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M Strand

Publications and source records attributed to M Strand.

At least 37 records · Page 2Linked to original sources
PubMed1994

Developmentally regulated localization and phosphorylation of SmIrV1, a Schistosoma mansoni antigen with similarity to calnexin.

Potential molecular targets of a protective humoral immune response against schistosomiasis have previously been identified based on their enhanced immunogenicity in mice vaccinated with irradiated cercaria as compared to chronically infected mice. One of these antigens, IrV1, has been molecularly cloned and its sequence shown to be similar to the molecular chaperone calnexin. In this investigation, we partially characterized IrV1 from different developmental stages of the schistosome. Immunoprecipitation studies with antibodies raised against a portion of recombinant IrV1 demonstrated its presence in cercaria, schistosomula, and adult worms with an apparent molecular mass on SDS-polyacrylamide gel electrophoresis of 90 kDa. There was an approximate 6-fold increase in protein expression level during the cercaria to schistosomula transformation. Consistent with a potential role as a molecular chaperone, IrV1 was associated with several metabolically labeled proteins in co-immunoprecipitation studies with the adult worm tegumental fraction. Similar to calnexin, IrV1 was metabolically labeled with 32P in adult worms on serine and threonine residues and was one of the major phosphoproteins of this stage. This phosphorylation was developmentally regulated and coincided with the transformation of cercaria into schistosomula. The localization was also stage-specific as IrV1 was transported from internal regions of cercaria to the outer tegumental layer of schistosomula. The presence of IrV1 on the surface of schistosomula, an unprecedented localization for this family of endoplasmic reticulum proteins, supports additional studies of the immunoprophylactic potential of this molecule.

Amino Acid Sequence↗
PubMed1994

Schistosoma mansoni: Sm23 is a transmembrane protein that also contains a glycosylphosphatidylinositol anchor.

Sm23, a surface protein of the human parasite Schistosoma mansoni, belongs to the family of "cysteine-rich, hydrophobic proteins," which are expressed on mammalian hematopoietic cells or tumor cells. Sm23 shares the highly conserved hydrophobicity profile of these proteins, which predicts four transmembrane segments, but is in addition linked to the membrane by a glycosylphosphatidylinositol (GPI) anchor. Our results suggest that Sm23 uses both the potential transmembrane domains and the GPI anchor for membrane insertion: (a) Sm23 was not released from the surface after cleavage with phosphatidylinositol-specific phospholipase C (PIPLC). (b) In a Triton X-114 phase-separation system, native [3H]ethanolamine- or [35S]methionine-labeled Sm23 partitioned into the detergent phase. Upon removal of the GPI anchor by PIPLC, the majority of the molecules stayed in the detergent-phase as expected of a transmembrane protein. (c) When full-length recombinant Sm23 was transcribed and translated in vitro, the polypeptide chain was inserted into microsomal membranes: Sm23 stayed associated with the membranes when they were incubated with carbonate buffer at pH 11.5, and membrane bound Sm23 was protected from digestion with proteinase K. (d) Recombinant Sm23, when expressed in the baculovirus expression system, was transported to the surface of infected insect cells, and similarly to the native protein it was not released from these cells after cleavage with PIPLC.

Animals↗
PubMed1994

Structural and functional similarities between synthetic HIV gp41 peptides and defensins.

Synthetic peptides derived from the putative immunosuppressive region of HIV-1 gp41 were examined to identify the amino acids required for suppressive activity. The active sequence was similar to the pseudosubstrate regulatory peptide of protein kinase C and to a sequence from the cyclic immune peptides known as defensins. The activity of the gp41 peptides appeared to be dependent upon a structural conformation, possibly a looped motif like that found in defensins. This similarity to defensins, which can insert themselves into lipid bilayers, was further strengthened by demonstrating that gp41 peptides have the ability to associate with liposomes and lymphocyte membranes. These results suggest that the immunosuppressive region of gp41 may participate in the membrane interactions required for viral fusion and infectivity.

Amino Acid Sequence↗
PubMed1994

Schistosoma mansoni: immunolocalization of two different fucose-containing carbohydrate epitopes.

We have used two monoclonal antibodies, 128C3/3 and 504B1, to immunolocalize their carbohydrate epitopes in different developmental stages of Schistosoma mansoni. Both epitopes contain fucose: mAb 128C3/3, as we have shown previously, recognizes fucose in a novel, possibly internal linkage (Levery et al. 1992) while mAb 504B1, as we show here, bound to the Le(x) epitope, which contains fucose alpha 1-->3 linked to N-acetyl-glucosamine. The tissue expression of these epitopes was strikingly different and both elicit an immune response in infected hosts. The mAb 128C3/3-defined epitope was exposed on the surface of all larval stages but not on adult worms; however, it was found in the excretory system of adult worms of both sexes. In contrast, surface expression of the Le(x) epitope was initiated after the transformation of cercariae to schistosomula and was maintained throughout the adult life in both sexes.

