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S H Pincus

Publications and source records attributed to S H Pincus.

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

Human eosinophils stimulate DNA synthesis and matrix production in dermal fibroblasts.

Fibrosis is frequently found in diseases exhibiting tissue eosinophilia, such as some parasitic worm infections, eosinophilia-myalgia syndrome, and eosinophilic fasciitis. Previously, eosinophil extracts have been shown to induce proliferation in neonatal foreskin fibroblasts in vitro. To determine if living eosinophils can induce synthesis of DNA and components of the extracellular matrix in dermal fibroblasts, we cultured purified human eosinophils for 2 or 7 days in the presence of the eosinophil-active cytokines granulocyte/macrophage colony stimulating factor, interleukin-3, or interleukin-5, and added eosinophil-conditioned medium to cultures of dermal fibroblasts. Using flow cytometry, we found that eosinophil-conditioned medium increased by two-fold the percentage of fibroblasts in S-phase. This stimulation of fibroblast DNA synthesis was corroborated using a standard tritiated thymidine assay and the two methods were shown to correlate well with each other. Eosinophil-conditioned medium stimulation of DNA synthesis was dose dependent and conditioned medium from eosinophils treated with any one of the three cytokines induced increased DNA synthesis. Treatment of fibroblasts with cytokines alone did not induce enhanced DNA synthesis. Eosinophil-conditioned medium also affected fibroblast matrix production. Eosinophil-conditioned medium induced a two-fold increase in soluble and cell-associated fibroblast glycosaminoglycan production and a 76% increase in collagen production. These observations support the concept that eosinophils may be active contributors to the pathophysiology of eosinophil-associated fibrotic disease.

Cell Division↗
PubMed1994

Epitope mapping and topology of baculovirus-expressed HIV-1 gp160 determined with a panel of murine monoclonal antibodies.

To define protein folding patterns of HIV-1 Env subunit vaccines, we have isolated a set of 30 monoclonal antibodies (MAbs) from BALB/c mice immunized with a recombinant gp160 vaccine (rgp160) expressed in a baculovirus system. This article describes epitope mapping for the MAb panel and topology of the epitopes for rgp160 and a recombinant gp120 (rgp120) also expressed in a baculovirus system. The following results are reported: (1) rgp160 harbors a minimum of 4 antigenic domains, 3 mapping to the C1, C2, and C3/V4 regions of gp120 and 1 mapping to the cytoplasmic tail of gp41; (2) there are at least 3 adjacent or overlapping epitopes in each antigenic domain; (3) a minimum of 14 independent epitopes were mapped, all of which are continuous sites; (4) each of the epitopes is exposed on rgp160 without prior manipulation of the protein; and (5) by contrast, 6 of the 8 epitopes mapping to the C1, C2, and C3/V4 regions are not exposed on rgp120, but become exposed when the protein is denatured. Taken together, these results show that rgp160 and rgp120 are folded differently, illustrating the use of this MAb panel to compare epitope topographies of recombination HIV-1 Env proteins. This MAb panel may aid in the refinement of HIV-1 Env subunit vaccines.

AIDS Vaccines↗
PubMed1994

Effect of nonprotective vaccination on antibody response to subsequent human immunodeficiency virus infection.

We have investigated the systemic anti-HIV antibody response in chimpanzees who were immunized with live vaccinia containing either the HIV envelope glycoprotein (gp160IIIB) or a control antigen (herpes simplex virus glycoprotein D) and then challenged with either a high dose (300,000 TCID50) or low dose (100 TCID50) of HIVIIIB. HIV was subsequently isolated from all animals, indicating failure of the vaccination to protect against HIV infection. Serum antibody responses were evaluated before immunization, at the time of challenge with HIV, and at multiple time points in the 9 mo after challenge. Immunization resulted in a more rapid rise of antibody to gp160 in both high and low dose animals. Antibodies to the V3 loop induced upon infection were unaffected by immunization. In low dose animals, neutralizing antibody rose more rapidly and to higher levels in the immunized animals as compared with the control. There was no difference in neutralizing antibodies between immunized and control chimpanzees in the high dose group. Epitope mapping of the anti-gp 160 response indicated that immunization with gp160 vaccinia induced a postinfection antibody response to a region of gp41 (amino acids 718-743) that was not immunogenic in control-vaccinated animals. These data indicate that failed vaccination with the HIV envelope can alter both the timing and epitope specificity of the subsequent anti-HIV antibody response. These studies also define the evolution and fine specificity of the antibody response during the critical period immediately postinfection.

AIDS Vaccines↗
PubMed1994

Temporal analysis of the antibody response to HIV envelope protein in HIV-infected laboratory workers.

