Proliferation Of Peripheral Blood Lymphocytes Research Articles

Nasal Polyp Cell Populations and Fungal-Specific Peripheral Blood Lymphocyte Proliferation in Allergic Fungal Sinusitis

Background Allergic fungal sinusitis (AFS) is considered a different disease from other polypoid chronic rhinosinusitis diseases (CRS) with eosinophilic mucus (EM) termed eosinophilic mucus chronic rhinosinusitis (EMCRS). To substantiate this, studies on cellular responses to fungi and sinus mucosal inflammatory cell populations in AFS and other EMCRS diseases are. required. This study was designed to examine polyp inflammatory cell populations and peripheral blood fungal-specific T-cell responses in AFS, other EMCRS subgroups (defined later), and polypoid CRS without EM. Methods A prospective study was performed. Clinical characteristics, including CRS symptoms, sinus computed tomography (CT) scans, allergy status, intraoperative endoscopy, presence of EM, and fungal culture results were used to define patient groups. Polyps and peripheral blood were examined for populations of eosinophils, lymphocytes (CD4+, CD8+ T cells, natural killer cells, and B cells), and neutrophils using immunohistochemistry, cytospin preparations and flow cytometry. Fungal-specific peripheral blood lymphocyte proliferation was examined in AFS patients, other EMCRS patients, CRS patients, and controls. Results There was no significant difference in the percentage of cell populations and fungal-specific lymphocyte proliferation between AFS and other EMCRS diseases. However, AFS and other EMCRS polyps had a higher percentage of eosinophils and CD8+ T cells whereas CRS polyps had higher CD44 T cells. Fungal-specific lymphocyte proliferation was significantly greater in AFS and other EMCRS patients regardless of fungal allergy, whereas in CRS and controls, higher proliferation was observed in fungal-allergic individuals. Conclusion These findings question the basis for differentiating AFS from, other EMCRS diseases based on fungal allergy and fungi in EM. Fungal-specific cellular response was present in AFS and other EMCRS diseases, different from, that associated with fungal allergy, suggesting a nonallergic fungal immune response. Increased CD8+ T cells in EMCRS polyps signify a different type of inflammation to CRS that may be driven by CD8+ T cells.

The myth of the subclinical rejection: Is it real

Introduction: Subclinical rejection (SCAR) of renalallografts refers to graft lymphocytic infiltrationtaking acute rejection histologic pattern despitestable renal function. There are no data to suggestthat subclinical tubulointerstitial inflammation isregulatory or in any way beneficial to the graft. Wehave investigated whether C57BL/6 CD8 T cellshome to long term engrafted (LTE) DBA/2 skinallografts and if it is protecting or rejecting. Methods and results: We transplanted two groupsof B6 CD4 KO mice, 6 mice each, with MHCmismatched DBA/2 skin. Only the 1st group wastreated with Rapamycin (RPM) as reported. After100 days of LTE, we challenged RPM treated hostswith a 2nd DBA/2 skin graft. The 2nd but not the 1stgraft was rejected. Then we investigated thefunctional effects of graft inflitrating CD8 T cells.DBA/2 skin grafts were harvested 100 daysposttransplantation from (i) RPM treated B6 CD4ko mice (N=5) and (ii) skin autografts (N=5) inDBA/2 recipients. LTE DBA/2 allografts or controlDBA/2 autografts were then transplanted ontoC57BL/6-Rag KO hosts, and peripheral bloodlymphocytes (PBL) samples were collected 30 dayspost skin transplantation. CD8T cells can not bedetected in PBL of nae RAG-/- mice. 4.6 % CD8T cells are detected in PBL of RAG-/- recipients ofLTE allografts, but not in recipients of syngeneicgrafts. To test the protective function of the grafthoming CD8 T cells (from LTE RPM mice) thatexpanded by homeostatic proliferation and arepresent in PBL of the RAG-/-, 0.2 x 106 CD8 Tcells from naive CD4KO mice were adoptivelytransferred into the RAG-/- hosts bearing the LTEDBA/2 allografts or DBA/2 autografts 30 daysfollowing skin transplantation. Survival of LTEDBA/2 skin allografts transplanted onto RAG-/-mice were significantly prolonged. Conclusion: Graft infiltrating CD8 T cells areregulatory and functionally active to protectallograft from rejection.

