Leprosy: clinical and immunopathological characteristics

Abstract Leprosy, a disease caused by Mycobacterium leprae, has polymorphic neurocutaneous manifestations strongly correlated with the host immune response. Peripheral neural damage can lead to sensory and motor losses, as well as deformities of the hands and feet. Both innate and acquired immune responses are involved, but the disease has been classically described along a Th1/Th2 spectrum, where the Th1 pole corresponds to the more limited presentations and the Th2 to the multibacillary ones. The aim of this review is to discuss this dichotomy in light of the current knowledge of the cytokines, T helper subpopulations, and regulatory T cells involved in each presentation of leprosy. The text will also address leprosy reactions related to increased inflammatory activity in both limited and multibacillary presentations, leading to exacerbation of chronic signs and symptoms and/or the development of new ones. Despite the efforts of many research groups around the world, there is no standardized serological test/biological marker for diagnosis so far, even in endemic areas, which could contribute to the eradication of leprosy.

Self-assembling, pH-responsive nanoflowers for inhibiting PAD4 and neutrophil extracellular trap formation and improving the tumor immune microenvironment

Self-assembling carrier-free nanodrugs are attractive agents because they accumulate at tumor by an enhanced permeability and retention (EPR) effect without introduction of inactive substances, and some nanodrugs can alter the immune environment. We synthesized a peptidyl arginine deiminase 4 (PAD4) molecular inhibitor, ZD-E-1M. It could self-assembled into nanodrug ZD-E-1. Using confocal laser scanning microscopy, we observed its cellular colocalization, PAD4 activity and neutrophil extracellular traps (NETs) formation. The populations of immune cells and expression of immune-related proteins were determined by single-cell mass cytometry. ZD-E-1 formed nanoflowers in an acidic environment, whereas it formed nanospheres at pH 7.4. Accumulation of ZD-E-1 at tumor was pH-responsive because of its pH-dependent differences in the size and shape. It could enter the nucleus and bind to PAD4 to prolong the intracellular retention time. In mice, ZD-E-1 inhibited tumor growth and metastasis by inhibiting PAD4 activity and NETs formation. Besides, ZD-E-1 could regulate the ratio of immune cells in LLC tumor-bearing mice. Immunosuppressive proteins like LAG3 were suppressed, while IFN-γ and TNF-α as stimulators of tumor immune response were upregulated. Overall, ZD-E-1 is a self-assembling carrier-free nanodrug that responds to pH, inhibits PAD4 activity, blocks neutrophil extracellular traps formation, and improves the tumor immune microenvironment.

Construction of CAR-T cells targeting CS1 and analysis of their antitumor activity in vitro / 生物工程学报

We constructed the CS1-targeted second- and third-generation CAR-T cells with genetic engineered 4-1BB or/and ICOS as a costimulatory signaling molecule by use of lentiviral platform. The CS1-targeted second-generation CAR-T cells with ICOS or 4-1BB had similar anti-neoplastic activity. When effector/target ratio was 1:1, the CAR-T cells with ICOS showed better killing effect on IM9-lucgfp cells than those with 4-1BB. However, The CS1-targeted third-generation CAR-T cells exihibited lower cytolytic capacity against IM9-lucgfp cells than the CS1-targeted second-generation CAR-T cells when the ratio of effector/target was 1:1, 2:1 or 5:1. When the ratio of effector/target was 10:1, the killing efficacy of both the second- and third-generation CAR-T cells against IM9-lucgfp cells was more than 85%, significantly higher than that of the control T cells. Taken together, both the CS1-targeted second- and third-generation CAR-T cells with ICOS or/and 4-1BB could efficiently kill CS1-positive multiple myeloma cells, but the CS1-targeted second-generation CAR-T cells had more potent killing effect on CS1-positive multiple myeloma cells than the CS1-targeted third-generation CAR-T cells.

