Studies on the pathogenesis of acute inflammation. IX. The influence of hyperosomolality secondary to hyperglycemia upon the acute inflammatory response induced by thermal injury to ear chambers of rabbits.

The inflammatory reaction induced in ear chambers of rabbits by heat injury was studied in nondiabetic animals made hyperglycemic with continuous infusions of glucose. Hypertonic solutions of glucose administered intravenously to rabbits induced the following triad: hyperglycemia, hyperosmolality, and metabolic lactic acidosis. It was found that relatively short periods of this metabolic abnormality were associated with a significant reduction in the intensity of the inflammatory reaction. There was no evidence microscopically of circulatory impairment within ear chambers; and since neither arterial hypotension nor oxygen deficit was recorded during experiments, it appeared most likely that decreased leucocytic margination per se best explained the inhibited exudative response. Hyperglycemia seemed the dominant factor responsible for this anti-inflammatory effect. It was found subsequently that hyperglycemia and hyperosmolality without metabolic acidosis impaired cellular exudation just as well.

Autoantibodies in healthy subjects of different age groups.

Autoantibody determinations in 1284 healthy Caucasian subjects of various age groups were made by indirect immunofluorescence for anti-nuclear, anti-gastric, and anti-thyroglobulin antibodies. A sex-dependent relationship between age and prevalence of anti-gastric and anti-thyroglobulin antibodies was found. No association was found between age and anti-nuclear antibodies.

Association between fluorescent antinuclear antibodies, capillary patterns, and clinical features in scleroderma spectrum disorders.

Antinuclear antibody and in vivo capillary patterns were studied in 33 patients with Raynaud's phenomenon only and in 68 patients with scleroderma spectrum disorders; the results were correlated with clinical and laboratory findings. In addition, antinuclear antibody results in the groups with Raynaud's phenomenon only and scleroderma spectrum disorders were compared with those found in 70 patients with systemic lupus erythematosus (SLE). Distinct antinuclear antibody profiles were observed in the three diagnostic groups. Comparison of patients with anticentromere antibodies with others in the group with scleroderma spectrum disorders demonstrated that anticentromere antibody-positive patients tended to have a milder disease: less skin and visceral involvement, less frequent presence of hypertension, anemia, and elevated sedimentation rate. These differences did not, however, reach statistical significance. Comparison of patients with scleroderma spectrum disorders according to in vivo capillary patterns revealed that those with an "active" pattern had significantly more extensive skin involvement than those with a "slow" pattern. Visceral involvement tended to be greater in all organ systems in the group with an "active" pattern and reached statistical significance for muscle and kidney. Hypertension was also significantly more frequent in the group with an "active" pattern than in the group with a "slow" one. The latter was positively correlated with the presence of anticentromere antibody.

An ultrastructural method for the use of polyvinylpyrrolidone and dextrans as electron opaque tracers.

A simple method has been developed whereby polymers of polyvinylpyrrolidone and dextran can be visualized at the ultrastructural level by postfixing tissue with osmium tetroxide partially reduced by potassium ferrocyanide. Further enhancement of polymer electron opacity is obtained by counterstaining thin sections with bismuth subnitrate chelated by alkaline tartrate. These tracers remain adequately dispersed in serum and, depending on molecular size and concentration, are easily detected at the ultrastructural level within vascular lumena, basement lamina, apical vacuoles of proximal and distal tubules and endothelial vesicles. This technique offers the potential of studying nephron and cell permeability with families of tracers, the members of which are similar in chemical affinities, charge density and molecular shape.

Gm allotypes in blacks with systemic lupus erythematosus.

Serum samples were collected from 328 healthy American Blacks and from 61 American Blacks with systemic lupus erythematosus (SLE). Sera were typed for the Gm1,2,3,5,6,13,17, and 21 allotypes as well as for the Km(1) allotype. The frequency of Gm phenotype 1,17;5,6,13 was significantly increased in the SLE patients (p = 0.0001, RR = 3.19, EF = 0.29). Our data suggest the existence of at least two immunoglobulin allotype associated genes that somehow interact to increase susceptibility to SLE in Blacks. To our knowledge, this is the first report of an association of Gm and SLE in Blacks.

Pulmonary function analysis in the rabbit following bronchochallenge to causative agents and mediators of the acute byssinotic response.

