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S L James

Publications and source records attributed to S L James.

At least 91 records · Page 5Linked to original sources
PubMed1986

Defective vaccine-induced resistance to Schistosoma mansoni in P strain mice. III. Specificity of the associated defect in cell-mediated immunity.

In contrast to many other strains, inbred P strain mice fail to develop significant levels of resistance to challenge Schistosoma mansoni infection as a result of prior vaccination with radiation-attenuated cercariae. In this study, the relationship between defects in resistance and development of cell-mediated immune reactivity was examined. Although splenocytes from immunized P mice demonstrated deficiencies in production of macrophage-activating lymphokine(s) in response to either antigenic or mitogenic stimulation, other aspects of T lymphocyte responsiveness including blastogenesis, production of interleukin 2, interleukin 3 and macrophage chemotactic factor, as well as helper cell function for secondary plaque-forming cell response to a T-dependent antigen and allospecific cytolytic T cell reactivity, appeared to be comparable with those of C57BL/6 mice, a strain that is protected by vaccination against S. mansoni. FACS comparison revealed no significant deficits in percentages of Thy-1+, Lyt-1+, or L3T4+ splenocytes in vaccinated P mice. The P-associated defect in production of macrophage-activating factor appeared to be at the level of the T cell rather than the antigen-presenting cell, because macrophages from P mice could reconstitute the lymphokine-producing capacity of T-enriched splenocytes from immunized, resistant (C57BL/6 X P) F1 or B10.P mice, whereas the converse was not true. These results indicate that vaccinated P mice have a selective defect in T cell function for production of macrophage-activating lymphokine, which is manifested as a failure to produce activated larvicidal macrophages at the site of specific antigen challenge in vivo and may be associated with the failure of this strain to become resistant to S. mansoni.

Animals↗
PubMed1986

Immunochemical characterization and purification of Sm-97, a Schistosoma mansoni antigen monospecifically recognized by antibodies from mice protectively immunized with a nonliving vaccine.

Mice protected against Schistosoma mansoni infection by intradermal (i.d.) vaccination with nonliving schistosomula or soluble extracts of larval or adult schistosomes (SCHLAP and SWAP, respectively) produce antibodies that react by Western blot analysis with one antigen of Mr (X 10(-3)) 97 in SWAP prepared in the presence of protease inhibitors and two antigens of Mr (X 10(-3)) 95 and 78 in SWAP prepared in their absence. Vaccine antibodies also immunoprecipitated a single 97k molecule, with a pI of 5.5, from detergent extracts of [35S] methionine-labeled schistosomes. Three hybridomas, produced from spleen cells of i.d. immunized mice, all recognized both the 95k/78k doublet and the 97k antigen, indicating that the two lower Mr components are degradation products of the same 97k molecule. The 97k/95k/78k complex (Sm-97) was purified by affinity chromatography and found to constitute 0.5% of the total protein in SWAP. 125I-concanavalin A bound weakly to purified Sm-97, indicating that this antigen is minimally glycosylated. By indirect immunofluorescence, Sm-97 was localized to regions just below the tegumental and gut syncitia of adult worms. Mice protected by i.d. vaccination produced high titers (1:10,240) of anti-Sm-97 antibodies, whereas chronically infected mice responded at a much lower level (titer 1:640). In contrast, mice protectively immunized with irradiated cercariae and mice nonprophylactically inoculated by the i.v. route failed to produce detectable anti-Sm-97 antibodies. Competitive radioimmunoassays performed with 125I-labeled monoclonal antibodies and purified antigen defined at least two distinct epitopes on Sm-97. Antibodies from i.d. vaccinated mice recognized both monoclonal antibody-defined epitopes, whereas anti-Sm-97 antibodies in chronic infection sera recognized neither. Finally, purified Sm-97 was shown to elicit delayed-type hypersensitivity in i.d. vaccinated mice, suggesting that this molecule is also capable of evoking cell-mediated responses, a finding consistent with its proposed function as a vaccine immunogen.

Animals↗
PubMed1986

Identification of paramyosin as schistosome antigen recognized by intradermally vaccinated mice.