Eukaryotic expression, protein purification and biological effects research of human CS1-Fc fusion protein / 生物工程学报

Signaling lymphocyte activation family 7 (SLAMF7/CS1) is a cell surface glycoprotein that is highly expressed in multiple myeloma cells. CS1 is a sensitive and specific biomarker for multiple myeloma. CAR-T cell immunotherapy is a new method for the treatment of multiple myeloma. CS1 CAR-T cell immunotherapy has good effect on relapsed refractory multiple myeloma. To detect the expression efficiency of CS1 CAR on CS1 CAR-T cells and to find an auxiliary means to CAR-T cell immunotherapy, we prepared a CS1-Fc fusion protein. First, the extracellular domain of CS1 was amplified from the existing plasmid by PCR and ligated with human IgG1-Fc fragment by overlap extension PCR. The recombinant fragment was ligated into pMH3 eukaryotic expression vector. After restriction enzyme digestion and DNA sequencing, the pMH3-CS1-Fc-his recombinant plasmid was successfully constructed. The recombinant plasmid was transfected into Chinese hamster ovary cell (CHO-S) by liposome. The expression of the CS1-Fc fusion protein in CHO-S cells was identified by flow cytometry after G418 pressure screening. Next, the CS1-Fc fusion protein was purified by nickel column. Western-blot analysis showed that molecular weight of the fusion protein was about 70 kDa was identified by Western blotting. The CS1-Fc fusion protein couldeffectively detect the expression rate of CS1 CAR and promote the activation, proliferation andcytokines secretion of the CS1 CAR-T cells. The results will lay the experimental foundation for the in vitro detection and potentiation of CAR-T cells in multiple myeloma treated with CS1 CAR-T cell.

Research advances in biological function of lymphocyte activation gene-3 (LAG-3) molecule and clinical application of antibody drugs targeting LAG-3 / 中国药理学与毒理学杂志

Lymphocyte activation gene-3 (LAG-3, CD223) is a class of immunosuppressive receptors, mainly expressed on the cell surface of activated T and natural killer (NK) cells. As with programmed death-1 (PD-1), T cell immunoglobulin domain and mucin domain-3 (TIM-3) and other immune check-points, the expression of LAG-3 in activated T cells is upregulated to prevent the occurrence of autoimmune diseases. In the tumor microenvironment, persistent antigenic stimulation can induce T cells to overexpress LAG-3 and other suppressor molecules, causing local immune suppression in the tumor microenvironment. However, the mechanism by which LAG-3 inhibits downstream signals of T-activation or interacts with other immunosuppressive molecules is still unknown. Preliminary clinical data has shown that anti-LAG-3 and anti-PD-1 antibodies possess synergistic effect on the treatment of tumors. Moreover, LAG-3 has different intracellular domains which are completely different from those of any other immunological negative regulatory molecules, suggesting that LAG-3 might have a unique molecular mechanism and prospect of application. Currently, there are at least 7 kinds of antibody drugs targeting LAG-3 and one kind of LAG-3-Fc fusion protein in clinical research, which indicates that anti-LAG-3 targeted drugs have a broad prospect of clinical application.

Antiepileptic drug-induced severe cutaneous adverse reactions and HLA alleles: A report of five cases with lymphocyte activation test

Antiepileptic drugs (AEDs) can induce severe cutaneous adverse reactions (SCARs) such as Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), and drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome. We performed HLA genotyping and lymphocyte activation tests (LATs) for five AED-induced SCAR patients (three males and two females; aged 40–66 years old). Three patients were treated with carbamazepine (CBZ) for pain control, one was treated with phenytoin (PHT) for seizure prevention, and one was treated with valproic acid (VPA) for seizure prevention. One patient was diagnosed with CBZ-induced DRESS syndrome and the remaining patients were diagnosed with SJS. All patients recovered from SCARs after stopping suspicious drugs and supportive care. LATs were conducted to confirm the culprit drug responsible for inducing SCARs; and LAT results were positive for the suspected culprit drugs, in all except in one case. HLA-A,

Progress of Epstein-Barr virus and lymphoma transformation / 白血病·淋巴瘤

Epstein-Barr virus (EBV) is one of the most common human herpesviruses, presenting a latent infection in more than 95% of healthy adults. EBV can regulate the differentiation, proliferation and colony formation of infected lymphocytes by coding viral proteins, and it is associated with Burkitt lymphoma, NK/T cell lymphoma, Hodgkin lymphoma, and vascular immunoblastic lymphoma. This article reviews the research progress of EBV in lymphoma transformation.

