Granuloma Volume Research Articles

Immunization with SmIg, a novel tegument protein fromSchistosoma mansoni, fails to induce protection in mice but reduces liver pathology

Proteins associated with the schistosome tegument are of great importance for the development of new intervention strategies since they may be exposed on the surface of the parasite. Herein, we have isolated a cDNA clone encoding for the Schistosoma mansoni SmIg and its recombinant protein was tested as a potential vaccine candidate. Initially, its amino acid sequence was analysed by bioinformatics and shown to possess an N-terminal signal peptide, a C-terminal transmembrane helix, 4 glycosylation sites, an immunoglobulin conserved domain and 73% similarity with a hypothetical S. japonicum protein of unknown function. SmIg was produced by E. coli as a recombinant protein (rSmIg) and its protective effectiveness was evaluated against S. mansoni infection with 100 cercariae in a murine model. Mice immunized with rSmIg induced an immune response characterized by dominant IgG1 isotype and significant levels of IFN-gamma, TNF-alpha, IL-10 and IL-4. Although immunogenic, the recombinant vaccine failed to induce worm burden reduction when compared to the infected control group. However, rSmIg-immunized mice had significant reductions of liver granuloma volume and fibrosis content by 31.8% and 49%, respectively. In conclusion, SmIg is a new tegument protein from S. mansoni that plays an important role in reducing pathology induced by parasite infection.

Immunomodulatory effects of curcumin treatment on murine schistosomiasis mansoni

Curcumin is a polyphenol derived from the dietary spice turmeric. It has been shown to regulate numerous transcription factors, cytokines, adhesion molecules, and enzymes that have been linked to inflammation. In addition to inhibiting the growth of a variety of pathogens, curcumin has been shown to have nematocidal activity. The present study was designed to evaluate the schistosomicidal activity of curcumin in vivo as well as immunomodulation of granulomatous inflammation and liver pathology in acute schistosomiasis mansoni. Mice were infected each with 80 Schistosoma ( S.) mansoni cercariae and injected intraperitoneally with curcumin at a total dose of 400 mg/kg body weight. Curcumin was effective in reducing worm and tissue-egg burdens, hepatic granuloma volume and liver collagen content by 44.4%, 30.9%, 79%, and 38.6%, respectively. Curcumin treatment restored hepatic enzymes activities to the normal levels and enhanced catalase activity in the liver tissue of infected mice. Moreover, hepato-spleenomegaly and eosinophilia induced by S. mansoni infection were largely improved with curcumin treatment. Infected mice treated with curcumin showed low serum level of both interleukin (IL)-12 and tumor necrosis factor alpha (TNF-α), but IL-10 level was not significantly altered. Specific IgG and IgG1 responses against both soluble worm antigen (SWAP) and soluble egg antigen (SEA) were augmented with curcumin treatment, but IgM and IgG2a responses were not significantly changed. In conclusion, curcumin treatment modulates cellular and humoral immune responses of infected mice and lead to a significant reduction of parasite burden and liver pathology in acute murine schistosomiasis mansoni.

No difference in early cellular response of the pseudo-synovial membrane after total hip arthroplasty: Comparison of 3 combinations of bearing materials

Background Wear-resistant bearing materials may hypothetically reduce chronic inflammation in the pseudosynovial membrane as compared to less wear-resistant bearing materials such as polyethylene. We assessed the foreign body response in the pseudosynovial membrane in vivo after total hip replacement.Methods 37 patients from a larger prospective randomized trial of 225 patients had biopsies taken arthroscopically from the artificial hip joint (i.e. the pseudosynovial membrane) 1 year after insertion of the implant. All patients had an identical hip prosthesis (Bimetric-RingLoc) except for the bearing materials, which consisted of polyethylene on zirconia, CoCr on CoCr, or alumina on alumina.Histological quantification was performed on 2-μm-thick semi-thin plastic sections or paraffin sections by point counting technique to compare the volume fraction of macrophages, granulomas and endothelial cells in biopsies of the pseudosynovial membrane.Results The median macrophage volume fractions for polyethylene-on-zirconia bearing material (n = 15), CoCr-on-CoCr (n = 9), and alumina-on-alumina (n = 11) were 0.02, 0.04, and 0.004, respectively. The median granuloma volume fractions for polyethylene-on-zirconia (n = 13), CoCr-on-CoCr (n = 9), and alumina-on-alumina (n = 13) were 0.02, 0.04, and 0.02, respectively. The median endothelial cell volume fractions for polyethylene-on-zirconia (n = 15), CoCr-on-CoCr (n = 9), and alumina-on-alumina (n = 11) were 0.03, 0.02, and 0.05, respectively.Statistical analysis showed no significant differences between the three groups with the different bearings with respect to volume fraction of macrophages, granulomas and endothelial cells.Interpretation Our study demonstrated that a granulomatous inflammation is a common finding in non-loose implants as early as 1 year after the operation not demonstrating a difference in macrophages and granuloma formation with the various bearing materials. Thus a high volume fraction of macrophages was found in the osteoarthritis control group compared to the operated group.