Mice immunized intradermally with extracts of Schistosoma mansoni in combination with the adjuvant BCG are significantly protected against subsequent infection with living larval forms of the parasite. Remarkably, these vaccinated animals produce antibodies predominantly against a single parasite protein of molecular weight 97 kilodaltons (Sm-97). A complementary DNA that encodes about half of the Sm-97 molecule has now been cloned and sequenced. Analysis of the deduced amino acid sequence reveals a protein containing periodic repeats of hydrophobic amino acids characteristic of an alpha-helical coiled-coil structure. The deduced amino acid composition of the cloned gene and several properties of the native protein are similar to that of paramyosin, an alpha-helical protein that forms the core for myosin filaments in invertebrate muscle. Paramyosin was isolated from Schistosoma mansoni adult worms and antibodies to Sm-97 were shown to react with this molecule as well as with a known paramyosin from molluscan muscle.

Amino Acid Sequence↗
PubMed1986

Identification of a genetic locus, Rsm-1, controlling protective immunity against Schistosoma mansoni.

Mice of most inbred strains develop moderate to high levels of resistance to challenge infection on vaccination with radiation-attenuated cercariae of Schistosoma mansoni. P strain mice, however, fail to display significant protective immunity after exposure to the same vaccine. To examine the genetic basis of this polymorphism in host immunity, vaccine-induced resistance was examined in (C57/BL6J X P/N)F1, F2, and reciprocal backcross generations. The defective immunity which characterizes the P strain parent was found to be inherited in a fully recessive manner and to be controlled by a single genetic locus, which we have designated Rsm-1. Linkage analyses revealed that Rsm-1 is not genetically associated with the major histocompatibility complex (chromosome 17), the immunoglobulin heavy chain locus (chromosome 12), or a single locus influencing defective anti-schistosomulum IgM antibody responses in the P parental stock. These data provide the first example of single gene control of vaccine-induced immunity against a helminth infection. Because P mice are also defective in their capacity to develop tumoricidal macrophages and in their immunity to Leishmania major, genes encoded by the Rsm-1 locus may play a general role in resistance to infection and malignancy.

Animals↗
PubMed1986

Induction of protective immunity against Schistosoma mansoni by a nonliving vaccine. II. Response of mouse strains with selective immune defects.

The efficacy of a new vaccination procedure against Schistosoma mansoni, involving intradermal injection of nonliving antigen combined with the bacterial adjuvant Mycobacterium bovis strain bacillus Calmette Guérin, was tested in several strains of mice. Development of protection against subsequent infection was compared with in vivo skin test reactivity and in vitro humoral reactivity to soluble and surface-associated schistosome antigens. Significant levels of resistance and immune response were displayed by many inbred mouse strains, including C57BL/6J, C3H/HeN, and CBA/J, as well as outbred Swiss-Webster mice. However, no definite correlation was observed between the level of any particular immune response and the level of resistance to challenge S. mansoni infection. Development of protective immunity was also examined in mice with various immune defects, to determine whether these responses are relevant to resistance in this model. Animals with defective specific immediate hypersensitivity response due to deficiencies in IgE (SJL/J) or mast cell (W/Wv) production displayed strong resistance as a result of immunization. Likewise, mice bearing the lpsd (C3H/HeJ) or xid (CBA/N) mutations, affecting cellular or humoral response to certain thymus-independent antigens, developed significant levels of resistance after immunization. A/J mice, with defects in cellular recognition of bacterial endotoxin as well as deficiencies in natural killer cell activity and complement function, also showed significant protective immunity. Thus, these reactivities do not appear to be essential to the resistance against S. mansoni induced by the nonliving vaccine. Two nonresponder strains were identified, P and BALB/c. P mice were defective in specific delayed hypersensitivity response as well as resistance to infection. However, BALB/c mice showed no obvious immune deficiencies at the time of challenge. These results agreed with previous findings in mice immunized by exposure to radiation-attenuated cercariae with one exception; BALB/c mice were protected by vaccination with irradiated cercariae but not by the nonliving vaccine. Thus, further examination of immune response in mice identified in this study as high and low responder strains should allow characterization of critical immune resistance mechanisms induced by the nonliving vaccine, as well as immune mechanisms operating in common between these two models of resistance to S. mansoni.

Animals↗
PubMed1986

Induction of protective immunity against Schistosoma mansoni by a nonliving vaccine. III. Correlation of resistance with induction of activated larvacidal macrophages.

