Search PubMed⌕ Search

Biomedical subjects

S H Pincus

Publications and source records attributed to S H Pincus.

At least 73 records · Page 4Linked to original sources
PubMed1989

Elevated plasma interleukin-1 levels in humans following ultraviolet light therapy for psoriasis.

Interleukin-1 (IL-1) is a potent cytokine with a wide range of biologic activities including induction of several acute phase responses. Ultraviolet radiation (UVR) is widely used as a therapeutic modality to treat many chronic skin diseases, including psoriasis. In the present study, we investigated whether ultraviolet B (UVB) radiation induced circulating IL-1 in the plasma of patients undergoing chronic UVB therapy. In order to remove plasma proteins which inhibit IL-1-induced T-cell proliferation, each plasma sample was chromatographed and each fraction was assayed for IL-1 activity. There was no detectable IL-1 before and 1 h after UVB radiation; IL-1 appeared 4 h after treatment and was absent after 24 h. Plasma IL-1 was neutralized by antibodies to recombinant human IL-1 beta and alpha. The anti-IL-1 alpha, but not anti-IL-1 beta, antibodies partially neutralized the IL-1 activity present in a keratinocyte cell line supernate. These results demonstrate that UVB therapy induces circulating IL-1 and that this IL-1 may originate from both keratinocyte and non-keratinocyte sources.

Antibodies↗
PubMed1989

In situ localization of interleukin-1 in normal and psoriatic skin.

Interleukin-1 (IL-1) is a family of polypeptides that mediates a wide range of inflammatory and immune responses. In human skin, unstimulated keratinocytes produce a large amount of such cytokines. Although the precise role of IL-1 in the skin is unknown, there is experimental evidence supporting involvement of IL-1 in the pathogenesis of inflammatory skin diseases. In this study, we investigated in situ localization of IL-1 alpha and IL-1 beta in normal and psoriatic skin. Using polyclonal antibodies and the avidin-biotin peroxidase complex, we demonstrated the presence of both IL-1 alpha and IL-1 beta in normal and psoriatic formalin-fixed paraffin-embedded tissues. In both cases, IL-1 alpha was more prominent. However, the distribution of IL-1 alpha differed between normal and psoriatic skin. In normal skin, IL-1 alpha distribution was predominantly intercellular, whereas IL-1 alpha distribution was predominantly within the cytoplasm in psoriatic skin. These studies confirm that IL-1 alpha is the predominant form of IL-1 in the skin and provide further support for the hypothesis that IL-1 participates in the pathogenesis of psoriasis.

Adult↗
PubMed1989

Modulation of human peripheral blood eosinophil function by tumor necrosis factor-alpha.

Eosinophils are important effectors in helminthic parasitic infection. Tumor necrosis factor (TNF-alpha) has been implicated as a mediator in the host response to parasitic infection and enhances eosinophil-mediated helminthotoxicity. We have examined the direct effects of recombinant human (rh) TNF on eosinophil functions of degranulation and oxidative metabolism. This report describes the minimal effects of rhTNF-alpha on eosinophil superoxide anion generation and enzyme secretion, which do not satisfactorily explain the observed increases in helminthotoxicity. In contrast to other cell types, eosinophils are unique in their differential responses to interleukin-1 beta and TNF.

Arylsulfatases↗
PubMed1988

Protective efficacy of IgM monoclonal antibodies in experimental group B streptococcal infection is a function of antibody avidity.

We have produced and characterized six mAb directed against group B streptococci (GBS). All antibodies are IgM. We have previously shown that some of these antibodies are highly protective in the treatment of experimental infections in neonatal rats, whereas others do not appear to have any protective efficacy. Using an ELISA, we demonstrate the specificity of both protective and nonprotective antibodies. Two antibodies, binding different epitopes, are directed against antigenic structures present on all GBS; two are specific for type III carbohydrate determinants; one binds to a protein Ag present on all type I and II GBS; and one appears to bind to type Ia GBS only. Quantitative absorption assays provide evidence that the difference between protective antibodies and nonprotective antibodies is the avidity that the antibody demonstrates for the epitope recognized on the surface of the bacteria; 10 to 15 times as much protective antibody binds to GBS as does nonprotective antibody. Direct binding experiments with radiolabeled antibody confirm this conclusion.