Three laboratory workers have been infected with the IIIB strain of HIV; their antibody response to HIV has been studied in serial serum specimens. Because the infecting virus is known, the fine specificity of the antibody response was studied on the homologous strain of HIV. Anti-p17, anti-p24, anti-gp160, CD4/gp120 blocking and neutralizing antibodies developed in parallel. Epitope mapping of the anti-gp160 response indicated several regions that consistently induced an antibody response. Serum contained antibody which reacted with V3-specific peptides corresponding to the very tip of the loop and crossreactivity was seen with V3 loop peptides from other sequence divergent strains of HIV. Antibody to the V1 loop was produced at levels comparable with that seen for the V3-loop. Anti-V1 neutralized HIV with a titration curve equivalent to an anti-V3 monoclonal antibody. Because the infecting virus is known and serial reisolates have been obtained, we explored the relationship between production of antibody to a given epitope and mutation in the virus. The data suggest that an association exists, but do not clearly indicate that antibody drives the selection for mutant viruses. The findings presented here provide a fine specificity analysis of the evolution of the antibody response to HIV in greater detail than has previously been performed.

Amino Acid Sequence↗
PubMed1993

Studies on antigen binding by intact and hinge-deleted chimeric antibodies.

A matched set of chimeric IgG1 and IgG4 antibodies were used to investigate the role of the IgG hinge in binding to Ag with differing space between the epitopes. Antibodies bearing identical V regions and either IgG1 or IgG4 C regions were engineered with and without hinges. We measured the binding of these antibodies to the peptide CYYYEEEEY and to CYYYEEEEY-BSA conjugates with decreasing numbers of peptides per BSA molecule. We earlier showed that V region differences in antibodies could affect Ag binding patterns in solid-phase but not solution-phase assays; however, both types of assay yielded similar results for the hinge-deleted antibodies. Binding of CYYYEEEEY-BSA by hinge-deleted and intact IgG1 was similar, but intact IgG1 bound free peptide better than did hinge-deleted IgG1. Intact IgG4 antibody bound less well to CYYYEEEEY and CYYYEEEEY-BSA than did IgG1 but, surprisingly, hinge-deleted IgG4 showed better binding than did intact IgG4 and was more like the IgG1 antibodies in binding affinity. Thus, the IgG4 hinge may impart a structural constraint that prevents high affinity binding to Ag. The hinge-deleted IgG4 antibody did not activate C, although it bound Ag similarly to IgG1. This study is the first to address the effect of the IgG hinge on Ag binding by using well defined Ag with different epitope densities. Our results may provide an explanation for the apparent low affinity of IgG4 antibodies.

Amino Acid Sequence↗
PubMed1993

Soluble CD4 enhances the efficacy of immunotoxins directed against gp41 of the human immunodeficiency virus.

Monoclonal antibodies specific for the gp120 or gp41 portions of the human immunodeficiency virus (HIV) envelope protein gp160 were conjugated to ricin A chain, and their immunotoxic activities against HIV-infected cells were evaluated in the presence or absence of soluble CD4 (sCD4). Immunotoxin activity was measured in vitro as cytotoxicity and inhibition of secretion of infectious HIV. The efficacy of anti-gp41 immunotoxins was enhanced at least 30-fold in the presence of sCD4. This effect was specific for HIV-infected cells, but not for uninfected cells, and was seen at concentrations of sCD4 as low as 0.1 micrograms/ml. Anti-gp120 immunotoxins were marginally inhibited at higher concentrations of sCD4. Flow cytometry analyses showed that sCD4 increased the expression of gp41 on the surface of infected cells and increased internalization of gp120 and gp41. These data suggest that sCD4 alters the cellular trafficking of HIV envelope proteins. These findings also have important implications for the therapeutic use of anti-HIV immunotoxins and may be generalizable to other immunotoxins as well.

Animals↗
PubMed1993

Interaction of group B streptococcal opacity variants with the host defense system.

Group B streptococci (GBS) demonstrate high-frequency phase variation of colony opacity. Colony opacity is a function of chain length, with opaque colonies consisting of GBS that form longer chains. Because opaque variants do not grow on standard streptococcal media, the role of opacity variation in GBS infection has not been studied. We have isolated stable variants from type III GBS that are either transparent (variants 1.2 and 1.3) or opaque (variants 1.1 and 1.5). In this study, we evaluated the interactions of these variants with different components of the host immune system both in vitro and in vivo. Opaque GBS were less immunogenic than transparent GBS. Opaque GBS were more susceptible to killing by polymorphonuclear neutrophils (PMNs) and could induce a chemiluminescent response of PMNs in the absence of antibody (Ab) or complement. Transparent GBS did not induce neutrophil chemiluminescence in the absence of Ab and complement. However, in the presence of Ab and complement, transparent GBS induced a stronger chemiluminescent response than did opaque GBS. Scanning electron micrographs of PMNs and GBS demonstrated differences in the attachment and engulfment of the different variants by the PMNs as well as different effects of the GBS on the PMNs themselves. Interactions with complement were affected by GBS opacity as well, with opaque variant 1.1 initiating complement activation in the absence of any Ab. The virulence of the GBS opacity variants was studied in vivo by inoculation of graded numbers of GBS into newborn mice. Transparent variants 1.2 and 1.3 were most virulent, with variant 1.1 intermediate and variant 1.5 minimally virulent. However, in mixed infections, variant 1.5 greatly enhanced the virulence of small numbers of transparent GBS. These results indicate that the opacity status of GBS can influence the interaction between the GBS and the host immune system.

