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Biomedical subjects

R C Kennedy

Publications and source records attributed to R C Kennedy.

At least 109 records · Page 6Linked to original sources
PubMed1990

Inhibition of lymphokine-activated killer activity during HIV infection: role of HIV-1 gp41 synthetic peptides.

Lymphokine-activated killer (LAK) activity was analyzed in 31 human immune deficiency virus 1 (HIV-1)-infected patients. It was found to be reduced in all groups of patients, being more pronounced in those with acquired immune deficiency syndrome (AIDS) and AIDS-related complex compared to HIV-1-seropositive, asymptomatic individuals. Only high doses of interleukin-2 were able to restore LAK activity comparable to that of normal controls. In addition, HIV-1 gp41 synthetic peptide sequences 735-752 and 846-860 were able to significantly inhibit normal LAK activity at all the effector:target ratios tested. HIV-1-positive serum and the supernatant fluids from cultured peripheral-blood mononuclear cells from HIV-1-infected patients had the same inhibitory effect on normal LAK activity. These data provide evidence that (1) LAK activity appears to be impaired during the course of HIV-1 infection and (2) HIV-1-positive serum and HIV-1 components could exert a profound inhibition of this functional activity.

Adult↗
PubMed1989

Strategies for developing vaccines against acquired immunodeficiency syndrome.

There exists an abundance of literature on the prospects for developing vaccines against the etiologic viral agent of acquired immunodeficiency syndrome (AIDS). Excellent reviews from a variety of investigators are available on this subject. It is our intention to review the literature relative to potential strategies for developing a successful AIDS vaccine and to impart some of our concerns on the different types of vaccine that are being tested. We have also focused on some vaccine strategies that have not received much attention in previous review articles. Finally, the role of animal models in assessing vaccine safety and efficacy against AIDS has been briefly described.

Acquired Immunodeficiency Syndrome↗
PubMed1989

Anti-idiotype modulation of the in vitro immune response to hepatitis B surface antigen (HBsAg) following remote infection by hepatitis B virus in man.

An antigen-inhibitable Ab-2 that exhibits internal image activity will selectively stimulate the in vitro production of anti-HBs in individuals with remotely established immunity to hepatitis B virus. This response is seen (1) in the absence of a polyconal increase in total IgG, (2) with the F(ab')2 component of the Ab-2, (3) in cultures depleted of T-cells, and (4) in the absence of stimulation by antigen. This observation demonstrates that the Ab-2-mediated stimulation of specific IgG production may be an important regulatory function in man.

Adult↗
PubMed1989

Methods for generating reagents to examine idiotype networks within antiviral immune responses.

The use of anti-idiotypic antibodies to examine and/or modulate the immune response to various viral antigens has the potential to be of use in many diverse systems. This paper details the method and immunologic parameters used in our laboratory to generate and characterize anti-idiotypic antibodies (anti-Id or Ab-2) with specificity for antibodies directed against viral antigens. These anti-Id reagents have been used in our laboratory for studies involving the immune responses to hepatitis B virus and simian virus 40, which we describe here, as well as herpes simplex virus, and the human immunodeficiency virus. We have utilized these anti-Id reagents to examine the fine specificity of the idiotypes on antiviral antibodies in these systems and have attempted to modulate or induce specific antiviral immune responses. It is anticipated that the methods described herein will be helpful in analyzing the immune response in other viral systems including studies involving viral-receptor interactions.

Animals↗
PubMed1989

Expression and characterization of hepatitis B virus surface antigen polypeptides in insect cells with a baculovirus expression system.

The baculovirus Autographa californica nuclear polyhedrosis virus was used as an expression vector to produce hepatitis B virus surface antigen with and without the pre-S domain. The S gene product was expressed as both fusion and nonfusion polypeptides. No difference was observed in the posttranslational modification of the fusion and nonfusion polypeptides. The S proteins were not secreted into the medium but were inserted into the endoplasmic reticulum, glycosylated, and partially extruded into the lumen of the endoplasmic reticulum as 22-nm lipoprotein particles. The oligosaccharide chains on the insect cell-derived S protein were of the N-linked high-mannose form, in contrast to the complex-type oligosaccharides detected on plasma-derived hepatitis B virus surface antigen. The pre-S-S polypeptides were inserted into the endoplasmic reticulum, glycosylated, and modified by fatty acid acylation with myristic acid. A procedure was developed to purify the S protein from cellular membranes by using detergent extraction and immunoaffinity chromatography. The purified S protein was in the form of protein-detergent micelles and was highly antigenic and immunogenic.

