This study was purposed to explore the changes in biological functions of human peripheral blood derived NK Cells after ex vivo expansion with different combinations of interleukin IL2 and/or IL12, IL15. According to different combination of cytokines, cultured NK cells were divided into 4 groups: group IL2, group IL2 + IL12, group IL2 + IL15 and group IL2 + IL15 + IL12. The group in which NK cells were cultured without cytokines was used as control. The cytotoxicity of cultured NK cells to target K562 cells was determined by using cell counting kit-8; the level of IFN-gamma in supernatants of NK cell culture was detected by ELISA; the perforin and granzyme B mRNA expressions were assayed by competitive quantitative RT-PCR. The results showed that the cytotoxicity of expanded NK cells in groups cultured with cytokines at different E:T ratio was significantly higher than that in group without cytokines (p < 0.01), although the cytotoxicity of NK cells in IL2 + IL15 + IL12 group seem to be slightly higher than that in IL2 + IL15 group, but there was no statistic difference (p > 0.05). The IFN-gamma levels in the supernatants of NK cell culture in the presence of cytokines significantly increased, and the IFN-gamma levels in IL2 + IL15 + IL12 group and IL2 + IL12 group were significantly higher than that in others (p < 0.01). The expressions of perforin and granzyme B mRNA of expanded NK cells in groups cultured with cytokines was significantly higher than that in control group (p < 0.01), and was consistent with cytotoxicity of NK cells. It is concluded that there are differences in the functions of NK cells cultured with different cytokines. IL2 and IL15 have synergistic effect on strengthening cytotoxicity of NK cells and promoting cell expansion. However, the main function of IL12 promotes NK cells to secrete IFN-gamma, which plays a role in immunoregulation.
Humans ; Interferon-gamma ; secretion ; Interleukin-12 ; administration & dosage ; pharmacology ; Interleukin-15 ; administration & dosage ; pharmacology ; Interleukin-2 ; administration & dosage ; pharmacology ; K562 Cells ; Killer Cells, Natural ; drug effects ; immunology

AIMTo observe effect of acute normovolemic hemodilution(ANH) with HAES-balanced solution as diluting agent on levels of cytokines including IL-1, IL-2, IL-6 and TNF-alpha in rabbit serum so as to provide theoretical basis for clinical application.

METHODSA total of 20 healthy adult rabbits were enrolled in the study and randomly divided into two groups (10 rabbits per group), i.e., control group (Group C) and HAES group (Group H). Under anesthesia of the rabbits, we performed incision of trachea, high-frequency jet ventilation and liberation of femoral artery and femoral veins. Group C was free from hemodilution. Group H was injected with dilution (2-fold of blood letting volume) via femoral veins during blood letting of the femoral artery. 6% HAES-steril plus compound solution of sodium lactate, with crystal/gel ratio of 2:1, blood letting volume = TBV x (Ho-Hf)/Hav. All blood was transfused back 60-120 min after blood letting. Venous blood was collected before blood letting (T0) and 30 min (T1), 60 min (T2), 120 min (T3) and 24 h(T4) after blood letting to detect Hb and Hct and measure level of IL-1, IL-2, IL-6 and TNF-alpha in serum.

RESULTSIn Group H, levels of IL-1, IL-2, IL-6 and TNF-alpha in serum were increased from T1 after ANH, reached peak at T3 but showed decrease at T4, with significant difference compared with Group C at T1, T2, T3 and T4 (P < 0.01) and significant difference compared with those before ANH (P <0.01). In Group C, there was no significant difference upon IL-1, IL-2, IL-6 and TNF-alpha in serum at different time points.

CONCLUSIONANH with HAES-balanced solution as diluting agent can up-regulate the levels of cytokines IL-1, IL-2, IL-6 and TNF-alpha in rabbit serum. In the meantime, ANH may arouse eustress with low intensity and short action time, which exerts effect of enhancing immune function of the organisms.

Animals ; Female ; Hemodilution ; methods ; Interleukin-1 ; blood ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Male ; Plasma Substitutes ; administration & dosage ; Rabbits ; Random Allocation ; Tumor Necrosis Factor-alpha ; blood

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Humans ; Interferons ; administration & dosage ; Interleukin-2 ; administration & dosage ; Leukemia, Myeloid, Acute ; drug therapy ; Recurrence ; Thalidomide ; administration & dosage