Mesenchymal Stem Cells Derived from Human Gingiva Are Capable of Immunomodulatory Functions and Ameliorate Inflammation-Related Tissue Destruction in Experimental Colitis

Aside from the well-established self-renewal and multipotent differentiation properties, mesenchymal stem cells exhibit both immunomodulatory and anti-inflammatory roles in several experimental autoimmune and inflammatory diseases. In this study, we isolated a new population of stem cells from human gingiva, a tissue source easily accessible from the oral cavity, namely, gingiva-derived mesenchymal stem cells (GMSCs), which exhibited clonogenicity, self-renewal, and multipotent differentiation capacities. Most importantly, GMSCs were capable of immunomodulatory functions, specifically suppressed peripheral blood lymphocyte proliferation, induced expression of a wide panel of immunosuppressive factors including IL-10, IDO, inducible NO synthase (iNOS), and cyclooxygenase 2 (COX-2) in response to the inflammatory cytokine, IFN-gamma. Cell-based therapy using systemic infusion of GMSCs in experimental colitis significantly ameliorated both clinical and histopathological severity of the colonic inflammation, restored the injured gastrointestinal mucosal tissues, reversed diarrhea and weight loss, and suppressed the overall disease activity in mice. The therapeutic effect of GMSCs was mediated, in part, by the suppression of inflammatory infiltrates and inflammatory cytokines/mediators and the increased infiltration of regulatory T cells and the expression of anti-inflammatory cytokine IL-10 at the colonic sites. Taken together, GMSCs can function as an immunomodulatory and anti-inflammatory component of the immune system in vivo and is a promising cell source for cell-based treatment in experimental inflammatory diseases.

Autologous mesenchymal stem cell treatment increased T regulatory cells with no effect on disease activity in two systemic lupus erythematosus patients

Mesenchymal stem cells (MSCs) exert suppressive effects in several disease models including lupus prone mice. However, autologous MSC therapy has not been tested in human systemic lupus erythematosus (SLE). We evaluate the safety and efficacy of bone marrow (BM)-derived MSCs in two SLE patients; the suppressor effect of these cells in-vitro and the change in CD4+CD25+FoxP3+ T regulatory (Treg) cells in response to treatment. Two females (JQ and SA) of 19 and 25 years of age, fulfilling the 1997 American College of Rheumatology (ACR) criteria for SLE were infused with autologous BM-derived MSCs. Disease activity indexes and immunological parameters were assessed at baseline, 1, 2, 7 and 14 weeks. Peripheral blood lymphocyte (PBL) subsets and Treg cells were quantitated by flow cytometry, and MSCs tested for in-vitro suppression of activation and proliferation of normal PBLs. No adverse effects or change in disease activity indexes were noted during 14 weeks of follow-up, although circulating Treg cells increased markedly. Patient MSCs effectively suppressed in-vitro PBL function. However, JQ developed overt renal disease 4 months after infusion. MSC infusion was without adverse effects, but did not modify initial disease activity in spite of increasing CD4+CD25+FoxP3+ cell counts. One patient subsequently had a renal flare. We speculate that the suppressive effects of MSC-induced Treg cells might be dependent on a more inflammatory milieu, becoming clinically evident in patients with higher degrees of disease activity.

In Vitro Genotoxicity of Wastewaters from the Town of Settat, Morocco

In recent years, the town of Settat has seen a considerable industrial growth, which has resulted in increased environmental pollution. This includes pollution by household and industrial wastewaters, which are released into the Boumoussa River without any preliminary treatment. The river valley crosses the community of Mzamza 8 km to the north of the town. Years of drought forced members of the community to use this polluted ground water for irrigation and put themselves and the environment at risk.The aim of this study was to determine the physicochemical and metal profile of Settat wastewaters and to assess their impact on the water table. The second objective was to investigate the genotoxic potential of wastewater on human peripheral blood lymphocytes in vitro, using the micronucleus test and cellular proliferation index.This study demonstrated significant pollution of Boumoussa valley groundwater and of the local wells. Sampled water induced a clear increase in the frequency of micronucleated cells and a lower cell proliferation in human peripheral blood lymphocytes in vitro.

