proliferation in response to NH36 and F1 was accompanied by a significant increase of IFN-γ and TNF-α secretion, which was 42–43% higher, in response to F1 than to NH36. The interleukin 17 (IL-17) secretion was stronger in asymptomatic subjects, in response to F1, as well as in cured cutaneous leishmaniasis after NH36 stimulation. While no IL-10 secretion was determined by F1, a granzyme B increase was detected in supernatants from cured patients after stimulation with either NH36 or F1. These data demonstrate that F1 is the domain of NH36 that induces a recall cellular response in individuals with acquired resistance to the infection by L. (L.) infantum. In addition, F1 and NH36 discriminated the IgG3 humoral response in patients with active visceral leishmaniasis due to L. (L.) donovani (Ethiopia) and L. (L.) infantum (Spain) from that of endemic and non-endemic area controls. NH36 showed higher reactivity with sera from L. (L.) donovani-infected individuals, indicating species specificity. We conclude that the F1 domain, previously characterized as an inducer of the Th1 and Th17 responses in cured/exposed patients infected with L. (L.) infantum chagasi, may also be involved in the generation of a protective response against L. (L.) infantum and represents a potential vaccine candidate for the control of human leishmaniasis alone, or in combination with other HLA epitopes/antigens.

Evaluation of Immunostimulatory Effects of N-(2-Hydroxy) Propyl-3-Trimethylammonium Chitosan Chloride for Improving Live Attenuated Hepatitis A Virus Vaccine Efficacy.

This study was to evaluate the immunostimulatory effects of N-(2-hydroxy) propyl-3-trimethylammonium chitosan chloride (HTCC) as an adjuvant for improving a commercial live attenuated hepatitis A virus (HAV) vaccine efficacy in mice. Mice in the experimental group were intraperitoneally immunized with a solution of HTCC and live attenuated HAV vaccine. And for those injected with sterile water, HTCC or live attenuated HAV vaccine were treated as mock group, negative group, and positive group in turn. The serum HAV-specific IgG titers and the ratios of the serum HAV-specific IgG2a/IgG1 in the experimental group were significantly increased (p = 0.00042 and p = 0.040, respectively). Splenocyte proliferation stimulation index in experimental group was higher than positive group (p = 0.021), and significantly higher than mock group and negative group (p = 0.0078 and p = 0.0050, respectively). The percentages of CD4+ T lymphocytes in the peripheral blood in experimental group were significantly higher than positive group, negative group, and mock group (p = 0.012, p = 0.012, and p = 0.045, respectively). Compared to the other three groups, experimental group showed a slightly higher ratio of CD4+/CD8+, but there were no significant differences (p > 0.05). In the percentages of CD8+ T lymphocytes, there were no significant differences among the four groups (p > 0.05). HTCC can enhance live attenuated HAV vaccine to generate stronger humoral responses and induce a Th1-biased immune response, as well as IgG2a class switching, compared with the live attenuated HAV vaccine alone. This study validated an important concept for further development of a safe and potent vaccine adjuvant.

Changes in some parameters of immune response under the effect of adrenoceptor blockers

We studied the effect of α-and β-adrenoceptor blockers on humoral and cell immune response in (CBA×C57Bl/6)F1 mice under normal conditions and during stress. The suppressive effect of α-adrenoceptor blocker dihydroergotamine on humoral immune response against the background of both immunization and combined effects of immobilization and injection of a thymus-dependent antigen was more pronounced compared to that of β-adrenoceptor blocker propranolol. Dihydroergotamine suppressed proliferative activity of T cells, stimulated proliferation of B cells at late terms after immunization, but inhibited it against the background of stress and antigen treatment. Propranolol stimulated proliferative activity of T and B cells at early terms after both immunization alone and stress with antigen treatment, but decreased the proliferation stimulation index of lymphoid cells at late terms after treatment (days 7–10).

