Darker female pigeons transmit more specific antibodies to their eggs than do paler ones

Melanin‐based coloration is widespread among vertebrates, yet the adaptive significance of such pigments remains elusive, particularly with regard to the link between melanin and immune‐mediated maternal effects. The aim of this study was to investigate whether melanin‐based coloration could signal the ability of mothers to mount a humoral response and to transfer maternal antibodies (Ab) to their young. We injected differently coloured (pale and dark) female feral pigeons (Columba livia) with Chlamydiae (a natural antigen) and Keyhole Limpet Haemocyanin (KLH, an artificial antigen), and found no significant difference in humoral response between differently coloured females. However, darker females transferred more Ab against Chlamydiae into their eggs than paler ones, despite similar circulating levels of Ab. In addition to this, melanin‐based coloration showed a high heritability value. This suggests that a genetically based coloured trait might be linked to the ability of females to transfer specific Ab against Chlamydiae (but not against KLH) to their offspring, independent of their ability to produce Ab. This suggests that transmission of maternal Ab is antigen dependent, and that melanin‐based coloration might signal female ability to transmit specific Ab against natural pathogens. © 2013 The Linnean Society of London     

Glycomics-driven discoveries in schistosome research

Schistosome glycans and glycoconjugates play a prominent role in the parasite's biology, in particular in the interaction with the human host. A large amount of structural data regarding glycosylation of different schistosome life stages and glycoconjugate subsets has been collected in the last decade, but many significant gaps in our knowledge of the schistosomal glycome remain. Here we will present a concise review of the already available data guided by a selection of recently generated stage-specific glycan profiles, and discuss implications and prospects of glycomics studies of schistosomes.     

In vivo acetylation of CheY, a response regulator in chemotaxis of Escherichia coli

CheY, the excitatory response regulator in the chemotaxis system of Escherichia coli, can be modulated by two covalent modifications: phosphorylation and acetylation. Both modifications have been detected in vitro only. The role of CheY acetylation is still obscure, although it is known to be involved in chemotaxis and to occur in vitro by two mechanisms—acetyl-CoA synthetase-catalyzed transfer of acetyl groups from acetate to CheY and autocatalyzed transfer from AcCoA. Here, we succeeded in detecting CheY acetylation in vivo by three means—Western blotting with a specific anti-acetyl-lysine antibody, mass spectrometry, and radiolabeling with [¹⁴C]acetate in the presence of protein-synthesis inhibitor. Unexpectedly, the level and rate of CheY acetylation in vivo were much higher than that in vitro. Thus, before any treatment, 9-13% of the lysine residues were found acetylated, depending on the growth phase, meaning that, on average, essentially every CheY molecule was acetylated in vivo. This high level was mainly the outcome of autoacetylation. Addition of acetate caused an incremental increase in the acetylation level, in which acetyl-CoA synthetase was involved too. These findings may have far-reaching implications for the structure-function relationship of CheY.     

Topology of the 10 subunits within the Decamer of KLH, the hemocyanin of the marine gastropod Megathura crenulata

Immunoelectron microscopy has been performed using negatively stained immune complexes of keyhole limpet hemocyanin isoform 1 (KLH1) decamers and a functional unit-specific monoclonal antibody anti-KLH1-c1. The antibody links hemocyanin molecules at both the collar and the collarless edge of the decamer, indicating a peripheral localization of functional units c. In isoform 2 (KLH2) the positions of functional units c have been identified with the peanut agglutinin (PNA), which has previously been shown to exclusively bind to KLH2-c. Ferritin linked to PNA was used to visualize labeled molecules electron microscopically. The pattern of labeling also indicates a peripheral localization of the c functional units. The data presented in this paper support only one of two possible models for the subunit orientation within the hemocyanin decamer.     

抗原物质引起滤泡的形成:一次投入与多次投入的比较

实验以不同方式投入抗原物质,即一次投入与分次投入,并观察了新产生滤泡数,维持的滤泡数,实验用小鼠２４ 只, 于足底注入铝和钥孔　血蓝素附合物（ＡＫＬＨ）, 分一次注入与三次注入组; 三次注入组又分间隔５ 日及间隔二周注入。注入后第３ 周与第１２ 周分别取出腘淋巴结,应用免疫组化法,第三周末观察可见一次投入产生的滤泡多, 而第十二周发现分次投入维持的滤泡数多。可见反应性滤泡的形成, 不仅与刺激物的性状和投入量有关, 而且与投入的方法有关     

Peptide and glycopeptide dendrimers. Part I

Recent progress in peptide and glycopeptide chemistry make the preparation of peptide and glycopeptide dendrimers of acceptable purity, with designed structural and immunochemical properties reliable. New methodologies using unprotected peptide building blocks have been developed to further increase possibilities of their design and improve their preparation and separation. Sophisticated design of peptide and glycopeptide dendrimers has led to their use as antigens and immunogens, for serodiagnosis and other biochemical uses including drug delivery. Dendrimers bearing peptide with predetermined secondary structures are useful tools in protein de novo design. This article covers synthesis and applications of multiple antigen peptides (MAPs), multiple antigen glycopeptides (MAGs), multiple antigen peptides based on sequential oligopeptide carriers (MAP‐SOCs), glycodendrimers and template‐assembled synthetic proteins (TASPs). Part I deals with the development of various structural forms of MAPs as well as their application as antigens, immunogens, and for immunodiagnostic and biochemical purposes. Copyright © 1999 European Peptide Society and John Wiley & Sons, Ltd.     

