川楝子的化学成分、药理作用及其毒性研究进展

川楝子作为传统中药,广泛应用于驱虫,是天然的杀虫剂。川楝子化学成分丰富,主要含有柠檬苦素型三萜、木脂素、黄酮、甾体、有机酸等成分,其具有抗肿瘤、抗氧化、抗菌、消炎镇痛、抗病毒、驱虫等广泛的药理作用,但由于川楝子有肝毒性、妊娠毒性等使其应用受到一定的限制。对川楝子的化学成分、药理作用和毒性研究进展进行综述,为其进一步的开发研究和临床应用提供科学依据。     

Effects of peripheral blood mononuclear cells on Haemonchus contortus larval motility in vitro

The objective of this experiment was to determine effects of peripheral blood mononuclear cells (PBMC), derived from parasite‐resistant St. Croix (STC) and parasite‐susceptible Suffolk (SF) sheep, on motility of Haemonchus contortus L₃ stage larvae in vitro. Peripheral blood mononuclear cells were collected from 10 lambs of each breed, 5 naïve and 5 which had received a priming infection with H. contortus. Larval motility was quantified using a MIF Nikon Sweptfield microscope and NIS Elements AR software, and measurements included path length (PL) (μm), velocity (VEL) (μm/s) and acceleration (ACC) (μm/s²). After 18 h of incubation, PL and VEL were greatest in larvae cultured with SF‐derived PMBC and were significantly different from all other groups (P < 0·01). No difference was observed in PL or VEL between larvae exposed to naïve or primed STC‐derived PBMC and primed‐SF PBMC. Differences in ACC were detected between larvae cultured with primed STC‐derived PBMC (10·91 μm/s²) and naïve SF‐derived PBMC (45·7 μm/s²) (P = 0·035). These data indicate an innate ability of STC‐derived PBMC to severely inhibit larval motility.     

Whole parasite vaccines for the asexual blood stages of Plasmodium

After many decades of research, an effective vaccine for malaria is still not available. Most research efforts have focused on identifying a key target antigen and then using powerful adjuvants to generate specific antibodies that can block parasites from entering host cells (hepatocytes, red blood cells). However, the inability to generate sufficiently potent antibody responses has led to significant disappointment with current vaccine programs. An additional challenge for sub-unit vaccines is that key vaccine antigens are highly polymorphic. These challenges have spurred radically different approaches to malaria vaccine development. Many of these involve the use of “whole parasites”—either extracted from mosquitoes or cultured. With these, every parasite molecule for that particular strain is included in the vaccine. This strategy is showing great promise following several clinical trials with irradiated sporozoites. However, a whole-parasite approach to a blood stage vaccine has not advanced as quickly. This article outlines the history, the different approaches that are being taken and the challenges associated with whole parasite blood stage vaccines and discusses recent exciting developments as these vaccines now move into the clinic.     

海南省农村儿童肠道寄生虫感染的调查

本文报道全省抽查的５个县、市１５个点的１－６岁农村儿童肠道寄生虫感染概况，按全国人体寄生虫分布调查实施细则的方法，共调查１－６岁儿童１３８７人，查见寄生虫感染者１２３７例，寄生虫总感染率为８９．２％。共查见寄生虫２３种，其中线虫１０种、原虫８种、吸虫４种、绦虫１种。以蛔虫、鞭虫、钩虫和蛲虫感染率较高，依次为６７．１％、５８．７％、２９．１％和３３．４％。其次为贾第虫和粪类圆线虫、其感染率分别为８．２％和２．６％。多虫感染者达７４．２％，１人同时感染３种寄生虫以上者占感染人数３７．１％，表明海南１－６岁农村儿童肠道寄生虫感染的普遍性和严重性。     

Trypanosoma cruzi infection induces a massive extrafollicular and follicular splenic B-cell response which is a high source of non-parasite-specific antibodies

Acute infection with Trypanosoma cruzi, the aetiological agent of Chagas’ disease, results in parasitaemia and polyclonal lymphocyte activation. It has been reported that polyclonal B‐cell activation is associated with hypergammaglobulinaemia and delayed parasite‐specific antibody response. In the present study we analysed the development of a B‐cell response within the different microenvironments of the spleen during acute T. cruzi infection. We observed massive germinal centre (GC) and extrafollicular (EF) responses at the peak of infection. However, the EF foci were evident since day 3 post‐infection (p.i.), and, early in the infection, they mainly provided IgM. The EF foci response reached its peak at 11 days p.i. and extended from the red pulp into the periarteriolar lymphatic sheath. The GCs were detected from day 8 p.i. At the peak of parasitaemia, CD138⁺ B220⁺ plasma cells in EF foci, red pulp and T‐cell zone expressed IgM and all the IgG isotypes. Instead of the substantial B‐cell response, most of the antibodies produced by splenic cells did not target the parasite, and parasite‐specific IgG isotypes could be detected in sera only after 18 days p.i. We also observed that the bone marrow of infected mice presented a strong reduction in CD138⁺ B220⁺ cells compared with that of normal mice. Hence, in acute infection with T. cruzi, the spleen appears to be the most important lymphoid organ that lodges plasma cells and the main producer of antibodies. The development of a B‐cell response during T. cruzi infection shows features that are particular to T. cruzi and other protozoan infection but different to other infections or immunization with model antigens.     