The effects of different stimulators on the activation of human lymphocytes / 药学学报

The research aims to study the effects of different stimulants on the activation of human lymphocytes. Human peripheral blood mononuclear cells were prepared by density centrifugation. The blood's sample was provided by National Institutes for Food and Drug Control and approved by its Ethics Committee. The expressions of CD69 in CD3+CD4+ and CD3+CD8+ human T cells were detected by flow cytometry after administrated with CD3/CD28 antibody, phytohaemagglutinin (PHA), Staphylococcus auresus enterooxin B (SEB), interleukin (IL27) and PMA plus ionomycin for 24 h. The proliferation of lymphocyte was detected by CellTiter-Glo kit. The secreted IFNγ in supernatant of medium was examined by ELISA kit. The proliferation of lymphocytes had no change after exposed of CD3/CD28 antibody, SEB, IL27 and PMA plus ionomycin for 24 h. However, the CD69 expressions in CD3+CD4+ and CD3+CD8+ T cells and IFNγ productions were significantly increased by CD3/CD28 antibody, SEB, IL27 and PMA plus ionomycin at 24 h, indicating that CD3+CD4+ and CD3+CD8+ T cells were activated under above-mentioned stimulated condition. CD3/CD28 antibody, SEB, IL27 and PMA plus ionomycin were valid stimulants for T cell activation.

Lymphocyte-Activation Gene-3 Expression and Prognostic Value in Neoadjuvant-Treated Triple-Negative Breast Cancer / 한국유방암학회지

PURPOSE: In this study, we aimed to evaluate lymphocyte-activation gene-3 (LAG-3) expression and its prognostic value in neoadjuvant-treated triple-negative breast cancer (TNBC). METHODS: LAG-3, programmed death-1 (PD-1), programmed death ligand-1 (PD-L1), and CD8⁺ tumor-infiltrating lymphocyte (TILs) levels were examined using immunohistochemistry in 148 preand 114 post-neoadjuvant chemotherapy (NACT) specimens of human TNBC tissue. Correlations between expression levels and clinicopathological features were analyzed. Prognostic values for combined detection in TNBC following NACT were evaluated. RESULTS: In pre-NACT specimens, LAG-3 expression showed a significant association with pathological complete response (pCR, p=0.038) and was correlated with PD-1 (p<0.001) and PD-L1 (p=0.008). In post-NACT specimens, high expression of LAG-3 showed significant effects on nodal status (p=0.023) and PD-1 (p<0.001). The expression of immune markers on TILs significantly increased following NACT. Multivariate analysis indicated that only nodal status (odds ratio [OR], 0.226; 95% confidence interval [CI], 0.079–0.644; p=0.005) and high quantities of CD8⁺TILs (OR, 3.186; 95% CI, 1.314–7.721; p=0.010) are independent predictors of pCR. Nodal status (hazard ratio [HR], 2.666; 95% CI, 1.271–5.594; p=0.010), CD8⁺TILs (HR, 0.313; 95% CI, 0.139–0.705; p=0.005), and the LAG-3-high/PD-L1-high group (HR, 2.829; 95% CI, 1.050–7.623; p=0.040) provided prognostic values for patients with TNBC following NACT. CONCLUSION: CD8+TILs were sensitive predictive markers in response to NACT. High expression of LAG-3 in residual tissues, especially in combination with PD-L1, was associated with poor prognosis.

