Panscleritis After Blunt Ocular Trauma in A Child with Epididymitis / 中国医学科学杂志(英文版)

We reported an 8-year-old boy with panscleritis in left eye and right epididymitis after falling on the ground. Etiologic diagnosis played a key role in this case. Systemic examinations ruled out systemic autoimmune diseases, tumors, and infections as the cause of scleritis and suggested that the disease was caused by a local delayed-type hypersensitivity (DTH) induced by ocular trauma and was non-infectious. Still, the right epididymitis was infectious. Both conditions were treated successfully using steroids and antibiotics, respectively. Thus, early etiologic diagnosis and reasonable treatment are crucial to prevent visual loss.

Formulation and Validation of Recombinant Antigens CFP10 and ESAT6 for Tuberculosis Diagnosis

Abstract The rapid and accurate diagnosis of tuberculosis (TB), especially considering limited resources, is still a challenge. Development of new methodologies and tests are needed to overcome several disadvantages of the available standard tests. We evaluated the diagnostic potential of two antigens specific for Mycobacterium tuberculosis, the CFP10 and ESAT6 recombinant proteins, and developed stable formulations thereof. Sensitivity and specificity of the delayed-type hypersensitivity (DTH) skin testing and the induction of gamma interferon production (IFN-γ) by lymphocytes, as a non-invasive test, were evaluated using the CFP10 and ESAT6 protein formulations. The recombinant proteins produced by our group presented a high DTH response and the ability to differentiate between tuberculosis infection, BCG vaccination, and the contact with non-tuberculous mycobacteria (NTM). The production of IFN-γ by stimulation with individual and combined proteins was detected in a panel of 40 individuals and showed a specificity of 100% and a sensitivity of 90% when the two proteins were used together. Lyophilized formulations were stable under all conditions, while soluble formulations were stable under freezing at -20 ºC and -80 ºC. The proposed formulations containing the ESAT6 and CFP10 recombinant antigens constitute satisfactory tools for TB testing, suitable to be developed and implemented in a large-scale trial.

Salivary gland antigens of laboratory-bred Phlebotomus sergenti and their immunogenicity in human volunteers in laboratory condition

To investigate Phlebotomus (P.) sergenti Parrot, 1917 (Diptera: Psychodidae) salivary gland antigens and their immune response in human. Methods: Human volunteers were exposed to sand flies' bites in the laboratory, and following each exposure the size of induration was recorded. The mean protein concentration of salivary gland lysate and specific anti-P. sergenti saliva IgG was measured. Sand fly salivary proteins were separated by SDS-PAGE and their immunoreactivity was examined by Western blotting assays. Results: Individuals exposed to P. sergenti salivary gland lysate for 8 months showed both antibody and delayed type hypersensitivity responses, although exposure for one month did not provoke any immune responses. The trend of antibody fluctuated during the exposure time and dropped by the end of antigen loading. The mean protein content was (0.36?0.08) ug in each pair salivary glands. Salivary gland lysate showed 11 to 12 major protein bands and 3 to 6 of them were immunoreactive. Conclusions: Our study showed that the salivary gland components of P. sergenti provoked both cellular and humoral immune responses in human. Furthermore, there are some immunogenic proteins in P. sergenti saliva which could be subjected for further investigation as vector-based vaccine candidate/s against anthroponotic cutaneous leishmaniasis.

Salivary gland antigens of laboratory-bred Phlebotomus sergenti and their immunogenicity in human volunteers in laboratory condition

To investigate Phlebotomus (P.) sergenti Parrot, 1917 (Diptera: Psychodidae) salivary gland antigens and their immune response in human. Methods: Human volunteers were exposed to sand flies' bites in the laboratory, and following each exposure the size of induration was recorded. The mean protein concentration of salivary gland lysate and specific anti-P. sergenti saliva IgG was measured. Sand fly salivary proteins were separated by SDS-PAGE and their immunoreactivity was examined by Western blotting assays. Results: Individuals exposed to P. sergenti salivary gland lysate for 8 months showed both antibody and delayed type hypersensitivity responses, although exposure for one month did not provoke any immune responses. The trend of antibody fluctuated during the exposure time and dropped by the end of antigen loading. The mean protein content was (0.36?0.08) ug in each pair salivary glands. Salivary gland lysate showed 11 to 12 major protein bands and 3 to 6 of them were immunoreactive. Conclusions: Our study showed that the salivary gland components of P. sergenti provoked both cellular and humoral immune responses in human. Furthermore, there are some immunogenic proteins in P. sergenti saliva which could be subjected for further investigation as vector-based vaccine candidate/s against anthroponotic cutaneous leishmaniasis.