Animals↗
PubMed1994

Cross-reactivity of anti-human immunodeficiency virus type 1 gp41 antibodies with human astrocytes and astrocytoma cell lines.

An antigen expressed by astrocytes in human brain tissue and by various human astrocytoma cell lines was shown to cross-react with a monoclonal antibody generated against amino acids (aa) 584 to 609 of the transmembrane protein gp41 of human immunodeficiency virus type 1 (HIV-1). This region is an immunodominant segment of gp41, and high levels of antibodies against this epitope have been detected in both serum and cerebrospinal fluid of HIV-infected individuals at all stages of HIV infection. Immunohistochemistry with this monoclonal antibody demonstrated the presence of a cross-reactive antigen in human brain tissue, with an increased frequency and intensity of staining in HIV-positive individuals when compared with HIV-negative controls. By using a panel of HIV-positive and -negative sera, we show that antibodies in HIV-positive serum specifically bound to the surfaces of human astrocytoma cells. HIV-positive sera depleted of antibodies recognizing gp41 aa 584 to 609 showed a significant diminution in cell surface binding. Conversely, the serum antibodies that bound to and were eluted from the aa 584 to 609 peptide also bound to the astrocyte cell surface. To identify the target antigen, the immunoreactivity of three astrocytoma cell lines was examined. By immunoprecipitation of metabolically labeled cell lysates and Western blot (immunoblot) analysis, we identified a protein of approximately 100 kDa as the target antigen. Cross-reactive antibodies between HIV proteins and astrocyte epitopes, such as this 100-kDa protein and others previously reported, suggests that an autoimmune response against these target antigens may disrupt the normal functions of astrocytes.

AIDS Dementia Complex↗
PubMed1993

Protective immunity in baboons vaccinated with a recombinant antigen or radiation-attenuated cercariae of Schistosoma mansoni is antibody-dependent.

Mice vaccinated with radiation-attenuated cercariae of Schistosoma mansoni exhibit high levels of resistance to challenge infection. We have previously shown that sera from these mice recognize polypeptides that are expressed on the surface of newly transformed schistosomula. We have cloned and sequenced a cDNA that encodes a 62-kDa portion of one of these polypeptides. Vaccination of mice with this 62-kDa polypeptide (designated rlrV-5) elicits high antibody titers and significant resistance to challenge infection. We report here the results of a vaccination trial in baboons with the rlrV-5 or radiation-attenuated cercariae. rlrV-5 was presented either in the form of protein micelles or complexed with the outer membrane protein of meningococcus to form proteosomes. The level of protection achieved in these groups ranged from 0 to 54%, with a mean of 27.7%. In baboons exposed to radiation-attenuated cercariae the level of protection was very high, with a mean of 84%. The resistance observed after vaccination with rlrV-5 or radiation-attenuated cercariae was reflected in the overall histopathology. Vaccination of baboons with rlrV-5 or radiation-attenuated cercariae elicited an antibody response against epitopes exposed on the surface of newly transformed schistosomula. In the case of baboons vaccinated with radiation-attenuated cercariae, this response was not limited to epitopes encompassed by rlrV-5. Analysis of individual baboon sera by ELISA demonstrated that there was a direct correlation between the anti-rlrV-5 titer and resistance to challenge worm burden, suggesting that the immunoprotective mechanism is antibody-dependent.

Animals↗
PubMed1993

Destabilization of tracts of simple repetitive DNA in yeast by mutations affecting DNA mismatch repair.

The genomes of all eukaryotes contain tracts of DNA in which a single base or a small number of bases is repeated. Expansions of such tracts have been associated with several human disorders including the fragile X syndrome. In addition, simple repeats are unstable in certain forms of colorectal cancer, suggesting a defect in DNA replication or repair. We show here that mutations in any three yeast genes involved in DNA mismatch repair (PMS1, MLH1 and MSH2) lead to 100- to 700-fold increases in tract instability, whereas mutations that eliminate the proof-reading function of DNA polymerases have little effect. The meiotic stability of the tracts is similar to the mitotic stability. These results suggest that tract instability is associated with DNA polymerases slipping during replication, and that some types of colorectal cancer may reflect mutations in genes involved in DNA mismatch repair.

DNA Repair↗
PubMed1993

Molecular cloning and expression of SmIrV1, a Schistosoma mansoni antigen with similarity to calnexin, calreticulin, and OvRal1.