Schistosomal granuloma modulation. III. Schistosma haematobium worms accelerate S. mansoni soluble egg antigen-induced hepatic granuloma formation in vivo.

Recurrent experimental evidence indicates that schistosomal egg granuloma formation at least in the murine model results from a host response generated against both egg- and worm-derived antigens. Further experiments aimed at identifying the existence in vivo of cross-sensitization between Schistosoma haematobium worms and S. mansoni-derived egg antigens were performed with respect to S. mansoni egg antigen-induced granuloma formation and fibrogenesis in the liver. Male OF1 mice bisexually infected with S. haematobium or S. mansoni were hepatically challenged (cecal vein injection) with S. mansoni SEA (soluble egg antigen)-coupled Sepharose beads at the end of prepatent infection (8-10 days prior to the start of egg deposition). The mean granuloma volume (MGV) of in-vivo-generated synchronized hepatic granulomas (8 days old) and the fibrotic response were estimated. Just like S. mansoni-infected rodents, mice carrying an S. haematobium infection generated an accelerated hepatic granulomogenesis [respective MGVs 4.72 +/- 0.56 and 5.41 +/- 0.75 x 10(6) microm3; P < 0.0001 versus unsensitized (MGV 3.00 +/- 0.40 x 10(6) microm3) mice] and an enhanced fibrotic response against S. mansoni SEA. They also had significantly enlarged spleens (P < 0.0001) and moderately enlarged livers (P = 0.02) as compared with S. haematobium-infected mice that were not challenged with SEA. From these observations we infer that in vivo, S. haematobium worms can positively modulate S. mansoni egg antigen-induced granuloma formation and hepatic fibrogenesis, resulting in more severe liver pathology.

Experimental paracoccidioidomycosis in hamsters (Mesocricetus auratus): gestational interactions.

Paracoccidioides brasiliensis is a dimorphic fungus presenting specific steroid hormone receptors, both in the yeast and mycelial forms and estrogen inhibits the transition from mycelium to yeast. In the acute phase, the disease occurs with equal frequency in both sexes but in adults, females are spared. Placental fungal infection has been reported, but references to fetal infection have not been confirmed. We used 78 Syrian female hamsters divided into 3 groups: GI consisted of 30 infected mated females, GII of 20 infected unmated females and GIII of 28 uninfected mated females. Animals of group I were mated 4 weeks after infection and half of them were submitted to cesarean section on day 15 after successful mating; the other half was maintained and submitted to cesarean section and sacrificed 14 weeks after infection. Half of the animals of group II were sacrificed seven weeks and the other half 14 weeks after infection. Uninfected animals of group III were treated the same as the animals of group I. The animals were infected with strain 18 of P. brasiliensis by the intracardiac route. We evaluated the disease by the volume of granulomas in different organs, number of fungi in liver and spleen and the immunologic responses [ELISA, Double Immunodifusion (DID), Delayed Hypersensitivity Skin Test (DHT) and Macrophage Migration Inhibition (MMI)]. We studied the infection through the gestation by evaluation of the abortions, morphologic and clinic examinations of the fetuses. Our results showed that the infection did not transfer to the fetus through the placenta, but the number of abortions was larger among infected females. The newborns of GI females were smaller, weighed less and showed little vitality. The disease was more severe and disseminated in infected mated females, especially in the second sacrifice 14 weeks after inoculation, when the total volume of granulomas in them (56.3 mm) was much greater than in the infected unmated females (12 mm).