Nasal polyp cell populations and fungal-specific peripheral blood lymphocyte proliferation in allergic fungal sinusitis.

Allergic fungal sinusitis (AFS) is considered a different disease from other polypoid chronic rhinosinusitis diseases (CRS) with eosinophilic mucus (EM) termed eosinophilic mucus chronic rhinosinusitis (EMCRS). To substantiate this, studies on cellular responses to fungi and sinus mucosal inflammatory cell populations in AFS and other EMCRS diseases are required. This study was designed to examine polyp inflammatory cell populations and peripheral blood fungal-specific T-cell responses in AFS, other EMCRS subgroups (defined later), and polypoid CRS without EM. A prospective study was performed. Clinical characteristics, including CRS symptoms, sinus computed tomography (CT) scans, allergy status, intraoperative endoscopy, presence of EM, and fungal culture results were used to define patient groups. Polyps and peripheral blood were examined for populations of eosinophils, lymphocytes (CD4+, CD8+ T cells, natural killer cells, and B cells), and neutrophils using immunohistochemistry, cytospin preparations and flow cytometry. Fungal-specific peripheral blood lymphocyte proliferation was examined in AFS patients, other EMCRS patients, CRS patients, and controls. There was no significant difference in the percentage of cell populations and fungal-specific lymphocyte proliferation between AFS and other EMCRS diseases. However, AFS and other EMCRS polyps had a higher percentage of eosinophils and CD8+ T cells whereas CRS polyps had higher CD4+ T cells. Fungal-specific lymphocyte proliferation was significantly greater in AFS and other EMCRS patients regardless of fungal allergy, whereas in CRS and controls, higher proliferation was observed in fungal-allergic individuals. These findings question the basis for differentiating AFS from other EMCRS diseases based on fungal allergy and fungi in EM. Fungal-specific cellular response was present in AFS and other EMCRS diseases, different from that associated with fungal allergy, suggesting a nonallergic fungal immune response. Increased CD8+ T cells in EMCRS polyps signify a different type of inflammation to CRS that may be driven by CD8+ T cells.

Effects of Dietary Zinc Level and an Inflammatory Challenge on Performance and Immune Response of Weanling Pigs

Two experiments were conducted to determine the effect of dietary zinc level on growth performance and immune function in normal (Experiment 1) and immunologically challenged (Experiment 2) weanling pigs. Treatments consisted of the following: i) a corn-soybean meal basal diet containing 36.75 mg/kg total Zn, ii) basal diet+60 mg/kg added Zn as ZnSO 4 , iii) basal diet+120 mg/kg added Zn as ZnSO 4 . Each diet was fed to six pens of four pigs per pen (Exp. 1) or six pens of three pigs per pen (Exp. 2). In Exp. 1, the dietary zinc level had no effect on average daily growth (ADG), average daily feed intake (ADFI), or feed conversion ratio (FCR). Concentrations of tissue and serum zinc were not affected. Peripheral blood lymphocyte proliferation (PBLP) was not affected by dietary treatments. Supplementation of 120 mg/kg Zn decreased (p<0.05) the antibody response to bovine serum albumin (BSA) on d 7 compared with pigs fed the basal diet, but not on d 14. In Exp. 2, LPS challenge had no effect on ADG ADFI and FCR in the entire trial (from d 0 to 21). LPS challenge significantly decreased ADG and ADFI (p<0.01) from d 7 to 14, but FCR was not affected. LPS challenge increased PBLP (p<0.05) and serum concentration of interleukin-1 (IL-1 ) (p<0.01), whereas the antibody response to BSA and serum concentration of interleukin-2 (IL-2) were not affected. Supplementation of Zn did not affect ADFI and FCR from d 7 to 14, but there was a trend for ADG to be enhanced with Zn supplementation (p<0.10). Supplementation of Zn tended to increase PBLP (p<0.10). Dietary treatment had no effect on the antibody response to BSA or concentrations of serum IL-1 and IL-2. Results indicate that the level of Zn recommended by NRC (1998) for weanling pigs was sufficient for optimal growth performance and immune responses. Zn requirements may be higher for pigs experiencing an acute phase response than for healthy pigs.

