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

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: 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

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

OBJECTIVE: To determine the relationship between intravoxel incoherent motion (IVIM) imaging derived quantitative metrics and serum soluble CD40 ligand (sCD40L) level in an embolic canine stroke model. MATERIALS AND METHODS: A middle cerebral artery occlusion model was established in 24 beagle dogs. Experimental dogs were divided into low- and high-sCD40L group according to serum sCD40L level at 4.5 hours after establishing the model. IVIM imaging was scanned at 4.5 hours after model establishment using 10 b values ranging from 0 to 900 s/mm². Quantitative metrics diffusion coefficient (D), pseudodiffusion coefficient (D*), and perfusion fraction (f) of ischemic lesions were calculated. Quantitative metrics of ischemic lesions were normalized by contralateral hemisphere using the following formula: normalized D = D(stroke) / D(contralateral). Differences in IVIM metrics between the low- and high-sCD40L groups were compared using t test. Pearson's correlation analyses were performed to determine the relationship between IVIM metrics and serum sCD40L level. RESULTS: The high-sCD40L group showed significantly lower f and normalized f values than the low-sCD40L group (f, p < 0.001; normalized f, p < 0.001). There was no significant difference in D*, normalized D*, D, or normalized D value between the two groups (All p > 0.05). Both f and normalized f values were negatively correlated with serum sCD40L level (f, r = −0.789, p < 0.001; normalized f, r = −0.823, p < 0.001). However, serum sCD40L level had no significant correlation with D*, normalized D*, D, or normalized D (All p > 0.05). CONCLUSION: The f value derived from IVIM imaging was negatively correlated with serum sCD40L level. f value might serve as a potential imaging biomarker to assess the formation of microvascular thrombosis in hyperacute period of ischemic stroke.
Animals ; CD40 Ligand* ; Diffusion ; Dogs ; Infarction, Middle Cerebral Artery ; Magnetic Resonance Imaging* ; Perfusion ; Stroke* ; Thrombosis

OBJECTIVETo construct a recombinant adenovirus vector expressing hCD40L gene and explore it in the use of anti-tumor gene therapy.

METHODS1,900 bp gene fragment was obtained form plasmid pORF-hCD40L by Xho I/Swa I cutting and then cloned directionally into the pShuttle plasmid, finally, the resultant plasmid was digested by restriction endonnuclease PmeI and subsequently cotransformtion into BJ5183 cells with the adenoviral backbone pAdEasy-1 to obtain the homologous recombinant and then the recombinant was packaged in the 293 cells. Some methods such as PCR and endonulease digestion were employed to identify the recombinant adenovirus.

RESULTSThe evidences of endonulease digestion and PCR analysis confirmed that recombinant hCD40L gene was correctly inserted into adenovirus vector.

CONCLUSIONThe adenoviral vector which expressed hCD40L gene was constructed. It provides an experimental basis for studies on it expression in the mammalian cells and in tumor gene therapy.

Adenoviridae ; Adenoviruses, Human ; Animals ; CD40 Ligand ; Genetic Therapy ; Genetic Vectors ; Humans ; Plasmids ; Polymerase Chain Reaction

Chemokine is a class of soluble active peptides that attract white blood cells to the inflammatory site. CD40 ligand (CD40L) involves in synthesis of proinflammatory mediators. Accumulation of chemokine and CD40L can induce non-hemolytic reaction after transfusion, transfusion-related acute lung injury (TRALI) and autoimmune disease during blood component storage. Pre-storage leucocyte deletion can prevent the release of leucocyte-derived chemokines, but not prevent the accumulation of platelet-derived chemokines. γ-irradiation or ultraviolet β irradiation is effective in preventing the increase of chemokines in the storage of platelet, thus prevents non-hemolytic febrile reaction after platelet transfusion. In this review, the recent advance in research of accumulation of chemokines and CD40L during blood component storage, and its effect on blood transfusion, as well as preventive measures are summarized.
Blood Platelets ; metabolism ; Blood Preservation ; Blood Transfusion ; CD40 Ligand ; blood ; Chemokines ; blood ; Humans

CD40 Ligand ; pharmacology ; HL-60 Cells ; drug effects ; Humans ; fas Receptor ; metabolism

OBJECTIVE: To explore the effect of soluble CD40 ligand (sCD40L) on the proliferation, apoptosis, and cell cycle of human non-Hodgkin lymphoma (NHL) cells, and analyze its possible mechanism. METHODS: NHL CA46 cell and Raji cell were treated with different concentrations of sCD40L for 48 h, CCK-8 was used to detect the effect of sCD40L on cell proliferation in vitro, flow cytometry on apoptosis and cycle of NHL cells, and Western blot on the expression of PTEN, BCL-2, and BAX in NHL cells. RESULTS: Compared with the control group, 4 and 8 μg/ml sCD40L could significantly inhibit the proliferation of lymphoma Raji cell and CA46 cell (P<0.05). The test results of flow cytometry showed that 4 μg/ml sCD40L could significantly promote the apoptosis of CA46 and Raji cells, and significantly inhibit the S phase proportions (P<0.05). Western blot results showed that sCD40L could promote the expression of PTEN and BAX, while inhibit the expression of BCL-2 (P<0.05). CONCLUSION sCD40L can promote the apoptosis and inhibit the proliferation of NHL cells through the PTEN signaling pathway.
Apoptosis ; CD40 Ligand ; Cell Line, Tumor ; Cell Proliferation ; Family ; Humans ; Lymphoma, Non-Hodgkin

To obtain chicken CD40L protein, the cDNA was prepared from chicken splenic cells and used as a template to clone and amplify CD40L by PCR. The target gene was cloned into pFastBac vector to construct a pFastBac-chCD40L donor plasmid. Recombinant plasmid was transformed into DH10Bac and recombinant Bacmid-chCD40L was obtained. The Bacmid-chCD40L plasmid was transfected into sf9 insect cells to obtain His-chCD40L protein. In addition, the target gene was cloned into pQM01 vector to construct a pQM01-chCD40L plasmid, recombinant plasmid was transfected into HEK 293T cells to obtain Strep-chCD40L protein. The chCD40L protein was purified by affinity chromatography, and the concentration of purified chCD40L protein was determined to be 0.01 mg/mL. Primary cells were isolated from the bursal tissue of 3-week old SPF chickens, and the chCD40L protein was added to the culture medium to stimulate cells. The chCD40L could bind to CD40 on B cells as examined by Western blotting, indirect immunofluorescence assay and flow cytometry, suggesting that chCD40L protein is biologically active. We successfully obtained chicken CD40L protein of biological activity, which laid the foundation in the in vitro culture of primary B lymphocytes for the isolation and diagnosis of virulent IBDV.
Animals ; Baculoviridae/genetics* ; CD40 Ligand/genetics* ; Chickens ; Cloning, Molecular ; Genetic Vectors/genetics* ; Recombinant Proteins/genetics*