Immune development in HIV-exposed uninfected children born to HIV-infected women

ABSTRACT Immunological and clinical findings suggestive of some immune dysfunction have been reported among HIV-exposed uninfected (HEU) children and adolescents. Whether these defects are persistent or transitory is still unknown. HEU pediatric population at birth, 12 months, 6-12 years were evaluated in comparison to healthy age-matched HIV-unexposed controls. Plasma levels of LPS, sCD14, cytokines, lymphocyte immunophenotyping and T-cell receptor excision circles (TREC) were assessed. HEU and controls had similar LPS levels, which remained low from birth to 6-12 years; for plasma sCD14, IL-2, IL-6, IL-7, IL-10, IL-12p70, IL-13, IL-17, IFN-γ, TNF-α, G-CSF, GM-CSF and MCP-1, which increased from birth to 12 months and then decreased at 6-12 years; and for TREC/106 PBMC at birth in HEU and controls. By contrast, plasma MIP-1β levels were lower in HEU than in controls (p=0.009) at 12 months, and IL-4 levels were higher in HEU than controls (p=0.04) at 6-12 years. Immune activation was higher in HEU at 12 months and at 6-12 years than controls based on frequencies of CD38+HLA-DR+CD8+T cells (p=0.05) and of CD38+HLA-DR+CD4+T cells (p=0.006). Resting memory and activated mature B cells increased from birth to 6-12 years in both groups. The development of the immune system in vertically HEU individuals is comparable to the general population in most parameters, but subtle or transient differences exist. Their role in influencing clinical incidences in HEU is unknown.

The role of interleukin - 21 in hepatitis B virus infection / 临床肝胆病杂志

Interleukin -21 (IL -21)is a cytokine produced by activated CD4 + T cells and can affect the function of various lymphocytes. It should be noted that IL -21 - mediated cellular and humoral immune responses play an important role in determining the outcome of viral infec-tion. This article introduces the research advances in the important role of IL -21 in hepatitis B virus (HBV)infection. IL -21 secreted by vi-rus - specific CD4 + T cells plays key roles in sustaining CD8 + T lymphocyte response and promoting B - lymphocyte response,which is vital for effective viral control. However,as an inflammatory mediator,IL -21 is also involved in the development of hepatitis B - related liver cir-rhosis and aggravation of liver injury. Therefore,IL -21 is considered as an immunoregulatory cytokine in HBV infection. Immunotherapeutic strategies aim at optimizing the beneficial effects of IL -21 in HBV infection,which may be a serious challenge in the future.

Effect of different stimulants on the LAG3 expression and function of spleen lymphocytes in mice / 中国生化药物杂志

Objective To investigate the effect of different stimulants on the LAG3 expression and function of spleen lymphocytes in mice. Methods The spleen lymphocytes from mice were isolated by density centrifugation.The LAG3 expressions in T cell subsets after exposure to conA, PMA, PHA or anti-CD3/28 antibodies for 24 h or 72 h were analyzed by Flow cytometry.The IFN-γsecretions of conditional medium were detected by ELISA kit.The proliferation of lymphocytes was examined by MTT analysis.Results Treatment with conA for 24 h or 72 h dose-dependently increased LAG3 +CD3 +and LAG3 +CD4 +CD3 +T cell percentages.Similarly, an exposure of anti-CD3/28 antibodies for 72 h significantly increased LAG3 +CD3 +and LAG3 +CD4 +CD3 + T cell percentages.Meanwhile, conA and anti-CD3/28 antibodies increased the IFN-γsecretion of lymphocytes in a time-dependent manner.Furthermore, Treatment with conA, PMA, PHA or anti-CD3/28 antibodies for 72 h could enhance the proliferation of lymphocyte. Conclusion conA and anti-CD3/28 antibodies are effective activators of T cells, and both of them could promote the expression of LAG3 and IFN-γsecretion of lymphocytes.

