Monoclonal antibody modulates human neutrophil chemotaxis to N-formyl-methionyl-leucyl-phenylalanine (fMLP).

Utilizing standard hybridoma techniques and a neutrophil chemotaxis assay for screening we produced a mouse monoclonal IgM antibody, 59/4, selected for specific inhibition of human neutrophil chemotaxis to the N-formyl-methionyl-leucyl-phenylalanine peptide (fMLP). Antibody 59/4 inhibited neutrophil chemotaxis to fMLP, but not human C5a or leukotriene B4. The antibody exhibited specific homogeneous binding to PMNs, heterogeneous binding to monocytes, and did not bind to lymphocytes in a pattern similar to the profile of N-formyl peptide binding in flow cytometric analysis. The antibody did not inhibit the binding of fluorescein-conjugated fMLPK or fML(3H)P ligands to neutrophils in flow cytometric or competitive binding assays. Other neutrophil functions including myeloperoxidase release and rosette formation with immunoglobulin or immunoglobulin C3b-coated sheep erythrocytes were not affected in the presence of antibody 59/4. These results suggest that 59/4 specifically inhibits chemotaxis to fMLP.

Dermatologic complications associated with administration of 2',3'-dideoxycytidine in patients with human immunodeficiency virus infection.

We describe a distinctive mucocutaneous eruption that occurred in 14 of 20 (70%) patients with human immunodeficiency virus infection while they were being treated with a new therapeutic agent, 2'3'-dideoxycytidine. A maculopapular eruption developed in these patients on day 10 or 11 of treatment. Seven of 14 patients, especially those receiving higher-dose therapy, also had systemic symptoms. In addition, oral ulcers developed in 9 of 14 patients on days 4 to 6 of therapy. The occurrence of the cutaneous and oral lesions correlated with dose, route, and schedule of administration of 2'3'-dideoxycytidine. In most instances the mucocutaneous lesions resolved even with continuation of therapy.

Rabbit model of disseminated syphilis: immunoblot and immunohistologic evidence for a role of specific immune complexes in lesion pathogenesis.

Circulating immune complexes (CIC) containing Treponema pallidum proteins have been preliminarily implicated as inducers of a neutrophilic vascular reaction in early human cutaneous lesions of secondary syphilis. To clarify the role of specific CIC in producing cutaneous and renal lesions, 12 rabbits were studied at the following intervals after induction of disseminated syphilis: 20 days (4 rabbits: biopsies of normal and lesional skin for direct immunofluorescence (IMF) for (IgG, IgM, IgA, Clq, C3, C4), fibrin, and T. pallidum proteins; routine histology; and immunoblots of serum for CIC containing T. pallidum proteins); 21 days (4 rabbits: as at 20 days without IMF for T. pallidum protein); 23 days (4 rabbits: as at 20 days without IMF); 30 days (same 12 rabbits restudied with routine histology of normal and lesional skin; kidneys from 4 rabbits removed for routine, IMF, and electron microscopy (EM). Treponemal polypeptide antigen (MW-87 kd) was demonstrated in CIC from rabbits. Routine cutaneous histology showed evolution of lesions from an early neutrophilic vascular reaction to the typical lymphoplasmacytic reaction. IMF showed vessel-based immunoreactants in 3 of the 4 rabbits tested at 20 days and 1 of 4 at 21 days, and T. pallidum proteins in 3 of 4 rabbits at 20 days. Routine histology, IMF, and EM studies of glomeruli showed glomerular abnormalities, but no evidence of immune deposits containing specific T. pallidum protein. Skin and kidney studies of 4 controls were all negative. These data indicate a role for specific immune complexes in the pathogenesis of cutaneous lesions in this rabbit model.

Phase I studies of 2',3'-dideoxycytidine in severe human immunodeficiency virus infection as a single agent and alternating with zidovudine (AZT).

Five dose regimens of 2',3'-dideoxycytidine (ddC) were administered, intravenously for 2 weeks then orally for 4 or more weeks, to 20 patients with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC). ddC was well absorbed from the gut and crossed the blood-brain barrier. 10 of the 15 patients who received 0.03-0.09 mg/kg every 4 h had increases in their absolute number of T4+ T cells at week 2 (p less than 0.05), though in many these rises were not sustained. 11 of 13 evaluable patients had a fall in their serum human immunodeficiency virus (HIV)p24 antigen by week 2 of therapy (p less than 0.01); in 4 patients the p24 antigen subsequently rose to baseline while in others the decline was sustained. Dose-related toxic effects included cutaneous eruptions, fever, mouth sores, thrombocytopenia, and neutropenia. A reversible painful peripheral neuropathy developed in 10 patients after 6-14 weeks' treatment. These results suggest that ddC has activity against HIV in vivo and has a different toxicity profile from that of zidovudine (AZT). 6 patients with AIDS or ARC were given an alternating regimen of oral AZT (200 mg every 4 h for 7 days) and oral ddC (0.03 mg/kg every 4 h for 7 days). The regimen was well tolerated, and the 5 patients who completed 9 or more weeks of treatment had sustained rises in their T4+ T cells and/or falls in p24 antigen.