We have investigated whether oral administration of ovalbumin (OVA) could prevent active systemic anaphylaxis to the antigen. Oral tolerance was induced by a single feecfing with 40 mg OVA before, but not after, sensitization characterized by diminished OVA-specific IgE and IgG responses. Feeding 15 mg OVA suppressed anaphylaxis and antibody responses to a lesser extent. Spleen cells from tolerant donors were incapable of transferring the tolerance to naive recipients. Pretreatment of cyclophosphamide (100 mg/kg) 2 days before OVA feeding did not restore the tolerance. Furthermore, in vitro cell mixing studies showed that the proliferation of spleen cells from OVA- sensitized donors was not inhibited by the addition of spleen cells from tolerant donors, arguing against the role of suppressor cells. Anergy was demonstrated by the ability to reverse the tolerant state after culturing tolerant cells in the presence of IL-2. These findings indicate that only a high-dose (40 mg) feeding OVA was found to be effective in inducing tolerance in this experimental system, and demonstrate anergy as the mechanism underlying oral tolerance to systemic anaphylaxis.
Administration, Oral ; Anaphylaxis* ; Antibody Formation ; Cyclophosphamide ; Humans ; Immunoglobulin E ; Immunoglobulin G ; Interleukin-2 ; Ovalbumin ; Ovum ; Spleen ; Tissue Donors

OBJECTIVETo investigate the effects of intensive insulin therapy on inflammatory response and prognosis of patients with severe trauma.

METHODSEighty severely injured patients were divided into intensive insulin therapy group (n = 40, IT) and routine therapy group (n = 40, RT) in random pair. At the time of admission, a continuous infusion of insulin (2-4 U/h) was pumped into the patients of IT group to maintain blood glucose level at 6-8 mmol/L. Patients in RT group were given routine treatment without administration of insulin. Fever, organ injury, and mortality of patients in 2 groups were recorded. Venous blood was drawn from patients of 2 groups on the morning of post treatment day (PTD) 1, 3, 5, and 7. Values of TNF-alpha, C-reactive protein (CRP), IL-2, and IL-10 in plasma were assayed.

RESULTSHigh fever appeared in 9 patients in IT group, and WBC exceeded 10.0 x 10(9) for more than 3 days in 17 patients in this group, versus 20 and 29 patients respectively in RT group. Dysfunction of 1 organ appeared in 31 patients in IT group and 30 patients in RT group. Dysfunction of 3 organs appeared in 10 patients in IT group and 19 patients in RT group. Dysfunction of 4 organs appeared in 7 patients in IT group and 12 patients in RT group. In IT group, 4 patients died within 3 post-injury day (PID), and 1 patient died after PID 3 (total case fatality: 12.5%). In RT group, 5 patients died within 3 PID, and 4 patient died after PID 3 (total case fatality: 22.5%). Plasma levels of TNF-alpha and CRP of patients in IT group were significantly lower than those of patients in RT group on PID 3-7 ( P < 0.05 or P < 0.01), while levels of IL-2 and IL-10 of patients in IT group were significantly higher than those of patients in RT group (P < 0.05 or P < 0.01). Plasma levels of TNF-alpha (1.3 +/- 0.6 microg/L) and CRP (55 +/- 16 mg/L) of patients in IT group on PTD 7 were lowered to the trough level, and they were significantly lower than those of patients in RT group (3.0 +/- 0.8 microg/L, 89 +/- 20 mg/L, respectively, P < 0.01).

CONCLUSIONSIntensive insulin therapy can mitigate systemic inflammatory response and improve prognosis of patients with severe trauma.

Adult ; Aged ; Female ; Humans ; Inflammation ; Insulin ; administration & dosage ; Interleukin-2 ; blood ; Male ; Middle Aged ; Prognosis ; Wounds and Injuries ; therapy ; Young Adult

Animals ; Antineoplastic Agents ; administration & dosage ; Electroporation ; methods ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Liver Neoplasms, Experimental ; immunology ; mortality ; pathology ; therapy ; Male ; Neoplasm Transplantation ; Rats ; Rats, Sprague-Dawley

The paper is focused on the clinical applications of Tanreqing injection after listing, detecting and analyzing the related blood indicators of patients with allergic reactions based on prospective, multi-center, large sample, registration-type clinical safety monitoring nested case-control study (NCCS) to explore the possible mechanisms of allergic reaction of Tanreqing injection, 3 006 patients cases used with Tanreqing injection were monitored, including 3 cases of adverse reactions and 2 cases of allergic reactions. Each patient of allergic reactions, according to the ratio of 1:4 matches four cases of not adverse reactions as a control group of patients, while 5 healthy and 5 cases of volunteers into the healthy group. We examined the correlation detection of cases of allergic reactions among groups such as T-IgE, IgA, IgG, C3, C4, IFN-g, IL-2, IL-4, IL-6, IL-10. Allergic reactions of Tanreqing injection may be mediated by IgE as type I based on existing research data. This results and conclusions will promote the justifiability and safety of clinical applications.
Adolescent ; Adult ; Case-Control Studies ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Humans ; Hypersensitivity ; diagnosis ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Male ; Prospective Studies ; Young Adult