Evaluation Of Immunomodulatory Activity Of Diosgenin In Rats

Background: Theimmune system is intrinsic to health. Modulation of the immune responses to alleviate the diseases by using herbal plants has been of interest for many years. Diosgenin, a naturally occurring steroid saponin mainly present in the seeds of fenugreek (Trigonella foenum graecum)and in the root tubers of wild yams (Dioscorea vil-losa). Activation of specific and nonspecific immunity results in stimulation of immune response. Diosgenin has the positive effects on both specific and nonspecific immunity.Aim: To study the immunomodulatory activity of Diosgen-in in rats. Method: The suspension of Diosgenin wasgiven orally at the dosage level of 50, 100 and 150 mg/kg for 21 days in a rat. The immunomodulatory activity on specific and non-specific immunity was studied by haemagglutina-tion antibody (HA) titer, delayed type hypersensitivity (DTH) response and carbon clearance test. Immunosuppres-sion in a rat was induced by using Cyclophosphamide (100 mg/kg, p.o.). Sheep red blood cells (SRBCs) were used as antigen (0.1ml 20% SRBCs) in haemagglutinating antibody titer and delayed type hypersensitivity response methods. Result: Diosgenin exhibited significant increase in the production of antibody titer in response to SRBC antigen. A significant increase in both primary and secondary HA titer was observed in immunosuppressed group treated with Diosgenin when compared with negative control. A significant increase in the DTH response was observed in immu-nosuppressed animals treated with Diosgenin, pre-sensitized with SRBCs antigen. Diosgenin exhibited significant in-crease in phagocytic index against control group, indicating the stimulation of the reticuloendothelial system. Con-clusion: The study indicates that Diosgenin triggers stimulatory effect on specific and nonspecific immune response. The immunostimulant effect of Diosgenin could be attributed due to its saponin glycoside.

Quantification of rutin and quercetin by HPTLC/HPLC and in vitro immunomodulatory and anticancer activities of Capparis moonii fruits extracts

Background: The current research was undertaken on dried fruits of Capparis moonii to screen its potential for immunomodulatory and cancer indications with identification of phytoconstituents by chromatographic techniques.Methods: Methanolic (MECN), hydro-methanolic (HMECN) and aqueous extracts (AQCN) of Capparis moonii were subjected to high performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC) after studying the total phenolic and flavonoid content by using rutin and gallic acid as standards respectively as well as undertaking powder characteristics and preliminary phytochemical screening. Immunomodulatory activities covered were hemagglutination antibody titre and delayed-type hypersensitivity reaction with the aid of sheep red blood cells (0.5×109) as antigens. The extracts were studied for antioxidant potential. Anticancer prospects were focusing on in vitro cell lines screening (MCF 7 and HCT 15) by Sulforhodamine B assay method and potato disc assay.Results: The total phenolic and flavonoid content of MECM, HMECM and AQCM fruits extracts were found to be 0.20, 0.11 and 0.47 mg of gallic acid/g and 78.3, 18.8 and 64.4 mg of rutin/g respectively. Rutin and quercetin were confirmed by HPTLC and HPLC showing well resolved peaks. IC50 values in antioxidant studies were found to be significant with all the extracts. Significant immunomodulatory effect was noticed at 200mg/kg in both models (high antibody titre levels and decrease paw volume after 48 h). Unsatisfactory results were observed with selected cell lines and disc assay.Conclusions: Thus, selected fruits may probably have immunomodulatory potential due to presence of flavonols (rutin and quercetin).

Carissa congesta Wight and Benincasa hispida (Thunb.) Cogn. as budding immunomodulatory agents.