A Case of Cycloserine-Induced Lichenoid Drug Eruption Supported by the Lymphocyte Transformation Test

Lichenoid drug eruption (LDE) is a rare form of delayed-type drug eruption. Among anti-tuberculosis (Tb) agents, cycloserine (CS) has been reported as a rare cause of LDE. Positive results on the lymphocyte transformation test (LTT) have not been reported in patients with LDE. In the present case, we performed LTT and a patch test, and successfully proved CS as the offending drug in this patient, who had been treated with multiple anti-Tb drugs. These observations suggest that CS should be considered a possible cause of LDE and that LTT can be an option for the diagnosis of LDE.

Allopurinol-induced severe cutaneous adverse reactions: A report of three cases with the HLA-B*58:01 allele who underwent lymphocyte activation test

Allopurinol-induced severe cutaneous adverse reactions (SCARs) such as Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), and drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome are reportedly associated with the HLA-B*58:01 genotype. Three patients who developed SCARs after allopurinol administration were subjected to HLA-B genotyping and lymphocyte activation test (LAT) to evaluate genetic risk and to detect the causative agent, respectively. All three patients given allopurinol to treat gout were diagnosed with DRESS syndrome. Symptom onset commenced 7-24 days after drug exposure; the patients took allopurinol (100–200 mg/d) for 2-30 days. HLA-B genotyping was performed using a polymerase chain reaction (PCR)-sequence-based typing (SBT) method. All patients had a single HLA-B*58:01 allele: HLA-B*13:02/*58:01 (a 63-year-old male), HLA-B*48:01/*58:01 (a 71-year-old female), and HLA-B*44:03/*58:01 (a 22-year-old male). Only the last patient yielded a positive LAT result, confirming that allopurinol was the causative agent. These findings suggest that patients with HLA-B*58:01 may develop SCARs upon allopurinol administration. Therefore, HLA-B genotyping could be helpful in preventing serious problems attributable to allopurinol treatment, although PCR-SBT HLA-B genotyping is time consuming. A simple genotyping test is required in practice. LAT may help to identify a causative agent.

The dynamic changes of regulatory T cells in the pathogenesis of experimental autoimmune uveitis / 中华实验眼科杂志

Background Studies show that regulatory T cells (Treg) are a kind of T cell subsets to negatively regulate immune response,and play an important role in maintaining immune homeostasis and immune tolerance.Autoimmune uveitis is an autoimmune disease,the regulation of Treg cells in pathogenesis and progression of autoimmune uveitis is not fully unelucidated.Objective This study was to observe the dynamic changes of Treg in experimental autoimmune uveitis (EAU) rats and explore the role of Treg cells in the pathological process of EAUrats.Methods Eighty four 6-8 week-old SPF Lewis rats were randomly divided into model group and control group.The mixed emulsifier of interphotoreceptor retinoid-binding protein (IRBP)1177-1191,tuberculin (TB),complete Freund adjuvant (CFA) and PBS (300 μl) was subcutaneously injected in double rear foot pad,abdominal side and back,and only equal amount of TB,CFA and PBS emulsifier was used in the same way in the control group.Ocular inflammation symptoms was examined at 9,13,18,23,28,35 and 48 days after modeling and scored based on the severity of the inflammatory.Six rats of each group were sacrificed in above time points respectively for the histopathological examination of iris,ciliary body and retinas by haematoxylin-eosin staining.The lymphocytes were isolated and cultured from rat spleens,and the proportion of Foxp3-labelled cells,a specific marker of Treg cells,was assayed by flow cytometry.The relative expression level of Foxp3 mRNA in the lymphocytes detected by using realtime quantitative PCR (RT-PCR).The use and care of the rats complied with the ARVO Statement.Results Eye inflammatory response appeared at 8 days after immunization,showing vasodilation and hyperemia of rat iris in the model group,and the response peaked at 13 days,with exudation and hypopyon in the anterior chamber.The highest inflammatory scores were 3.75±0.42 at day 13,and the ocular inflammation reaction was gradually relieved after that and disappeared at 23 days after immunity.A significant difference in ocular inflammatory scores of model rats was found among different time points (F =81.709,P ＜ 0.001);while no inflammatory symptom was observed in the control group.Histopathology examination showed obvious infiltration of inflammatory cells in the iris,ciliary body and retinas in model rats,including neutrophils,lymphocytes and mononuclear cells.The proportion of Foxp3-labelled cells in spleen lymphocytes was (5.50 ± 0.64)％,(13.36 ± 0.98)％,(10.34 ± 0.79)％,(9.58 ± 1.02)％,(6.73 ±0.81)％ and (5.58 ± 0.47) ％ in the model group on day 13,18,23,28,35,48 respectively,with statistically significant differences in comparison with (2.80 ± 0.38) ％,(3.36 ± 0.53) ％,(3.65 ± 0.57) ％,(3.37 ± 0.43) ％,(3.33±0.50)％ and (3.13±0.61)％ in the control group (t=-6.272,-15.556,-11.910,-9.753,-6.154,-5.491,all at P＜0.01).The change trend of Foxp3 mRNA expression was consistent to the dynamic change of the proportion of Foxp3-labelled cells.Conclusions The pathogenesis and development is closely associated with the dynamic changes of CD4+ CD25 + Foxp3+ Treg cells in EAU rats.