Dog Leukocyte Antigen (DLA)-Identical Sibling Cord Blood Transplantation (CBT) Following Myeloablative Total Body Irradiation (TBI).

Cord blood (CB) is increasingly used for hematopoietic cell transplantation (HCT) due to its rapid availability and less stringent HLA matching requirements. However, low cell dose of available CB units and delayed engraftment remain significant obstacles for increased use of CBT in adults. Recent clinical results with 2-unit CBT may help overcome these problems. We sought to develop a large animal model of CBT to improve the understanding of engraftment across histocompatibility barriers and cell dose limitations of multiple units of CBT. Since the outbred dog is a very well-established preclinical model of HCT, our first aim was to establish feasibility of canine CBT. We harvested and cryopreserved individual units of canine CB obtained from litters following Caesarian section at day 56 to 60 of gestation. We asked if multiple units of previously cryopreserved, DLA-identical sibling CB could engraft in DLA-identical recipient dogs after myeloablative total body irradiation (TBI). Five adult dogs (8–18 months of age) received 920 cGy TBI (7 cGy/min) followed by intravenous infusion of 2 to 4 units of thawed CB with a combined total nucleated cell dose range 0.4x107 − 2.6x107/kg. Transplanted total CD34+/kg cell dose range was 0.5−2.5x105/kg. Postgrafting immunosuppression was combined cyclosporine and mycophenolate mofetil for 35 and 28 days, respectively. G-CSF was given until recovery of neutrophil counts, and dogs received blood product transfusion support and intravenous and oral antibiotics. Four dogs engrafted and survived; they are currently 35 to 212 days after CBT. One dog died on day 13 due to neutropenic sepsis. All 4 surviving dogs engrafted with sustained neutrophil recovery >1000/mL between 29 – 35 days after CBT. Bi-weekly chimerism analysis was assessed by PCR using informative microsatellite markers. For each of the 4 surviving recipients all transplanted donor CB units were detected after engraftment. However, among the 3 dogs currently at 3 to 7 months after CBT, only 1 of the multiple transplanted CB units dominated hematopoiesis with 75–95% donor chimerism. For 2 of the 3 dogs, the CB unit with the highest CD34+ dose was the dominant donor. There was no evidence of acute or chronic graft-versus-host disease (GVHD). Among the 3 dogs at 3 to 7 months after CBT, immune reconstitution studies included normal T cell proliferation stimulation index (mixed lymphocyte reaction) and gradual recovery of the absolute number of CD4 and CD8 T-cell subsets. In summary, cryopreserved DLA-matched CB successfully engrafted and provided durable hematopoietic recovery in lethally irradiated dogs. The cell dose of transplanted CB units was comparable with CB units for adult human transplantation. These results establish that multiple units of canine CB with limited cell dose can engraft and initiate immune reconstitution without GVHD in DLA-identical dogs transplanted after marrow-lethal irradiation.

Comparative immunomodulatory effect of scopoletin on tumoral and normal lymphocytes

Some coumarins possess enhancing effects on lymphocyte mitogen responsiveness. In this investigation, the activity of scopoletin, a coumarin that has been isolated from different plants and in this case specifically from T. cordata Mill., was evaluated. For this purpose, normal T lymphocytes and a hyperproliferative T lymphoma cell line were used. Scopoletin was found to exert a dual action on tumoral lymphocytes exhibiting both a cytostatic and a cytotoxic effect. These effects varied with the concentrations analysed and the time of cell incubation (EC 50: 251 ± 15 μg/ml) and were associated to the induction of apoptosis. Scopoletin induced cell proliferation on normal T lymphocytes (Proliferation stimulation index: 1 μg/ml scopoletin: 1.26 ± 0.1; 10 μg/ml scopoletin: 3 ± 0.25; 100 μg/ml scopoletin: 1.86 ± 0.08); this stimulatory action was found to be due to the interaction with kinase C (PKC) protein. These results indicate that scopoletin could be a potential antitumoral compound to be used for cancer treatment.