Carissa congesta and Benincasa hispida are well-known medicinally important plants associated with diabetes, inflammation, protozal infections and cancer. Here, we emphasized up on the immunomodulatory potential of these plants as the source of lupeol, β-sitosterol and ursolic acid. Petroleum ether extracts of C. congesta roots and B. hispida seeds were subjected to acute toxicity studies. They were screened for its immunomodulatory prospective in rats by Haemagglutination Antibody (HA) titre and Delayed-Type Hypersensitivity (DTH) response using Sheep Red Blood Cells (SRBCs of-0.5×109) as antigens. Carbon Clearance test (Phagocytic Index) was estimated by Indian ink suspension. Complete Freund’s Adjuvant (CFA) induced arthritis model interpretation was done by paw edema, kene joint erosion (transverse section), body weights, arthritic index and biochemical levels (RBC, WBC and Hb levels). Both the extracts were found to be therapeutically safe up to 5000 mg/kg. Dosage of 100 mg/kg was not satisfactory; and 500 and 250 mg/kg showed significant immunostimmulation (HA Titre) and immunosuppression (DTH response, 48 h). Benincasa hispida seed and Carissa congesta root extracts showed phagocytic Index of 0.0163±0.003, 0.0145±0.003 and 0.0183±0.003, 0.0176±0.003 at 250 mg/kg and 500 mg/kg, respectively. CFA model revealed that the B. hispida seed and C. congesta root extracts decreased paw volume, knee joint erosion, increased body weights and biochemical parameters with an arthritic index of 1.31±0.12, 1.44±0.15 and 1. 16±0.09, 1.36±0.13 at 250 mg/kg and 500 mg, respectively. The results were interpreted by One-way ANOVA followed by Dunnett test. Extracts showed relevance as promising immunostimulators as compared to control.

Protective Effects ofDasangju Mixture on Liver Injury Induced by Delayed-type Hypersensitivity in Mice / 中国中医药信息杂志

Objective To investigate the protective effects ofDasangju Mixture on the liver injury induced by delayed-type hypersensitivity in mice; To discuss its mechanism of anti-inflammatory-immunity.Methods After enterocoelia intravenous injection of cyclophosphamide was used, 2,4,6-picryl chloride was used twice for exterior coating for skin of abdomen to cause sensation of skin. Then percutaneous transhepatic puncture was used to attack for modeling. Male Kunming mice were randomly divided into blank group, model group, high-, medium- and low-dose treatment groups. The high-, medium- and low-dose treatment groups were given Dasangju Mixture for gavage, once a day for 7 days. ELISA was applied to determine the serum levels of aspartate aminotransferase (AST), alamine aminotransferase (ALT) and tumor necrosis factor (TNF)-α. Pathological features of liver tissue were observed by HE staining.Results Compared with the blank group, the levels of AST, ALT and TNF-α in the model group increased significantly (P<0.05); Compared with the model group, the levels of AST, ALT and TNF-α in the high-, medium- and low-dose treatment groups decreased significantly (P<0.05,P<0.01,P<0.001); Compared with the model group, high-, medium- and low-dose treatment groups can better alleviate topical inflammation infiltration and edema of liver tissue.ConclusionDasangju Mixture has certain protective effects on the liver injury induced by delayed-type hypersensitivity in mice.

Effect of interferon gamma on immune factor and CD69 , CD107 a in patients with delayed type hypersensitivity / 中国生化药物杂志

Objective To investigate effect of interferon gemma on immune factor and CD69, CD107a in patients with delayed type hypersensitivity by interferon gamma.Methods 76 cases with delayed type hypersensitivity were selected and divided into 2 groups.38 cases in control group were treated conventional therapy, 38 cases in experiment group were treated by interferon gamma.Peripheral blood CD69, CD107a, immune factor, T cell subsets and the treatment efficiency were compared after the treatment.Results Compared with the control group, the serum CD69 and CD107a levels were lower in the experimental group (P<0.05), serum IgG and IgA levels were higher, the serum IgE level was lower (P<0.05), and the CD3 +,CD4 +,CD4 +/CD8 +level was higher, serum CD8 + was lower (P<0.05) .The effective rate of the treatment group was significantly higher than that of the control group (P<0.05) .Conclusion Interferon gamma has good clinical effect in the patients with delayed type hypersensitivity, and can effectively reduce the levels of CD69 and CD107a in the peripheral blood, and regulate the immune function of the body.

