Protective efficacy of Chlorophytum borivilianum root extract against murine visceral leishmaniasis by immunomodulating the host responses

Background: The available drugs for treating visceral leishmaniasis are limited. Moreover, the disease isassociated with suppression of immune function. Therefore, therapies with effective immunomodulatoryagents are needed which can decrease parasitic burden and enhance adaptive immunity.Objectives: The present study was planned to evaluate the antileishmanial efficacy of crude ethanolicextract of roots of Chlorophytum borivilianum (CBREE) against murine visceral leishmaniasis throughimmunomodulation.Materials and methods: The in vitro studies were carried out to check leishmanicidal activity againstpromastigote form and cytotoxicity against HeLa cells. The parasite load in liver smears, immunologicaland biochemical changes induced by 500 and 1000 mg/kg b.wt. of CBREE were assessed on 1, 7, 14 and 21post treatment days in infected and treated BALB/c mice.Results: CBREE showed inhibitory effect on growth of promastigotes with IC50 of 28.25 mg/mL andnegligible cytotoxicity. The extract was toxicologically safe in BALB/c mice when administered orally with5 g/kg b.wt. of extract. A significant reduction in parasite load was observed along with active immunomodulation through enhanced Th1 type of immune responses and suppressed Th2 type of immuneresponses.Conclusion: The treatment with both doses showed no toxic effect as evidenced by normal liver andkidney function tests and normal histological observations of liver and kidney. Therefore, it should befurther explored for its active components in pursuit of the new effective antileishmanial agents in theplant kingdom.© 2017 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Immunomodulatory activities of Acacia catechu, a traditional thirst quencher of South India

Background: Acacia catechu has been widely used in Ayurveda for treating many diseases. Its heartwoodextract is used in asthma, cough, bronchitis, colic, diarrhea, dysentery, boils, skin afflictions, sores and forstomatitis. The decoction of heartwood is used for drinking purpose in southern part of India especiallyin Kerala.Objective: The current study was carried out to evaluate immunomodulatory effects of heartwood extracts of A. catechu in Swiss albino mice.Material and methods: In vivo immunomodulatory activity was analyzed by hemagglutinating antibody(HA) titer, plaque forming cell assay and delayed type hypersensitivity (DTH). In vitro immunomodulatory potential of the extracts was studied using peritoneal macrophages and splenocytes from mice.Effect of extracts on phagocytic activity of macrophages was analyzed by nitroblue tetrazolium (NBT)reduction assay and cellular lysosomal enzyme assay. Anti-inflammatory activity was studied by nitricoxide (NO) assay and production of TNF-a and IL-10.Results: A dose dependent increase in antibody titer was observed with extracts treatment. Treatmentwith extracts produced an enhancement in the number of antibody producing cells in the spleen. DTHreaction was significantly decreased with extracts treatment. An increased phagocytic response wasshown by peritoneal macrophages on treatment with the extracts as evidenced by its effect on NBTreduction and cellular lysosomal enzyme activity. The extracts inhibited the release of pro-inflammatorycytokine TNF-a and the production of NO. IL-10 production was significantly increased after extracttreatment.Conclusion: The results of the present study indicate the immunomodulatory effects of A. catechu extracts on humoral, cell mediated and non-specific immune functions.© 2017 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Interpretation of Mantoux test in children

