Gastric cancer overexpressing the human epidermal growth factor 2 (HER2) protein has a poor outcome, although a combination of chemotherapy and the anti-HER2 antibody trastuzumab has been approved for the treatment of advanced gastric cancer. Vascular endothelial growth factor (VEGF) expression in gastric cancer is correlated with recurrence and poor prognosis; however, the anti-VEGF antibody bevacizumab has shown limited efficacy against gastric cancer in clinical trials. In this study, we evaluated the antitumor effects of trastuzumab; VEGF-Trap binding to VEGF-A, VEGF-B and placental growth factor (PlGF); and a combination of trastuzumab and VEGF-Trap in a gastric cancer xenograft model. Although trastuzumab and VEGF-Trap each moderately inhibited tumor growth, the combination of these agents exerted greater inhibition compared with either agent alone. Immunohistochemical analyses indicated that the reduction in tumor growth was associated with decreased proliferation and increased apoptosis of tumor cells and decreased tumor vascular density. The combined treatment resulted in fewer proliferating tumor cells, more apoptotic cells and reduced tumor vascular density compared with treatment with trastuzumab or VEGF-Trap alone, indicating that trastuzumab and VEGF-Trap had additive inhibitory effects on the tumor growth and angiogenesis of the gastric cancer xenografts. These data suggest that trastuzumab in combination with VEGF-Trap may represent an effective approach to treating HER2-overexpressing gastric cancer.
Animals ; Antibodies, Monoclonal, Humanized/administration & dosage/*therapeutic use ; Antineoplastic Combined Chemotherapy Protocols/*therapeutic use ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Mice ; Mice, Inbred BALB C ; Neovascularization, Pathologic/drug therapy ; Receptor, erbB-2/*antagonists & inhibitors ; Receptors, Vascular Endothelial Growth Factor/administration & dosage/*therapeutic use ; Recombinant Fusion Proteins/administration & dosage/*therapeutic use ; Stomach Neoplasms/*drug therapy/pathology ; Vascular Endothelial Growth Factor A/antagonists & inhibitors ; Xenograft Model Antitumor Assays

BACKGROUND: Toll-like receptors (TLRs) have been extensively studied in recent years. However, functions of these molecules in murine B cell biology are largely unknown. A TLR4 stimulant, LPS is well known as a powerful polyclonal activator for murine B cells. METHODS: In this study, we explored the effect of a murine TLR9 stimulant, M6-395 (a synthetic CpG ODNs) on B cell proliferation and Ig production. RESULTS: First, M6-395 was much more potent than LPS in augmenting B cell proliferation. As for Ig expression, M6-395 facilitated the expression of both TGF-beta1-induced germ line transcript alpha (GLTalpha) and IL-4-induced GLTgamma1 as levels as those by LPS and Pam3CSK4 (TLR1/2 agonist) : a certain Ig GLT expression is regarded as an indicative of the corresponding isotype switching recombination. However, IgA and IgG1 secretion patterns were quite different--these Ig isotype secretions by M6-395 were much less than those by LPS and Pam3CSK4. Moreover, the increase of IgA and IgG1 production by LPS and Pam3CSK4 was virtually abrogated by M6-395. The same was true for the secretion of IgG3. We found that this unexpected phenomena provoked by M6-395 is attributed, at least in part, to its excessive mitogenic nature. CONCLUSION: Taken together, these results suggest that M6-395 can act as a murine polyclonal activator but its strong mitogenic activity is unfavorable to Ig isotype switching.
B-Lymphocytes ; Cell Proliferation ; Germ Cells ; Immunoglobulin A ; Immunoglobulin Class Switching ; Immunoglobulin G ; Oligodeoxyribonucleotides ; Recombination, Genetic ; Toll-Like Receptors