Mice protected against Schistosoma mansoni infection by intradermal (i.d.) immunization with nonliving larval or adult worm antigens plus bacterial adjuvant developed 24-hr skin test responsiveness to schistosome antigens with the histologic features of delayed hypersensitivity. Intraperitoneal antigen injection elicited a mononuclear cell-enriched exudative population containing macrophages activated for direct cytotoxicity against schistosomula and tumor cell targets. This was likely to be due to in vivo exposure to macrophage-activating lymphokines, since these cells were unresponsive to further lymphokine stimulation in vitro and splenocytes from immunized mice reacted to specific in vitro antigen challenge by production of lymphokines capable of conferring larvacidal activity upon control macrophages. In contrast, mice treated with schistosome antigens by i.v. injection, which were not protected against challenge infection, failed to develop delayed hypersensitivity or activated macrophages in response to specific antigen challenge in vivo, and the titers of macrophage-activating lymphokine produced by in vitro antigen-stimulated splenocytes from these mice were threefold to fourfold lower than those produced by cells from animals immunized by the i.d. route. Thus, sensitization for cell-mediated immune responses including lymphokine production and macrophage activation correlated with induction of resistance to S. mansoni in this model of vaccination.

Animals↗
PubMed1986

Kinetic correlation of the acquisition of resistance to immune attack in schistosomula of Schistosoma mansoni with a developmental change in membrane potential.

When Schistosoma mansoni cercariae penetrate the skin of the mammalian host they rapidly pass from fresh water to a high salt physiologic environment and transform into schistosomula. Following this transition, the parasites migrate from the skin to the lungs during which time they change from being highly susceptible to immune attack to being refractory, as measured by in vitro cytotoxicity assays. In this study, in vivo or in vitro schistosomula of different ages were examined for developmentally linked changes in membrane function which might correlate with the attainment of the resistant state. In particular, alterations in the distribution of tetraphenylphosphonium (TPP+), a synthetic lipophilic cation which shows a potential dependent partition across membranes, were followed. Three-hour-old schistosomula, which are greater than 75% susceptible to antibody-dependent complement-mediated attack or lymphokine-activated macrophage-mediated cytotoxicity, acquired TPP+ at a similar rate and steady state level to 5-day-old lung worms, which were completely resistant to both these effector mechanisms. The addition of ouabain, a Na+/K+-ATPase inhibitor, caused a 50% decrease in both the rate and steady state of TPP+ uptake by 3 h parasites but had little effect on these parameters in lung worms. Valinomycin, a K+-ionophore, completely inhibited TPP+ influx in both stages. The characteristics of TPP+ efflux from 3-h and 5-day-old parasites preloaded with the cation were found to be dissimilar. Whereas 30% of acquired TPP+ was lost from lung worms within 2 h, only 10% of acquired cation was released from 3-h schistosomula during the same period.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗
PubMed1986

Post lung-stage schistosomula of Schistosoma mansoni exhibit transient susceptibility to macrophage-mediated cytotoxicity in vitro that may relate to late phase killing in vivo.

Studies of protective immunity against Schistosoma mansoni in immunized mice suggest that a proportion of challenge parasites may be eliminated after they have passed through the lungs of the host several days after infection; however, no potential immune effector mechanism of resistance against this stage of the parasite has yet been identified, since schistosomes have been shown to rapidly become resistant to antibody-dependent killing mechanisms. In this study, different development stages of S. mansoni were examined for their susceptibility to in vitro cytotoxicity by lymphokine-activated macrophages. As previously shown, newly transformed larvae were readily killed by lymphokine-treated peritoneal macrophages or the macrophage cell line IC-21 (80% mortality over 48 h in vitro), whereas 7 and 10 day old lung-stage parasites had become refractory to macrophage effects. However, after 2 to 2 1/2 weeks of development in vivo, juvenile parasites recovered from the liver were again susceptible to activated macrophage-mediated cytotoxicity (25-65% mortality). Ultrastructural studies of 2 1/2 week old parasites co-cultured with activated IC-21 cells revealed that damage was largely restricted to the areas beneath the parasite surface and gut syncitia; surface membrane disruption was not evident. This late stage of susceptibility was transient and by 4 to 6 weeks liver-stage worms had again become refractory to macrophage killing. The interaction of post lung-stage parasites with activated macrophages was antibody independent. Furthermore, schistosomes isolated from the portal circulation 2 1/2 weeks after infection showed no evidence of surface-bound immunoglobulin in a quantitative immunofluorescence assay, nor did antisera from chronically infected mice (CIS) or mice vaccinated with irradiated cercariae (VS) react with the surface of these parasites in vitro, making the possibility of direct antibody-dependent killing mechanisms unlikely. However, both CIS and VS did recognize excretory/secretory proteins synthesized by 2 1/2 week old liver-stage schistosomes, including a major antigen of approximate Mr (X 10(-3] 220 (220K). It is therefore possible that such antigens might participate in protective immunity, for example via immune complex formation or activation of sensitized T cells. These observations support the role of macrophages as immune effector cells in mice immunized against Schistosoma mansoni, and provide the first physiologically relevant mechanism whereby the immune system might recognize and kill post-lung stage schistosomes.

