Immunostimulant principles from Curculigo orchioides.

Curculigo orchioides Gaerten belongs to the family Amaryllidaceae. The rhizomes of the plants are used for the treatment of decline in strength, jaundice and asthma. Its methanolic extract has been shown to enhance phagocytic activity of macrophages. Present studies have led to the isolation of two phenolic glycosides and a purified glycoside fraction. These were studied for their effect on macrophage migration index (MMI), haemagglutination (HA) titre, plaque forming cell (PFC), PHA-induced blast transformation of lymphocytes (BTL) and delayed type hypersensitivity (DTH). Significant immunostimulant activity was found in purified glycoside-rich fraction isolated from the ethyl acetate extract. The exact structure of the active glycoside is yet to be determined. The enhancement of HA titre and PFC count on one hand and that of DTH response on the other indicates that glycoside fraction stimulates both humoral and Cell-mediated immune responses. Glycoside fraction stimulates immune response by acting both on macrophages and the lymphocytes.

Immunostimulant activity of dry fruits and plant materials used in indian traditional medical system for mothers after child birth and invalids.

Products of certain plants given to mothers after child birth or to invalids were studied for immunostimulant activity using the macrophage migration index (MMI) as a parameter of macrophage activation and cell-mediated immunity and haemagglutinating antibody (HA) titres and plaque-forming cell (PFC) counts as parameters of humoral immunity. Feeding of Prunus amygdalus (Almond(1)) and Buchanania lanzan (Chirronji(1)) significantly stimulated both CMI and humoral immunity in BALB/c mice as evidenced by the enhancement of MMI, HA titres, and PFC counts. Euryale ferox (Tel makhana(1)), Phoenix dactylifera (Chhohara(1)) and Zingiber officinale (Sonth(1)), however, stimulated humoral immunity to a greater extent than CMI. The observation provides scientific basis for feeding the products of above plants to mothers after child birth and to invalids with a relatively poor immune status.

Antigiardial and immunostimulatory effect of Piper longum on giardiasis due to Giardia lamblia.

Piper longum fruit, used in traditional remedies as well as in the Ayurvedic system of medicine against intestinal disorders, was tested for its efficacy against experimental infection of Giardia lamblia in mice. On in vitro test, an aqueous extract of P. longum fruit powder (PF) at 250 microg/mL and its ethanol extract at 125 microg/mL showed 100% giardicidal activity. A low order activity was found in the n-butanol extract. Further fractionation in hexane and chloroform resulted in a total loss of activity. The survival of-trophozoites in mice at 900 mg/kg body weight was 11.12 in PF, 8. 54 in aqueous extract, 5.81 in ethanol extract. The antigiardial activity of PF in hexane, chloroform and n-butanol soluble fractions was comparable to the drug-untreated control (47.63). Piper longum possessed a demonstrable immunostimulatory activity, both specific and nonspecific, as evident from the standard test parameters such as haemagglutination titre (HA), plaque forming cell (PFC) counts, macrophage migration index (MMI) and phagocytic index (PI). A maximum effect was found at 225 mg/kg body weight in mice. The effect was marginally reduced at higher doses of 450 and 900 mg/kg or the lower dose of 112.5 mg/kg.

Management of giardiasis by a herbal drug 'Pippali Rasayana': a clinical study.

Pippali Rasayana (PR), an Indian ayurvedic drug prepared from Palash (Butea monosperma (Lamk) Kuntze; Leguminaceae) and Pippali (Piper longum L.; Piperaceae), was administered at a dose of 1 g p.o. three times daily for a period of 15 days to patients (25 treated, 25 placebo controls) suffering from giardiasis with clinical signs and symptoms, and stools positive for trophozoites/cysts of Giardia lamblia. After 15 days of drug treatment there was a complete disappearance of G. lamblia (trophozoites/cysts) from the stools of 23 out of 25 patients. General signs and symptoms of ill health and abdominal discomfort, presence of mucus, pus cells and RBCs were significantly reduced. There was a marked improvement in the clinical and haematological profile of the patients. Spontaneous recovery in 20% cases was recorded in placebo controls.

Management of giardiasis by an immuno-modulatory herbal drug Pippali rasayana.

Pippali rasayana (PR), an Ayurvedic herbal medicine, prepared from Piper longum (Pippali) and Butea monosperma (Palash), and prescribed for the treatment of chronic dysentery and worm infestations was tested for anti-giardial and immuno-stimulatory activity in mice, infected with Giardia lamblia trophozoites. It produced up to 98% recovery from the infection. The rasayana had no killing effect on the parasite in vitro. It induced significant activation of macrophages as evidenced by increased macrophage migration index (MMI) and phagocytic activity. Enhancement of host resistance could be one of the possible mechanisms contributing towards the recovery of animals from the giardial infection.

