OBJECTIVE: To explore the expression of CD40/CD40L in multiple myeloma(MM) patients and its influence on prognosis. METHODS: Thirty patients with MM treated in Cangzhou People's Hospital from May 2016 to June 2017 were selected and divided into MM group, then 30 healthy people with a physical examination in our hospital at the same time were selected as the normal group. The serum CD40/CD40L levels of the patients in the two groups was detected by flow cytometry, and its correlation with the lymphocyte population, pathological grade and prognostic significance of MM patients was anaysis. RESULTS: The expression of CD40 in serum of the patients in MM group was significantly higher than those in normal group (P<0.05). The expression of CD40L in serum of the patients in MM group showed no significant difference as compared with those in normal group (P>0.05). The levels of CD40 and CD40L in the patients before and after chemotherapy showed no difference(P>0.05). The levels of Ts and NK cells in the patients of MM group were lower than those in normal group (P<0.05). The proportion of total B lymphocytes, Th and Th/Ts cells between the two groups showed no significant difference (P>0.05). The CD40 level was correlated with the serum total B lymphocyte level of the patients in MM group (r=0.877, P=0.005). There was a correlation with CD40L and Th cells in the serum of MM patients (r=-0.783, P=0.035). The expression of serum CD40 in the patients at phase III-IV was higher than those of the patients at phase I-II, the levels of serum CD40L in MM patients at different periods showed no significant difference(P>0.05). The survival rate of MM patients with high CD40 expression was lower than that of MM patients with low CD40 expression (χ CONCLUSION The increasing of CD40 level in MM patients is related to the pathological grade of the patients. Chemotherapy can reduce the level of CD40. The increasing of CD40 is an important factor for the poor prognosis of MM patients. CD40L level is not meaningful for MM treatment and prognosis.
B-Lymphocytes ; CD40 Antigens ; CD40 Ligand ; Humans ; Lymphocyte Subsets ; Prognosis

CD40 and its receptor CD40L are a very important pair of co-stimulating molecule in immune response, which have extensive biological effects. After stimulating CD40 signal, it can exert corresponding function through MAPK (JNK, ERK, p38) pathway, PI3K cascade, as well as NF-κB and STAT. The CD40 signal is closely related to tumor immunity, this moleculer has already become targeted-molecule for cancer treatment. Recently, there have been many anti-CD40 monoclonal antibodies displaying good anti-cancer effect, among which CHIR-12.12, SGN-40 and CP-870, 893 developed rapidly and successively have entered clinical research stage. This review focuses the status of anti-CD40 monoclonal antibody, including distribution of CD40, physiological function of CD40, CD40 and tumor immunity, anti-CD40 monoclonal antibodies and so on.
Animals ; Antibodies, Monoclonal ; CD40 Antigens ; immunology ; Humans ; Neoplasms

OBJECTIVETo explore effects and action mechanism of warm moxibustion on regulation of blood lipids and anti-atherosclerosis.

METHODSForty-one male Japanese big-ear white rabbits were randomly divided into a blank group (10 rabbits), a model group (10 rabbits), a moxibustion group (10 rabbits) and a medication group (11 rabbits). Normal diet was applied in the blank group while high-cholesterol diet combined with injection of bovine serum albumin were applied in the rest groups to establish rabbit model of atherosclerosis. After establishment, the model group was not intervened and warm moxibustion was applied in the moxibustion group at "Zusan-li" (ST 36) and "Shenque" (CV 8), 10 min per acupoint per day for continuous 4 weeks. The medication group was treated with intragastric administration of lovastatin capsule (3.6 mg/kg) for continuous 4 weeks. The level of blood lipids, such as total cholesterol (TC), triglyceride (TG), and content of CD40 ligand (CD40L), soluble CD40 ligand (sCD40L) and expression of nuclear factor NF-kappaB were tested after 4 weeks.

RESULTSCompared with the model group, the moxibustion group and medication group could effectively reduce the contents of TC and low density lipoprotein (all P < 0.05), lower the level of sCD40L [(8.310 +/- 1.221) ng/mL in the model group, (7.097 +/- 0.846) ng/mL in the moxibustion group and (7.354 +/- 0.631) ng/mL in the medication group], reduce expression of CD40L [(0.235 +/- 0.179) mm2 in the model group, (0.072 +/- 0.079) mm2 in the moxibustion group and (0.039 +/- 0.015) mm2 in the medication group] and NF-kappaB [(0.145 +/- 0.052)mm2 in the model group, (0.052 +/- 0.012) mm2 in the moxibustion group and (0.036 +/- 0.013) mm2 in the medication group], indicating the significant difference (P < 0.05, P < 0.001). There was no statistical difference between the moxibustion group and medication group (all P > 0.05).

CONCLUSIONThe warm moxibustion has great effect on regulation of blood lipids and anti-atherosclerosis, in which lowering expression of CD40-CD40L could be one of possible mechanisms to take effect of anti-atherosclerosis.