Synthetic adjuvants for vaccine formulations: evaluation of new phytol derivatives in induction and persistence of specific immune response

Terpenoids are ubiquitous natural compounds that have been shown to improve vaccine efficacy as adjuvants. To gain an understanding of the structural features important for adjuvanticity, we studied compounds derived from a diterpene phytol and assessed their efficacy. In a previous report, we showed that phytol and one of its derivatives, PHIS-01 (a phytol-derived immunostimulant, phytanol), are excellent adjuvants. To determine the effects of varying the polar terminus of PHIS-01, we designed amine and mannose-terminated phytol derivatives (PHIS-02 and PHIS-03, respectively). We studied their relative efficacy as emulsions with soluble proteins, ovalbumin, and a hapten-protein conjugate phthalate-KLH. Immunological parameters evaluated consisted of specific antibody responses in terms of titers, specificities and isotype profiles, T cell involvement and cytokine production. Our results indicate that these new isoprenoids were safe adjuvants with the ability to significantly augment immunogen-specific IgG1 and IgG2a antibody responses. Moreover, there was no adverse phthalate cross-reactive anti-DNA response. Interestingly, PHIS-01 and PHIS-03 influenced differentially T-helper polarization. We also observed that these compounds modulated the immune response through apoptotic/necrotic effects on target tumor cells using murine lymphomas. Finally, unlike squalene and several other terpenoids reported to date, these phytol derivatives did not appear arthritogenic in murine models.     

Effects of testosterone on cell-mediated and humoral immunity in non-breeding adult European starlings

One of the primary assumptions of the immunocompetence hypothesisis that testosterone is immunosuppressive. Although many studiesin birds and mammals have supported this assumption, conflictingresults have been reported in a variety of species. We investigatedthe effects of testosterone manipulation on both cell-mediatedand humoral immunity in adult songbirds, European starlings(Sturnus vulgaris). Male and female starlings were wild-caught,housed in the laboratory, and implanted with either empty silasticcapsules or capsules containing testosterone. Six weeks after implantation, humoral immunity was assessed by injecting thebirds with a novel antigen, keyhole limpet hemocyanin, andmeasuring specific antibody responses 10 and 15 days latervia an enzyme-linked immunosorbant assay. Cell-mediated immunitywas assessed 7 weeks after implantation via intradermal injectionof the T-cell mitogen phytohemagglutinin into the wing web and measuring the degree of swelling 24 h later. Antibody responsesto antigenic challenge were significantly suppressed in testosterone-treatedfemales 10 days post-injection and in both sexes 15 days post-injection.Furthermore, there was a significant inverse relationship betweenindividual variability in antibody responsiveness and plasmatestosterone concentrations. Cell-mediated responses to phytohemagglutininstimulation were also significantly suppressed in testosterone-treatedmales compared to same-sex controls. Testosterone treatmentsignificantly increased plasma corticosterone concentrations compared to controls, and the possibility of this effect mediatingthe immunosuppressive effects of testosterone is discussed.The present study is among the first to demonstrate testosterone-inducedsuppression of both cell-mediated and humoral immunity in aspecies of songbird.     

Keyhole limpet hemocyanin contains Gal(β1–3)-GalNAc determinants that are cross-reactive with the T antigen

Keyhole limpet hemocyanin (KLH) is widely used as a carrier molecule to enhance immune responses to administered antigens, and for immunotherapy of bladder and renal carcinoma. In the present study we show, using lectin and antibody binding studies, that native KLH contains Gal(1–3)GalNAc-bearing oligosaccharides, and that immunization with KLH in Lewis rats induces the production of anti-Gal(1–3)GalNAc antibodies. This might explain the beneficial effect of KLH in bladder cancers that express crossreactive Gal(1–3)GalNAc determinants or the T antigen.Supported by NIH grant NS11766 and by the William Rosenwald Family Fund Inc.     

A preclinical study comparing approaches for augmenting the immunogenicity of a heptavalent KLH-conjugate vaccine against epithelial cancers

Previously using a series of monovalent vaccines, we demonstrated that the optimal method for inducing an antibody response against cancer cell-surface antigens is covalent conjugation of the antigens to keyhole limpet hemocyanin (KLH) and the use of a saponin adjuvant. We have prepared a heptavalent-KLH conjugate vaccine containing the seven epithelial cancer antigens GM2, Globo H, Lewis(y), TF(c), Tn(c), STn(c), and glycosylated MUC1. In preparation for testing this vaccine in the clinic, we tested the impact on antibody induction of administering the individual conjugates plus adjuvant compared with a mixture of the seven conjugates plus adjuvant, and of several variables thought to augment immunogenicity. These include approaches for decreasing suppressor cell activity or increasing helper T-lymphocyte activity (low dose cyclophosphamide or anti-CTLA-4 MAb), different saponin adjuvants at various doses (QS-21 and GPI-0100), and different methods of formulation (lyophilization and use of polysorbate 80). We find that: (1). Immunization with the heptavalent-KLH conjugate plus GPI-0100 vaccine induces antibodies against the seven antigens of comparable titer to those induced by the individual-KLH conjugate vaccines, high titers of antibodies against Tn (median ELISA titer IgM/IgG 320/10240), STn (640/5120), TF (320/10240), MUC1 (80/20480), and globo H (640/40); while lower titers of antibodies against Lewis(y)()(160/0) and only occasional antibodies against GM2 are induced. (2). These antibodies reacted with the purified synthetic antigens by ELISA, and with naturally expressed antigens on the cancer cell surface by FACS. (3). None of the approaches for further altering the suppressor cell/helper T-cell balance nor changes to the standard formulation by lyophilization or use of polysorbate 80 had any impact on antibody titers. (4). An optimal dose of saponin adjuvant, QS-21 (50 microg) or GPI-0100 (1000 microg), is required for optimal antibody titers. This heptavalent vaccine is sufficiently optimized for testing in the clinic.