Immunostimulant activity of Nyctanthes arbor-tristis L.

Nyctanthes arbor-tristis L. (Oleaceae), a plant widely used in the traditional medicinal systems of India, has recently been reported to possess hepatoprotective, antileishmanial, antiviral and antifungal activities. In the present study strong stimulation of antigen specific and non-specific immunity, as evidenced by increases in humoral and delayed type hypersensitivity (DTH) response to sheep red blood cells (SRBC) and in the macrophage migration index (MMI), has been demonstrated in mice fed with 50% ethanolic extract of seeds, flowers and leaves of this plant. Maximum activity was found in the seeds in which the active principle(s) appear to be mainly associated with lipids. In flowers and leaves, however, the major activity was found in the aqueous fraction of the 50% ethanol extract. The immunostimulant substance(s) found in N. arbor-tristis L. are likely to play a role in its antiamoebic, antileishmanial, antiviral and certain other activities.

Immunostimulant activity of a novel lipopeptide and its protective action against Leishmania donovani.

Novel lipopeptides 84/201 and 86/450 synthesized in this laboratory stimulated antibody and delayed type hypersensitivity (DTH) response to ovalbumin in guinea pigs. Lipopeptide 86/450 also stimulated antibody and DTH responses in albino mice and enhanced nonspecifically macrophage migration index (MMI), phagocytic activity and incorporation of [14C] glucosamine in peritoneal macrophages of the treated animals. Proliferative response of splenocytes from lipopeptide 86/450 treated animals was significantly higher than that from untreated controls. Peritoneal macrophages from lipopeptide 86/450 treated mice were less susceptible to Leishmania donovani promastigote invasion when co-cultured in vitro. The treated animals on challenge with L. donovani promastigote/amastigote showed 80 to 90% lower intake of infection than the control animals.

Immunostimulant agents from Andrographis paniculata.

EtOH extract and purified diterpene andrographolides of Andrographis paniculata (Acanthaceae) induced significant stimulation of antibody and delayed type hypersensitivity (DTH) response to sheep red blood cells (SRBC) in mice. The plant preparations also stimulated nonspecific immune response of the animals measured in terms of macrophage migration index (MMI) phagocytosis of 14C-leucine labelled Escherichia coli and proliferation of splenic lymphocytes. The stimulation of both antigen specific and nonspecific immune response was, however, of lower order with andrographolide than with the EtOH extract, suggesting thereby that substance(s) other than andrographolide present in the extract may also be contributing towards immunostimulation.

Immunostimulant activity of Picroliv, the iridoid glycoside fraction of Picrorhiza kurroa, and its protective action against Leishmania donovani infection in hamsters.

Picroliv, a standardised fraction from root and rhizome of Picrorhiza kurroa, consisting of iridoid glycosides and shown to be responsible for its hepatoprotective activity, was studied for immunostimulant activity. Oral administration of Picroliv (10 mg/kg x 7 days) in mice prior to immunization with sheep red blood cells (SRBC) resulted in a significant increase in haemagglutinating antibody (HA) titre, plaque forming cells (PFC), and delayed hypersensitivity (DTH) response to SRBC. Picroliv enhanced the non-specific immune response characterized by an increase in macrophage migration index (MMI), [14C]-glucosamine uptake, phagocytosis of [14C]-leucine labelled Escherichia coli, chemiluminescence of peritoneal macrophages, and higher uptake of [3H]-thymidine in the lymphocytes of treated mice. It also induced a high degree of protection in golden hamsters against challenge infection with Leishmania donovani promastigotes.

Immunostimulant Activity of Picroliv, the Iridoid Glycoside Fraction of Picrorhiza kurroa, and its Protective Action against Leishmania donovani Infection in Hamsters1.

Picroliv, a standardised fraction from root and rhizome of PICRORHIZA KURROA, consisting of iridoid glycosides and shown to be responsible for its hepatoprotective activity, was studied for immunostimulant activity. Oral administration of Picroliv (10 mg/kg x 7 days) in mice prior to immunization with sheep red blood cells (SRBC) resulted in a significant increase in haemagglutinating antibody (HA) titre, plaque forming cells (PFC), and delayed hypersensitivity (DTH) response to SRBC. Picroliv enhanced the non-specific immune response characterized by an increase in macrophage migration index (MMI), [ (14)C]-glucosamine uptake, phagocytosis of [ (14)C]-leucine labelled ESCHERICHIA COLI, chemiluminescence of peritoneal macrophages, and higher uptake of [ (3)H]-thymidine in the lymphocytes of treated mice. It also induced a high degree of protection in golden hamsters against challenge infection with LEISHMANIA DONOVANI promastigotes.

An indirect fluorescent antibody (IFA) test for the serodiagnosis of Kala-Azar.