Animals↗
PubMed1988

Human immunoglobulin heavy-chain-variable (VH) region gene families defined by hybridization with cloned human and murine VH-genes.

The Southern DNA hybridization technique was used to determine the organization of human immunoglobulin heavy-chain-variable region (VH) gene families and the extent of polymorphism within these families. Human DNA was digested with a restriction enzyme and then hybridized to a cloned human or murine VH gene. Six human and six murine genes were used. Hybridization patterns seen in the human genes fell into three groups, which corresponded to the Kabat subgroup assignments based on sequence. When DNA from different individuals was compared, polymorphism was demonstrated in each of the three gene families. The VH II family demonstrated a higher degree of polymorphism than the others. Hybridization with the murine probes, each of which represented a different murine VH-gene family, revealed six distinct patterns of hybridization. Five of the probes detected all or a portion of one of the previously defined human families. The sixth probe, 36-60, yields a unique pattern of hybridization, suggesting a fourth VH-gene family. Dot hybridizations show that 36-60 does not hybridize with human VH-genes representing each of the known families, further supporting this possibility.

Animals↗
PubMed1988

A novel approach for the evaluation and treatment of psoriasis. Oral or topical use of 1,25-dihydroxyvitamin D3 can be a safe and effective therapy for psoriasis.

We investigated the effects of 1,25-dihydroxyvitamin D3 (1,25-(OH)2-D3) on cultured fibroblasts and keratinocytes from patients with psoriasis and treated 17 patients with psoriasis with orally or topically administered 1,25-(OH)2-D3. Cultured fibroblasts from three of five patients showed a normal response to the antiproliferative activity of a physiologic dose of 1,25-(OH)2-D3, whereas fibroblasts from the other two had a partial resistance to the drug. Cultured keratinocytes from two patients with psoriasis possessed nuclear receptors for 1,25-(OH)2-D3 and the drug caused a dose-dependent inhibition of proliferation and induction of terminal differentiation of these cells similar to effects in normal cultured keratinocytes. Ten of 14 patients with moderate to severe psoriasis who received oral 1,25-(OH)2-D3 showed significant clearing of their hyperkeratotic plaques. Three patients had complete clearing that was sustained with maintenance therapy, but four patients received little or no benefit from the therapy. By the administration of 1,25-(OH)2-D3 as a single oral dose at bedtime, larger doses of the drug could be tolerated without evidence of hypercalciuria or hypercalcemia. Three patients who received topical 1,25-(OH)2-D3 showed a rapid response with complete clearing after 6 weeks of therapy. Therefore, these preliminary findings suggest that orally or topically administered 1,25-(OH)2-D3 may be a safe and effective alternative therapy for the treatment of psoriasis.

Administration, Cutaneous↗
PubMed1988

Oxidative requirement for degranulation of human peripheral blood eosinophils.

Eosinophils possess both oxygen-dependent and oxygen-independent mechanisms for damaging helminthic parasites such as schistosomula. We have studied the release of the granular enzymes beta-glucuronidase and arylsulfatase to evaluate the oxidative requirement for degranulation. Both ionophore-mediated and immunoglobulin G-mediated release of granular enzymes were enhanced in the presence of oxygen (P less than or equal to 0.05). Calcium ionophore-mediated degranulation under aerobic conditions was reduced by the addition of the degradative enzymes catalase and superoxide dismutase, suggesting that active oxygen products enhance degranulation. In contrast, oxygen products did not appear to contribute to degranulation induced by immunoglobulin G-coated beads.

Aerobiosis↗
PubMed1988

Characterization of monoclonal antidigoxin antibodies immobilized to a solid support.