Animals↗
PubMed1993

Differences in the antibody response to human immunodeficiency virus-1 envelope glycoprotein (gp160) in infected laboratory workers and vaccinees.

Studies of the immune response to the human immunodeficiency virus (HIV) have been hampered by the antigenic diversity of the HIV envelope protein. In an effort to predict the efficacy of vaccination we have compared the systemic anti-envelope antibody response in seronegative volunteers immunized with recombinant gp160 (either in vaccinia or as soluble protein produced in baculovirus) derived from the HTLV-IIIB strain of HIV-1 and in two laboratory workers accidentally infected with the same strain. 11 of 14 vaccinees responded to immunization by producing anti-gp160 of similar titer and the same isotype as that seen in the laboratory workers. Four vaccinees also had antibody to the principal neutralizing domain (V3 loop) that was comparable in titer with that seen in the laboratory workers, but the fine specificity of anti-V3 antibody was qualitatively different in the two groups. Antibody that can block the interaction between CD4 and gp120 was present at comparable levels in three vaccines and the lab workers. Neutralizing antibody titers were markedly lower in the vaccinees than in the laboratory workers. In seven of the vaccinees, an immunodominant epitope was at amino acid 720-740. Analyses of monoclonal antibodies to this region indicate that they do not neutralize, bind to infected cells, nor function as immunotoxins. Although the anti-gp160 antibody response was of similar magnitude in both infected and vaccinated individuals, there were important qualitative differences.

AIDS Vaccines↗
PubMed1993

When the pregnant woman experiences itching.

It is important for dermatology nurses to know the presentation and outcome of the various dermatoses which are unique to pregnancy so that they may identify potential risks to the mother and fetus and evaluate future pregnancies. Differentiating features, adjunctive laboratory studies, and the medical implications regarding the risk of recurrence in a future pregnancy are examined in this article.

Female↗
PubMed1992

Effect of H chain V region on complement activation by immobilized immune complexes.

Activation of C by immune complexes (IC) in tissues and the inflammatory consequences are major determinants in the pathogenesis of many autoimmune disorders. To assess the factors involved in C activation by such IC, we examined the binding of C components by chimeric IgG1 antibodies bound to immobilized Ag. We previously reported that alterations in the H chain V regions can affect the binding of first component of C (C1q) and a major breakdown product of the third C component (C3b) when otherwise identical antibodies were bound to immobilized (Tyr, Glu)-Ala-Lys. To evaluate C activation of these antibodies in well defined IC, we utilized a 9-amino acid peptide conjugated to BSA as Ag. The peptide:BSA conjugate was bound similarly by the two IgG1 antibodies which differed mainly in the CDR3 regions, but also in 9 other amino acids in the H chain V region. When soluble IC were prepared with the two antibodies, they activated C similarly. However, C activation by solid phase Ag:antibody complexes differed; we found that antibody 10B bound more C1q and C3b than antibody B11 did, unless the Ag was present at high density on the plates. These data suggest that the variable region differences affect C activation by these antibody when they are bound to immobilized Ag. Furthermore, these results underscore the differences in C activation by the same antibody depending upon whether the IC are free in solution or immobilized.

Amino Acid Sequence↗
PubMed1992

Eosinophils as host cells for HIV-1.

Transient expression of HIV-1 p24 antigen was observed in eosinophils acutely infected with the HTLV-IIIB strain of HIV-1. PCR analysis of eosinophils isolated from 18 seropositive individuals showed HIV-1 sequences to be present in 2 subjects. These data suggest that eosinophils may act as host cells for HIV-1.

DNA, Viral↗
PubMed1992

A defect in the humoral immune response to protein antigens and haptens in immunoglobulin mu heavy-chain transgenic mice.

We have examined the antibody response in mice expressing a functionally rearranged mu Ig heavy chain derived from a hybridoma antibody with specificity for the hapten 4-hydroxy-3-nitrophenyl (NP). Transgenic mice and their normal littermates were immunized with the antigens NP-OVA, the synthetic polypeptide (Tyr,Glu)-Ala-Lys ((T,G)-A-L), or saline. The presence of serum antibodies to NP-BSA, OVA, (T,G)-A-L, and BSA was examined by ELISA. Sera were evaluated prior to immunization and at periods of up to 4 months following immunization. Prior to immunization, transgenic mice had high levels of IgM anti-NP antibody but no detectable antibody to the other antigens. Both the primary and secondary antibody responses of transgenic mice to NP, OVA, and (T,G)-A-L were depressed when compared with the response of non-transgenic mice. Because of reports that these transgenic mice have increased proportions of CD5 + B-cells, a subpopulation associated with the production of autoantibodies, we examined these mice for the production of both IgG and IgM rheumatoid factors and anti-DNA antibodies. Transgenic mice had a modest increase in the spontaneous production of IgM anti-DNA. These data demonstrate a functional defect in the humoral immune response of mu transgenic mice.

Animals↗
PubMed1992

The interplay of microbes and their hosts.

Microbes and their hosts exert considerable evolutionary pressure on one another. This brief report of a recent meeting describes the strategies and tactics, and highlights some of the key molecules involved in the complex host-parasite relationship.

Animals↗