OBJECTIVETo evaluate the immunotoxicological effects of genetically modified wheat with TaDREB4 gene in female BALB/c mice.

METHODSFemale mice weighing 18-22 g were divided into five groups (10 mice/group), which were set as negative control group, common wheat group, parental wheat group, genetically modified wheat group and cyclophosphamide positive control group, respectively. Mice in negative control group and positive control group were fed with AIN93G diet, mice in common wheat group, non-genetically modified parental wheat group and genetically modified wheat group were fed with feedstuffs added corresponding wheat (the proportion is 76%) for 30 days, then body weight, absolute and relative weight of spleen and thymus, white blood cell count, histological examination of immune organ, peripheral blood lymphocytes phenotyping, serum cytokine, serum immunoglobulin, antibody plaque-forming cell, serum half hemolysis value, mitogen-induced splenocyte proliferation, delayed-type hypersensitivity reaction and phagocytic activities of phagocytes were detected.

RESULTSNo immunotoxicological effects related to the consumption of the genetically modified wheat were observed in BALB/c mice when compared with parental wheat group, common wheat group and negative control group.

CONCLUSIONFrom the immunotoxicological point of view, results from this study demonstrate that genetically modified wheat with TaDREB4 gene is as safe as the parental wheat.

Animals ; Antibody-Producing Cells ; immunology ; Body Weight ; Cytokines ; blood ; Female ; Genes, Plant ; Hemolysis ; Hypersensitivity, Delayed ; Immune System ; drug effects ; Immunoglobulins ; blood ; Mice ; Mice, Inbred BALB C ; Organ Size ; Phagocytosis ; Plants, Genetically Modified ; toxicity ; Spleen ; immunology ; Thymus Gland ; immunology ; Triticum ; genetics

This study is to elucidate the immunoregulation mechanisms of artesunate (AST) on allergic contact dermatitis (ACD). Pharmacodynamics analyses, HE staining, semi-quantitative RT-PCR and Western blotting were used to explore the effects of AST on the related cytokines, transcription factor and signaling molecule of ACD respectively. The results indicated that topical administration of AST not only reduced the increase of ear swelling, spleen index and inflammatory cells infiltration in ACD mice, but also inhibited remarkably the expression of IFN-gamma, T-bet and NF-kappaB p65. It's suggested that AST could exhibit suppressive effects on inflammatory response and immune function of ACD, which indicates the possibility of developing AST as a novel immunoregulatory agent in the treatment of ACD and other immune-related diseases.
Administration, Topical ; Animals ; Artemisinins ; administration & dosage ; chemistry ; pharmacology ; Dermatitis, Allergic Contact ; immunology ; metabolism ; pathology ; Ear ; pathology ; Female ; GATA3 Transcription Factor ; genetics ; metabolism ; Hypersensitivity, Delayed ; drug therapy ; Immunosuppressive Agents ; administration & dosage ; chemistry ; pharmacology ; Interferon-gamma ; genetics ; metabolism ; Interleukin-4 ; genetics ; metabolism ; Mice ; Mice, Inbred ICR ; Molecular Structure ; NF-kappa B ; metabolism ; RNA, Messenger ; metabolism ; T-Box Domain Proteins ; genetics ; metabolism

Food allergy is defined as reaction to a food which has an immunologic mechanism. Its prevalence is increasing in children globally and is therefore of increasing clinical importance. A useful clinical approach is to distinguish food allergic reactions by the timing of clinical reaction in relation to food exposure and classified as immediate (generally IgE-mediated) and delayed (generally non-IgE-mediated), with the exception of eczema and eosinophilic gastrointestinal disease, which, when associated with food allergy may be associated with either mechanism. This review is aimed at providing the clinician with a Singaporean perspective on the clinical approach and management of these disorders.
Breast Feeding ; Child ; Child, Preschool ; Eczema ; diagnosis ; immunology ; Food Hypersensitivity ; diagnosis ; prevention & control ; Humans ; Hypersensitivity, Delayed ; diagnosis ; prevention & control ; Immunoglobulin E ; blood ; Infant ; Infant, Newborn ; Skin Test End-Point Titration

OBJECTIVETo study the role of dendritic cells (DCs) in initiating delayed-type hypersensitivity (DTH) to fluorescein isothiocyanate (FITC) after trauma-hemorrhage in mice.