Methazolamide-induced toxic epidermal necrolysis confirmed by lymphocyte activation test

Among various dermatological entities, toxic epidermal necrolysis (TEN) is a rare but potentially fatal delayed hypersensitivity reaction to numerous medications. A 38-year-old male presented with systemic hypersensitivity reaction, such as high fever, pain in the eyes, and diffuse pruritic erythematous maculopapular eruptions with multiple targetoid plaques that became vesicular and bullous. Oral mucosa and conjunctivae were involved. The first sign appeared about 1 week after taking methazolamide (50 mg twice a day) for the management of glaucomatous eyes. Although methazolamide was discontinued, blistering and skin denudation progressed to affect up to 80% of the body surface area and a positive Nikolsky sign was noted. High fever also persisted. Skin lesions started to improve after 2 weeks of management and fever subsided. Cutaneous lesions improved with minimal permanent sequele 2 months later. HLA-B*5901 was found by high-resolution genotyping. The lymphocyte activation test performed 6 months after remission showed a positive response to methazolamide challenge. This is the first case of methazolamide-induced TEN in which methazolamide was confirmed as a culprit drug by the lymphocyte activation test.

Expressions of activation antigens CD69 and HLA-DR in peripheral blood mononuclear cells and skin lesions of patients with psoriasis vulgaris / 中华皮肤科杂志

Objective To investigate changes in expressions of activation antigens CD69 and HLA-DR in CD3+T lymphocytes in peripheral blood and skin lesions in patients with psoriasis vulgaris. Methods Peripheral blood samples were obtained from 20 patients with psoriasis vulgaris and 20 healthy controls, and skin specimens from the lesions of 15 out of the 20 patients and 10 healthy controls. Flow cytometry was performed to quantify the expressions of CD69 and HLA-DR in peripheral blood CD3+T cells, and an immunohistochemical study to measure the expression of HLA-DR in skin specimens. Statistical analysis was carried out by a two-sample t-test and Pearson correlation analysis with the SPSS 19.0 software. Results Compared with the healthy controls, the patients with psoriasis vulgaris showed increased expression rates of CD69 (4.70%± 1.90%vs. 1.56%± 0.95%, t=6.629, P<0.01)and HLA-DR (8.97%± 1.79% vs. 3.02% ± 1.15%, t= 6.204, P< 0.01)in peripheral blood. Pearson correlation analysis revealed that the percentage of CD3+HLA-DR+cells in peripheral blood was positively correlated with the psoriasis area and severity index (PASI)score (r=0.5626, P<0.05). The expression rate of HLA-DR was significantly higher in the dermis (64.87%± 17.31%vs. 19.80%± 5.69%, t=7.916, P<0.01), but lower in the epidermis(11.80%± 5.55%vs. 27.40%± 8.61%, t=5.479, P<0.01)in the psoriatic specimens compared with the control specimens. Immunohistochemically, HLA-DR was widely expressed in the dermis of psoriatic lesions, but mainly distributed around blood vessels in the control skin. Conclusions There is an aberrant activation of CD3+T cells in peripheral blood and inflammatory cells in skin lesions in patients with psoriasis vulgaris, and the percentage of CD3 +HLA-DR+ cells in peripheral blood is correlated with the severity of psoriasis vulagaris.