Immunosuppressive and regulatory mechanism of artesunate / 中国药学杂志

OBJECTIVE: To determine the immunoregulatory mechanism of artesunate both in vitro and in vivo. METHODS: The immunosuppressive action of artesunate was investigated through lymphocyte proliferation and cytotoxicity was also detected in vitro. The delayed-type hypersensitivity (DTH) model in mice was used for the investigation of artesunate in vivo. The mRINA expression of T-bet, GATA-3, ROR-γt and IL-17 gene was determined by RT-PCR. ELISA assay was used to detect the contents of IL-17, and Western blot assay was applied to investigate the p38 mitogen-activated protein kinase (MAPK) activation. RESULTS: Artesunate could inhibit ConA-induced T lymphocyte and LPS-induced B lymphocyte proliferation in vitro significantly, which also showed lower toxicity than dexamethasone. Meanwhile, topical application of artesunate could both suppress the increase on ear thickness in DTH mice and inhibit the thymus index and spleen index. Furthermore, artesunate was found to regulate the expression level of transcription factor (T-bet in the GATA-3), and down-regulate ROR-γt and IL-17 expression. What's more, the production of IL-17 was decreased in ear tissue. Finally, artesunate was observed to decrease p38 mitogen-activated protein kinase (MAPK) activation. CONCLUSION: Artesunate, as a new immunomodulatory agents, might contribute to performing immunosuppression through modulating the balance of treg/Th17 and anti-inflammatory.

Safety and skin delayed-type hypersensitivity response in vervet monkeys immunized with Leishmania donovani sonicate antigen delivered with adjuvants / Segurança e reação de hipersensibilidade tardia na pele de macacos vervet imunizados com antígeno sonicado de Leishmania donovani junto com adjuvantes

In this study, we report on the safety and skin delayed-type hypersensitivity (DTH), responses of the Leishmania donovani whole cell sonicate antigen delivered in conjunction with alum-BCG (AlBCG), Montanide ISA 720 (MISA) or Monophosphoryl lipid A (MPLA) in groups of vervet monkeys. Following three intradermal injections of the inoculums on days 0, 28 and 42, safety and DTH responses were assessed. Preliminary tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) levels were also measured and these were compared with DTH. Only those animals immunized with alum-BCG reacted adversely to the inoculum by producing ulcerative erythematous skin indurations. Non-parametric analysis of variance followed by a post-test showed significantly higher DTH responses in the MISA+Ag group compared with other immunized groups (p < 0.001). The MPLA+Ag group indicated significantly lower DTH responses to the sonicate antigen compared with the AlBCG+Ag group. There was a significant correlation between the DTH and cytokine responses (p < 0.0001). Based on this study we conclude that Leishmania donovani sonicate antigen containing MISA 720 is safe and is associated with a strong DTH reaction following immunization.

Neste estudo reportamos segurança e resposta de hipersensibilidade tardia (DTH) do antígeno sonicado de células totais de Leishmania donovani introduzidos juntamente com alume-BCG (AIBCG) Montanide ISA 720 (MISA) ou lípide A monofosforilado (MPLA) em grupos de macacos vervet. Depois de três injeções intradérmicas do inóculo nos dias 0, 28 e 42 segurança e resposta DTH foram avaliados. Preliminarmente níveis de fator de necrose tumoral alfa (TNF-α) e interferon gama (IFN-γ) foram também medidos e comparados com o DTH. Somente os animais imunizados com alume-BCG reagiram de maneira diversa ao inóculo produzindo indurações ulceradas e eritematosas na pele. Análise não paramétrica de variação seguida por um teste posterior mostraram resposta significantemente mais alta do DTH no grupo MISA + Ag quando comparado com outros grupos imunizados (p < 0.001). O grupo MPLA + Ag demonstrou resposta DTH significantemente menor do antígeno sonicado comparado com o grupo AIBCG + Ag. Houve correlação significante entre o DTH e a resposta às citocinas (p < 0.0001). Baseados neste estudo concluímos que o antígeno sonicado de Leishmania donovani contendo MISA 720 é seguro e está associado com forte reação DTH após imunização.

Reacciones medicamentosas severas en piel / Severe skin drug reactions

Las drogas son sustancias químicas que pueden interferir con el sistema inmune y a veces conducen a reacciones inusuales y severas. Éstas pueden amenazar la vida, requerir hospitalizaciones prolongadas o dejar secuelas significativas. Cerca del 2 por ciento de las reacciones cutáneas inducidas por fármacos se consideran graves. Estas son el angioedema, el shock anafiláctico, el síndrome de Stevens-Johnson (SSJ), la necrolisis epidérmica tóxica (NET), y el síndrome de hipersensibilidad (DRESS), entre otros. Requieren atención especial ya que los síntomas clínicos son heterogéneos y pueden imitar diferentes enfermedades, lo que lleva a retardar el diagnóstico correcto.