METHODSInbred BALB/c mice (6-8 weeks old, male) were epicutaneously sensitized with FITC 12 hours, 1 day, 2 days, 4 days and 7 days after closed bilateral femur fractures combined with hemorrhage. And 5 days after sensitization, DTH was evaluated by ear swelling after a challenge of FITC. Draining lymph node cells were examined for the percentages of FITC-positive cells, cluster of differentiation (CD)11c-positive cells and major histocompatibility complex II (MHC II)-positive cells by means of flow cytometry. In vitro proliferative responses of syngeneic lymphocytes and in vivo passive transfer of DTH to naive recipients induced by isolated DCs from the draining lymph nodes were determined.

RESULTSThe time of DTH to FITC decreased more significantly in the mice with trauma-hemorrhage (12 hours to 4 days) than in the mice with sham injury. After sensitization, the relative percentages of FITC+ cells, FITC+/CD11c+ cells and FITC+/CD11c+/MHC II+ cells from the draining lymph nodes were all significantly reduced following injury. And the capacity of DCs from the draining lymph nodes in stimulating proliferative responses of lymphocytes and transferring DTH to naive recipients were also inhibited after injury.

CONCLUSIONSTrauma-hemorrhage induces repressive DTH in mice, which may be attributed, at least partially, to the reduced trafficking of DCs into the draining lymph nodes and insufficient maturation during DC migration.

Animals ; Dendritic Cells ; physiology ; Flow Cytometry ; Fluorescein-5-isothiocyanate ; Hemorrhage ; immunology ; Hypersensitivity, Delayed ; etiology ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; Wounds and Injuries ; immunology

Leishmania tropica and L. major are etiologic agents of human cutaneous leishmaniasis. Delayed type hypersensitivity (DTH) is an immunologic response that has been frequently used as a correlate for protection against or sensitization to leishmania antigen. In BALB/c mice, L. tropica infection results in non-ulcerating disease, whereas L. major infection results in destructive lesions. In order to clarify the immunologic mechanisms of these 2 different outcomes, we compared the ability of these 2 leishmania species in induction of DTH response in this murine model. BALB/c mice were infected with L. major or L. tropica, and disease evolution and DTH responses were determined. The results show that the primary L. major infection can exacerbate the secondary L. major infection and is associated with DTH response. Higher doses of the primary L. major infection result in more disease exacerbation of the secondary L. major infection as well as higher DTH response. L. tropica infection induces lower DTH responses than L. major. We have previously reported that the primary L. tropica infection induces partial protection against the secondary L. major infection in BALB/c mice. Induction of lower DTH response by L. tropica suggests that the protection induced against L. major by prior L. tropica infection may be due to suppression of DTH response.
Animals ; Disease Models, Animal ; Ear/pathology ; Female ; Foot/pathology ; *Hypersensitivity, Delayed ; Leishmania major/*immunology ; Leishmania tropica/*immunology ; Leishmaniasis, Cutaneous/*immunology/*parasitology/pathology ; Mice ; Mice, Inbred BALB C

OBJECTIVETo study the effects of hemorrhage combined with closed fracture on delayed type hypersensitivity (DTH) responses in mice and to explore the relevant mechanisms.

METHODSDTH responses were induced with 2, 4-dinitro-1-fluorobenzene (DNFB) or fluorescein isothiocyanate (FITC) skin painting after injury, and single cell suspensions from pooled inguinal lymph nodes were analyzed by flow cytometry for FITC+ cells and dendritic cells (DC). The ability of cells from pooled inguinal lymph nodes was tested 24 hours after skin painting with DNFB in transferring sensitization for DTH to DNFB.

RESULTSThe DTH responses after injury decreased significantly compared with that of sham-injured mice (P<0.01). Flow cytometry showed that FITC+ cells, FITC+/CD11c+ cells, and FITC+/CD11c+ / major histocompatibility complex II+ cells were all significantly decreased after trauma (P<0.01). The ability of cells to transfer sensitization for DTH to DNFB also declined (P<0.01).