Dexamethasone Protects Mice against Acute Inflammatory Liver Injury by Regulating T Cell Immune Response / 胃肠病学

Background:Dexamethasone can protect mice against the acute inflammatory liver injury by inhibiting innate immune cell function. However,the roles of T cell in this protective effect remain unknown. Aims:To investigate the regulatory effect of dexamethasone on T cell immune response in acute inflammatory liver injury. Methods:Six male C57BL/ 6J mice were randomly divided into 2 groups. One hour before induction of acute inflammatory liver injury by lipopolysaccharide, dexamethasone 5 mg/ kg and PBS were given intraperitoneally in experimental group and model group,respectively. All the mice were sacrificed 12 hours after model construction. The clinical score and liver function parameters were assessed;splenic mononuclear cells were isolated for measurements of T cell activation,as well as cytokine expression,secretion, and transcriptional factor expression for different T-cell subsets. Results:Clinical score and serum levels of transaminase were significantly lower in experimental group when compared with the model group. Meanwhile,percentage of CD44 +CD62L - T cells(i. e. activated or memory T cells)from spleen was significantly decreased in experimental group. Among splenic T cell population,expression and secretion of IFN-γ,a Th1-type cytokine,was decreased;expression and secretion of IL-4,a Th2-type cytokine,percentage of regulatory T cells(Treg cells),and ratios of Th2 / Th1 and Treg/ Th1 were increased;transcriptional factor specific for Th1 cells was down-regulated,and those for Th2 and Treg cells were up-regulated. Conclusions:Dexamethasone inhibits T cell activation and directs the reciprocal T cell lineage differentiation (repressing Th1 cell differentiation,promoting Th2 and Treg cell differentiation),which may contribute to the protection against acute inflammatory liver injury.

Effects of Euphorbia milii latex on mitogen-induced lymphocyte proliferation / Efeito do latex de Euphorbia milii sobre a proliferação de linfócitos induzida por mitogénio

The crude latex of "Crown-of-Thorns" (Euphorbia milii var hislopii, syn E.splendens) is a potent plant molluscicide. For this reason, toxicological studies have been performed to evaluate the health risks posed by its use in schistosomiasis control programs. The present study is part of a more comprehensive immunotoxicological evaluation of this molluscicide. Here, we investigated the effects of E. milii latex on the proliferation of human lymphocytes in vitro. Lyophilized latex of E. milii (0, 0.5, 5, 25 and 50 µg/ml) was incubated with whole blood in the presence of proliferation stimulators, i.e. lectins (phytohemagglutinin, concanavalin A and pokeweed mitogen), as well as with human monoclonal antibody against CD3 and tetanus toxoid. Cell proliferation was measured by ³H-thymidine incorporation, and the effects of latex on mitogen-induced cell proliferation were compared to the effects of 10 ng/ml of 12-O-tetradecanoylphorbol-13-acetate (TPA). Results showed that mitogen-induced cell proliferation was markedly enhanced by E. milii latex. This synergistic effect of latex on mitogen-induced lymphocyte proliferation may be due to the presence of TPA-like phorbol esters and/or to mitogenic plant lectins.