T cell expression of MyD88 is required for resistance to Toxoplasma gondii

Resistance to Toxoplasma gondii depends on dendritic cells to recognize this pathogen and secrete IL-12, in turn promoting IFN-gamma production from responding T cells. The adaptor protein, myeloid differentiation primary-response gene 88 (MyD88), is important for most Toll-like receptor (TLR) signaling, as well as IL-1R/IL-18R signals. There is considerable evidence that MyD88 is required for the innate sensing of T. gondii and IL-12 responses. Although Myd88(-/-) mice challenged with T. gondii have defective IL-12 and Th1 effector responses and succumb to disease, administration of IL-12 to Myd88(-/-) mice partially restores the Th1 response and yet fails to prolong survival. This finding suggested that MyD88 may mediate signals within T cells important for resistance to this pathogen. To evaluate the role of MyD88 in T cells under noncompetitive conditions, bone marrow chimeras were generated, in which the T cells lacked MyD88, but MyD88-dependent innate immune responses were intact. Upon challenge with T. gondii, these chimeric mice were more susceptible to disease, developing severe toxoplasmic encephalitis and succumbing within 30 days. Splenocytes and brain mononuclear cells isolated from infected chimeric mice produced less IFN-gamma when cultured with a T. gondii-derived antigen. The increase in susceptibility observed was independent of signals via the IL-1R and IL-18R, suggesting a role for TLRs in MyD88-mediated T cell responses to T. gondii. These observations show that, in addition to a role for MyD88 in innate responses, T cell expression of MyD88 is necessary for prolonged resistance to a pathogen.     

Immunoregulation of genetically controlled acquired responses to Leishmania donovani infection in mice: the effects of parasite dose, cyclophosphamide and sublethal irradiation

On a B10 (Lshs) genetic background, the development of acquired T cell mediated immunity to Leishmania donovani infection in mice is under H-2 linked genetic control. Following intravenous inoculation of 10(7) amastigotes three phenotypic patterns of recovery have been described: 'early cure' (H-2r,s), 'cure' (H-2b) and 'non-cure' (H-2d,q,f). In an attempt to determine the immunological basis for this H-2 linked genetic control the effects of varying parasite dose (5 x 10(3) to 5 x 10(7) amastigotes) and of pre-treatments with cyclophosphamide (50 or 200 mg/kg body weight CY) or sublethal irradiation (100 or 550 rad) on the course of infection, and on circulating anti-leishmanial IgG levels, were examined in strains representative of the three phenotypes: B10.D2/n (H-2d), C57BL/10ScSn (H-2b) and B10.RIII (H-2r). It was found that with low parasite doses (5 x 10(3) or 5 x 10(4)) 'non-cure' mice presented a 'cure' profile whilst raising the dose (5 x 10(7)) caused some perturbation of the normal self-curing response in 'cure' (but not 'early cure') mice. The highest dose did not, however, lead to progressive disease in the genetically non-cure strain. For the parasite dose experiments circulating anti-leishmanial IgG levels were higher in the early cure and cure strains than in the H-2d non-cure strain. The higher doses of CY and sublethal irradiation administered prior to infection had a clear prophylactic effect on the non-cure strain with some effect also observed in cure and early cure strains. This was thought to be due to deletion of the precursors of T suppressor (TS) cells suppressing cell-mediated immunity. Resolution of the liver parasite load in pre-treated mice took place despite minimal or undetectable levels of circulating anti-leishmanial IgG. Similarly, the earlier resolution of parasite load in pre-treated cure and early cure mice occurred even though the antibody response was severely reduced. This suggests that the high antibody responses observed in early cure and cure strains do not normally mediate cure and may simply reflect the independent effect of H-2 on T helper function or the humoral response.     

