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.

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.

The Changes of Cell Mediated Immunity Correlated with Severity of Head Injury

Severe head injury results in the suppression of cellular immunity associated with dysfunctioning of effector lymphocytes, such as helper T cells(CD4) (and cytotoxic T cells(CD8). Despite progress in the management of increased intracranial pressure following head injury, infection remains the most common complication and the primary cause of prolonged hospitalization and death. This study attempts to assess the cellular immune function following head injury according to the degree of severity, and to establish the clinically available parameters of cell mediated immune(CMI) function, which can then be used for coherent prediction of infection risk. Eighteem head injury patients without severe systemic injury, who divided into three subgroups depending on the severity of head injury, were estimated with the use of CMI multitest kit(Merieux Institute, France) to test delayed-type hypersensitivity(DTH) and enumerated the circulating lymphocyte subpopulation(pan T-cell marker CD3, helper T cell marker CD4, cytotoxic T cell marker CD8 and B-cell marker CD19) on the 1st, 7th, and 21th day of injury. Patients were monitored for evidence of infection for this period. Fourteen patients had no reaction to any antigens of the DTH skin test(anergy) and the remaining four patients had also some degree of anergy. Seven patients became infected and all of them were anergic. There were significant decrease of circulating effector T lymphocytes, both CD4-positive and CD8-positive cells, within 24 hours of injury in the mild as well as the moderate and severe head injury group. CD4-positive cells were nearly completely recovered by the 7th day of injury. CD8-positive cells had sustained significant decrease even after 3 weeks of injury. There was no significant change in pan T-cells(CD3-positive cells) and B-cells(CD19-positive cells). The results suggest that DTH skin test and effector T cell enumeration are both relatively simple and highly sensitive parameters for monitoring CMI function. Especially, anergy of DTH skin test can be used for indicator to predict risk of infection. Mild as well as moderate and severe head injuries may result in the suppression of cellular immunity associated with the dysfunctioning of effector T cell.

The preparation of type Ⅱ collagen induced arthritis model in mice / 中国药理学通报

AIM To establish the model of type Ⅱ collagen(CⅡ) induced arthritis in mice. METHODS The type Ⅱ collagen induced arthritits(CIA) mice were induced by intradermal injection with CⅡ and complete Freund adjnvant emulsion. RESULITS In comparison with control mice, the arthritic swelling and index were significant increased. The responses of CⅡ induced delayed type hypersensitivity were remarkably positive, and IgG anti bodies to CⅡ in serum could be detected in CIA mice. CONCLUSION The model CIA mice was successfully established.

REGULATORY EFFECTS OF CPP ON CELL-MEDIATED IMMUNITY IN MICE / 中国药理学通报

Codnopsispilosula polysaccharides (CPP) , which administered intra-peritoneally 200mg/kg?d-1 for 5 d or 8 d in mice, enhanced thephagocytic activity of peritoneal macrophages on chiken RBC, the erythrocyte (E)rosette formation of thymus T lymphocyte and the clearance rate of charcoal particles in normal mice and antagonized the inhibition of E rosette formation and of peritoneal phagocytic activity caused by cyclophosphamide ( CY ) or hydrocortisone ( HCT ) . It inhibited the delayed type hypersensitivity ( DT H ) induced by DNCB and enhanced DT H induced by dexamethasons ( DMS ) . It has regulatory important signification in resisting decrepitude.