Drugs are chemicals that can interfere with the immune system and may sometimes lead to unusual and severe reactions. These can be life threatening, requiring prolonged hospitalization or have significant sequelae. About 2 per cent of drug-induced skin reactions are considered serious. They are angioedema, anaphylactic shock, the Steven-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), and hypersensitivity syndrome (DRESS), among others. They require special attention because clinical symptoms are heterogeneous and can mimic different diseases, leading to a delay in the correct diagnosis.

Delayed-type hypersensitivity skin test responses to PPD and other antigens among BCG-vaccinated HIV-1-infected and healthy children and adolescents / Resposta de testes de hipersensibilidade tardia utilizando PPD e outros antígenos em crianças e adolescentes saudáveis e infectados pelo HIV-1 e vacinados com BCG

INTRODUCTION: Among HIV-1-infected patients, CD4+ T cell counts are well-established markers of cell-mediated immunity. Delayed-type hypersensitivity (DTH) skin tests can be used to evaluate in vivo cell-mediated immunity to common antigens. METHODS: DTH responses to tuberculin purified protein derivative (PPD), sporotrichin, trichophytin, candidin and streptokinase/streptodornase antigens were assessed. Thirty-six HIV-1-infected children/adolescents and 56 age- and sex-matched HIV-1/HIV-2-seronegative participants were tested. All participants had a BCG scar. Fisher's exact test was used to evaluate significant differences between groups (p<0.05). RESULTS: The main characteristics of the HIV-1 patients were as follows: median age 8.1 years; 20/36 were males; 35 were vertical transmission cases; 34 were AIDS cases under antiretroviral therapy; median viral load = 3.04 log10 copies/ml; median CD4+ T cell count = 701 cells/μl. A total of 25 percent (9/36) and 87.5 percent (49/56) of HIV-1-infected and healthy participants, respectively, displayed DTH reactivity to at least one antigen (p<0.001). Among HIV-1-infected participants, reactivity to candidin predominated (8/36, 22.2 percent), while PPD positivity prevailed among healthy participants (40/56, 71.4 percent). PPD reactivity in the HIV-1-positive group was 8.3 percent (p<0.01). The median PPD induration was 2.5mm (range: 2-5mm) in the HIV-1 group and 6.0 mm among healthy participants (range: 3-15mm). There was no correlation between PPD positivity and age. No correlation between CD4+ T cell counts and DTH reactivity was observed among HIV-1-infected patients. CONCLUSIONS: DTH skin test responses, including PPD reactivity, were significantly lower among HIV-1-infected participants compared to healthy controls, which likely reflects advanced disease and T cell depletion.

INTRODUÇÃO: A contagem de células CD4+ representa marcador da resposta imune celular em pacientes infectados pelo HIV-1. Testes cutâneos de hipersensibilidade tardia (DTH) podem ser empregados para avaliar in vivo respostas celulares a antígenos comuns. MÉTODOS: DTH para derivado proteico purificado de tuberculina (PPD), esporotriquina, tricofitina, candidina e estreptoquinase/estreptodornase foram realizados. Foram testados crianças/adolescentes infectados pelo HIV-1 (n=36) e indivíduos saudáveis (n=56), soronegativos para HIV-1/HIV-2 pareados por sexo-idade, todos com cicatriz vacinal por BCG. Teste exato de Fisher foi aplicado (p<0,05). RESULTADOS: Entre as crianças/adolescentes infectados pelo HIV-1, mediana de idade=8,1 anos; 20/36 eram do sexo masculino; 35 casos de transmissão vertical; 34 casos de AIDS sob terapia antirretroviral; mediana de carga viral = 3.04lc10 cópias/ml; mediana de contagem de células CD4+ = 701 células/μl. Entre os infectados e saudáveis a reatividade DTH a pelo menos um dos antígenos foi, respectivamente, 25 por cento (9/36) e 87,5 por cento (49/56) (p<0,001). Reatividade à candidina predominou nos infectados (8/36, 22 por cento) e ao PPD nos indivíduos saudáveis (40/56, 71,4 por cento). A reatividade ao PPD entre infectados foi de 8,3 por cento (p<0,01). A mediana da induração ao PPD foi 2,5mm (variação: 2-5mm) entre infectados e 6,0mm (variação: 3-15mm) entre os saudáveis. Não observamos correlação entre positividade ao PPD e idade. No grupo de infectados, não observamos correlação entre contagens de células CD4+ e reatividade ao DTH. CONCLUSÕES: Respostas DTH significativamente diminuídas, incluindo a reatividade ao PPD foram observadas em crianças/adolescentes infectados pelo HIV-1 comparadas com controles saudáveis, provavelmente refletindo doença avançada e supressão da imunidade mediada por células T.

