Schistosoma mansoni: circulating antigens and immune complexes in infected mice.

Circulating schistosome antigens (CSA) and circulating immune complexes (CIC) were investigated during the course of Schistosoma mansoni infection in mice. The radioimmunoprecipitation-polyethylene glycol (PEG) assay (RIPEGA) with [¹²⁵I]anti-S. mansoni antibodies or [¹²⁵I] anti-antigen “4” antibodies detected, respectively, total CSA and antigen “4” in serum and in 3% polyethylene glycol-precipitated CIC from infected mice. Complement fixation test and [¹²⁵I] C_1q-binding test revealed, respectively, an anticomplementary activity and the presence of C_1q-binding CIC. All these substances appeared in infected mice at approximately the same period, i.e., between the 40th- and the 55th- day postinfection. No correlation was observed between the detection of anticomplementary active substances and C_1q-binding CIC. In contrast, a close relationship was noticed between CSA and complement-activating material during the course of the infection. This suggests that substances with anticomplementary activity in serum from infected mice could be one or various CSA. A close correlation was also observed between C_1q-binding CIC and free or “complexed” antigen “4.” This observation supports well the possibility that antigen “4” is one of the major complexed circulating antigen present in schistosomiasis. The immunoglobulins G₁, G_2a, M, and A were also characterized in 3% PEG-precipitated CIC from infected mice during the period in which we detected C_1q-binding CIC. The roles played by specific S. mansoni CIC in either schistosomal nephropathy or protective mechanisms to a challenge infection in mice are discussed.     

Micromorphology of hairs and spines on ovary and fruit ofDaucus carota L. var.sativa (The cultivated carrot)

The primary ridges of the fruit wall bear hairs in two rows and the secondary ridges have spines in a single row. Scanning electron microscopic studies reveal the finer details of the morphology of hairs and spines. The development of hairs and spines is traced out to give a critical morphohistogenic explanation.     

Effect of the spleen extracts on primary immune response in mice]

Experiments were conducted on mice. Direct Jerne's test demonstrated a possibility of intensification of the primary immune response in the sexually mature mice under the effect of the splenic extracts. The significance of the extract dose and of the time of administration for the manifestation of the stimulating action was studied.     

Schistosoma mansoni heat shock protein 70 elicits an early humoral immune response in S. mansoni infected baboons

A study was undertaken to search for DNA recombinant Schistosoma mansoni proteins responsible for eliciting an antibody response from the host at a very early phase after infection. A S. mansoni adult worm cDNA expression library was screened using pooled sera from baboons with four weeks of infection. Based on their specific reactivity with the S. mansoni infected sera and no reactivity when tested against the pre-infection sera from the same baboons, four clones were selected for further studies. Sequence analysis revealed that they were homologous to the S. mansoni heat shock protein 70 (hsp70). The insert sizes of the four selected clones varied from 1150 to 2006 bp. The preliminary characterization for antibody reactivity against a panel of baboon sera showed that the longest clone was the most reactive, eight out of eight acute and three out of four chronic sera reacting positively to this clone. The shortest clone was the least reactive. Our results suggest that the S. mansoni hsp70 elicits an early and strong antibody response in baboons and that antibodies to this protein can be detected in chronically infected animals. Therefore S. mansoni hsp70 may be a valid target for immunodiagnosis. However further studies are needed to identify the portion of the hsp70 that best fits the requirements for a valuable diagnostic antigen.     

Effect of somatostatin on gastrointestinal contractility in Schistosoma mansoni infected mice

Schistosoma mansoni infection induces severe gastrointestinal motility disturbances which are characterised by hyperactivity of intestinal muscle, abdominal pain, diarrhoea, vomiting and nausea. During schistosomiasis, the neuropeptide somatostatin is generated within inflammatory granulomas. However, somatostatin is also an important inhibitory modulator of gastrointestinal motility. In the present study, we have investigated the potential of somatostatin to reduce schistosomiasis-induced hyperactivity of gastrointestinal smooth muscle. Organ bath experiments were performed to study the contractility of isolated smooth muscle strips of intestine from control mice and from mice that were infected with S. mansoni for 2, 4, 8 and 16 weeks. Electrical field stimulation (0.5-8 Hz) of enteric nerves induced frequency-dependent neurogenic contractions of cholinergic origin in all regions of the small intestine. Somatostatin (0.1-1 microM) concentration-dependently inhibited the contractions to enteric nerve stimulation in the small intestine from uninfected control mice and from acutely S. mansoni infected mice (2 and 4 weeks of infection). After 8 weeks of infection with S. mansoni, this inhibitory effect of somatostatin was less pronounced and after 16 weeks of infection it was completely abolished. Histology demonstrated that chronic infection of mice with S. mansoni was associated with significant alterations in the musculature of the small intestine. These alterations may be associated with physiological changes in the responsiveness to somatostatin and suggest that the somatostatin neuroregulatory circuit of enteric neurotransmission in the small intestine is disturbed during chronic schistosomiasis mansoni.     