The Ruthenium Complex cis-(Dichloro)Tetraammineruthenium(III) Chloride Presents Immune Stimulatory Activity on Human Peripheral Blood Mononuclear Cells

Ruthenium compounds in general are well suited for medicinal applications. They have been investigated as immunosuppressants, nitric oxide scavengers, antimicrobial agents, and antimalarials. The aim of this study is to evaluate the immunomodulatory activity of cis-(dichloro)tetraammineruthenium(III) chloride (cis-[RuCl(2)(NH(3))(4)]Cl) on human peripheral blood mononuclear cells (PBMC). The cytotoxic studies performed here revealed that the ruthenium(III) complex presents a cytotoxic activity towards normal human PBMC, only at very high concentration. Results also showed that cis-[RuCl(2)(NH(3))(4)]Cl presents a dual role on PBMC stimulating proliferation and interleukin-2 (IL-2) production at low concentration and inducing cytotoxicity, inability to proliferate, and inhibiting IL-2 production at high concentration. The noncytotoxic activity of cis-[RuCl(2)(NH(3))(4)]Cl at low concentration towards PBMC, which correlates with the small number of annexin V positive cells and also the absence of DNA fragmentation, suggest that this compound does not induce apoptosis on PBMC. For the first time, we show that, at low concentration (10-100 microg L(-1)), the cis-[RuCl(2)(NH(3))(4)]Cl compound induces peripheral blood lymphocytes proliferation and also stimulates them to IL-2 production. These results open a new potential applicability of ruthenium(III) complexes as a possible immune regulatory compound acting as immune suppressor at high concentration and as immune stimulator at low concentration.

Pharmacology of a Novel, Orally Active PDE4 Inhibitor

Background: Intracellular cyclic adenosine monophosphate (cAMP) in inflammatory cells and airway smooth muscle is critical to the modulation of inflammatory response generation. Phosphodiesterase 4 (PDE4), an enzyme that catalyzes cAMP degradation, is therefore being actively explored as a molecular target for the treatment of airway inflammation, particularly asthma and chronic obstructive pulmonary disease. The field has undergone major advances in optimizing generation of compounds with a safe therapeutic margin; however, most PDE4 inhibitors tested so far have unacceptable side effects, particularly nausea and vomiting. Methods: We evaluated NIS-62949 in a wide range of in vitro and ex vivo cell-based assays to ascertain its anti-inflammatory potential. The compound was evaluated in murine models of lipopolysaccharide-induced endotoxemia and pulmonary neutrophilia. Parameters of airway inflammation, airway hyperreactivity and bronchoconstriction were evaluated in a guinea pig model of antigen-induced allergy. In order to assess the emetic potential, the compound was evaluated biochemically for binding to high-affinity rolipram-binding site. Subsequently, the compound was tested in a surrogate model for emesis, and the results obtained were correlated directly to tests conducted in a Beagle dog model. Results: NIS-62949 is a potent, highly selective PDE4 inhibitor. The compound demonstrated potent ability to inhibit tumor necrosis factor-α release from human peripheral blood mononuclear cells, lymphocyte proliferation and cytokine production. The in vitro profile of NIS-62949 prompted further evaluation of the compound in vivo and the compound was found to be comparable to roflumilast in several experimental models of pulmonary inflammation. Importantly, NIS-62949 displayed a safer profile compared to roflumilast. Conclusions: Our results report the development of a promising, novel PDE4 inhibitor, NIS-62949, with a wider therapeutic window as compared to second-generation PDE4 inhibitors such as roflumilast.