Graft-versus-host disease (GVHD) is an uncommon but fatal complication of orthotopic liver transplantation (OLT). The pathogenesis and mechanism of GVHD after OLT remains unclear and the treatment is therefore largely empirical. The prevention of GVHD including avoidance of closely matched human leukocyte antigen donors and old-age recipients is of particular importance because effective treatment is lacking. To reduce immunosuppressants combined with high dose of methylprednisolone and IL-2 receptor antibodies can be applied as a reasonable regime.
Autoantibodies ; therapeutic use ; Graft vs Host Disease ; drug therapy ; prevention & control ; Humans ; Immunosuppressive Agents ; administration & dosage ; Liver Transplantation ; adverse effects ; Methylprednisolone ; administration & dosage ; Receptors, Interleukin-2 ; immunology

Ginsenoside Rh2 (Rh2) is one of the major bioactive ginsenosides in Panax ginseng. However, the oral bioavailability of Rh2 is low, with P-glycoprotein (P-gp) and CYP3A4 being reported to be the main factors. The purpose of the present study was to determine the enhancing effect of piperine on the oral bioavailability as well as bioactivity of Rh2. The inhibitory effect of piperine on P-gp and CYP3A4 was determined using a Caco-2 monolayer model and a recombinant CYP3A4 metabolic system, respectively. The pharmacokinetics of oral Rh2 (10 mg·kg) administered alone or in combination with piperine (10 and 20 mg·kg) was performed in rats. The immune boosting effect of Rh2 was assessed in rats by measuring IL-12 level after treated by Rh2 alone or co-administered with piperine. The results indicated that piperine significantly increased the permeability of Rh2 and inhibited the metabolism of Rh2. The pharmacokinetic study results showed that the AUC of Rh2 was significantly increased in combination with piperine at high dose (20 mg·kg) when compared to the control group, with relative bioavailability of 196.8%. The increase of Rh2 exposure led to increased serum levels of IL-12. In conclusion, piperine may be used as a bioenhancer to improve pharmacological effect of Rh2 when given orally.
Administration, Oral ; Alkaloids ; administration & dosage ; Animals ; Benzodioxoles ; administration & dosage ; Biological Availability ; Caco-2 Cells ; Cytochrome P-450 CYP3A ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Ginsenosides ; administration & dosage ; pharmacokinetics ; Humans ; Interleukin-2 ; metabolism ; Panax ; chemistry ; Piperidines ; administration & dosage ; Polyunsaturated Alkamides ; administration & dosage ; Rats ; Rats, Sprague-Dawley

EPSAH is an exopolysaccharide from Aphanothece halophytica GR02. The present study was designed to evaluate its toxicity and adjuvant potential in the specific cellular and humoral immune responses in ovalbumin (OVA) in mice. EPSAH did not cause any mortality and side effects when the mice were administered subcutaneously twice at the dose of 50 mg·kg(-1). Hemolytic activity in vitro indicated that EPSAH was non-hemolytic. Splenocyte proliferation in vitro was assayed with different concentrations of EPSAH. The mice were immunized subcutaneously with OVA 0.1 mg alone or with OVA 0.1 mg dissolved in saline containing Alum (0.2 mg) or EPSAH (0.2, 0.4, or 0.8 mg) on Day 1 and 15. Two weeks later, splenocyte proliferation, natural killer (NK) cell activity, production of cytokines IL-2 from splenocytes, and serum OVA-specific antibody titers were measured. Phagocytic activity, production of pro-inflammatory cytokines IL-1 and IL-12 in mice peritoneal macrophages were also determined. EPSAH showed a dose-dependent stimulating effect on mitogen-induced proliferation. The Con A-, LPS-, and OVA-induced splenocyte proliferation and the serum OVA-specific IgG, IgG1, and IgG2a antibody titers in the immunized mice were significantly enhanced. EPSAH also significantly promoted the production of Th1 cytokine IL-2. Besides, EPSAH remarkably increased the killing activities of NK cells from splenocytes in the immunized mice. In addition, EPSAH enhanced phagocytic activity and the generation of pro-inflammatory cytokines IL-1 and IL-12 in macrophages. These results indicated that EPSAH had a strong potential to increase both cellular and humoral immune responses, particularly promoting the development of Th1 polarization.
Adjuvants, Immunologic ; administration & dosage ; Animals ; Cyanobacteria ; chemistry ; Female ; Immunity, Cellular ; Immunity, Humoral ; Immunization ; Interleukin-12 ; immunology ; Interleukin-2 ; immunology ; Killer Cells, Natural ; immunology ; Mice ; Mice, Inbred ICR ; Ovalbumin ; immunology ; Polysaccharides ; administration & dosage ; immunology ; Rabbits ; Th1 Cells ; immunology ; Th2 Cells ; immunology