Effect of cultivation age and embryo size on specific oxygen uptake rate in developing somatic embryos ofDaucus carota L.

Summary The average specific oxygen uptake rate of carrot (Daucus carota L.) somatic embryos in batch culture decreased with cultivation time, coinciding with an increase in the size and maturity of the embryos. From experiments performed on three size classes obtained at 15 days of culture, larger embryos were found to have a lower oxygen uptake rate suggesting either a lower intrinsic metabolic activity and/or transport limitations leading to a reduction in the observed metabolic activity.     

Schistosoma mansoni: mice infected with different worm strains.

Infections with five geographical strains and substrains of Schistosoma mansoni were compared in mice. Two substrains (Lc-1 and Lt-1) were derived from the parent (L-1) St. Lucian strain on the basis of differing infectivity for various snail strains. The Puerto Rican strains (PR-1 and PR-2) were obtained with an interval of 25 years. Consistent differences among the lines were found in egg distribution and numbers of eggs in tissues and feces. One Puerto Rican strain (PR-2) and one St. Lucian substrain (Lc-1) had longer prepatent periods than the other strains. Mice infected with the PR-1 strain consistently had the highest egg accumulation in the tissues per worm pair. Relatively few eggs were passed in the feces of the Lt-1 strain. By Week 9 of infection, eggs were noted in the spleens of mice carrying each of the strains and substrains.     

Ultrastructural visualization of sites binding concanavalin a on the cell membrane ofDaucus carota

Summary Protoplasts fromDaucus carota showed differences in binding to Concanavalin A according to which of three enzyme preparations tested were used for their isolation. Protoplast bound Concanavalin A was visualized ultrastructurally using a peroxidase-diaminobenzidine staining. The variable amount of staining was supposed to be due to differences in the composition of contaminating membrane active enzymes in the crude enzyme preparations. Enzymatically removed binding sites for Concanavalin A in the plasmalemma were rapidly resynthesized when the isolated protoplasts were incubated in a sorbitol solution. At room temperature, Concanavalin A bound to the plasmalemma was found in clusters, while at 4 °C and on prefixed protoplasts the binding sites were homogeneously distributed. These results and the effects of the crude enzyme preparations on the cell membrane of the protoplasts will be discussed in relation to the fluid membrane model.     

Modification of immune response in nude mice infected with mouse hepatitis virus.

In nude mice experimentally infected with mouse hepatitis virus (MHV), the numbers of early and later plaque forming cells (PFC) to sheep red blood cells (SRBC) generated in the spleen were 7 to 20 times and 2 to 163 times, respectively, greater than those in non-infected nude mice, when SRBC were given at day 0 to day 21 postinfection. Splenic theta-positive lymphocytes in infected nude mice were shown to increase only at day 10 or more postinfection. PFC response to bacterial lipopolysaccharide, a T cell-independent antigen, was not modified in MHV-infected nude mice.     

Effect of neutrophilokines on the immune response in mice

The influence of different peptide fractions obtained from the intact and latex-stimulated neutrophils on the immune response was studied. It was demonstrated that neutrophils after stimulation synthesize the factors activating immune response, the intact neutrophils synthesize the suppressor factors of peptide nature.     

Effect of Curcuma longa or parziquantel on Schistosoma mansoni infected mice liver--histological and histochemical study

Effect of drug praziquantel (PZQ) and C. longa extract on S. mansoni infected mice is reported. The level of glycogen, alkaline and acid phosphatases (ALP and ACP respectively), and body weight, liver weight and liver weight/body weight ratio were studied in mice infected with S. mansoni. ALP level was increased after infection. C. longa treated mice showed marked reduction in ALP level more than after PZQ-treatment. C. longa enhanced the concentration of glycogen after being reduced by infection, while PZQ-treatment revealed more reduction. C. longa caused enhancement in body weight while PZQ treatment had no effect. The formation of granuloma around schistosome eggs in the liver produced inflammation. C. longa extract and PZQ were effective in reducing granuloma size in infected mice.     

Schistosoma mansoni: control of hepatotoxicity and egg excretion by immune serum in infected immunosuppressed mice is schistosome species-specific, but not S. mansoni strain-specific.