Protective immunity against schistosomiasis induced by vaccination of mice with irradiated cercaria can be passively transferred to uninfected mice with sera or IgG. Antigens that are uniquely or more strongly recognized by such protective sera compared with sera from infected unprotected mice have been identified previously. Two genes, SmIrV1 and SmIrV5, were selected from an adult worm cDNA library by screening with antibodies raised against these candidate vaccine proteins. Active immunization with the SmIrV5 protein induces high levels of protection in mice. We report here the molecular cloning and sequencing of SmIrV1 which contains a deduced amino acid sequence of 582 residues with similarity to three proteins: calnexin, calreticulin, and OvRal1, a surface antigen of the filarial nematode Onchocerca volvulus. SmIrV1 can be divided into three regions: a neutral N-terminal region with a putative signal sequence, followed by a proline- and tryptophan-rich P region in which two sets of sequences are repeated four times and a C-terminal region which is highly acidic with an isoelectric point of 4.7. We expressed the P and C regions of SmIrV1 and showed that this polypeptide reacts with sera of immunized as well as chronically infected mice.

Amino Acid Sequence↗
PubMed1993

Assessment of the antibody response to the immunosuppressive/immunodominant region of HIV gp41 in a 5-year longitudinal study.

The antibody response of HIV-infected individuals to the 581-609 amino acid (aa) region of HIV-1 gp41 containing the putative immunosuppressive and immunodominant sequences was examined. Sera collected every 5 to 6 months over a period of 5 years from 50 HIV-1-infected homosexual and bisexual men, 25 of whom progressed to AIDS during the collection period, were monitored for changes in ELISA reactivity against synthetic peptides encompassing aa581-609 of gp41. The data obtained in this blinded, historical prospective study were analyzed with respect to changes in mean ELISA absorbance over time and differences in absorbance between patient groups (those who progressed to AIDS and those who did not). No correlation was found between time or disease state and the presence of antibodies to the aa581-597 immunosuppressive sequence. In contrast, ELISA absorbance against the aa598-609 immunodominant sequence continued to increase over time in both the AIDS and non-AIDS groups. The rate of increase in absorbance was similar for both groups; however, the AIDS group had a significantly higher mean absorbance level at the outset of the study and maintained this difference throughout the study. This region of gp41 has been proposed to play a role in complement-mediated antibody-dependent enhancement of infection.

Acquired Immunodeficiency Syndrome↗
PubMed1993

Schistosoma: a 200-kDa chemotherapeutic target antigen is differentially localized in African vs Oriental species.

A 200-kDa protein of the African schistosome Schistosoma mansoni has been identified as a target of antibodies that act in synergy with praziquantel. Treatment of worms with praziquantel exposes selective epitopes of the 200-kDa protein on the surface of S. mansoni and transfer of a monoclonal antibody recognizing the 200-kDa protein at the time of drug treatment restores the effectiveness of praziquantel against infections in B-cell-depleted mice. In the present study, a cross-reactive 200-kDa protein was identified in the three major schistosome species by Western blot analysis using a polyclonal rabbit antiserum recognizing the 200-kDa protein from S. mansoni. Surprisingly, three monoclonal antibodies generated against immunogenic epitopes of the 200-kDa protein did not recognize the 200-kDa protein of the Oriental species Schistosoma japonicum, although they did recognize the 200-kDa protein of another African species Schistosoma haematobium. Furthermore, in the case of S. japonicum, treatment with praziquantel did not expose the 200-kDa protein on the worm surface. Even acetone treatment, which makes surface epitopes more accessible, did not expose the 200-kDa protein on the surface of S. japonicum. In contrast, the 200-kDa protein of S. haematobium was exposed following treatment with praziquantel, and acetone fixation resulted in significantly increased reactivity of the rabbit alpha 200-kDa antiserum with the surface of both S. mansoni and S. haematobium worms.

Animals↗
PubMed1993

Schistosoma mansoni: immunoreactivity of human sera with the surface antigen Sm23.

Sm23, a surface antigen of Schistosoma mansoni, elicits an immune response in patients infected with the parasite, although to a lesser degree than Sj23, the homologue of Sm23 in Schistosoma japonicum. To characterize the host immune response to Sm23, we have expressed full-length Sm23 using the baculovirus expression system and have also expressed the N-terminal 133 amino acids and the 85 C-terminal amino acids by means of the prokaryotic vector pGEX. A total of 70 sera from patients from Sudan and Egypt were examined for alpha-Sm23 antibodies in an ELISA and in Western blot analysis using both the C-terminal polypeptide and the full-length protein. A subset of these sera was also tested for reactivity with the N-terminal polypeptide. The alpha-Sm23 antibody titers in infected patients varied widely and were not correlated with egg counts or age of the individuals. Most of the seroreactivity was directed against the C-terminal polypeptide.

Age Factors↗
PubMed1993

Detection and partial characterization of glycosylphosphatidylinositol-specific phospholipase activities from Fasciola hepatica and Schistosoma mansoni.