In vitro proliferative response of human peripheral blood mononuclear cells to concanavalin A

The proliferation response of human peripheral blood mononuclear cells (PBMC) to concanavalin A (con A) was tested in a medium with or without addition of fetal calf serum (FCS) or a serum substitute. The time profile of the proliferative response of PBMC to con A was different in cells cultured in a medium supplemented with or without FCS. This different pattern occurred for the con A-induced DNA-synthesis as well as for the RNA- or protein-synthesis. The peak proliferation rate and the stimulation index of proliferation of human PBMC to con A was also higher when the cells were cultured in a medium with 10% FCS compared to cells cultured in a FCS-free medium. Substitution of the fetal calf serum by a serum substitute even induced a profound inhibition of the de novo synthesis of DNA in human PBMC. The results indicate that a lymphocyte proliferation response to con A can still be obtained in a culture medium where no FCS was added, although a weaker stimulation occurred in comparison to a culture medium with FCS. However, addition of a serum substitute caused a marked inhibition of the lymphocyte proliferation rate.

Immunologic mechanisms in mosquito allergy: correlation of skin reactions with specific IgE and IgG antibodies and lymphocyte proliferation response to mosquito antigens.

Allergic reactions to mosquito bites are a common problem. Although IgE-mediated hypersensitivity has been reported, other immunologic mechanisms may be involved. To study the relationship between skin bite reactions and immunologic parameters. Forty-one subjects were experimentally exposed to mosquito (Aedes vexans) bites. Immediate and delayed skin reactions were traced at 20 minutes and 24 hours, respectively, after the bites. Sera were analyzed for mosquito salivary gland-specific IgE (mosquito-IgE) and IgG (mosquito-IgG) by ELISA. Lymphocyte proliferation assays with mosquito extract were also performed. One of 41 subjects had only a delayed skin reaction to the bite, 23 had both immediate and delayed reactions, 6 had only immediate reactions, and 11 had no reaction. The mean mosquito-IgE and -IgG concentrations were higher in the subjects with immediate reactions than in those without immediate reactions (P < .007). The mean lymphocyte proliferation stimulation index was higher in the subjects with delayed reactions than in those without delayed reactions (P < .015). Further, both mosquito-IgE and -IgG levels correlated with skin immediate and delayed reactions (P < .04), while lymphocyte proliferation indices only correlated with skin delayed reactions (P < .006). Inverse correlations were found between the size of skin reactions and the number of years lived in Canada (P < .04), but not with age. These results indicate that IgE-, lymphocyte- and, probably, local IgG immune-complex-mediated hypersensitivities are involved in mosquito allergy. Naturally acquired desensitization to mosquito bites occurs during long-term exposure.

Immunosuppression and Intracellular Calcium Signaling in Splenocytes from Chicks Infected with Chicken Anemia Virus, CL-1 Isolate

Hematocrits, histopathology, concanavalin A-induced lymphocyte proliferation, intracellular calcium signaling, and lymphocyte subpopulations were analyzed over a 6-week period in individual chicks inoculated with the CL-1 isolate of chicken anemia virus. Lymphoid depletion/atrophy was present in the thymus and bone marrow by 11 days post-infection (PI). Anemia was present at 14 days PI. The mean lymphocyte proliferation stimulation index (SI) of the inoculated group was significantly lower than that of the control group at 11 days PI. This response was reversed at 18 days PI, when the SI of the inoculated group was significantly higher than that of the controls; values subsequently returned to baseline. The increase in intracellular calcium levels in CAV-infected chicks and controls paralleled the proliferative response. Percentages of CD3-, CD4-, CD8-, and natural-killer-positive-staining cells decreased significantly at 18 and 25 days PI. The most dramatic decrease occurred in the CD8-positive-staining cell population at 18 and 25 days PI.