A high-affinity monoclonal antidigoxin antibody, produced by somatic cell fusion, was amplified by the formation of ascites. Purification from ascites was accomplished by affinity chromatography by passing the ascites over a digitoxin-amine-agarose column. Affinity-purified antidigoxin antibody was coupled to a pellicular microbead at concentrations of 10, 25, 50, and 100 mg/g bead. The immobilized antibody was characterized for binding affinity, for specificity to other cardiac glycosides, and for binding capacity. There were no changes in the binding affinity observed for the immobilized antibody when compared to that of the antibody grown in culture media. Binding capacities for the immobilized antibody were decreased from calculated theoretical values. Saturating the microbead with increasing concentrations of antibody lowered the binding efficiency of the antibody from 32 to 22% of theoretical values. Attempts to improve the binding capacity by immobilizing antibodies to the microbead at the immunoglobulin carbohydrate by periodate oxidation were unsuccessful. These data demonstrate that antidrug antibodies immobilized on solid supports remain functional and may have the capability of removing drug from biological fluids passed over the support.

Antibodies, Monoclonal↗
PubMed1987

Activated human eosinophils synthesize new proteins.

In order to study the biochemical consequences of prolonged in vitro activation of human blood eosinophils, aqueous whole cell lysates, cell-free supernatants from resting eosinophils, and cells activated with opsonized zymosan, calcium ionophore (A23187), N-formylmethionylleucylphenylalanine (fMet-Leu-Phe), and phorbol 12-myristate 13-acetate (PMA) were analyzed by polyacrylamide gel electrophoresis (PAGE). In comparison to resting eosinophils, opsonized zymosan-activated eosinophil extracts demonstrated altered protein composition on both the native PAGE and sodium dodecyl sulfate (SDS) -PAGE. Three new polypeptides of apparent molecular mass 24 kDa, 43 kDa and 60 kDa appeared on SDS-PAGE gels when opsonized zymosan-activated eosinophil extracts were electrophoresed. In contrast, extracts from fMet-Leu-Phe, A23187, and PMA-activated eosinophils demonstrated neither altered polypeptide composition nor new polypeptides. Opsonized zymosan also induced the incorporation of L-[35S]methionine into eosinophil proteins and this was completely blocked by pretreating the cells with cycloheximide. This finding suggests that eosinophils activated by certain stimuli synthesize new proteins. These newly synthesized proteins, which are freely secreted into the medium during cell activation, may possess important immunological functions.

Autoradiography↗
PubMed1987

Phosphorylation and dephosphorylation of soluble proteins in human eosinophils.

The effect of phorbol 12-myristate 13-acetate (PMA), calcium ionophore (A23187), opsonized zymosan (OZ), and N-formylmethionyl-leucyl-phenylalanine (f-Met-Leu-Phe) on protein phosphorylation was examined in purified eosinophils (eos) isolated from human peripheral blood. Eos were prelabeled with [32P]orthophosphate, stimulated with several activating agents for varying periods of time. The soluble proteins were then analyzed by one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography. In resting eos, there was phosphorylation of endogenous soluble proteins with molecular weights of 12, 16, 21, 40, and 66 kilodaltons (kDa). PMA, a potent activator of oxidative metabolism, induced phosphorylation of 19-, 40-, and 67-kDa proteins. A23187, a strong degranulating stimulus, caused phosphorylation of 40-, 53-, and 67-kDa proteins. OZ, a relatively weak stimulus for eos function, caused phosphorylation of 30-34-, 59-, 67-, and 93-kDa proteins. In addition, all the above stimuli caused a time-dependent dephosphorylation of 21-kDa protein. In contrast, f-Met-Leu-Phe caused neither phosphorylation of new proteins nor dephosphorylation of preexisting eos proteins. These findings demonstrate that selected stimuli affect phosphorylation of soluble eos protein. These results also suggest that phosphorylation of specific proteins in eos is an intermediary step in external stimulus-induced cell activation, which may involve many different cell functions.

Blood Proteins↗
PubMed1987

Characterization of monoclonal antibodies with specificity for DNA and the synthetic polypeptide antigen (T,G)-A-L.