Animals ; Atherosclerosis ; blood ; therapy ; CD40 Antigens ; blood ; CD40 Ligand ; blood ; Humans ; Lipids ; blood ; Male ; Moxibustion ; methods ; Rabbits

BACKGROUND: We studied the role for expression of CD40 and CD40L by CD4 and CD8 T cells in the generation of CD8 T cell response to minor histocompatibility antigen, H60. H60 is a cellular antigen to which CD8 responses require CD4 T cell help. METHODS: CD40- or CD40L-deficient mice were adoptively transferred with normal CD4 or CD8 T cells or with memory CD4 or CD8 T cells, and were immunized with male H60 congenic splenocytes to induce CD8 T cell response to H60. Peripheral blood CD8 T cell from the immunized mice were stained with the H60 tetramer. RESULTS: CD8 T cell response to H60 was not induced in both CD40- and CD40L-deficient mice. Adoptive transfer of CD40(+/+) CD8 T cells into CD40-deficient mice did not compensate the defect in inducing CD8 T cell response to H60, while the H60-specific CD8 T cells were activated in the CD40-deficient mice that were adoptively transferred with CD40(+/+) CD4 T cells. Adoptive transfer of CD40L(+/+) CD4 T cells into CD40L-deficient mice induced primary CD8 T cell response for H60 and the presence of CD40L(+/+) CD4 T cells was required even for memory CD8 T cells response to H60. CONCLUSION: Our results suggest that the CD40-CD40L interaction mediates the delivery of CD4 T cell help to na?ve and memory H60-specific CD8 T cells. While the expression of CD40L by CD4 T cells is essential, signaling through CD40 on CD8 T cells is not required for the induction of CD8 T cell response to H60.
Adoptive Transfer ; Animals ; CD40 Ligand* ; Histocompatibility Antigens ; Humans ; Male ; Memory ; Mice ; T-Lymphocytes

BACKGROUND/AIMS: The expression of CD40 in gastric cancer has not been studied. The aims of this study were to determine the expression of CD40 in gastric cancer and to investigate the effect of CD40 on the apoptosis of gastric cancer cells. METHODS: We examined the expression of CD40 by immunohistochemistry and flow cytometry. CD40 mRNA in 5 gastric cancer cell lines was analyzed by RT-PCR. To assess the effect of CD40 on the viability of gastric cancer cells, we performed MTT assay. The effect of CD40 signaling on the apoptosis of gastric cancer cells was examined by annexin V affinity assay. RESULTS: Twelve of twenty human gastric cancer tissues demonstrated positive staining for CD40. Among 5 gastric cancer cell lines, AGS cell line expressed membrane-bound CD40 antigen and CD40 mRNA. In AGS cells, CD40 stimulation significantly reduced the cell viability. CD40 ligation significantly increased the apoptosis in AGS cells compared to the control. CONCLUSIONS: CD40 is expressed in human gastric cancer tissues and gastric cancer cell line, and induces apoptosis in gastric cancer cells. These results suggest that CD40 expression in gastric cancer may play an important role in host defense mechanism against the gastric cancer.
Antigens, CD40/*analysis/physiology ; *Apoptosis ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Stomach Neoplasms/*chemistry/pathology

BACKGROUND: Immunotherapy using dendritic cells (DC) loaded with tumor antigens may represent a potentially effective method for inducing antitumor immunity. We evaluated the effectiveness of DC-based antitumor immune response in various conditions. METHODS: DC were cultured from peripheral blood mononuclear cells (PBMNC) in myelogenous leukemia (ML) and lysates of autologous leukemic cells are used as tumor antigen. The effectiveness of interleukin-12 (IL-12) and CD40L (CD154) on the antigen presenting function of lysates-loaded DC was analyzed by proliferation, cytokine production, and cytotoxicity tests with activated PBMNC (mainly lymphocytes). For generating antigen-loaded DC, direct fusion of DC with ML was studied. RESULTS: Antigen loaded DC induced significantly effective antitumor immune response against autologous leukemic cells. Administration of IL-12 on the DC based antitumor immune response showed higher proliferation activity, IFN-gamma production, and cytotoxic activity of PBMNC. Also, fused cell has a potent antitumor immune response. CONCLUSION: We conclude that lysates-loaded DC with IL-12 may be effectively utilized as inducer of antitumor immune reaction in ML and in vivo application with DC-based antitumor immunotherapy or tumor vaccination seems to be feasible.
Antigens, Neoplasm ; CD40 Ligand ; Cell Fusion ; Dendritic Cells* ; Humans ; Immunotherapy ; Interleukin-12* ; Leukemia ; Leukemia, Myeloid* ; Vaccination

