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

OBJECTIVETo explore the immune response of silicotic rats to sheep red blood cells(SRBC).

METHODSSilicotic rats were immunized with SRBC by tracheal instillation(Group 1) or intraperitoneal injection (Group 2), and non-silicotic rats were immunized by tracheal instillation as normal control(Group 3). The levels of serum hemolytic index(HC50) were measured on 7, 12, 20, 25, and 32 days after primary immunization and 5, 12, 15 days after the second immunization. Special anti-SRBC IgG was measured with ELISA(A490 nm) on 12, 20, 25, 32 days and 5, 12, 15, 27 days respectively. Delayed-type hypersensitivity(DTH) to SRBC was measured 20 days after second immunization and DTH reaction was determined at 24, 48, 72, and 96 h after administration. Total cell count and cell populations in the bronchoalveolar lavage fluid(BALF), lung associated lymph node(LALN) and spleen weight, special IgG secreted from spleen cells were measured at the end of the experiment.

RESULTSThe HC50 of Group 1(47.4 +/- 1.0, 52.2 +/- 4.6, 31.1 +/- 11.9, 43.8 +/- 3.5, 33.6 +/- 16.8, 49.0 +/- 2.3, 92.9 +/- 20.2, 87.7 +/- 5.2) were statistically higher than those of Group 3(40.4 +/- 10.6, 2.8 +/- 2.5, 0.8 +/- 0.6, 6.6 +/- 5.8, 1.4 +/- 0.1, 36.5 +/- 16.5, 53.0 +/- 33.2, 2.6 +/- 2.2). The special anti-SRBC IgG response in Group 1(1.67 +/- 0.19, 1.98 +/- 0.36, 1.12 +/- 0.50, 1.38 +/- 0.30, 2.75 +/- 0.15, 2.60 +/- 0.28, 2.86 +/- 0.10, 2.50 +/- 0.20) were much stronger than those in Group 3 (0.59 +/- 0.30, 0.56 +/- 0.21, 0.21 +/- 0.16, 0.22 +/- 0.01, 0.81 +/- 0.25, 0.74 +/- 0.25, 0.69 +/- 0.26, 1.38 +/- 0.41). Furthermore, the results of DTH showed positive response and the ratios for diameter of skin rash > 5 mm at 24, 48, 72, 96 h were 16/16, 16/16, 16/16, 15/16 respectively in Group 1, while those in Group 3 were 8/15, 1/15, 1/15, 1/15 respectively. Total cell count in the BALF, LALN and spleen weight, and special IgG secreted from spleen cells in Group 1 were higher too. Group 2 expressed almost of the same but with mild immunologic responses as Group 1.

CONCLUSIONSilicosis-induced extremely strong DTH and over-response of humoral immunity to some antigens may contribute to the likelihood of silicosis complicated with tuberculosis.

Animals ; Erythrocytes ; immunology ; Hypersensitivity, Delayed ; etiology ; Immunization ; Immunoglobulin G ; blood ; Rats ; Sheep ; Silicosis ; immunology

In the present study, normal guinea pigs were used to investigate the possible pathogenic role of cell-mediated immunity in Sjogren's syndrome. The effects of anti-salivary gland antibodies on circulating lymphocytes, various organs including salivary glands, thymus and the reticuloendothelial system, and on delayed hypersensitivity were studied. Our study demonstrated that anti-salivary gland antibodies directly affected circulating lymphocytes. There was a 60-80% decrease in the lymphocyte count from the original level with a maximum effect at 5 hours after the introduction of the antibodies. When antibodies were injected repeatedly, the recovery to the pre-injection level of lymphocytes was delayed. We also found that antisalivary g1and antibodies were not organ-specific and were cross-reactive with various organs that are often involved in Sjogren's syndrome. Direct immunofluorescent study showed antibody deposits in the thymus-dependent areas of lymph nodes. These results suggest that antisalivary gland antibodies are lymphocytotoxic and have an anti-T cell property. The anti-salivary gland antibodies prepared in this experiment did not produce any pathological lesions such as those found in Sjogren's syndrome. The amount of antiserum or the period of administration might not have been long enough to produce pathological changes. Another possibility is that the anti-salivary gland antibodies might be species-specific. On the basis of these results, it appears that impaired cell-mediated immunity is not the primary pathogenic factor responsible for Sjogern's syndrome but rather that deranged immunity is secondary to the development of anti-salivary gland antibodies which occur in Sjogern's syndrome.
Animal ; Antibodies* ; Guinea Pigs ; Hypersensitivity, Delayed ; Leukocyte Count ; Lymphocytes/immunology* ; Rabbits ; Reticuloendothelial System/immunology ; Salivary Glands/immunology* ; Sjogren's Syndrome/immunology ; T-Lymphocytes/immunology