Enhancement of Peripheral Blood CD8+T Cells and Classical Swine Fever Antibodies by Dietaryβ-1,3/1,6-glucan Supplementation in Weaned Piglets

The experiment was conducted to evaluate the effect of dietary supplementation with 0, 50, 100 and 200 mg/kg of beta-1,3/1,6-glucan extracted from Saccharomyces cerevisiae on cellular immune responses and classical swine fever virus (CSFV) antibody production of weaned piglets. Thirty-two (Landrace x Large White; 26 days old; 7.05+/-0.25 kg body weight) castrated male piglets were randomly allocated to four treatments, eight pigs per treatment. All piglets were vaccinated with live-attenuated Chinese strain cell vaccines at 26 days of age. The results showed that dietary supplementation with beta-1,3/1,6-glucan could significantly (P<0.01) enhance peripheral blood lymphocyte proliferation activity in weaned piglets after 14 days administration, whereas no significant effect (P>0.05) was observed on day 28. Dietary beta-1,3/1,6-glucan supplementation could significantly enhance CSFV antibody blocking rate (P<0.05) on days 14 and 21 after inoculation. Although dietary beta-1,3/1,6-glucan supplementation did not significantly influence (P>0.05) lymphocyte subgroups on day 14, high levels of beta-1,3/1,6-glucan (100 and 200 mg/kg) increased the percentage of cytotoxic/suppressor lymphocytes (CD8(+)) significantly (P<0.05), decreased the ratio of CD4(+)/CD8(+) significantly (P<0.05) and increased the percentage of total T lymphocytes (CD3(+)) on day 28. These results suggest that beta-1,3/1,6-glucan may have a beneficial effect on the immune system of swine, and dietary beta-1,3/1,6-glucan supplementation had a transient impact on lymphocyte proliferation activity. The CSFV antibody blocking rate and the percentage of peripheral blood CD8(+) T cells could be enhanced by continuous beta-1,3/1,6-glucan supplementation. These findings support the potential application of beta-1,3/1,6-glucan as a prophylactic agent in increasing animal immune functions and controlling CSF disease.

Effects of soybean glycinin on performance and immune function in early weaned pigs

Glycinin, a major storage protein in soybean, has been identified as an important food allergen. The present study was conducted to investigate the effects of soybean glycinin on the local and systemic immune responses using a swine model specific for glycinin allergy. Performance, peripheral blood lymphocyte proliferation and CD4+ and CD8+ subsets in the plasma of naive and sensitised piglets were determined. In addition, the concentrations of IgA, IgG, IgM, interleukin-4 and interleukin-6 in the jejunum mucosa were measured. Our results showed that dietary supplementation of glycinin reduced piglet performance (p < 0.05), while increasing lymphocyte proliferation and CD4+/CD8+ ratio (p < 0.01). Intestinal mucosal immune responses to glycinin were enhanced with high levels of IgA (p < 0.01) as well as interleukin-4 and interleukin-6 in the jejunum mucosa in glycinin-treated piglets compared with the control (p < 0.01). There were no differences due to treatment in the densities of IgG and IgM. In conclusion, it was found that glycinin stimulated local and systemic immune responses in allergic piglets and had negative effects on piglet performance. The severity of the immune reactions depends on the dose of glycinin with higher doses causing more severe symptoms.

Effects of Achyranthes Bidentata Polysaccharide on Growth Performance, Immunological, Adrenal, and Somatotropic Responses of Weaned Pigs Challenged with Escherichia coli Lipopolysaccharide

A study was conducted with 48 weaned barrows (28±3 d, 8.45±0.14 kg) to determine the effect of Achyranthes bidentata polysaccharide (ABPS) supplementation on pig performance, immunological, adrenal and somatotropic responses following Escherichia coli lipopolysaccharide (LPS) challenge. The experiment was a 2x2 factorial design; the main factors included diet (supplementation with 0 or 500 mg/kg ABPS) and immunological challenge (LPS or saline). On d 14 and 21 of the trial, pigs were given an intraperitoneal injection with either 100 μg/kg BW of LPS or an equivalent amount of sterile saline. Blood samples were obtained 3 h after injection for analysis of tumor necrosis factor-a (TNF-a), prostaglandin E 2 (PGE 2 ), cortisol, growth hormone (GH), insulin-like growth factor (IGF)-I and immunoglobulin G (IgG). On d 2 after LPS challenge, peripheral blood lymphocyte proliferation (PBLP) was measured. LPS administration decreased average daily feed intake (ADFI) (p<0.05), had a tendency to decrease average daily gain (ADG) (p<0.10) during both the first and second challenge periods and increased (p<0.05) feed:gain ratio only during the first challenge period. ABPS tended to improve ADG (p<0.10) during the first challenge period, and improved ADG (p<0.05) and tended to improve ADFI (p<0.10) during the second challenge period. ABPS did not affect feed:gain ratio. An interaction (p<0.05) between LPS challenge and diet was observed for the plasma concentrations of TNF-a, PGE 2 and cortisol after both LPS challenges such that, among LPS-treated pigs, pigs fed the ABPS diet were lower for these indices than those receiving the control diet. In contrast, pigs fed the ABPS diet had higher IGF-I (p<0.05) compared with those fed the control diet. No effect of diet, LPS challenge or both on GH and IgG was observed after both LPS administrations. LPS challenge increased PBLP when these cells were incubated with 8 μg/ml of LPS during both the challenge periods, and did likewise when incubated with 8 μg/ml of concanavalin A only after the first challenge. ABPS had no effect on PBLP. These data demonstrate that ABPS alters the release of pro-inflammatory cytokines following an immunological challenge, which might enable pigs to achieve better performance.