As CD40 transduces activation signals involved in inflammatory and immune disorders, we explored the expression and response to CD40 engagement in human glioma cell lines in this study. The CD40 expression in BT-325 and U251 cells was flow cytometrically detected. The cells were incubated with srhCD40L for 72 h to assess its effects on cell growth in vitro. TNF-α expression was quantified by real-time PCR, and protein expression was analyzed by ELISA. The I-κb mRNA was detected by RT-PCR. I-κB expression decreased after stimulation with 1 μg/mL srhCD40L, but it was upregulated after the cells were pretreated with CD40 antibody. srhCD40L significantly inhibited the proliferation of the CD40+ human glioma cells. The stimulation of CD40+ glioma cells with soluble CD40L (CD154) up-regulated the expression of TNF-α at both mRNA and protein levels. We are led to conclude that CD40L/CD40 could inhibit human glioma cells through I-κb signaling pathway. Interferon-γ can augment CD40 expression and the inhibitory effect of CD40 ligand on cell growth in vitro. These results suggest that srhCD40L may benefit the therapy strategy of glioma.
CD40 Antigens ; metabolism ; CD40 Ligand ; metabolism ; Cell Proliferation ; Cells, Cultured ; Glioma ; genetics ; metabolism ; pathology ; Humans ; NF-kappa B ; metabolism ; Signal Transduction ; physiology ; Tumor Cells, Cultured

The interaction of CD40 with its cognate ligand, CD40L (CD154), plays important roles in immune responses. Blockade of CD40-CD40L signal pathway can protect the progression of antibody- and cell-mediated autoimmune diseases, and reduce allograft rejection thus prolonging graft survival, even engendering long-lived antigen-specific tolerance. The present study aims to enhance the binding activity of CD40 by incorporating an isoleucine zipper (IZ) trimeric motif into CD40 ectodomain to promote the formation of soluble CD40 trimers, which would be useful for blocking CD40-CD40L interaction. A prokaryotic expression vector for soluble human CD40 ectodomain fused with an IZ motif and a hexa-histidine (His6) tag at its carboxyl terminus (sCD40IZ) was constructed by multiple round PCR using cloned CD40 cDNA as a template. The recombinant sCD40IZ protein was expressed highly in Escherichia coli (E. coli) with a molecular weight of 27kD, which is consistent with its theoretical value. It mainly existed in inclusion bodies. After refolding from inclusion bodies, soluble sCD40IZ protein was purified by gel filtration. Its molecular weight in solution was about 91kD when determined by gel filtration, suggesting that it most probably existed in the form of trimers. Moreover, this protein could bind to CD40L expressed on Jurkat T cells and its binding activity was significantly higher than that of soluble CD40 without an IZ motif. These results suggest that incorporation of an IZ motif at the carboxyl terminus of soluble CD40 can facilitate the formation of trimers and enhance its binding activity with CD40L. Thus, the trimeric CD40 protein may be used to block CD40-CD40L signal pathway, suggesting that it may have potential application in preventing autoimmune diseases and transplantation rejection.
CD40 Antigens ; biosynthesis ; genetics ; metabolism ; CD40 Ligand ; metabolism ; Escherichia coli ; genetics ; metabolism ; Humans ; Isoleucine ; biosynthesis ; genetics ; Leucine Zippers ; genetics ; Protein Multimerization ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; metabolism

Adult ; Blood Platelets ; chemistry ; CD40 Antigens ; blood ; CD40 Ligand ; blood ; Cholesterol ; blood ; Female ; Humans ; Hypercholesterolemia ; blood ; drug therapy ; Male ; Middle Aged ; Simvastatin ; therapeutic use

OBJECTIVEThe pathogenesis of bronchiolitis has not been fully identified. Immune function abnormality following virus infection may be associated with the pathogenesis. CD40 and CD40 ligand (CD40L) is a pair of co-stimulatory molecules in immunoreaction. They might play an important role in the development of bronchiolitis. This study aimed to investigate the expression of CD40 and CD40L in peripheral blood mononuclear cells (PBMCs) in children with bronchiolitis and explore their possible roles in the disease.

METHODSThirty children with bronchiolitis, 26 children with bronchopneumonia and 30 healthy children (control) were enrolled. Flow cytometry was used to detect CD40 and CD40L expression in PBMCs. Total serum IgE level was measured using ELISA.

RESULTSCompared with the control group, CD40L expression significantly increased in the bronchiolitis and bronchopneumonia groups (P< 0.05). The CD40L expression in the bronchiolitis group was significantly higher than that in the bronchopneumonia group (P< 0.05). A significantly increased CD40 expression was also found in the bronchiolitis group when compared with the bronchopneumonia and the control group (P< 0.01). Total serum IgE level in the bronchiolitis group was significantly higher than the bronchopneumonia and the control groups (P< 0.01). CD40 and CD40L expression was positively correlated with serum IgE level in the bronchiolitis group (r=0.607, r=0.819, respectively; P< 0.01).

CONCLUSIONSCD40 and CD40L expression in PBMCs and serum IgE level increased and there is a positive correlation between CD40 and CD40L expression and serum IgE level in children with bronchiolitis. Over-expression of CD40 and CD40L may play an important role in the development of bronchiolitis.

Bronchiolitis ; etiology ; immunology ; CD40 Antigens ; blood ; physiology ; CD40 Ligand ; blood ; physiology ; Female ; Humans ; Immunoglobulin E ; blood ; Infant ; Leukocytes, Mononuclear ; chemistry ; Male