Animals↗
PubMed1985

Deaf children's revision behaviors in conversations.

This study was designed to investigate school-age deaf children's responses to a listener's indication of communication breakdown and to determine if the deaf children's responses were related to their syntactic or semantic abilities and/or typical mode of communication. Subjects were 14 deaf children between the ages of 5 yr, 10 mo and 8 yr, 11 mo. Approximately once every 3 min during a 60-min conversation, the experimenter said, and simultaneously signed, "What?" The child's responses to the "What?" probe were classified as Revision, Repetition, and No Response. Revisions were categorized into eight subcategories involving changes in sentence constituents and supraconstituents. Revisions were the most frequent responses when communication broke down. Most of the children's revisions involved changes in sentence constituents. The frequency of revisions, repetitions, and no responses was related to the children's level of syntactic production.

Child↗
PubMed1985

Comparison of immune responses between high and low responder strains of mice in the concomitant immunity and vaccine models of resistance to Schistosoma mansoni.

Mice of the inbred P strain fail to develop significant resistance to challenge Schistosoma mansoni infection at 6 weeks after either low-grade primary infection or vaccination with attenuated homologous parasites, in contrast to other strains such as C57Bl/6, and thus provide a model for comparison of potential immune resistance mechanisms in low versus high responder animals. In this study, the antigen-specific cellular responses found to correlate with resistance in these strains were delayed cutaneous hypersensitivity, production of macrophage activating lymphokine and macrophage larvicidal activity, all of which were greater in infected or vaccinated C57Bl/6 mice than in similarly immunized but non-resistant P mice. Humoral responses correlating with resistance were IgM reactivity to schistosomula surface antigens in both infected and vaccinated animals, as well as both IgM and IgG reactivity to soluble schistosome antigens in infected mice. Immune responses that showed no relationship with resistance included IgG reactivity to larval surface antigens and immediate hypersensitivity to soluble worm antigens. In infected mice, neither granuloma size nor extent of hepatic fibrosis correlated positively with resistance to challenge infection. Thus, similarities exist between patterns of resistance and immune response at this early time after immunization with either viable or attenuated parasites. These observations suggest that common immune effector mechanisms could be involved, with activated macrophages playing a central role in resistance.

Animals↗
PubMed1985

Ultrastructural studies of the killing of schistosomula of Schistosoma mansoni by activated macrophages in vitro.

Immunologically activated murine macrophages have been shown elsewhere to kill skin stage schistosomula of Schistosoma mansoni in vitro, in a manner analogous to the extracellular killing of tumour cell targets. In this study, the kinetics of the interaction between activated macrophages and larval targets and the resultant ultrastructural changes in parasite morphology that culminated in death have been analysed in detail. Unlike granulocyte-mediated schistosomular killing, macrophage-mediated cytotoxicity did not appear to be directed against the surface tissues of the parasite. Macrophages adhered only transiently following initiation of the cultures, yet changes in the subtegumental mitochondria and muscle cells of the larva were detected within the first hour of incubation. Progressive internal disorganisation followed rapidly, but the tegument and tegumental outer membrane remained intact, to form a 'shell' that maintained the general shape of the parasite. Such changes were recognised irrespective of whether the effector cell population comprised peritoneal macrophages activated by lymphokine treatment in vitro, or by infection with Mycobacterium bovis (strain BCG), or S. mansoni in vivo. That macrophages rather than contaminating granulocytes or lymphocytes, had mediated the observed damage was demonstrated by the use of a lymphokine treated macrophage cell line, IC-21. The observation that macrophage cytotoxicity is directed against internal organelles rather than the tegumental outer membrane of this multicellular target, may help to elucidate the general mechanism of extracellular killing by these cells.

Animals↗
PubMed1985

Induction of protective immunity against Schistosoma mansoni by a nonliving vaccine is dependent on the method of antigen presentation.

A preparation of nonliving parasite antigens containing both soluble and particulate components of frozen-and-thawed invasive larvae was used to immunize C57BL/6J mice against challenge Schistosoma mansoni infection. The method of antigen presentation was observed to be critical to the ability of this preparation to induce protective immunity, because intradermal administration in conjunction with a bacterial adjuvant (BCG) resulted in strong protection against challenge parasites (51% reduction in worm burden in six experiments), whereas i.v. injection of the same antigenic preparation was completely ineffective. Induction of resistance was accompanied by specific immune responsiveness toward schistosome antigens. Protection correlated more closely with sensitization for specific delayed hypersensitivity than with elicitation of circulating antibodies to larval surface antigens or immediate hypersensitivity in these models. These results suggest that it will be possible to design a defined vaccine against S. mansoni infection, but that identification of the route of antigen presentation that most effectively elicits relevant immune effector mechanisms will be crucial to the success of any vaccination protocol involving nonliving antigens.