The tegument of trematodes serves as a dynamic host-parasite interface where surface antigens are shed in a process of immune evasion. Phospholipases, which could provide an enzymatic mechanism for release of glycosylphosphatidylinositol (GPI)-anchored proteins, were detected in detergent extracts of adult worms of Fasciola hepatica and cercaria and adult worms of Schistosoma mansoni. The enzymatic activities were partially characterized from both adult worm species and demonstrated a preference for [3H]GPI substrate over [3H]PI. Lipase activities from both species were sensitive to sulfhydryl-modifying reagents and the detergents CHAPS and n-octylglucoside. The presence of 1 M ammonium sulfate increased the enzyme activity in adult worms of both species by 8-11-fold and in cercaria by 146-fold, whereas other conditions of high ionic strength were inhibitory. Such stimulation suggested dissociation of a negative inhibitor which is prominent in the cercarial stage. The schistosome extract, which was partially sensitive to cation chelators and o-phenanthroline, contained a GPI-phospholipase D activity. In contrast, the F. hepatica extract contained a cation-independent phospholipase C activity which was partially purified and shown by gel filtration to have a molecular mass of 30,000-80,000.

Animals↗
PubMed1992

Induction of protective immunity in mice using a 62-kDa recombinant fragment of a Schistosoma mansoni surface antigen.

Mice exposed to radiation-attenuated cercariae of Schistosoma mansoni are highly resistant to challenge infection, and sera from these mice can confer partial resistance when transferred to naive recipients. These sera recognize Ag present in schistosomular and adult worms, among them an Ag of 200 kDa. A cDNA encoding a 62-kDa portion of this Ag was cloned; the deduced amino acid sequence of this cDNA clone shares homology with myosins of other species. To assess the immunoprophylactic potential, we carried out vaccination trials in mice using the recombinant polypeptide expressed as a fusion protein with beta-galactosidase presented in the form of proteosome complexes with the outer membrane protein of meningococcus. The level of protection achieved was 32%, and this level could be increased to 75% by removal of those amino acids included in the fusion protein that were derived from the vector to yield a polypeptide, designated rIrV-5. A similar level of protection was achieved when mice were immunized with the same dose of rIrV-5 in the form of protein complexes but without outer membrane protein, suggesting that protection did not require the use of adjuvant. However, at least three immunizations were necessary to achieve protection. Using mAb and sera from mice vaccinated with rIrV-5, we demonstrated that the native protein recognized by antibodies against rIrV-5 is a 200-kDa protein that is expressed on the surface of newly transformed schistosomula. The protection achieved with rIrV-5 in mice encourages additional studies of its potential as a vaccine candidate for the prevention of schistosomiasis.

Amino Acid Sequence↗
PubMed1992

Effective alpha-particle-mediated radioimmunotherapy of murine leukemia.

The specificity, toxicity, and efficacy of alpha-particle-mediated radioimmunotherapy of murine erythroleukemia was assessed by use of tumor-specific monoclonal antibody 103A labeled with 212Bi. Forty % of the injected dose/g tissue targeted to neoplastic spleens within 1 h after i.v. injection. When 212Bi-103A was injected on day 13 of disease, a dose-dependent response was achieved, as measured by a reduction in splenomegaly and absence of liver metastasis. Mice treated with 212Bi-103A on day 8 of disease showed no histological evidence of erythroleukemia on day 22 and survived significantly longer (median, 118 days) than mice treated with 212Bi-control IgG (78 days) or untreated mice (63 days), indicating successful specific radioimmunotherapy.

Alpha Particles↗
PubMed1992

Characterization of a series of novel fucose-containing glycosphingolipid immunogens from eggs of Schistosoma mansoni.

Lipid extracts of eggs, worms, and cercariae of the parasitic trematode Schistosoma mansoni have been shown to contain a large number of highly immunogenic glycolipids (Weiss, J. B., Magnani, J. L., and Strand, M. (1986) J. Immunol. 136, 4275-4282). Three fractions of schistosome egg glycolipids were selected on the basis of their reactivity with an anti-schistosome monoclonal antibody (128C3/3), which recognizes a developmentally regulated carbohydrate epitope present on both glycolipid and glycoprotein antigens from S. mansoni. These fractions were purified by silica gel chromatography and preparative high performance thin layer chromatography and characterized by monosaccharide, fatty acid, and linkage analysis with gas chromatography-mass spectrometry, as well as by positive and negative ion fast atom bombardment-mass spectrometry. The immunogens were shown to be glycosphingolipids having homologous structures based on a highly novel extension of glucosylceramide. Monosaccharide inhibition studies indicated that the epitope recognized by 128C3/3 residues in an outer region of the immunogens consisting of Fuc2GlcNAc (where Fuc is fucose) repeating units. The largest antigen characterized may have the following structure, based on the evidence presented in this paper. [sequence: see text] The evidence indicated the existence of a series of glycan structures created by deletions of one or more Fuc1----3 side chains from the above structure.

Animals↗