Effects of Liquid dl-2-Hydroxy-4-Methylthio Butanoic Acid on Growth Performance and Immune Responses in Broiler Chickens

An experiment was conducted to determine the effects of different doses of liquid dl-2-hydroxy-4-methylthio butanoic acid (LMA) on growth performance and immune response in broiler chickens. In an arrangement with 4 graded levels of LMA to meet 80, 100, 120, and 140% of methionine requirements of broilers recommended by Chinese feeding standards for chickens, 256 one-day-old Arbor Acres male broiler chickens were randomly divided into 4 treatments with 8 replicates of 8 birds each. Growth performance, cellular immunity, and humoral immunity were determined. Results from increasing LMA levels were as follows. There were no significant differences (P > 0.05) in body weight gain and feed intake among the treatments, but the ratio of feed to gain was linearly decreased and significantly greatest (P < 0.05) in the group fed at 80% of methionine requirement. Serum globulin levels on d 21 and 42 were linearly increased significantly (P < 0.05); phagocytosis of neutral red of peripheral blood lymphocyte was quadratic and was lowest in the deficient group (P < 0.05). The proliferation of peripheral blood lymphocytes in response to lipopolysaccharide was quadratically influenced, and that of the 120% group on d 21 and the 100% group on d 42 was significantly greater than in the other groups (P < 0.05). Antibody titers to Newcastle disease virus on d 4 after the first inoculation of the vaccine were quadratically increased, anti-bovine serum albumin antibody production on d 13 after the second immunization was quadratic, and antibody titers were greatest in the groups fed at 100 or 120% of methionine requirement. In conclusion, methionine deficiency resulted in decreased feed utilization and decreased humoral and nonspecific immuno-competence of broiler chickens. The use of LMA to correct a methionine deficiency corrected these problems.

Induction of suppressor cells byMycobacterium paratuberculosisantigen in inflammatory bowel disease

We studied the M. paratuberculosis-induced proliferation and suppressor cell generation by peripheral blood lymphocytes from patients with inflammatory bowel disease. Peripheral blood lymphocytes were separated from 33 patients with Crohn's disease, 18 with ulcerative colitis, nine with other intestinal diseases, and five with autoimmune disorders. Proliferation of peripheral blood lymphocytes from normal individuals in response to 10 micrograms/ml of M. paratuberculosis antigen was reduced by depletion of CD4+ T cells. The ability of M. paratuberculosis antigen to suppress concanavalin A-induced proliferation (expressed as a percentage suppression) was reduced by depletion of CD8+ T cells. This suppression was the same whether peripheral blood lymphocytes were from normal individuals, patients with intestinal diseases other than inflammatory bowel disease, or patients with autoimmune disorders (47 +/- 14%, 44 +/- 24%, and 30 +/- 26%, respectively). In contrast, the suppression induced by M. paratuberculosis for patients with Crohn's disease and ulcerative colitis (66 +/- 22% and 67 +/- 22%) was much greater than that for normal individuals (P less than 0.001). In particular, lymphocytes from patients with active Crohn's disease demonstrated little proliferation in response to this antigen but marked suppressor activity (79 +/- 13%). How the immunomodulatory effects of this antigen relate to the pathogenesis of the inflammatory bowel diseases remains to be determined.