Schistosomal granuloma modulation. I. Schistosoma mansoni worm antigens CAA and CCA prime egg-antigen-induced hepatic granuloma formation.

Adult Schistosoma mansoni worms can positively modulate soluble egg antigen (SEA)-induced granulomas formed around SEA-coupled beads implanted in the liver. In this study, our aim was to further unravel the immunopathological characteristics of S.mansoni-worm-derived antigens in vivo. (a) Adult worm antigen (AWA)-coupled Sepharose beads, implanted into the liver, induced granulomas, containing numerous eosinophilic granulocytes and elicited marked periparticular fibrosis (composed of interstitial matrix proteins and basement membrane components). (b) Quantitative morphological analysis demonstrated that in naive mice, AWA-induced hepatic granuloma formation peaked in volume 16 days after injection of the beads. An accelerated response against AWA-coupled particles (peak volume at 8 days) was observed in mice carrying a single-sex, male S. mansoni infection. (c) When the granuloma volume induced by SEA-coupled beads in unisexually S. mansoni infected mice was compared to granulomas induced by beads laden with both SEA and AWA in unsensitized mice, no significant differences in granuloma volume were seen, indicating the existence of in vivo egg/worm antigen cross-sensitization. (d) Naive mice, sensitized with the worm antigens circulating anodic antigen (CAA) or circulating cathodic antigen (CCA), mounted a strongly accelerated response towards SEA-coupled beads implanted in the liver. We infer that, in vivo, worm antigens cross-sensitize with egg antigens and have both granulomogenic and fibrogenic characteristics. The S. mansoni soluble worm antigens CCA and CAA prime hepatic egg-antigen-induced granuloma formation possibly through the presence of immunogenic carbohydrates. These mechanisms lead to an accelerated response against SEA.

Adhesion molecules in intestinal Schistosoma mansoni infection.

Adhesion molecules constitute essential elements in inflammation, mediating various cellular interactions. We investigated the expression of adhesion molecules mediating cell-cell [intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1)] and cell-matrix interactions [very late antigen-4 (VLA-4), VLA-6, and syndecan-1] in intestinal granulomas of mice infected with the parasite Schistosoma mansoni. Up-regulation of ICAM-1, LFA-1, and VLA-4 was seen in ileal and colonic granulomas, at both the acute (8 weeks postinfection) and the chronic stage (13-16 weeks postinfection). Up-regulation of VLA-6 was absent in all intestinal granulomas. Syndecan-1 immunoreactive (antigen-driven) B-lymphocytes were seen in the proximity of egg-antigen-laden macrophages in the inner part of ileal and colonic granulomas, although B-cells are considered to be absent in ileal granulomas. Estimation of intestinal granuloma volumes demonstrated the lack of down-modulation observed in ileal granulomas. From our results we infer that adhesion molecules constitute important elements in schistosomal intestinal granuloma formation. Organ-related differences between hepatic and intestinal granulomas exist (e.g., granuloma volume), but these differences are not morphologically reflected in a differential expression of the adhesion molecules ICAM-1, LFA-1, and VLA-4. Syndecan-1 immunoreactive B-lymphocytes also appear to be involved in ileal granuloma formation.

Differential regulation of granuloma size and hepatic fibrosis in schistosome infections.

Granuloma size is the variable most frequently used to evaluate the immunopathogenesis of schistosome infections. However, hepatic fibrosis is at the least an equally relevant variable. Hepatic fibrosis and the size of circumoval granulomas are frequently dissociated in experimental murine Schistosoma mansoni and S. japonicum infections. Virtually nothing is known of the immunoregulation of schistosomal hepatic fibrosis. This review notes many of the studies which have found discrepancies in granuloma volume and hepatic fibrosis, attempts to put them in perspective and to evaluate different methods of calculating changes in collagen synthesis or content.

Suppressive effect of interleukin-4 neutralization differs for granulomas around Schistosoma mansoni eggs injected into mice compared with those around eggs laid in infected mice.