Cell-free antigens from precocious Paracoccidioides brasiliensis culture induce a typical delayed-type hypersensitivity reaction

Cell-free antigens (CFAg) derived from Paracoccidioides brasiliensis have typically been used in immunodiffusion reactions for serodiagnosis or therapeutic follow-up of paracoccidioidomycosis patients. Thus, we investigated the usefulness of CFAg obtained from cultures at different ages, to evaluate cellular immunity by the footpad test, in experimental murine paracoccidioidomycosis. Male mice infected with P. brasiliensis 265 strain were challenged in the footpad with CFAg obtained from four- (4d CFAg) or 11-day-old cultures (11d CFAg). The increase in footpad swelling provoked by 4d CFAg and 11d CFAg was similar and showed significant difference in relation to control groups. However, the infiltrate pattern was strikingly different: 4d CFAg induced a predominant mononuclear infiltrate whereas 11d CFAg provoked a predominant polymophonuclear infiltrate. These different inflammatory patterns were associated with distinct electrophoretic characteristics. By comparison with 11d CFAg, 4d CFAg showed more numerous and intense bands, including a strong one of 43 kDa (gp43). These results suggest that CFAg derived from Pb 265 isolate can be used as a reagent to evaluate cellular immunity; however, the culture's age is critical because only young cultures are able to induce a typical mononuclear infiltrate. The efficacy of this new paracoccidioidin to assay the cellular immunity in infections caused by other P. brasiliensis isolates is under investigation.

Effects of uric acid sodium salt on antibody response, dendritic cells and delayed-type hypersensitivity in BALB/c mice / 中国病理生理杂志

AIM: To investigate the effects of uric acid sodium salt (UANa) as adjuvant on humoral and cellular immune response in BALB/c mice. METHODS: BALB/c mice were immunized with trichosanthin (TCS) as antigen together with UANa suspension as adjuvant. The antibody titers of IgG were detected by enzyme-linked immunosorbent assay. Dendritic cells (DC) were induced in vitro, the phenotypes of DC were analyzed by flow cytometry and the effect of UANa on DC maturity was evaluated. A delayed-type hypersensitivity (DTH) model was used to analyze the effect of UANa on cellular immune responses in vivo. The in vitro proliferation of lymphocytes was determined by ConA stimulation. RESULTS: Freunds adjuvant greatly enhanced the antibody response of mice to TCS, while UANa adjuvant failed to promote the antibody response but significantly reduced the antibody response as compared to TCS only. No effect of UANa on the expression of CD11c and CD83 in DC was observed by flow cytometry analysis. However, UANa significantly enhanced the expression of MHC II molecule. In the DTH model, UANa enhanced the degree of allergen-induced ear swelling and promoted the ability of lymphocyte proliferation in vitro. CONCLUSION: UANa suspension as adjuvant significantly enhances the cellular immune response but inhibits the humoral immune response to a certain degree, suggesting that UANa has potential application in the vaccine research.

Boosting the suppressive effects of Ascaris suum components in IFN-γ-deficient mice / Potencialização do efeito supressor de componentes do Ascaris suum em camundongos deficientes em IFN-γ

High molecular weight components from Ascaris suum extract suppress ovalbumin-specific immunity in mice. In IFN-γ-deficient mice, ovalbumin-specific delayed-type hypersensitivity reactions are more strongly downregulated by these suppressive components. Here, the cellularity of the delayed-type hypersensitivity reaction in IFN-γ-deficient mice and the increased downregulation induced by Ascaris suum components were analyzed. IL-12p40-dependent neutrophilic influx was predominant. Suboptimal doses of the suppressive fraction from this nematode completely inhibited the hypersensitivity reaction, thus indicating intensification of the immunosuppression under conditions of intense recruitment of IFN-γ-independent neutrophils.