Because of evidence for structural similarity of variable region genes of anti-DNA and anti-(T,G)-A-L antibodies, polyspecific interactions of monoclonal anti-DNA and anti-(T,G)-A-L antibodies were investigated. Of 20 monoclonal antibodies from C57BL/10 mice with (T,G)-A-L binding, two bound DNA as determined by ELISA. In contrast, two of five anti-DNA monoclonal antibodies from MRL-lpr/lpr mice bound (T,G)-A-L. For both sets of antibodies, antigen binding was shown to be the activity of the same antibody by cross-inhibition studies. To determine whether such polyspecific antibodies were expressed during autoimmune disease, sera of autoimmune MRL-lpr/lpr mice were tested for anti-(T,G)-A-L activity. This analysis demonstrated minimal elevations of anti-(T,G)-A-L in comparison to BALB/c controls. These studies thus confirm predictions about the binding activity of anti-(T,G)-A-L and anti-DNA antibodies based on structural analysis of variable region genes. They further indicate, that while anti-(T,G)-A-L and anti-DNA antibodies may have overlapping specificity, polyspecific antibodies of this kind are not preferentially expressed during autoimmunity.

Animals↗
PubMed1987

Primary sclerosing cholangitis and low-dose oral pulse methotrexate therapy. Clinical and histologic response.

Two patients with sclerosing cholangitis responded to low-dose methotrexate treatment. A 40-year-old man who had previously undergone total colectomy for ulcerative colitis presented with refractory erythroderma and sclerosing cholangitis. Both disorders were alleviated and have remained in remission on methotrexate, 5 mg every 12 hours three times each week (15 mg/wk). Liver function improved, bile duct scarring did not worsen, and repeat liver biopsy samples have shown striking improvement. A 60-year-old man with long-standing ulcerative colitis and repeated exacerbations of sclerosing cholangitis had a similar response to low-dose methotrexate, 2.5 mg every 12 hours three times each week (7.5 mg/wk), during a 6-year period. Recurrent episodes of cholangitis have disappeared, liver function has become normal, bile duct scarring has not worsened, and liver histologic findings have become normal. Because of the potential hepatotoxicity of methotrexate, we suggest that a prospective, randomized trial be done.

Administration, Oral↗
PubMed1987

Eosinophils stimulate fibroblast DNA synthesis.

Fibrosis complicates a number of chronic inflammatory diseases and occurs in some conditions following chronic hypereosinophilic syndromes. We assessed whether eosinophils might be a source of fibrogenic factors. Extracts of human and guinea pig cell populations enriched for eosinophils contained substances that stimulated tritiated thymidine incorporation by human fibroblasts. Supernatants derived from resting eosinophils and extracts prepared from eosinophil granules also contained fibrogenic factors. Our findings demonstrate a new potential role for eosinophils and suggest a causal relationship between tissue eosinophilia and scar formation in certain parasitic conditions.

Animals↗
PubMed1986

Variable regions of antibodies to synthetic polypeptides. II. Analysis of variable region genes encoding antibodies specific for (T,G)-A--L.

Monoclonal antibodies specific for the synthetic polypeptide antigen (T,G)-A--L have been produced in two strains of mice, C57BL/10 and C3H.SW. The genes encoding the variable (V) regions of these antibodies have been studied by using the DNA hybridization technique of Southern, as well as by gene cloning and sequencing. Hybridization of DNA from 14 different cell lines with a kappa-chain probe revealed that the different cell lines used one of two different gene rearrangements to encode the recombined V region gene. There was a perfect correlation between light chain rearrangement, idiotype expression, and fine specificity. Hybridization analyses of the heavy chain revealed a more complex pattern. Seven hybridomas had the rearranged heavy chain V region genes on a 4.4 kb EcoRI restriction fragment. Others were found on restriction fragments that differed in length by several hundred base pairs. The recombined heavy chain V region genes were cloned from three different hybridoma cell lines secreting anti-(T,G)-A--L antibodies, all of which express the same idiotype and fine specificity pattern. Restriction mapping and sequencing indicate that all three utilize the same V gene, identified as the 186-2 germline gene. However, different D and J genes are used to encode each of the antibodies. In contrast to the results seen in other antigen systems, heavy chain D and J genes do not have a major influence on idiotype expression and fine specificity of antibodies to the synthetic polypeptide (T,G)-A--L.

Animals↗
PubMed1986

Interaction of IL 1 and TPA in modulation of eosinophil function.