Co-infection with Trypanosoma brucei brucei prevents experimental autoimmune encephalomyelitis in DBA/1 mice through induction of suppressor APCs

The immune system has co-evolved with the infectious agents that challenge it, and in response pathogens have developed different mechanisms to subvert host immunity. A wealth of evidence suggests that infections are important components in the development of a functional immune system, and understanding the modulation of the host immune system by pathogens may offer new therapeutic strategies in a non-infectious setting. We investigated how infection with the protozoan parasite Trypanosoma brucei brucei (Tbb) modulates the autoimmune response to recombinant myelin oligodendrocyte glycoprotein (rMOG) in DBA/1 mice. Mice harbouring a Tbb infection did not develop experimental autoimmune encephalomyelitis (EAE) induced by immunization with rMOG in CFA, an animal model for the human autoimmune disease multiple sclerosis. Additionally, mice infected with the parasite at the time of immunization or 1 week later developed less severe EAE than uninfected controls. Protected mice displayed a markedly diminished rMOG-specific proliferation and IFNgamma production in lymph node cells and had correspondingly low titres of serum anti-rMOG IgG. Antigen-presenting cells (APCs) from spleens of Tbb-infected mice presented rMOG less efficiently to rMOG-specific T cells in vitro than did splenic APCs from uninfected mice and could also inhibit antigen-specific proliferation in control in vitro cultures. This suppressive effect is at least in part due to increased release of IL-10. Transfer of splenic APCs from Tbb-infected mice into mice immunized with rMOG-CFA 7 days previously abrogated disease significantly. These findings indicate that infections can prevent autoimmunity and that APCs might be used as immunomodulants.     

Co‐operative suppression of inflammatory responses in human dendritic cells by plant proanthocyanidins and products from the parasitic nematode Trichuris suis

Interactions between dendritic cells (DCs) and environmental, dietary and pathogen antigens play a key role in immune homeostasis and regulation of inflammation. Dietary polyphenols such as proanthocyanidins (PAC) may reduce inflammation, and we therefore hypothesized that PAC may suppress lipopolysaccharide (LPS) ‐induced responses in human DCs and subsequent T helper type 1 (Th1) ‐type responses in naive T cells. Moreover, we proposed that, because DCs are likely to be exposed to multiple stimuli, the activity of PAC may synergise with other bioactive molecules that have anti‐inflammatory activity, e.g. soluble products from the helminth parasite Trichuris suis (TsSP). We show that PAC are endocytosed by monocyte‐derived DCs and selectively induce CD86 expression. Subsequently, PAC suppress the LPS‐induced secretion of interleukin‐6 (IL‐6) and IL‐12p70, while enhancing secretion of IL‐10. Incubation of DCs with PAC did not affect lymphocyte proliferation; however, subsequent interferon‐γ production was markedly suppressed, while IL‐4 production was unaffected. The activity of PAC was confined to oligomers (degree of polymerization ≥ 4). Co‐pulsing DCs with TsSP and PAC synergistically reduced secretion of tumour necrosis factor‐α, IL‐6 and IL‐12p70 while increasing IL‐10 secretion. Moreover, both TsSP and PAC alone induced Th2‐associated OX40L expression in DCs, and together synergized to up‐regulate OX40L. These data suggest that PAC induce an anti‐inflammatory phenotype in human DCs that selectively down‐regulates Th1 response in naive T cells, and that they also act cooperatively with TsSP. Our results indicate a novel interaction between dietary compounds and parasite products to influence immune function, and may suggest that combinations of PAC and TsSP can have therapeutic potential for inflammatory disorders.     

Identification of the first Toll-like receptor gene in passerine birds: TLR4 orthologue in zebra finch (Taeniopygia guttata)

Toll-like receptors (TLRs) are the basic components of the vertebrate pathogen recognition system. Despite uniform general structure, remarkable variability in domain composition can be found in individual TLRs among species. Knowledge of interspecific differences is of particular importance to our understanding of selective pressures on TLRs. Currently, most TLRs are characterized only in a limited number of model species, including domestic chicken as a universal avian model. Here, we describe structure and expression pattern of TLR4 in zebra finch, a widely used passerine model species. The tgTlr4 gene consists of three exons (204, 167 and 3033–3043 bp) that are transcribed into messenger RNA with a relatively long 3'-untranslated region (788 bp). Predicted protein is composed of 842 amino acids (aas) forming extracellular domain with nine leucine-rich repeat (LRR) motives flanked at the carboxy-terminal end by leucine-rich repeat carboxy-terminal domain, transmembrane domain and cytoplasmic toll/interleukin-1 receptor domain. The overall structure is similar to other known TLR4 molecules with 32%–49% aa identity to various mammals and 74% to chicken. Although the position of most of the domains in zebra finch TLR4 resembles their position in chicken, there is one extra LRR at the aa position 207–229 in tgTLR4 and one LRR known in chTLR4 is missing. The gene is highly expressed in the bone marrow and in the spleen, intermediately in the gut and low expression was found in the liver and lungs. For the first time in birds, expression of tgTLR4 in peritoneal macrophages was found to be enhanced by the Escherichia coli lipopolysaccharide treatment.