Animals↗
PubMed1985

Induction of protective immunity against Schistosoma mansoni by a non living vaccine. I. Partial characterization of antigens recognized by antibodies from mice immunized with soluble schistosome extracts.

A single intradermal injection of frozen and thawed schistosomula in conjunction with the bacterial adjuvant Mycobacterium bovis strain Bacille Calmette Guerin, Phipps substrain (BCG) induced significant levels of resistance to challenge Schistosoma mansoni infection in C57BL/6 mice. Immunization with the aqueous fraction remaining after 100,000 X G centrifugation of the larval lysate was also protective under these conditions, suggesting that some immunogenic determinants may not be membrane associated. Frozen-thawed cercariae and soluble components of adult worms also protected against challenge infection in these experiments. These observations indicate that soluble immunogens are present in both early and late developmental stages of the parasite, and therefore may be good candidate antigens for an immunochemically defined vaccine against schistosomiasis. Induction of humoral reactivity against soluble or membrane antigens was examined in mice protected against cercarial challenge by prior exposure to frozen-thawed larvae, soluble larval, or soluble adult antigens plus BCG. Animals that were immunized with frozen-thawed larvae produced low but significant levels of antibodies against larval surface antigens when examined by indirect immunofluorescence or by immunoprecipitation of surface-labeled schistosomula. Mice immunized with soluble antigens, however, showed negligible antibody reactivity against surface membrane antigens. Because mice immunized with soluble antigens were resistant to challenge infection, these results strongly suggest that anti-surface membrane reactivity is not required in the mechanism of protective immunity in this model. Sera from mice immunized with either total freeze-thaw larval lysate or soluble schistosome extracts all showed strong reactivity against soluble antigens, as detected by ELISA. Western blot analysis showed these antisera to react with a restricted number of high m.w. antigens that were present both in schistosomula and in adult worms. These antigens are therefore likely to play a major role in the development of resistance in this model as immunogens and/or as targets of protective immune response.

Adaptation, Physiological↗
PubMed1984

Genetic control of macrophage activation for killing of extracellular targets.

Activated peritoneal macrophages from Corynebacterium parvum-treated mice of most inbred strains, including C57BL/6J (B), are cytotoxic to adherent 1023 sarcoma target cells as well as to larvae of Schistosoma mansoni. Macrophages from A/J (A) strain mice, on the other hand, are defective in this function. Segregation analysis of these two traits was compatible with the hypothesis that effective activation is, in each case, controlled by a single, dominant, autosomal gene. Typing of individual animals of the segregating backcross progeny and of AXB/BXA recombinant inbred strains for the expression of macrophage activation for tumoricidal and schistosomulicidal activity indicates that the genetic control of these two traits is closely linked or identical.

Animals↗
PubMed1984

Activation of macrophages to kill rickettsiae and Leishmania: dissociation of intracellular microbicidal activities and extracellular destruction of neoplastic and helminth targets.

Mononuclear phagocytes undergo dramatic changes during differentiation from bone marrow stem cells to resident tissue macrophages. Throughout differentiation, cells lose or acquire numerous morphologic, metabolic and functional capacities such that mature, resident macrophages of one tissue often bear little resemblance to resident cells of another. Superimposed on the intrinsic continuum of mononuclear phagocyte differentiation are the reactive changes in macrophages induced by endogenous and exogenous stimuli: the ability of mononuclear phagocytes to respond to a particular stimulus may also change with cell differentiation. This dynamic interaction of cell differentiation and response to a micro-environment, and the resulting heterogeneity among mononuclear phagocytes for many functional characteristics, is clearly illustrated by the effector activities of activated macrophages that we describe in this report. Despite the common regulatory events for induction and expression of transient nonspecific cytotoxic reactions effective against such diverse targets as rickettsiae, leishmania, schistosomula, and neoplastic cells, these effector functions can be dissociated by the cells that perform the effector activity, and the signals that regulate these activities. The differential susceptibility of the various targets to particular killing mechanisms induced by LK in responsive populations only adds to the complexity of these in vitro analyses. The details of effector functions of activated macrophages are unique for each target.

Animals↗