The tumor co-promotor TPA is believed to enhance a wide variety of cellular processes by interacting with protein kinase C. Interleukin (IL 1) is a family of highly active molecules which augments the host response to infection. We have explored the interactions of these activators of cell function on the modulation of selected eosinophil functions. The effects of purified monocyte-derived IL 1 on the eosinophil functions of oxidative metabolism (as measured by superoxide anion production) and degranulation (as measured by release of the granular enzymes arylsulfatase and beta-glucuronidase) have been examined. Superoxide anion production by eosinophils stimulated with standard doses of the stimulant phorbol myristic acetate (TPA) (1 microgram/ml) was augmented approximately 20% by preincubation with IL 1. However, IL 1 alone had no effect on superoxide anion production. At suboptimal doses of TPA, there was a dose-dependent inhibition of superoxide anion production in the presence of IL 1. Calcium ionophore (2 X 10(-7) M) markedly enhanced superoxide anion production elicited by 0.1 ng/ml of TPA, but had only modest effects in the absence of TPA. When IL 1 was added to eosinophils stimulated by TPA in the presence of calcium ionophore, there was a dose-dependent increase in superoxide anion production. In contrast to other cell types, degranulation as measured by the release of arylsulfatase and beta-glucuronidase was not elicited by the addition of TPA (1 microgram/ml). Although calcium ionophore (2 X 10(-6) M) caused enzyme release (24.2% release of beta-glucuronidase, 29.4% release of arylsulfatase), this release was inhibited by the addition of TPA. The addition of IL 1 alone caused an approximate twofold increase in enzyme release, but pretreatment with IL 1 (1 U) reduced ionophore-mediated degranulation (p less than or equal to 0.05). Studies employing purified monocyte IL 1 were confirmed by recombinant IL 1-beta. These studies demonstrate for the first time that eosinophil function is modulated by IL 1. IL 1 may also modify the response of eosinophils to other stimuli such as ionophore and TPA. Because TPA is known to act by direct binding to protein kinase C, these studies also demonstrate that, in eosinophils, activation of protein kinase C by phorbol esters may augment one cellular function (oxidative metabolism) while inhibiting another cellular function (degranulation). Similarly, phorbol esters may act synergistically with calcium ionophore in regulation of one function (oxidative metabolism) and act antagonistically with another function (degranulation). The concept that IL 1 uniformly enhances cell function may need to be re-evaluated.

Calcimycin↗
PubMed1986

Lectin binding by eosinophils.

Lectins which identify carbohydrates and glycoproteins have been used to characterize specific components of the surface of guinea pig peritoneal exudate eosinophils. Agglutination of eosinophils purified by discontinuous metrizamide gradients was scored microscopically. Wheat germ agglutinin (WGA) was most effective (0.05 micrograms/ml). However, higher concentrations of soybean lectin and concanavalin A (Con A) were also effective. No differences in lectin binding were noted between eosinophils harvested from uninfected animals, Trichinella spiralis-infected animals, or animals receiving weekly intraperitoneal injections of polymyxin B. Neuraminidase pretreatment to remove surface sialic acid reduced agglutination by WGA. Eosinophils did not adhere to WGA-coated Sepharose beads; however, they did adhere to Con A-coated beads. Pretreatment with neuraminidase did not affect the adherence of eosinophils to plastic surfaces, suggesting that sialic acid does not play an important role in adherence. Formation of lectin-inhibitor complexes within the incubation mixture complicated interpretation of studies of binding to plastic surfaces. These studies demonstrate that lectin binding sites are present on the surface of eosinophils. Lectin-type binding may be important in interactions between eosinophils and noningestible parasites.

Agglutination↗
PubMed1986

Circulating HLA-DR bearing T cells: correlation with genetic rather than clinical variables.

Elevated levels of circulating HLA-DR bearing (activated) T cells have been reported in a variety of conditions. The role played by these cells in the pathogenesis of disease is not established. We found elevated levels of activated T lymphocytes in patients with rheumatoid arthritis (RA), dermatitis herpetiformis, gluten sensitive enteropathy, as well as presumed healthy persons with HLA-B8 antigen. We could not show any change in the concentration of these cells with disease activity, therapy, or in a case of dermatitis herpetiformis, challenge with the presumed antigen, gluten. Our studies do not suggest an apparent causal relationship between activated T lymphocytes and the above disorders.

Arthritis, Rheumatoid↗