Animals↗
PubMed1988

Autoantibodies and common idiotypes in men and women with sperm antibodies.

Antisperm antibodies have been implicated as a causative factor of infertility and pregnancy wastage. Since concomitant autoimmune phenomena were reported in men with antisperm antibodies, we investigated known antisperm antibody-positive sera from 25 women, 27 men, and the respective seminal plasma samples. The investigated autoimmune panel included a search for antinuclear antibodies, autoantibodies (in IgG, IgM and IgA isotypes) to seven phospholipids (cardiolipin, phosphatidylserine, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidic acid), to four histone subfractions (H1, H2A, H3, H4), and to four polynucleotides [ssDNA, dsDNA, poly(I), and poly(dT)], total immunoglobulin levels, and sperm antibody titers. The sera were also evaluated for the presence of a common anti-deoxyribonucleic acid antibody, and anticardiolipin antibody idiotypes. Levels of sperm antibody titers were significantly lower in women than in men. Both men and women with antisperm antibodies demonstrated elevated total IgG levels compared with those of normal control subjects. Only women showed elevated levels of total IgM. Sera from 24% of women and 11% of men with antisperm antibodies demonstrated antinuclear antibody titers greater than 1:40. The most striking autoantibody abnormalities were found among antiphospholipid antibodies. Sera from women with antisperm antibodies demonstrated higher autoantibody production than was found in their male counterparts. A significant correlation was found between antisperm antibodies and IgM anticardiolipin and IgA anti-phosphatidylinositol in women and between sperm antibodies and IgA phosphatidylserine antibodies in men. The presence of anticardiolipin and anti-deoxyribonucleic acid antibody idiotypes was significantly more frequent in women than in men. By means of discriminant analysis and variables selected by this mathematical model, the identification of 24 of 25 women and 26 of 27 men with antisperm antibodies was correctly predicted. These results suggest that women and men respond differently to sperm antigens. The apparent cross-reactivity between sperm antibodies and other autoantibodies, usually associated with autoimmune disease, suggests that a polyclonal B cell activation, similar to that seen in autoimmune diseases, occurs in patients with sperm antibodies.

Adult↗
PubMed1988

Synthetic peptides homologous to HIV transmembrane glycoprotein suppress normal human lymphocyte blastogenic response.

Human immunodeficiency virus (HIV) envelope glycoprotein is synthesized as a polyprotein precursor of 160 kDa (gp 160) and is subsequently cleaved into an amino terminus subunit, gp 120, and a carboxyl terminus transmembrane subunit, gp 41. Two synthetic peptides corresponding to amino acid sequences 735-752 and 846-860, respectively, as deduced from the nucleotide sequence of HTLV-IIIB gp 160 were synthesized and used to assess their effects on normal human lymphocyte blastogenesis. Peptides 735-752 and 846-860 conjugated to protein carriers, but not free peptides, exerted a pronounced suppression of the normal human lymphocyte proliferative response to concanavalin A (Con A), phytohemagglutinin (PHA), pokeweed mitogen (PWM), and alloantigens. A synthetic peptide homologous to a 17 amino acid sequence of the gene product of HIV trans-acting transcriptional (TAT III) gene had no suppressive effects. Peptides 735-752 and 846-860 also inhibited the IL-2-dependent proliferation of the murine CTLL-2 cell line and the PHA-induced proliferation of normal mouse spleen cells. HIV peptide-induced suppression of human blastogenesis required a 2- to 3-day incubation of responder cells with peptides, suggesting that binding of peptides to the cell membrane was not sufficient for suppression. These results suggest that, in addition to the selective cytopathic effects of HIV, the etiological agent of the acquired immunodeficiency syndrome (AIDS), on the T-helper/inducer lymphocyte subset, viral peptide-mediated immunosuppression may also play an important role in the pathogenesis of the disease. Moreover, these data clearly indicate the need to address the potential immunosuppressive property of HIV antigens in the effort to select and develop effective prophylactic means against AIDS.