Immunosuppressed mice infected with Schistosoma mansoni suffer from an acute hepatotoxicity reaction, and they fail to excrete as many parasite eggs as comparably infected immunologically intact control animals. The hepatotoxicity was shown here to be preventable, and egg excretion rates were enhanced, by transfer of serum from donors with chronic S. mansoni infections, but not by serum from donors with heterologous infections of Schistosoma haematobium, Schistosoma bovis, or Schistosoma japonicum. The effects of the transferred sera are considered to be due to specific antibody, but the possibility of cytokine involvement is discussed. A high degree of serological cross-reactivity was found between sera from mice infected with the different schistosome species and unfractionated egg homogenate (SEA) in ELISA. Cross-reactivity of the heterologous sera was, however, reduced against CEF6, a partially purified fraction of S. mansoni eggs that contains the putative hepatotoxin and has serodiagnostic potential. S. mansoni isolates from Puerto Rico, Brazil, Egypt, and Kenya shared similar characteristics with respect to the immune dependence of egg excretion and hepatotoxicity in immunosuppressed mice. The S. mansoni geographic isolates were also indistinguishable serologically, in terms of both the capacity of respective infection sera to neutralize hepatotoxicity and in their capacity to promote egg excretion of the other isolates in vivo. Complete immunological cross-reactivity of the geographically distinct isolates was also observed in ELISA with both CEF6 and SEA. Utilization of CEF6 for serodiagnosis of schistosomiasis mansoni is therefore unlikely to be restricted by geographical considerations.     

Alterations of the intestinal innervation in mice infected with Schistosoma mansoni.

Schistosomiasis mansoni is a parasitic disease in which granulomas form around schistosome eggs in the liver and intestines. The purpose of this study was to determine the alterations in the intrinsic innervation of the distal ileum and proximal colon resulting from schistosomiasis. Using murine schistosomiasis mansoni, we examined light microscopic preparations stained with osmium-zinc iodide or the dihydronicotinamide adenine dinucleotide: nitro BT oxidoreductase (NADH) method. We also examined specific populations of peptidergic nerves (vasoactive intestinal polypeptide and substance P) using an avidin-biotin complex (ABC) immunohistochemical technique. We found that granulomas focally destroyed the enteric nerves. Occasionally nerves were found within granulomas, particularly at the periphery of the lesions. Nerve cell bodies close to granulomas had altered staining, which included increased staining for vasoactive intestinal polypeptide. The distribution of nerve injury varied between the 2 enteric segments studied. In the distal ileum, the principal injury was to the myenteric plexus; whereas, the submucous and mucosal plexuses were predominantly damaged in the proximal colon. The physiologic significance of this injury to the enteric nerves requires elucidation.     

Stimulation by adjuvants of cell-mediated immune response in Plasmodium berghei infected mice

Immunological adjuvants (alum, liposomes and saponin) were utilized to stimulate cell-mediated immune response in Plasmodium berghei infected Balb/c mice. It was shown that malaria antigen mixed with adjuvant induced appreciably delayed type hypersensitivity and production of migration inhibition factor compared to antigen alone.     

Effect of anthelminthic treatment on the immune response of mice

Immunologic function was tested both in vivo and in vitro in mice undergoing prophylactic anthelminthic therapy with three agents to assess whether these drugs affected immune responses. This study was performed because investigators often are concerned about the effect of drug treatment on the induction of specific immune responses. While helminthic infestation is recognized as deleterious to the host, it is unclear whether anthelminthic treatment might be immunosuppressive. The effects of piperazine or trichlorphon administered to drinking water or fenbendazole administered in feed were insignificant in BALB/c mice. The induction of allospecific cytolytic T lymphocytes (CTLs) in vitro, influenza specific memory T cells in vivo, influenza specific antibody secretion in vivo, or influenza-specific helper T cells and CTLs in vitro were examined. Results of this study indicate that anthelminthic treatments did not interfere with immune responses.     

Neuronal activation and plasticity in Schistosoma mansoni infected mice

Schistosomiasis leads to structural and functional changes which may result from unbalanced release of some inflammatory mediators. The aim of the study was to investigate the effect of intestinal parasitic infection on nitric oxide release and to evaluate the neural plasticity that leads to motility disturbance. Experiments were performed in Swiss mice 8- and 12-weeks following infection with Schistosoma mansoni compared to untreated controls. Jejunal motility was assessed using a Trendelenburg preparation to study aboral directed peristaltic pressure waves. Histological examination was used to determine the pathological characteristics of inflammation.Parasitic infection produces diffuse inflammatory infiltrate in both 8- and 12-weeks infected animals. Inflammation had significant effect on peristaltic pressure waves amplitude and intervals at 8-weeks compared to control; whereas, in 12-weeks post infection there was a significant decrease in peristaltic pressure waves amplitude and interval compared to 8- weeks and control.Nitric oxide synthase inhibitor (L-NAME 100 μM) induced a significant increase in amplitude and decrease in intervals in control, 8- and 12- weeks infected animals. In conclusion, parasitic infection leads to disturbance in the release of the inflammatory mediators. This study indicated the role of nitric oxide in developing granulomatous inflammation and participating in motility disturbance.