CONCLUSIONHemorrhage combined with closed fracture decreases the DTH responses in mice, which may be attributed to the reduced antigen-presenting capacity of DC in the injured mice.

Animals ; Dendritic Cells ; immunology ; Fractures, Closed ; complications ; immunology ; Hemorrhage ; complications ; immunology ; Hypersensitivity, Delayed ; immunology ; Mice

The protective effect of the Synadenium carinatum latex lectin (ScLL), and the possibility of using it as an adjuvant in murine model of vaccination against American cutaneous leishmaniasis, were evaluated. BALB/c mice were immunized with the lectin ScLL (10, 50, 100 microgram/animal) separately or in association with the soluble Leishmania amazonensis antigen (SLA). After a challenge infection with 10(6) promastigotes, the injury progression was monitored weekly by measuring the footpad swelling for 10 weeks. ScLL appeared to be capable of conferring partial protection to the animals, being most evident when ScLL was used in concentrations of 50 and 100 microgram/animal. Also the parasite load in the interior of macrophages showed significant reduction (61.7%) when compared to the control group. With regard to the cellular response, ScLL 50 and 100 microgram/animal stimulated the delayed-type hypersensitivity (DTH) reaction significantly (P < 0.05) higher than SLA or SLA plus ScLL 10 weeks after the challenge infection. The detection of high levels of IgG2a and the expression of mRNA cytokines, such as IFN-gamma, IL-12, and TNF-alpha (Th1 profiles), corroborated the protective role of this lectin against cutaneous leishmaniasis. This is the first report of the ScLL effect on leishmaniasis and shows a promising role for ScLL to be explored in other experimental models for treatment of leishmaniasis.
*Adjuvants, Immunologic ; Animals ; Antibodies, Protozoan/immunology ; Antibody Formation ; Antigens, Protozoan/immunology ; Cytokines/genetics/immunology ; Euphorbiaceae/*chemistry ; Hypersensitivity, Delayed/immunology ; Immunization ; Immunoglobulin G/immunology ; Latex/chemistry ; Leishmania/immunology ; Leishmaniasis, Cutaneous/*immunology/pathology ; Mice ; Mice, Inbred BALB C ; Nitric Oxide Synthase Type II/genetics/immunology ; Plant Lectins/*immunology/isolation & purification ; Protozoan Vaccines/immunology/pharmacology ; Skin/pathology

AIMTo investigate the effect of cetirizine hydrochloride on the expression of neuropeptide substance P (SP) in IgE-dependent triphasic cutaneous reaction induced by dinitrofluorobenzene (DNFB) in the ears of BALB/c mice.

METHODSBALB/c mice were passively sensitized by intravenous infection of anti-DNP IgE monoclonal antibody 24 h before DNFB challenge. Skin reaction was elicited by applying DNFB to both sides of each ear of sensitized mice. Mice were treated with cetirizine (1 and 10 mg x kg)-1), ig). The ears were removed for pathohistological examination and immunohistochemical staining of SP at different designated times after challenge. The contents of SP in the skin of mouse ear were determined by radioimmunoassay (RIA).

RESULTSThe mice exhibited a triphasic cutaneous reaction with an immediate-phase response (IPR) at 1 h, a late-phase response (LPR) at 24 h and a very late-phase response (vLPR) at 7 days after challenge with DNFB. The expression of SP in different phases increased gradually. Cetirizine (1 and 10 mg x kg(-1)) was shown to significantly inhibit the ear swellings induced by the IPR (P < 0.01), while no obvious effect on the vLPR. The SP contents in ear skin of triphasic cutaneous reaction were decreased by cetirizine.

CONCLUSIONSP is considered to be involved in the pathogenesis of allergic dermatitis. Cetirizine hydrochloride can inhibit the expression of SP in IgE-dependent triphasic cutaneous reaction. It might be part of the mechanisms of anti-anaphylaxis of cetirizine.