Glycoproteins↗
PubMed1988

Inhibition of normal human natural killer cell activity by human immunodeficiency virus synthetic transmembrane peptides.

The inhibitory effect on normal natural killer (NK) cell activity of two synthetic peptides corresponding to amino acid sequences 735-752 and 846-860, respectively, as deduced from the amino acid sequences of HTLV-IIIB gp160, was assessed. Sequences 735-752 and 846-860 correspond to regions located within the HIV transmembrane gp41, the carboxy terminus of HIV gp160. These two synthetic peptides have been shown previously to suppress the mitogen- and alloantigen-induced normal human lymphocyte blastogenic responses. Peptides 735-752 and 846-860 conjugated to protein carriers exerted a significant inhibition on the normal NK cell activity assayed against K562 tumor target cells in an in vitro 51Cr-release cytoltoxicity assay. At variance, control peptides similarly conjugated had no effect on NK activity. Addition of exogenous recombinant human interleukin-2 (IL-2) resulted in a partial restoration of the suppression of NK cell activity exerted by both peptides. Binding experiments indicated that peptides 735-752 and 846-860 did not affect the formation of effector cell-target cell conjugates, suggesting inhibitory effect(s) subsequent to the formation of the lytic complex as one potential mechanism of the observed NK suppression. These results suggest that peptides 735-752 and 846-860 homologous to sequences within the HIV transmembrane gp41 may play an important role in the pathogenesis of the defective NK cell activity observed in patients with acquired immunodeficiency syndrome (AIDS).

Adult↗
PubMed1988

Neutralization of diverse HIV-1 strains by monoclonal antibodies raised against a gp41 synthetic peptide.

Three IgM monoclonal antibodies raised against synthetic peptide analogs of a hydrophilic region of the gp41 transmembrane env protein of HIV-1 neutralize different HIV-1 isolates but not HIV-2 isolates, as determined by HIV titration and by syncytial inhibition assays. VSV (HIV-1) pseudotypes, however, were not neutralized, indicating that gp41 was not accessible to these antibodies on the pseudotype particles. The antibodies affect early steps in adsorption and penetration of HIV-1.

Antibodies, Monoclonal↗
PubMed1988

Anti-idiotypic antibodies as immunogens: idiotype-based vaccines.

Numerous studies have documented that antibodies may regulate the immune system and form the basis of vaccines, namely anti-idiotype vaccines. Antibodies carry individual idiotype antigenic determinants against which antibodies can be formed. When the anti-idiotype recognizes the same site that recognizes the primary antigen, a mirror image or combining site antibody may be generated. Other anti-idiotypes which recognize non-combining antigenic determinants have also been used. The evidence is reviewed for the existence of a broad range of anti-idiotypes and details are given of how an anti-idiotype vaccine based on the hepatitis B surface antigen has protected against virus challenge in the most relevant animal model system, namely the chimpanzee. Furthermore, the definition of the CD4 molecule as the conserved binding site for all known human and similar immunodeficiency viruses, (in marked contradiction to their varied neutralizing properties) has led to the raising of anti-idiotypes in mice based on the CD4 receptor which have the capacity to neutralize a broad range of isolates.

Acquired Immunodeficiency Syndrome↗
PubMed1987

Neutralisation of HIV isolates by anti-idiotypic antibodies which mimic the T4 (CD4) epitope: a potential AIDS vaccine.

Polyclonal anti-idiotypic antibodies were raised in mice against anti-Leu3a, a mouse monoclonal anti-human T4 (CD4) antibody that blocks the in-vitro binding of human immunodeficiency virus (HIV) to the CD4 molecule. The anti-idiotypes recognized anti-Leu3a but not OKT4, an anti-human T4 antibody that does not inhibit HIV binding to CD4. The anti-idiotypes specifically reacted with the HIV envelope glycoprotein in solid-phase immunoassays. More importantly, the anti-idiotypes neutralised three distinct isolates of HIV-1 and one isolate of HIV-2 in a syncytial inhibition assay. These results have implications for a potential AIDS vaccine of anti-CD4 preparations to induce an anti-idiotypic response with the capacity to bind HIV at its receptor site.