The IFA test developed using Leishmania donovani promastigote and amastigote antigens showed very high antibody titre in clinically and parasitologically established cases (30) of kala-azar, the geometrical mean reciprocal titre (GMRT) being 870 +/- 5.4 and 5370 +/- 1.80 respectively with the two antigens. In contrast, the GMRT of normal subjects of endemic area was only 12.44 +/- 6.19 and 80.35 +/- 1.66 respectively with these antigens. None of the subjects from non-endemic area suffering with other parasitic diseases, such as malaria, filaria, amoebiasis, leprosy or tuberculosis (20 cases each) gave a positive response to any of the antigen. The test holds promise in the diagnosis of Kala-azar.

Macrophage migration as an index of immune status.

Peritoneal macrophages from mice and guinea pigs pretreated with Freund's complete adjuvant (FCA) or any other immunostimulant when packed in a glass capillary and placed in a migration chamber migrated to a larger area than macrophages from normal untreated animals. The extent of migration could be correlated with the dose of FCA and the period of treatment. Under optimum conditions the ratio between the areas of migration of macrophages from FCA-treated animals and of macrophages from untreated animals was above 3.0. A close correlation was observed between macrophage migration and delayed type hypersensitivity (DTH) response in animals sensitized with ovalbumin or sheep red blood cells. The macrophages of immunostimulant-treated animals had relatively higher phagocytic activity. The macrophage migration index (MMI) appears to be a close correlate of macrophage activation and possibly also of the status of cell mediated immune response.

Adjuvant activity of some nonpyrogenic hydrophobic analogues of muramyl dipeptide in enhancing the primary humoral and cell mediated immune responses in guinea pig model.

Five muramyl dipeptide analogues synthesized by derivatization of gamma-carboxyl of D-isoglutamine residue of MDP into alkyl amides or incorporation of lysine residue at the site via epsilon-NH2 function were evaluated for immuno-adjuvant activity. Derivatization of gamma-carboxyl of D-isoglutamine into butyl, octyl and dibutyl residues stimulated delayed type of hypersensitivity (DTH) response, the maximum stimulation being observed with octyl amide. Introduction of lauryl amide residue abolished DTH response. The antibody response was impaired with all the alkyl amide analogues except for the lysyl amide derivative with which the response was higher than MDP. Correlation was observed between DTH response and macrophage migration.

Gamma-glutamyl-transpeptidase in lymphatic tissues of Mastomys natalensis during an infection with Acanthocheilonema viteae.

During Acanthocheilonema viteae infection, the specific activity of gamma-glutamyltranspeptidase (gamma-GT) increased in peritoneal exudate cells and bone marrow and decreased in lymph nodes of Mastomys natalensis throughout the course of infection. However, though there was an increase in specific activity of gamma-GT in thymus and spleen during the prepatent phase of A. viteae infection, the level either returned to normal or decreased during the latent phase of infection. A close correlation was observed between the host's immune status during A. viteae infection and the level of gamma-GT in lymphoid tissues.

Enzyme-linked immunosorbent assay in the diagnosis of kala-azar in Bhadohi (Varanasi), India.

An enzyme-linked immunosorbent assay (ELISA) developed using Leishmania donovani promastigote soluble antigen gave positive responses in practically all the clinically and parasitologically diagnosed patients with kala-azar in a preliminary study. Healthy control subjects from non-endemic and endemic areas gave negative results. No cross-reaction was observed with patients having leprosy, filariasis, malaria, tuberculosis, or amoebiasis. Blood samples collected on filter paper were adequate for the test. The test appears promising in the diagnosis of kala-azar.

Adaptogenic activity of Indian Panax pseudoginseng.

The crude extract and saponins of Indian pseudoginseng and saponins of Korean ginseng have been studied using a battery of biological tests in rats and mice. Indian pseudoginseng saponins were found to exhibit better activity than the Korean ginseng saponins in several tests employed. The results indicate a need for in-depth study of Indian pseudoginseng as an adaptogenic agent, after cultivation of the plant under controlled conditions.

Characterization of an immunodominant Onchocerca volvulus antigen with patient sera and a monoclonal antibody.

Adult Onchocerca voluvlus and infective larvae, but not microfilariae contain an immunodominant antigen (33,000 and 21,000 Mr in females, 39,000, 33,000, and 21,000 Mr in males, 133,000 Mr in infective larvae) which is recognized by an Onchocerca-specific mAb. The component is part of the reproductive organs and muscles. 96.2% of onchocerciasis sera contained antibodies detectable by immunoblotting against it. Antigen purified by immunoaffinity chromatography was specifically recognized in immunoblots by onchocerciasis sera, but not by sera from other filarial infections. The high immunogenicity, the specificity, and the occurrence in infective larvae of this antigen indicate an immunodiagnostic potential and a possible role in the immunobiology of the parasite.