Nevus sebaceus associated with major ophthalmologic abnormalities.

Nevus sebaceus rarely occurs as part of a syndrome consisting of central nervous system and ophthalmologic abnormalities. We describe a case of nevus sebaceus associated with an epibulbar complex choristoma and colobomas of the optic disc and peripapillary choroid, and review the dermatologic, ophthalmologic, and neurologic literature on the nevus sebaceus syndrome. When associated with other developmental abnormalities, nevus sebaceus and epidermal nevus have erroneously been considered to be a single entity. In this article, we delineate and emphasize the ophthalmologic abnormalities associated with the nevus sebaceus syndrome.

Phialophora verrucosa-induced subcutaneous phaeohyphomycosis. Fine needle aspiration findings.

A 34-year-old woman on immunosuppressive therapy presented with a subcutaneous, cystic lesion on the dorsum of the right foot. Cytologic examination of material obtained by fine needle aspiration (FNA) revealed a mixture of acute and granulomatous inflammation as well as brown-pigmented fungi in the form of budding yeast, pseudohyphae and septate hyphae. The findings suggested subcutaneous phaeohyphomycosis (phaeomycotic cyst). Culture grew Phialophora verrucosa. The cytologic, histologic and cultural findings are given. This case demonstrates that phaeohyphomycosis can be diagnosed by FNA but that fungal culture is necessary to establish the identity of the etiologic agent. This appears to be the first case of P. verrucosa-induced subcutaneous phaeohyphomycosis reported in the Western Hemisphere.

Role of circulating immune complexes in human secondary syphilis.

Studies in animal models and in the glomerulonephritis of human secondary syphilis and results from in vitro assays have suggested a role for circulating immune complexes (CICs) in human secondary syphilis. Nine adult subjects with early secondary syphilis were studied. All patients tested had CICs on C1q-binding or Raji cell assays. Proteins previously described as Treponema pallidum-specific antigens were detected by radioimmunoblot techniques in CICs from all five subjects tested. Biopsy of early cutaneous lesions revealed immunoreactants (IgG, C3, and/or C1q) in three of nine subjects and treponemal antigen in six of eight subjects tested. Histamine was injected intradermally as a trap for CICs, and biopsy of these injection sites revealed immunoreactants in four of nine subjects and treponemal antigen in five of eight subjects tested. A neutrophilic vascular reaction consistent with CIC-mediated vessel damage was seen in three of nine lesions and six of nine histamine injection sites. Normal controls did not show these changes.

Primary idiopathic cutaneous pustular vasculitis.

Pustular cutaneous vasculitis results from a heterogeneous group of disorders characterized by pustules on purpuric bases. Although the cause of this group of conditions is diverse, the histopathologic picture of the lesions is the same, showing a Sweet's-like or leukocytoclastic vasculitis. These distinctive lesions may occur in patients with Behçet's syndrome, bowel-associated dermatosis-arthritis syndrome, or chronic gonococcemia. We describe, for the first time, a patient with primary idiopathic cutaneous pustular vasculitis. This patient had evidence of both circulating immune complexes and serum enhancement of neutrophil migration. Extensive evaluation failed to reveal any underlying systemic disease. A classification of the pustular vasculitides is proposed.

Cutaneous secondary syphilis: preliminary immunohistopathologic support for a role for immune complexes in lesion pathogenesis.

A circulating immune complex-mediated pathogenesis for lesions of secondary syphilis has been postulated. Textbook descriptions of a lymphoplasmacytic histopathologic picture have contradicted a role for circulating immune complexes in lesion pathogenesis. Four patients with early cutaneous lesions of secondary syphilis were studied. All four patients had serum Raji cell and/or Clq binding assay evidence for circulating immune complexes. Three patients showed a neutrophilic vascular reaction on histologic study of early lesions. The patients studied had immunofluorescence microscopic evidence of immunoreactant deposition in dermal blood vessels (4 hours) and/or a neutrophilic vascular reaction (24 hours) after intradermal histamine injection. Dieterle staining of lesional tissue from all patients showed the presence of treponemal organisms in dermal blood vessels. This new preliminary evidence adds some support to a circulating immune complex-mediated pathogenesis of cutaneous lesions in human secondary syphilis.

Characterization of the antigenic determinants and host components in immune complexes from patients with secondary syphilis.