Transforming growth factor-B1 (TGF-B1) is well known to be one of the most potent Immunosuppressive cytokines. To determine whether TGF-B1 secreted in the latent form can be immunoregulatory, TGF-B1 cDNA driven by the human -actin promotor was transfected into a murine thymoma cell line, EL4 cells. The transfectants (ELJ4) secreted a latent torm of TGF-B1 at a concentration of 5 ng/ml under the influence of TPA. Transfected TGF-b1 transcripts was readily detected by RT-PCR in ELJ4 cells regardless of the presence of TPA, but not in EL4 cells. In addition, we found the degree of Thy-B1 expression, IL-2 secretion and the proliferation rate are not altered by the transfection. Finally, EL4 and ELJ4 cells were injected into C57BU6 mice (syngenic strain), subcutaneously. Tumor cell masses derived from both cell populations survived longer than 1 wk, and the size of tumor derived from ELJ4 was three times larger (2.5 cm of diameter) than that from EL4. Virtually, there was no histopathological difference between two tumors. Taken together, the results from the present study indicates that EL4 thymomas transfected with TGF-1 secretes a latent form of TGF-B1 which may suppress host immune defence system.
Humans ; Mice ; Animals

Sialolithiasis is a condition characterized by the obstruction of a salivary gland or its duct due to the formation of calcareous material or sialoliths resulting in salivary ectasia and even provoking the subsequent dilation of the salivary gland or salivary duct. The most difficult cases involve sialoliths in the posterior part of the Stensen's duct, the presence of multiple stones with stenosis of the distal part of the duct. We report on a case of Stensen's duct abscess with multiple sialolithiasis in a 46-year-old man. The patient's cheek was swollen, and showed no evidence of any other lesions. The patient was treated with surgical removal of stones by the intraoral approach, sialodochoplasty and antibiotics therapy. There has been no recurrence nor duct problem during the 12 months period of follow up.
Abscess ; Anti-Bacterial Agents ; Cheek ; Constriction, Pathologic ; Dilatation, Pathologic ; Follow-Up Studies ; Humans ; Middle Aged ; Parotid Gland ; Recurrence ; Salivary Duct Calculi ; Salivary Ducts ; Salivary Gland Calculi ; Salivary Glands

Aluminum hydroxide (alum) is the most widely used adjuvant in human vaccines. Nevertheless, it is virtually unknown whether alum acts on B cells. In the present study, we explored the direct effect of alum on Ig expression by murine B cells in vitro. LPS-activated mouse spleen B cells were cultured with alum, and the level of isotype-specific Ig secretion, IgG1 secreting cell numbers, and Ig germ-line transcripts (GLT) were measured using ELISA, ELISPOT, and RT-PCR, respectively. Alum consistently enhanced total IgG1 production, numbers of IgG1 secreting cells, and GLTgamma1 expression. These results demonstrate that alum can directly cause IgG1 isotype switching leading to IgG1 production.
Alum Compounds ; Aluminum Hydroxide ; Animals ; B-Lymphocytes ; Cell Count ; Enzyme-Linked Immunosorbent Assay ; Enzyme-Linked Immunospot Assay ; Humans ; Hydroxides ; Immunoglobulin Class Switching ; Immunoglobulin G ; Mice ; Spleen ; Vaccines

During chronic inflammatory response, mono- cytes/macrophages produce 92-kDa matrix metalloproteinase-9 (MMP-9), which may contribute to their extravasation, migration and tissue remodeling. Activation of peroxisome proliferator- activated factor receptor-gamma (PPAR-gamma) has been shown to inhibit MMP-9 activity. To evaluate whether ox-LDL, a PPAR-gamma activator, inhibits PMA-induced MMP-9 expression and activity, and if so, whether CD36 and PPAR-gamma are involved in this process, we investigated the effect of ox-LDL on MMP-9 expression and activity in PMA-activated human monocytic cell line U937. PMA-induced MMP-9 expression and activity were suppressed by the treatment with ox-LDL (50 micrigram/ml) or PPAR-gamma activators such as troglitazone (5 micrometer), ciglitazone (5 micrometer), and 15d- PGJ2 (1 micrometer) for 24 h. This ox-LDL or PPAR-gamma activator-mediated inhibition of micrometer P-9 activity was diminished by the pre-treatment of cells with a blocking antibody to CD36, or PGF2a (0.3 micrometer), which is a PPAR-gamma inhibitor, as well as overexpression of a dominant-negative form of CD36. Taken together, these results suggest that ox-LDL suppresses PMA-induced MMP-9 expression and activity through CD36-mediated activation of PPAR-gamma.
Antibodies, Blocking/pharmacology ; Antigens, CD36/immunology/*physiology ; Cells, Cultured ; Chromans/pharmacology ; Gelatinase B/antagonists & inhibitors/genetics/*metabolism ; Humans ; Lipoproteins, LDL/pharmacology/*physiology ; Monocytes/drug effects/*enzymology/metabolism ; NF-kappa B/antagonists & inhibitors ; PPAR gamma/*metabolism ; Prostaglandin D2/*analogs & derivatives/pharmacology ; RNA, Messenger/analysis/metabolism ; Research Support, Non-U.S. Gov't ; Tetradecanoylphorbol Acetate/antagonists & inhibitors/pharmacology ; Thiazolidinediones/pharmacology ; Transcription, Genetic/drug effects