Tuberculosis is the most common infectious cause of death worldwide. Young children especially infants usually are more susceptible to tuberculous infection. Disease usually develops within 1 year of infection. The present study has been conducted upon 41 children in whom Mantoux test was positive (n=41). These children were coming to our outpatient department, in a Teaching Hospital, Nizamabad, Telangana State with mild fever, cough, night sweats, anorexia, and loss of weight. The age group selected for this study was 1 to 12 year, and the period of study was from August, 2013 to December, 2015. Among the 41 children under study 13 children were 1 to 4 year and 28 were 5 to 12 year. Among the total 41 cases of Mantoux positive, 16 (39%) were found to be suffering from tuberculous disease and anti tuberculous drugs were started. In these cases 7 were 1 to 4 year (n=13), and 9 were 5 to 12 year (n=28). After 3 months follow up, all the children who were on ATT became free from their symptoms. It shows association between mantoux positivity and tuberculosis is more in children between 1 and 4 year than children between the age of 5 and 12 year. Most of the other children needed antipyretics, other symptomatic drugs and antibiotics only. But in 3 cases symptoms were not relieved either with antibiotics or with trial ATT. The factors for deciding tuberculosis are history of contact, clinical picture, CBP, ESR, chest- x- ray, Mantoux test, sputum for AFB, antibiotic trial, follow up, trial ATT. All these factors are put together and then only we will decide whether to start ATT or not.

Immunological response to re-infections with clones of the Colombian strain of Trypanosoma cruzi with different degrees of virulence: influence on pathological features during chronic infection in mice

Re-infections with Trypanosoma cruzi are an aggravating factor for Chagas disease morbidity. The Colombian strain of T. cruzi represents multiclonal populations formed by clonally propagating organisms with different tropisms and degrees of virulence. In the present study, the influence of successive inoculations with clones of the Colombian strain, exhibiting different degrees of virulence, on chronic myocarditis and the humoral and cellular immune responses (Col-C1 high virulence, Col-C8 medium virulence and Col-C5 low virulence) were demonstrated. Mice from three groups with a single infection were evaluated during the acute (14th-30th day) and chronic phases for 175 days. An immunofluorescence assay, ELISA and delayed type hypersensitivity (DTH) cutaneous test were also performed. Mice with a triple infection were studied on the 115th-175th days following first inoculation. The levels of IgM and IgG2a were higher in the animals with a triple infection. DTH showed a higher intensity in the inflammatory infiltrate based on the morphometric analysis during a 48 h period of the triple infection and at 24 h with a single infection. The histopathology of the heart demonstrated significant exacerbation of cardiac inflammatory lesions confirmed by the morphometric test. The humoral responses indicate a reaction to the triple infection, even with clones of the same strain.

Effect comparison on intradermal injection and scratch inoculation of Brucella vaccine / 中国人兽共患病学报

ABSTRACT:The guinea pigs were immunized by the Brucella vaccine through intradermal injections and the skin scratch respectively ,and then the immune effects of the two ways were evaluated .Serum samples were collected one month after the last injection and detected for the total IgG titer by interval ELISA .Cell‐mediated immune was evaluated by late‐onset hyper‐sensitivity .The guinea pigs were challenged with Brucella melitensis M5 ,and then were killed to isolated M5 from spleen of each guinea pig to compare the protective effects of two methods of immunization .The ELISA results showed that both of the two methods of immunization could induce strong humoral immune response ,and DTH response to Br‐PPD antigen were 100%in both methods .No significant difference in the immune protective effect of two methods was detected .Results of humoral im‐munity ,cellular immunity and protective effect showed the same effect by intradermal injections and skin scratches .

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.

Evaluation of Mycobacterium tuberculosis specific RD antigens for delayed type hypersensitivity responses in guinea pig.

Tuberculin skin test (TST), an age old method is based on measuring delayed-type hypersensitivity (DTH) response to purified protein derivative (PPD). However, inspite of simplicity, ease and cost effectiveness, the usefulness of PPD test is limited due to its inability to distinguish among a protective immune response, latent infection and active tuberculosis disease. On the other hand, a skin test based on RD antigens would add advantages of a high specificity of antigens with the logistics of a skin test. However, except few reports, in vivo data of intradermal use of RD antigens for skin testing is limited. Therefore, in the present study, four M. tuberculosis (Mtb) specific antigens (ESAT6, CFP10, CFP21 and MPT64) were evaluated for their diagnostic utility based on DTH response. These antigens alone and their multiple combinations induced strong DTH response in Mtb infected guinea pigs and the response was negligible in BCG vaccinated and sham immunized animals.