Acquired Immunodeficiency Syndrome↗
PubMed1987

Immune response to human immunodeficiency virus. In vivo administration of anti-idiotype induces an anti-gp160 response specific for a synthetic peptide.

Anti-idiotypic antibodies (anti-Id) to chimpanzee antibodies directed against a synthetic peptide corresponding to a native epitope associated with gp41 of human immunodeficiency virus (HIV) envelope glycoprotein were produced in rabbits. The peptide was analogous to amino acid sequences 735 to 752 from the human T cell leukemia virus-IIIB isolate of HIV. Characteristics of the anti-Id preparation included: 1) detection of a shared determinant present on a second chimpanzee and one of three rabbit antibody preparations directed against the synthetic peptide, 2) failure to recognize an idiotype (Id) in BALB/c mouse antisera to the peptide, and 3) partial inhibition of the homologous chimpanzee Id preparation from binding either peptide or a recombinant HIV gp160 preparation. Immunization of BALB/c mice with the anti-Id induced an antipeptide response which bound a recombinant gp160 preparation without subsequent peptide or gp160 exposure. The anti-gp160 containing sera from mice immunized with anti-Id were able to inhibit the Id-anti-Id reaction indicating that an Id-positive antibody response was induced. This Id is not normally expressed in the murine anti-gp 160 immune response to the synthetic peptide and suggests that this anti-Id may activate normally silent clones. This study indicates that Id networks may be operational during the immune response to HIV epitopes. Alternatively, anti-Id may be useful in altering the serologic characteristics of an antibody response to HIV and may offer potential for modulating the immune response in this viral infection.

Animals↗
PubMed1987

Relation of the Escherichia coli dnaX gene to its two products--the tau and gamma subunits of DNA polymerase III holoenzyme.

The Escherichia coli DNA polymerase III holoenzyme 71.1 kDa tau subunit is a 643 amino acid protein encoded by the dnaX gene. This gene also encodes the holoenzyme 56.5 kDa gamma subunit. The tau factor (as a tau'-LacZ' fusion protein) has been isolated and shown to be cleaved in vitro to form gamma and a 135 kda C-terminal cleavage product. The tau'-LacZ' fusion protein, gamma, and the C-terminal cleavage product have been isolated. N-terminal sequencing has demonstrated that tau and gamma share the same N-terminal sequences and that tau is proteolytically cleaved in vitro between residues 498 and 499 to form gamma. In addition, residues 420-440 were shown to be present in both tau and gamma by use of antibody specific for a synthetic peptide corresponding to that sequence. Some mechanism functions in vivo to ensure that tau and gamma are synthesized in a ratio of about one-to-one, as shown by radioimmune precipitation of tau and gamma from cellular extracts.

Bacterial Proteins↗
PubMed1987

Induction of an anti-hepatitis B surface antigen response in mice by noninternal image (Ab2 alpha) anti-idiotypic antibodies.

Anti-idiotype antibodies to a mouse monoclonal antibody A-12 directed against HBsAg were produced in rabbits. The anti-Id consisted of an Ab-2 alpha preparation that did not display any detectable internal image activity. Immunization of BALB/c mice with the anti-Id (Ab-2 alpha) coupled to KLH induced an anti-HBs response without subsequent HBsAg exposure. No anti-HBs was detected in control groups of mice immunized with other rabbit anti-Id-KLH preparations. The anti-HBs containing sera from mice immunized with the Ab-2 alpha were able to inhibit the Id-anti-Id reaction, indicating that an Id-positive, anti-HBs response was induced. This idiotype is not normally expressed during the murine immune response to HBsAg and suggests that noninternal image anti-Id activates silent clones. This study, along with our previous results obtained with the use of internal image anti-Id, suggests that there is more than one Id network operational during the BALB/c murine immune response to HBsAg.

Animals↗