The principal pathological manifestation of murine Schistosoma mansoni infection is the egg-induced granuloma. Synchronous pulmonary granulomas forming around intravenously injected schistosome eggs are widely used to study the immunopathology of schistosomiasis. A number of anticytokine antibody treatments have a remarkable effect in modulating granulomas in this model but little effect on the size of hepatic granulomas around laid eggs during experimental infection. To examine this discrepancy, we examined the effects of anticytokine antibodies on liver and lung granulomas around injected eggs and around eggs laid during infection in both locations. Anti-interleukin-4 (IL-4) treatment greatly reduced the volume of granulomas around eggs injected into the liver via the portal vein and around eggs injected into the lung via the tail vein. On the contrary, granulomas around eggs laid by worms in either the liver or the lung during the course of infection were not significantly decreased in size by anti-IL-4 treatment. Thus, site is not important for the disparate effects of anti-IL-4 in granuloma formation around injected versus laid eggs. This effect is seen in naive and sensitized animals and is most probably due to differences in the quality of injected eggs versus those laid in situ by the worms.

Relationship between schistosomicidal drug efficacy, gonadal steroid hormonal changes and state of disease immunopathology in murine Schistosomiasis mansoni

The schistosomicidal efficacy of praziquantel (2 × 500 mg/kg), oxamniquine (1 × 100 mg/kg) and combined one-third the curative dose of each of them (333 mg/kg praziquantel+33 mg/kg oxamniquine), were correlated to gonadal steroid hormonal changes and state of disease immunopathology, in mice infected with 100 Schistosoma mansoni (S. mansoni) cercariae. Maximum efficacy recorded with combination regimen particularly 4 weeks after treatment, was accompanied by maximum reduction in granuloma volume (65%), least fall in testosterone level (−56·2 and −60·2% in male and female mice respectively) and increased progesterone level (+ 33 · 9 and + 81 · 5% in male and female mice respectively). These findings revealed a potentiated effect of combination therapy on mature infection and the possible involvement of gonadal steroid hormones in affecting the efficacy of schistosomicidal drugs and state of disease immunopathology.

Effect of praziquantel on hepatic fibrosis in experimental Schistosomiasis mansoni

The effect of specific chemotherapy on hepatic fibrosis was studied in Swiss albino mice infected with Schistosoma mansoni. The effect of treatment with praziquantel at 8, 12, and 20 weeks postinfection on fibrosis was assessed by determination of total hepatic collagen content, ratio of type III I + III collagen, granuloma volume, and histopathological examination of liver section. Total collagen content was reduced in mice treated at the 8th week of infection compared to respective infected controls. The ratio of type III I collagen was within normal limits. The decrease in collagen deposition was not observed when treatment was given 12 or 20 weeks postinfection. Early specific treatment of schistosomiasis may be important in the therapeutic approach to the control of morbidity in schistosomiasis.

Schistosomiasis mansoni, haematobia, and japonica in hamsters: Liver granuloma measurements

Differences in the granulomatous response of hamsters infected with Schistosoma mansoni, Schistosoma haematobium, or Schistosoma japonicum were studied by measuring granulomas formed around eggs in the livers at 1, 3, 5, 11, and 19 week intervals after patency of the infections. For each species, the mean diameters of both granulomas containing only 1 egg and granulomas selected at random were determined. The mean volume of single egg granulomas in S. mansoni-infected hamsters was greater than in those with other species at all time intervals studied. The decrease in single egg granuloma size with time occurred more gradually in S. mansoni infections. The mean volume of granulomas measured at random was greatest in S. japoracum-infected animals at 1, 3, and 11 weeks after patency; was greatest in S. haematobium-iufected. hamsters at 5 and 19 weeks; and was least in S. mansoni-infected hamsters at all time periods. During the 19 week period following patency, the mean volume of randomly measured granulomas increased with time in S. haematobium infections, decreased gradually in S. mansoni infections, and decreased markedly in S. japonicum infections. Multi-egg granulomas represented 2–6% of all granulomas measured in S. mansoni infections, 38–68% in S. haematobium infections, and 20–53% in S. japonicum infections. An estimated proportion of the liver occupied by granulomas, i.e., a lesion-to-tissue ratio, was computed. There was no consistent difference in this ratio among the 3 species when the data were grouped in comparable intervals of mean number of eggs per g of liver. The lesion-to-tissue ratio increased with increasing numbers of eggs per g of liver. The host response of hamsters to these 3 species of schistosomes differed markedly even at comparable levels of eggs per g of liver. This study provides further evidence of an intrinsic qualitative difference in the eggs of the 3 species. The species of eggs present in tissues apparently has greater influence on the host response than does the quantity of eggs present.