Componentes de alto peso molecular do extrato de Ascaris suum suprimem a imunidade específica à ovalbumina em camundongos. Em camundongos geneticamente deficientes de IFN-γ a reação de hipersensibilidade tardia específica para ovalbumina foi mais fortemente prejudicada por estes componentes supressivos. Aqui, a celularidade da reação de hipersensibilidade tardia em camundongos deficientes de IFN-γ e o incremento na supressão induzida por componentes do Ascaris suum foram analisados. Influxo neutrofílico, dependente de IL-12p40, foi predominante. Dose sub-ótima da fração supressiva do nematódeo inibiu completamente a reação de hipersensibilidade, indicando uma intensificação da imunossupressão em condições de recrutamento intenso de neutrófilos independente de IFN-γ.

Cytokine profile in Montenegro skin test of patients with localized cutaneous and mucocutaneous leishmaniasis / Perfil de citocinas na intradermorreação de Montenegro em doentes de leishmaniose cutânea localizada e cutâneo-mucosa

American tegumentary leishmaniasis presents as two major clinical forms: localized cutaneous leishmaniasis (LCL) and mucocutaneous leishmaniasis (MCL). The immune response in leishmaniasis is efficiently evaluated by the response to Leishmania antigen through the Montenegro skin test (MST). Both LCL and MCL present positive response to MST, indicating that the patients present cell-mediated immunity against the parasite - Leishmania. In spite of the presence of immunity in MCL, this is not sufficient to stop disease progression and prevent resistance to treatment. In this study we demonstrated interleukin (IL) 2, 4, 5 and interferon (IFN) gamma expression in biopsies of MST of ten patients with American tegumentary leishmaniasis. The obtained results were compared between LCL (n = 5) and MCL (n = 5) patients. The MST of MCL patients displayed a higher expression of IL-2, IL-4 and IL-5, in comparison to LCL. There was no significant difference in IFN-gamma expression between groups. The obtained results suggest the role of IL-4 and IL-5 in the maintenance of the immunopathogenic mechanism of the destructive lesions that characterize MCL.

A leishmaniose tegumentar americana apresenta duas formas clínicas mais comuns: a leishmaniose cutânea localizada e a leishmaniose cutâneo-mucosa. A imunidade da leishmaniose é avaliada pela resposta ao antígeno Leishmania através da Intradermorreação de Montenegro. Estas duas formas apresentam resposta positiva, indicando que o paciente apresenta imunidade celular contra o parasita Leishmania. Apesar da presença da imunidade celular na leishmaniose cutâneo-mucosa, esta não é suficiente para barrar a progressão da doença e a resistência ao tratamento. Neste estudo, detectamos quatro citocinas por imunohistoquímica, IL-2, IL-4, IL-5 e IFN-gama nas biópsias da intradermorreação de Montenegro de pacientes com leishmaniose tegumentar americana (n = 10), cinco com leishmaniose cutânea e cinco com cutâneo-mucosa. Os resultados mostraram uma alta expressão significativa de IL-2, IL-4, IL-5 na leishmaniose cutâneo-mucosa comparada com a leishmaniose cutânea localizada, mas sem diferença significante na expressão do IFN-γ entre os grupos. Estes resultados sugerem a importância da participação da citocina IL-4 e IL-5 na manutenção do mecanismo imunopatogênico das lesões destrutivas da forma cutâneo-mucosa.

American tegumentary leishmaniasis: langerhans cells in montenegro skin test / Leishmaniose tegumentar americana: células de Langerhans na reação intradérmica de Montenegro

This work analyzed the histopathology and epidermal Langerhans cells (LC) of Montenegro skin test (MST) in patients with American tegumentary leishmaniasis (ATL) in order to in situ characterize and compare the immunological reaction of the two major clinical forms of ATL, localized cutaneous leishmaniasis (LCL) and mucocutaneous leishmaniasis (MCL). MST histopathology of both LCL and MCL showed superficial and deep perivascular inflammatory infiltrate composed mainly of lymphocytes and histiocytes. Epidermal LC population was higher in MST biopsies taken from LCL patients when compared to MCL group, at 48 and 72 hours after antigen inoculation. Increased number of epidermal LC displayed in MST biopsies of LCL patients represents specific cellular immunity against parasites. The decrease of LC in MST biopsies of MCL patients does not necessarily indicate a worse specific cellular immunity in this clinical form of leishmaniasis.