Cellular immunity in tuberculous pleural effusions: evidence of spontaneous lymphocyte proliferation and antigen-specific accelerated responses to purified protein derivative (PPD).

The kinetics of in vitro cellular proliferation against a PPD of Mycobacterium tuberculosis or streptococcal antigen (streptokinase-streptodornase) was evaluated in pleural fluid and peripheral blood mononuclear cells (PBMC) from patients with tuberculous and non-tuberculous pleuritis. The peak proliferative response to PPD by mononuclear cells from pleural fluid occurred earlier (day 3) in 65% of patients with tuberculosis, a finding not seen in non-tuberculous effusions. Spontaneous lymphocyte proliferation of both peripheral blood lymphocytes and pleural effusion lymphocytes was frequently observed, irrespective of etiology. However, 20 of 21 tuberculous patients manifesting spontaneous lymphocyte proliferation had accelerated kinetics of proliferation to PPD, which was antigen-specific. These results suggest that spontaneous lymphocyte proliferation occurs as a response to antigen stimulation at the site of disease, and is not a non-specific response to inflammation. Furthermore, enhanced reactivity against mycobacterial antigen, manifested by accelerated kinetics of proliferation, has diagnostic potential in patients with pleural effusions.

Parasite antigens recognized by patients with cutaneous leishmaniasis

Humoral and cell-mediated responses to crude and purified parasite antigens were examined in patients with active cutaneous leishmaniasis caused by Leishmania major. The patients had serum antibody titres against parasite lysates ranging from 1/500 to 1/10,000 and recognized multiple components by Western blotting with molecular weights between 5000 and greater than 200,000. Several components, particularly at 5 and 50 kD, were recognized by most of the patients. The lymphoproliferative responses to two pure antigens, promastigote surface protease and lipophosphoglycan, both considered potential candidates for the development of a human vaccine were measured. No response to promastigote surface protease was observed even at 10 micrograms/ml; however, weak proliferation to lipophosphoglycan was consistently present. T cell blots examining peripheral blood lymphocyte proliferation, showed antigen-specific responses to a 72-82 kD component in promastigote lysates.

Evaluation of Immunomodulatory Activity of Methanolic Extract of Piper betel

Many of the disorders today are based on the imbalances of immunological processes. This necessitates the search for newer and safer immunomodulators. Thus, the objective of the present study was to explore the immunomodulatory activity of the methanolic extract of Piper betel L. (MPb) (Family: Piperaceae). The MPb consists of mixture of phenols, flavonoids, tannins and polysaccharides. Both in vitro as well as in vivo evaluation was carried out. The effects of MPb on lymphocyte proliferation, interferon-gamma receptors and the production of nitric oxide were measured in vitro. Further, the extract at different dose levels was studied in vivo for the humoral and cellular immune responses on mice immunized with sheep red blood cells. P. betel significantly suppressed phytohaemagglutinin stimulated peripheral blood lymphocyte proliferation in a dose-dependent manner. The decrease in antibody titre and increased suppression of inflammation suggests possible immunosuppressive effect of extract on cellular and humoral response in mice. Thus, the MPb could be explored extensively as a therapeutic agent to treat various immune disorders including autoimmune disorders.

Effect of Dietary 棺-1,3/1,6-glucan Supplementation on Growth Performance, Immune Response and Plasma Prostaglandin E2, Growth Hormone and Ghrelin in Weanling Piglets