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 immunosuppressive effect of rapamycin on lymphocytic proliferation and DTH、GVHR / 中华器官移植杂志

Objective To manufacture rapamycin (RPM)-a new type immunosuppressant made in our country and investigate its immunosuppressive mechanism.Methods The effect of RPM on mouse splenocyte (Tc) proliferation induced by ConA,splenocyte (Bc) proliferation induced by LPS,DTH induced by DNCB and rat GVHR were studied.Results RPM obviously restrained the proliferation of Tc with IC50=1 nmol/L,significantly lower than IC50(10 nmol/L) of CsA (P＜0.05).RPM evidently repressed the proliferation of Bc with IC50=1 nmol/L significantly lower than IC50＞10 nmol/L of CsA(P＜0.05).RPM inhibited murine DTH to DNCB with ED50:1.8 mg/kg,significantly lower than ED50＞30mg/kg of CsA(P＜0.05).RPM controlled rat's GVHR with ED50=3 mg/kg,significantly lower than ED50≥30 mg/kg of CsA(P＜0.05).Conclusions RPM can inhibit lymphocytic proliferation and DTH,GVHR more markedly than CsA and they have combined efforts.

Effect of vernoniaanthelmintica wild on the immune function of mice / 中国免疫学杂志

Objective:To explore the effect of the vernoniaanthelmintica willd injection on the immune function of mice. Methods: The action of vernoniaanthelmintica willd injection on spleen T, B cell proliferation and antibody production response by [3H]-TdR incorporation and antibody forming cell assays, respectively. Results: Vernoniaanthelmintica willd injection inhibited splenocyte proliferation of normal mouse in vitro and in vivo, the antibody forming cell in vivo, and delayed type of hypersensitivity (DTH) Conclusion: Vernoniaanthelmintica willd injection could inhibit the cellular immune function and humoral immune function.

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.

Effects of Weiankang on immune function in mice / 中成药

Objective:To study the effects of WAK on immune function in mice.Methods: After different doses of WAK were given to mice ig, its effects on delayed type hypersensitivity (DTH), hemolysin production, NK and LAK cell killing activity were observed.Results: WAK could enhance DTH and hemolysin production in immunosuppressive mice and improve NK and LAK cell killing activity in mice.Conclusions: WAK could exhibit its antitumor effect by increasing immune function.

Effects of selenium-enriched yeast on DTH in immunosuppressive mice / 中国药理学通报

The effects of selenium-enriched yeast (Sey) on DTH in immunosuppressive mice have been investigated. It was found that Sey (30, 60, 90 mg?kg-l?d-1?6 d, ig) was able to antagonize lowered DTH by cyclophospha-mide (Cy); Meanwhile, the thymus and spleen weights were increased by Sey (30, or 60 mg?kg-1?d-1) in immunosuppressive mice. Sey also potentiated the lowered ConA and LPS-indu-ced proliferation of splenocytes in DTH mice at the doses of 30, 60, or 90 mg ?kg-1 and 60 or 90 mg?kg-1 respectively. These results suggested that Sey could enhance the functions of lymphocytes.

POTENTIATING EFFECTS OF ROYAL JELLY ON IMMUNO LOGICAL FUNCTION IN MICE / 中国药理学通报

Royal jelly (2.5g/kg/d igX8d) increased the clearance rate of iv charcoal particles in normal mice, and completely antagonized the decrease of clearance rate caused by cortisone acetate. It also potentiated the delayed type hypersensitivity ( DTH ) reaction of footpad induced by SRBC in normal mice, and entirely antagonized the inhibition of DTH reaction caused by eye lophosphamide. Royal jelly (2.5 g/kg/d ig X 7 d ) had no marked influence on hemolysin formation in normal mice and immunosuppressed mice caused by eye lophosphamide.

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.