Animals ; Anti-Allergic Agents ; pharmacology ; Cetirizine ; pharmacology ; Dose-Response Relationship, Drug ; Ear ; Edema ; metabolism ; Female ; Hypersensitivity, Delayed ; metabolism ; Hypersensitivity, Immediate ; metabolism ; Immunoglobulin E ; immunology ; Mice ; Mice, Inbred BALB C ; Passive Cutaneous Anaphylaxis ; drug effects ; Substance P ; metabolism

OBJECTIVETo investigate the immune responses and protection from virus challenge, induced by the coinjection of IL-2cDNA with herpes simplex virus type 1 (HSV-1) glycoprotein-D (gD) DNA vaccine.

METHODSTwo DNA vaccines (pgD and pIL-2) were constructed by inserting the gD gene and IL-2 cDNA into the eukaryotic expression vector pcDNA3.1, respectively. The BALB/c mice were inoculated intramuscularly three times at 2-week intervals. Two weeks after the final immunization, mice were bled for antibody assay and spleen cells were separated for Th cell proliferation and cytokine assays. Delayed type hypersensitivity (DTH) response was detected by the pinna-swelling test. Corneal protection under HSV-1 virus challenge was continuously observed with slit-lamp microscope.

RESULTSIL-2 cDNA coinjection remarkably enhanced the specific IgG2a level when compared with gD plasmid vaccination alone. Th cell proliferation and secretion of cytokines (IL-2 and IFN-gamma) were significantly increased by IL-2 cDNA coinjection. However, the production of IL-10 was inhibited. The DTH response was also enhanced by IL-2 coinjection. When the mice were challenged with HSV-1, the cornea epithelial lesions were significantly alleviated by IL-2 coinjection as compared with gD vaccination alone.

CONCLUSIONIL-2 cDNA can enhance both the humoral and cellular immune responses, and thus increase the vaccine potency.

Animals ; Antibodies, Viral ; blood ; COS Cells ; Cell Proliferation ; Cercopithecus aethiops ; DNA ; genetics ; Female ; Herpesvirus 1, Human ; pathogenicity ; Hypersensitivity, Delayed ; immunology ; Immunization ; Immunoglobulin G ; blood ; Interferon-gamma ; blood ; Interleukin-2 ; biosynthesis ; genetics ; Mice ; Mice, Inbred BALB C ; Random Allocation ; Th1 Cells ; cytology ; Transfection ; Vaccines, DNA ; immunology ; Viral Envelope Proteins ; biosynthesis ; genetics ; Viral Vaccines ; immunology

BACKGROUNDAnterior chamber associated immune deviation (ACAID) is characterized by a Th2 cell response. GATA-3 has been shown to be necessary for the activation of Th2 cells. This study was designed to examine the expression of GATA-3 in the development of ACAID.

METHODSACAID was induced by injection of 50 microg interphotoreceptor retinoid binding protein (IRBP) into the anterior chamber (AC) of Wistar rats. Delayed-type hypersensitivity (DTH) was evaluated on day 3, 7, 14, 21, 28 after IRBP inoculation. GATA-3 expression was detected using immunohistochemical staining. The expression of GATA-3 mRNA at different time points after AC injection of IRBP was assayed by reverse transcriptase polymerase chain reaction (RT-PCR).

RESULTSA significant DTH reaction was observed in Wistar rats on day 3 and 5 after IRBP inoculation. The DTH reaction was decreased 7 days after IRBP inoculation. GATA-3 expression was weak at both mRNA and protein levels in the normal spleen, but was significantly increased on day 5, 7, 14, and 21 after AC injection of IRBP.

CONCLUSIONThe expression of GATA-3 is increased during ACAID, suggesting that GATA-3 may be involved in the development of ACAID.

Animals ; Anterior Chamber ; immunology ; Eye Proteins ; immunology ; Female ; GATA3 Transcription Factor ; analysis ; genetics ; physiology ; Hypersensitivity, Delayed ; immunology ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Retinol-Binding Proteins ; immunology ; Spleen ; metabolism ; Th2 Cells ; immunology