O látexbrutoda "Coroa de Cristo" (Euphorbia miliivarhislopii, syn E.splendens) é um potente moluscicidavegetal. Neste sentido, são necessários estudos toxicológicosque visemavaliar possíveis riscos à saúdeassociados ao uso em larga escala desta espécie em áreas endêmicas para esquistossomose. O presente estudo é parte deuma avaliação mais abrangentesobre o potencial tóxico destemoluscicida. Foram investigados in vitro osefeitos dolátex da E.miliisobre a proliferação delinfócitoshumanos. O látexliofilizado (0; 0,5;5;25 e 50 µg/ml)foi incubado comsangue totalna presençade agentes mitogênicos, tais como lectinas(fitohemaglutinina, concanavalina Ae pokeweed), anticorpomonoclonalhumano anti-CD3etoxóide tetânico. A proliferação celularfoi quantificada atravésincorporaçãode ³H-timidina eos efeitos do látexnaproliferação celular induzida por agentes mitogênicosforam comparados comos efeitos de10 ng/mlde12-O-tetradecanoilforbol-13-acetato (TPA). Os resultados demonstram quea proliferação celular induzida poragentes mitogênicosfoimarcadamenteaumentada na presença do látex daE.milii.Oefeito sinérgico observado pode ser devidoà presença deésteres de forbol, como o TPA, e/oude lectinas com ação mitogênica presentes nesta espécie vegetal.

Cytokine detection for the diagnosis of chromium allergy / Deteccao de citocinas no diagnostico de pacientes alergicos ao cromo

BACKGROUND: Patch testing remains the gold standard method for the identification of the etiologic agent of allergic contact dermatitis. However, it is a subjective, time-consuming exam whose technique demands special care and which presents some contraindications, which hamper its use. In a recent study, we showed that the proliferation assay can suitably replace patch testing for the diagnosis of chromium allergy, which had been previously demonstrated only for nickel allergy. In this study, we try to refine the method by reducing the incubation period of cultures for lymphocyte proliferation assays in response to chromium. OBJECTIVE: Develop an alternative or complementary diagnostic test for chromium allergic contact dermatitis. METHODS: We compared the production of 9 cytokines (IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, IL-17 and RANTES) between 18 chromium-allergic patients and 19 controls. RESULTS: Chromium increased the production of IFN-y, IL-5, IL-2 and IL-13 in allergic patients, but only IL-2 and especially IL-13 helped discriminate allergic patients from controls. The sensitivity, specificity and accuracy found with IL-13 were about 80%. CONCLUSIONS: IL-13 and IL-2 detection may be used to diagnose chromium allergy in 2-day cultures. However, in general, the 6-day cultures seem to be superior for this purpose. .

FUNDAMENTOS: O teste de contato permanece como padrão ouro para a identificação do agente causal da dermatite de contato alérgica, mas é um exame subjetivo, que demanda considerável tempo do paciente e do medico, exige cuidados na sua técnica e apresenta algumas contra-indicações que dificultam o seu uso. Em um estudo recente demonstramos que o teste de proliferação pode adequadamente substituir o teste de contato no diagnóstico de alergia ao cromo, algo previamente demonstrado apenas para o níquel. Neste estudo tentamos refinar o método reduzindo o período de incubação das culturas do teste de proliferação para o cromo. OBJETIVO: Desenvolver um método alternativo ao teste de contato para o diagnóstico dermatite de contato alérgica ao cromo. MÉTODOS: Comparamos o estímulo provocado pelo cromo na produção de nove citocinas (IFN-γ, IL2, IL-4, IL-5, IL-10, IL-12, IL-13, IL-17 e RANTES) no sobrenadante das culturas de células do sangue periférico de 18 pacientes alérgicos ao cromo e 19 controles. RESULTADOS: O cromo aumentou a produção de IFN-y, IL-5, IL-2 e IL-13, mas apenas as citocinas IL-2 e principalmente IL-13 foram capazes de discriminar pacientes de controles. A sensibilidade, especificidade e acurácia encontradas com a IL-13 foram de aproximadamente de 80%. CONCLUSÕES: Concluímos que a detecção de IL-2 e IL-13 podem ser útil para o diagnóstico de alergia a cromo na cultura de 2 dias. Todavia, as culturas de 6 dias parecem, de um modo geral, superiores as de 2 dias para esse fim. .