To evaluate the in vivo effect of autologous serum including antibodies to house dust mite in atopic individuals, we observed the immediate (15 mins) and late (6 hours) skin reactions (ISR, LSR) on intradermal (ID) test of serially diluted Dermatophagoides farinae antigens (DFa, Allergopharma, Germany) mixed with autologous sera (DFa-S) and diluent alone (DFa-D). We tested 34 DFa-skin reactive atopic individuals including 12 asthmatics (BA), 8 asthmatics on immunotherapy with DFa (IT), and 14 healthy atopic controls (AC). We observed complete inhibition of ISR in the lowest allergen dose of DFa-S in 7 (58.3%) of 12 BA, 3 (37.5%) of 8 IT, and 2 (14.3%) of 14 AC. In BA, the inhibition of ISR was more frequent than AC (p< 0.05). We observed larger late reactions in half of LSR positive cases on ID test by DFa-S than by DFa-D (> or = 1.5 X size; accentuation of LSR). Accentuation of LSR were shown more frequently by DFa mixed with larger amount of serum (25% in 1:1 mix; 80% in 1:3 mix, p< 0.05). But there were no differences of DFa-specific IgE and IgG subclass antibodies regardless of the inhibition of ISR or the accentuation of LSR. In conclusion, some autologous sera from DFa-sensitive individuals showed the inhibition of ISR and the accentuation of LSR on DFa-ID test.
Animal ; *Blood Physiology ; Dermatitis, Atopic/blood/*immunology ; Human ; Hypersensitivity, Delayed/*immunology ; Hypersensitivity, Immediate/*immunology ; Immunoglobulin E/metabolism ; Immunoglobulin G/metabolism ; Intradermal Tests ; Mites/*immunology ; Skin/*immunology ; Support, Non-U.S. Gov't

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 investigate the specific cell-mediated immune efficacy of the an HPV16 prophylactic vaccine.

METHODSC57BL/6 mice were randomly divided into 3 groups: experimental group I (treated with pcDNA L1), control group II (treated with pcDNA3.1) and control group III (treated with PBS buffer). The mice were immunized three times during a three-week interval. Ten to fourteen days after the third inoculation, a footpad swelling test was used to detect delayed-type hypersensitivity (DTH) responses. Antigen-specific splenocyte proliferation assay and quantitation of IFN-gamma cells in splenocytes were performed by FACS assay.

RESULTSIn the experimental group, splenocytes actively proliferated after stimulation with HPV16 VLP, and had developed a markedly larger amount of CD8(+) IFN-gamma(+) cells, which is an index for special CTL. Also, the footpad was significantly thickened upon inoculation with HPV16 VLP.

CONCLUSIONNaked DNA vaccine of HPV16 L1 can induce specific cell-mediated immune responses in mice, which should be considered for evaluation of HPV16 DNA vaccine feasibility.

Animals ; Hypersensitivity, Delayed ; etiology ; Immunization ; Interferon-gamma ; biosynthesis ; Lymphocyte Activation ; Mice ; Mice, Inbred C57BL ; Papillomaviridae ; immunology ; Spleen ; cytology ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology

BACKGROUND: Atopic dermatitis(AD) exibit multiple immune akno malities including elevated serum IgE levels and impaired cell mediated immunity, but basic immunology defect is largely unknown. OBJECTIVE: The purpose of this study was to evaluate the immnologic changes after treatment with Cyclosporin A(CsA) in AD patients. MEHTODS: Eight patients of severe AD were treated with CsA(4mg/kg body weight per day) for 5 weeks, and evaluated clinical and immunological parameters before aid after treatment. RESULTS: Peripheral blood eosinophil counts were significantly decreased after treatment(P<0.05), and serum IgE level was decressed but not significantly. In immediate hypersensitivity reaction by pinprick test, antigen score was.significantly decreased, and in the evaiuation of cell mediated immunity to recall antigens, total induration score was significantly increased(p<0.05). The number of helper T cell and helper/suppressor 7 cell ratio in peripheral blood, and the niimber of helper T and suppressor T cell in skin lesion were both significantly decreased after treatment(P<0.05). Clinical state after treatrnent was good in all of patients. CONCLUSION: The results indicate that CsA act on peripheral blood osinophil and immediate and delayed hypersensitivity reaction as well as helper T-cells in peripheral blood and skin lesion, and it may by useful in the treatrnent of severe AD.
Allergy and Immunology ; Body Weight ; Cyclosporine* ; Dermatitis, Atopic* ; Eosinophils ; Humans ; Hypersensitivity, Delayed ; Hypersensitivity, Immediate ; Immunity, Cellular ; Immunoglobulin E ; Skin ; T-Lymphocytes, Helper-Inducer

OBJECTIVETo determine whether B7-2 could improve hepatitis B virus (HBV)-specific immunity induced by inoculation of a plasmid encoding HBV preS2+S HBV DNA vaccine.

METHODSB7-2 expression plasmids were inoculated with the DNA vaccine in the gastrocemius muscles. Then the cytolytic T Lymphocyte activity (CTL), the delayed-type hypersensitivity (DTH) response and the titers of anti-HBs were analysed.

RESULTSThe DTH response and CTL activity were significantly enhanced when B7-2 expression plasmids were coinoculated with the DNA vaccine (P<0.01), but the titers of anti-HBs were unaffected (P>0.05).

CONCLUSIONSThe results suggest that B7-2 expression plasmid could improve HBV specific cell-mediated immunity induced by HBV DNA vaccine, and the gene-based coinoculation strategy using HBV viral antigen and B7-2 costimmulating could be a powerful means of combating HBV infection.

Adjuvants, Immunologic ; Animals ; Antigens, CD ; immunology ; B7-2 Antigen ; Hepatitis B Antibodies ; biosynthesis ; Hepatitis B Vaccines ; immunology ; Hypersensitivity, Delayed ; immunology ; Male ; Membrane Glycoproteins ; immunology ; Mice ; Mice, Inbred C57BL ; Plasmids ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, DNA ; immunology

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

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