OBJECTIVE: To investigate the prognostic values of somatosensory evoked potential (SEP) and electric motor evoked potential (eMEP) studies according to the varying spinal cord injury by incremental height of weight-drop impactor and progress of functional recovery METHOD: Thirty Sprague-Dawley rats (300+/-50 grams, male) were used. The spinal cord injury was made by weight-drop device from 12.5, 25.0 and 50.0 mm height at T10 cord segment. The three groups of each drop-height (n=10) and laminectomized sham group (n=10) were subjected to functional analysis using inclined plane test and Basso Beattie Bresnahan (BBB) locomotor scales at the 1, 3, 5, 7, 14, 21 and 28th day after the contusive injury. SEP by sciatic nerve stimulation and eMEP at the gastrocnemius muscle were recorded. RESULTS: Maximal angle of inclination and BBB scales had an inverse relation with the contusion severity (p <0.05). There were significant correlations among the changes of peak latencies and amplitudes of SEP, contusion severity, and the motor recovery (p <0.05). The changes of onset latencies and amplitudes of eMEP were significantly correlated with the contusion severity and the motor recovery (p <0.05). CONCLUSION: The SEP and eMEP studies had significant values according to the contusion severity and functional recovery in contusive rat model of the spinal cord.
Animals ; Contusions ; Evoked Potentials* ; Evoked Potentials, Motor ; Evoked Potentials, Somatosensory ; Models, Animal* ; Muscle, Skeletal ; Rats* ; Rats, Sprague-Dawley ; Sciatic Nerve ; Spinal Cord Injuries ; Spinal Cord* ; Weights and Measures

B cell-activating factor belonging to the TNF family (BAFF) is primarily expressed by macrophages and stimulates B cell proliferation, differentiation, survival, and Ig production. In this study, we explored the effect of lactoferrin (LF) on BAFF expression by murine macrophages. We determined the level of BAFF expression at the transcriptional and protein levels using RT-PCR and ELISA, respectively. LF markedly enhanced BAFF expression in mouse macrophages at both the transcriptional and protein levels. Overexpression of Smad3/4 further increased LF-induced BAFF transcription while DN-Smad3 abolished the LF-induced BAFF expression. These results demonstrate that LF can enhance BAFF expression through Smad3/4 pathway.
Animals ; Cell Proliferation ; Enzyme-Linked Immunosorbent Assay ; Humans ; Lactoferrin ; Macrophages ; Mice ; Transforming Growth Factor beta1

It is well established that TGF-beta1 and retinoic acid (RA) cause IgA isotype switching in mice. We recently found that lactoferrin (LF) also has an activity of IgA isotype switching in spleen B cells. The present study explored the effect of LF on the Ig production by mouse peritoneal B cells. LF, like TGF-beta1, substantially increased IgA production in peritoneal B1 cells but little in peritoneal B2 cells. In contrast, LF increased IgG2b production in peritoneal B2 cells much more strongly than in peritoneal B1 cells. LF in combination with RA further enhanced the IgA production and, interestingly, this enhancement was restricted to IgA isotype and B1 cells. Similarly, the combination of the two molecules also led to expression of gut homing molecules alpha4beta7 and CCR9 on peritoneal B1 cells, but not on peritoneal B2 cells. Thus, these results indicate that LF and RA can contribute to gut IgA response through stimulating IgA isotype switching and expression of gut-homing molecules in peritoneal B1 cells.
Animals ; B-Lymphocytes ; Immunoglobulin A* ; Immunoglobulin Class Switching ; Immunoglobulin G ; Lactoferrin* ; Mice ; Spleen ; Transforming Growth Factor beta1 ; Tretinoin*