Este trabalho analisou e quantificou as células de Langerhans e as características histopatológicas da reação de Montenegro nos pacientes com leishmaniose tegumentar americana (LTA) para caracterizar seu comportamento imunológico nas duas formas clínicas mais comuns da LTA, a leishmaniose cutânea localizada (LCL) e a leishmaniose cutâneo-mucosa (LCM). O exame histopatológico apresentou infiltrado inflamatório perivascular superficial e profundo, com predomínio de histiócitos e linfócitos, sem diferença significante entre as duas formas da doença. O resultado da quantificação das CL apresentou aumento das CL na LCL e diminuição na LCM em 48 e 72 horas após a inoculação do antígeno (p < 0,001). O aumento das células de Langerhans epidérmicas na reação de Montenegro da LCL demonstra a presença de imunidade celular específica, enquanto a diminuição das mesmas células na LCM não necessariamente demonstra uma diminuição da imunidade celular específica.

Coptis chinensis Extract Inhibits the Production of Inflammatory Mediators and Delayed Type Hypersensitivity in Mice

BACKGROUND: Coptis chinensis rhizome has been used as a medicinal herb in traditional Oriental medicine. We investigated the effects of Coptis chinensis extract on inflammatory mediators and delayed type hypersensitivity in mice. METHODS: The inhibitory effect of ethanolic extract of Coptis chinensis (CCE) on cell proliferation was evaluated using MTS assay. The lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and the Con A-activated mouse splenocytes were cultured with various concentrations of CCE. Total nitric oxide (NO) production was determined by Griess reaction. The amounts of secreted prostaglandine E2 (PGE(2)), interleukin (IL)-2 and IFN-gamma were measured by ELISA. To investigate the in vivo anti-inflammatory effect of CCE, oxazolone-induced delayed type hypersensitivity (DTH) model was used. RESULTS: The CCE at 100 microgram/ml significantly blocked the LPS-induced production of pro-inflammatory mediators (NO and PGE) in RAW264.7 macrophages. Also, it significantly inhibited cell proliferation and cytokine (IL-2 and IFN-gamma) production in splenocytes. Furthermore, when splenocytes from CCE fed mice (200 mg/kg for 2 weeks) were activated with Con A, cell proliferation and cytokine production were significantly inhibited. In addition, CCE decreased in vivo inflammation in oxazolone-induced DTH model mice. CONCLUSION: We suggest that Coptis chinensis can be used as an anti-inflammatory drug by exerting an inhibitory effect in inflammatory mediator- and cell-mediated inflammation.

Coptis chinensis Extract Inhibits the Production of Inflammatory Mediators and Delayed Type Hypersensitivity in Mice

BACKGROUND: Coptis chinensis rhizome has been used as a medicinal herb in traditional Oriental medicine. We investigated the effects of Coptis chinensis extract on inflammatory mediators and delayed type hypersensitivity in mice. METHODS: The inhibitory effect of ethanolic extract of Coptis chinensis (CCE) on cell proliferation was evaluated using MTS assay. The lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and the Con A-activated mouse splenocytes were cultured with various concentrations of CCE. Total nitric oxide (NO) production was determined by Griess reaction. The amounts of secreted prostaglandine E2 (PGE(2)), interleukin (IL)-2 and IFN-gamma were measured by ELISA. To investigate the in vivo anti-inflammatory effect of CCE, oxazolone-induced delayed type hypersensitivity (DTH) model was used. RESULTS: The CCE at 100 microgram/ml significantly blocked the LPS-induced production of pro-inflammatory mediators (NO and PGE) in RAW264.7 macrophages. Also, it significantly inhibited cell proliferation and cytokine (IL-2 and IFN-gamma) production in splenocytes. Furthermore, when splenocytes from CCE fed mice (200 mg/kg for 2 weeks) were activated with Con A, cell proliferation and cytokine production were significantly inhibited. In addition, CCE decreased in vivo inflammation in oxazolone-induced DTH model mice. CONCLUSION: We suggest that Coptis chinensis can be used as an anti-inflammatory drug by exerting an inhibitory effect in inflammatory mediator- and cell-mediated inflammation.