Sera from patients with secondary syphilis were evaluated for abnormal levels of circulating immune complexes (IC), immunoglobulins (Ig), and complement components. Clq-solid-phase assays (Clq-SPA) that made use of monoclonal and polyclonal antibodies directed against IgG subclasses indicated that human IC were composed primarily of IgG3 and IgG1; these findings appeared consistent with subclass profile responses of electrophoretically transferred blots (Western blots) of Treponema pallidum reacted with syphilitic sera. Complexes were isolated from reactive sera by polyethylene glycol precipitation followed by either anti-Clq column chromatography or protein A-Sepharose chromatography. Although qualitative and quantitative differences were noted, all purified materials contained a treponemal polypeptide antigen with a m.w. of approximately 87,000. Subsequent analysis of this polypeptide, which was also present in purified IC from rabbits with experimental syphilis, suggests that it may represent the fibronectin receptor of the organism. The 76,000 and 66,000 materials, earlier identified in purified rabbit IC, appeared to represent C-terminal degradation products of fibronectin presumably of host origin, rather than treponemal antigens. Although fibronectin binds avidly to Clq and could represent a co-precipitable contaminant throughout the isolation procedure, anti-fibronectin antibodies in the sera of patients detectable by radioimmunoassay and the present of antibodies to 76,000 and 66,000 dalton fibronectin fragments in the globulin fractions of disassociated complexes argues against such a conclusion.

Cystic fibrosis. II. The urinary mucociliary inhibitor.

In the current study, the cystic fibrosis cationic mucociliary inhibitor has been purified from urine by ion exchange chromatography, gel filtration, lectin affinity chromatography, isoelectric focusing, and high performance liquid chromatography. The molecular size of the cationic mucociliary inhibitor was estimated to be in the range of 4,000 to 13,500 MW, by its elution on Sephadex G-50, and between 7,500 and 2,750 MW, by urea-sodium dodecyl sulfate polyacrylamide gel electrophoresis. In addition to the cationic mucociliary inhibitor, an anionic mucociliary inhibitor was also detected in the urinary fraction isoelectrically focused between pH 4.5 and 4.9. The identity of the mucociliary inhibitor as a glycoprotein was established in the current study by affinity chromatography on Phaseolus lunatus lectin, by radiolabeling the carbohydrate with galactose oxidase and tritiated sodium borohydride, and by determining the presence of a large concentration of glucosamine and small amounts of galactosamine by amino acid analysis. The amino acid analysis of the purified major component of the cationic mucociliary inhibitor reveals that the glucosamine concentrations represents a high percentage of the composition of the glycoprotein.

Studies of cystic fibrosis utilizing mucociliary activity in oyster gills.

Crassostrea virginica, the oyster native to the gulf coast, has served as a source of ciliated epithelium for studies on the inhibitory factor in cystic fibrosis. In these studies, protein molecules with biological activity, obtained from serum, urine, saliva, and cells from cystic fibrosis homozygotes and heterozygotes, were detected, purified, and characterized. Properties of these factors are similar to those detected by other ciliated systems in mussel gills and rabbit trachea. Although none of the ciliated assays in their present stage of development offer a reliable means for heterozygote screening or for prenatal diagnosis, they represent powerful tools for characterizing biologically active molecules related to cystic fibrosis. The importance of purifying and characterizing the various cystic fibrosis factors described by several laboratories is based on evidence that their biological activities observed in vitro mimic some of the expressions of the disease observed in vivo and that the concentration of the mucociliary inhibitor in serum and fibroblast medium preparations from cystic fibrosis heterozygotes is approximately one-half of that in serum and fibroblast medium preparations from homozygotes.

Cystic fibrosis: studies with the oyster ciliary assay.

Bioassays using ciliary systems have detected a factor or factors in cystic fibrosis (CF) sera and tissue culture medium derived from CF cells. The typical shortcomings of an assay measuring biological activity have been studied, and the means to overcome the weaknesses of the oyster gill cilia assay have been established. The presence of the cystic fibrosis mucociliary inhibitor (CFMI) in experimental fractions may be determined by accepting data from only those assays in which authentic CF and normal (non-CF) fractions give defined reactions, by measuring the reaction of each sample at least three times, and by examining each experimental sample at a protein concentration greater than the minimum established in this study. The relative concentrations of the CFMI present in the first steps of purification of serum and medium have been calculated in terms of units of inhibition. Generally, the units of inhibition present in serum and medium fractions from heterozygotes are close to one-half of that in fractions from homozygous sources. Analogous fractions concentrated from a normal (non-CF) source never inhibited mucociliary activity, even when tested at nearly 100 times the CF concentration. Ciliary assays utilizing oyster gills are essential for monitoring fractionation procedures aimed at purifying the CFMI, and have been shown to be capable and reliable enough to do so.