The experiment was conducted to evaluate the effect of β-1,3/1,6-glucan on growth performance, immunity and endocrine responses of weanling piglets. One hundred and eighty weanling piglets (LandracexLarge White, 7.20±0.25 kg BW and 28±2 d of age) were randomly fed 1 of 5 treatment diets containing dietary β-1,3/1,6-glucan supplemented at 0, 25, 50, 100 and 200 mg/kg for 4 wks. Each treatment was replicated in 6 pens containing 6 pigs per pen. On d 14 and 28, body weight gain, feed consumption and feed efficiency were recorded as measures of growth performance. Peripheral blood lymphocyte proliferation and serum immunoglobulin G (IgG) were measured to study the effect of dietary β-1,3/1,6-glucan supplementation on immune function. Plasma prostaglandin E 2 (PGE 2 ), growth hormone (GH) and ghrelin were measured to investigate endocrine response to β-1,3/1,6-glucan supplementation. Our results suggest that average daily gain (ADG) and feed efficiency had a quadratic increase trend with dietary β-1,3/1,6-glucan supplementation from d 14 to 28, whereas it had no significant effect on average daily feed intake (ADFI). The treatment group fed with 50 mg/kg dietary β-1,3/1,6-glucan supplementation showed a numerical increase in ghrelin, a similar change trend with ADG and no significant effect on GH. Lymphocyte proliferation indices, serum IgG and plasma PGE 2 concentrations varied linearly with dietary supplementation levels of β-1,3/1,6-glucan on d 14. Higher levels of β-1,3/1,6-glucan may have a transient immuno-enhancing effect on the cellular and humoral immune function of weanling piglets via decreased PGE 2 . Taking into account both immune response and growth performance, the most suitable dietary supplementation level of β-1,3/1,6-glucan is 50 mg/kg for weanling piglets.

The nucleocapsid protein of severe acute respiratory syndrome coronavirus inhibits cell cytokinesis and proliferation by interacting with translation elongation factor 1alpha.

Severe acute respiratory syndrome coronavirus (SARS-CoV) is the etiological agent of SARS, an emerging disease characterized by atypical pneumonia. Using a yeast two-hybrid screen with the nucleocapsid (N) protein of SARS-CoV as a bait, the C terminus (amino acids 251 to 422) of the N protein was found to interact with human elongation factor 1-alpha (EF1alpha), an essential component of the translational machinery with an important role in cytokinesis, promoting the bundling of filamentous actin (F-actin). In vitro and in vivo interaction was then confirmed by immuno-coprecipitation, far-Western blotting, and surface plasmon resonance. It was demonstrated that the N protein of SARS-CoV induces aggregation of EF1alpha, inhibiting protein translation and cytokinesis by blocking F-actin bundling. Proliferation of human peripheral blood lymphocytes and other human cell lines was significantly inhibited by the infection of recombinant retrovirus expressing SARS-CoV N protein.

The immunomodulator PSK induces in vitro cytotoxic activity in tumour cell lines via arrest of cell cycle and induction of apoptosis

BackgroundProtein-bound polysaccharide (PSK) is derived from the CM-101 strain of the fungus Coriolus versicolor and has shown anticancer activity in vitro and in in vivo experimental models and human cancers. Several randomized clinical trials have demonstrated that PSK has great potential in adjuvant cancer therapy, with positive results in the adjuvant treatment of gastric, esophageal, colorectal, breast and lung cancers. These studies have suggested the efficacy of PSK as an immunomodulator of biological responses. The precise molecular mechanisms responsible for its biological activity have yet to be fully elucidated.MethodsThe in vitro cytotoxic anti-tumour activity of PSK has been evaluated in various tumour cell lines derived from leukaemias, melanomas, fibrosarcomas and cervix, lung, pancreas and gastric cancers. Tumour cell proliferation in vitro was measured by BrdU incorporation and viable cell count. Effect of PSK on human peripheral blood lymphocyte (PBL) proliferation in vitro was also analyzed. Studies of cell cycle and apoptosis were performed in PSK-treated cells.ResultsPSK showed in vitro inhibition of tumour cell proliferation as measured by BrdU incorporation and viable cell count. The inhibition ranged from 22 to 84%. Inhibition mechanisms were identified as cell cycle arrest, with cell accumulation in G0/G1 phase and increase in apoptosis and caspase-3 expression. These results indicate that PSK has a direct cytotoxic activity in vitro, inhibiting tumour cell proliferation. In contrast, PSK shows a synergistic effect with IL-2 that increases PBL proliferation.ConclusionThese results indicate that PSK has cytotoxic activity in vitro on tumour cell lines. This new cytotoxic activity of PSK on tumour cells is independent of its previously described immunomodulatory activity on NK cells.