New Zealand White rabbits were acutely bronchochallenged for 5 min to ascertain airway responsiveness with six potential byssinogenic agents and mediators: 0.1 g/mL cotton dust extract (CDE), 0.1 g/mL cotton bract extract (CBE), 1 mg/mL endotoxin, 1 mg/mL n-formyl methionyl peptide (n-fMet), 10 mg/mL 5-hydroxytryptamine (5-HT), and 1 mg/mL prostaglandin F2 alpha (PGF2 alpha). Methacholine (MC), 10 mg/mL, was used as a control bronchoconstrictor. Clinically objective criteria were established using increases in resistance values compared to those obtained with saline controls. Animals were classified as: mild responders (Mi) = 125-149%; moderate responders (Mo) = 150-199%; or severe responders (S) = greater than 200%. Three of five (2Mo, 1S) rabbits showed increased pulmonary resistance to CDE bronchochallenge, 3/5 (1Mi, 1Mo, 1S) to CBE, 1/5 (Mo) to purified endotoxin, 4/5 (1Mo, 3S) to n-fMet, 3/5 (1Mi, 1Mo, 1S) to 5-HT, and 2/5 (1Mo, 1S) to PGF2 alpha. All five rabbits (1Mo, 4S) responded to MC bronchochallenge. Rabbits responded minimally to saline, the common solvent of all test agents; however, when challenged with methacholine, a known bronchoconstrictor, rabbits showed significant overt symptoms of acute respiratory distress with immediate and substantial increases in resistance over saline controls. CDE, CBE, and n-fMet inhalation challenge resulted in a majority or all animals showing increased resistance. 5-HT contained in CDE and CBE, exhibited similar resistance increases; however, endotoxin, also found in cotton dust, showed little airway reactivity. The rabbit is useful for characterizing changes in pulmonary function parameters seen in the acute byssinotic reaction. This study has demonstrated that bronchochallenge in the rabbit with potential byssinogenic agents (CDE, CBE, endotoxin, and n-fMet) and mediators (5-HT and PGF2 alpha) result in measurable changes in airway function, particularly increased resistance. Since bronchoconstriction is the major clinical manifestation of the acute byssinotic reaction in man and animals, it is likely that bronchoconstriction observed in cotton mill workers may be in part or totally the result of inherent dust constrictor substances or secondarily released mediators.

Diabetic glomerulonephropathy: histopathologic, immunofluorescent, and ultrastructural studies of 16 cases.

Sixteen cases of diabetic glomerulopathy are reported. Direct immunofluorescent and ultrastructural studies of renal biopsy tissues demonstrated that two patients had linear deposits of IgM and C'3 in the absence of IgG, four diabetic patients had sclerosis-induced entrapment of immunoglobulins and complement, and one patient had granular immune complexes in the subepithelial and intramembranous portion of the glomerular basement membrane. In one patient, who had nodular glomerular lesions, diffuse fibrillar deposits of electron-dense material were observed in the mesangium. In this mesangial infiltrate, light microscopy revealed the absence of amyloid and direct immunofluorescence revealed the absence of all immunoglobulins, complement components, and fibrinogen. Our study suggests that the morphologic alterations observed in diabetic glomerulopathy might be mediated by either immune mechanism or by abnormal biochemical or functional factors, such as impairment of the mesangial IgA clearance mechanism.

Membranous glomerulonephritis in a child asymptomatic for hepatitis B virus. Concomitant seropositivity for HBsAG and anti-HBs.

The presence of hepatitis B surface antigen (HBsAg) in association with immunoglobulins and complement components within the glomerular basement membranes of adults having chronic active hepatitis has been well documented. In addition, investigators in Poland have demonstrated HBsAg immune complexes in glomeruli of children who did not have clinical evidence of hepatitis. More recently, a single case of childhood membranous glomerulonephritis in an asymptomatic carrier of hepatitis B virus was cited by observers in Canada. Reported here is the deposition of HBsAg immune complexes in the glomerular basement membranes of a 13-year-old black boy who had membranous glomerulopathy but not clinical evidence of hepatitis. This may be the first reported case in the United States of HbsAg-associated membranous glomerulonephritis in a child asymptomatic for hepatitis B virus, and only the second such case in North America. However, unlike previous studies of childhood glomerulopathy in association with hepatitis B virus, this patient is seropositive for both HBsAg and anti-HBs (antibody for hepatitis B surface antigen). Similar "rare" serologic findings were found for the patient's eldest male sib.

Cell-mediated immunity in vitro in atopic dermatitis.

The function of T cells in atopic dermatitis was studied by leukocyte migration agarose and lymphocyte transformation tests. We found that phytohemagglutinin (PHA)- and PPD-induced release of leukocyte migration inhibition factor (LIF) from lymphocytes is significantly decreased (P less than .001) in patients with atopic dermatitis as compared with healthy controls. Blastogenic response of lymphocytes induced by PPD was also decreased in atopic patients as compared with controls (P less than .01). No differences were found in spontaneous blastogenesis or in blastogenic response of lymphocytes in vitro to PHA, concanavalin A (ConA), or streptokinase-streptodornase (SK-SD) between patients with atopic dermatitis and controls.

Anticentromere antibody and immunoglobulin allotypes in scleroderma.

Fifty-five unrelated whites with disorders in the scleroderma spectrum who had both antinuclear antibodies and Raynaud's phenomenon (RP) were studied. Of the 22 patients with anticentromere antibody (ACA), three had diffuse scleroderma; 16 had the complete or incomplete syndrome of calcinosis, RP, esophageal dysmotility, sclerodactyly, and telangiectasia (CREST syndrome); and three had RP only. Thirty-three patients with other nuclear patterns all had systemic scleroderma (28 diffuse scleroderma, five CREST syndrome). Patients with ACA had less organ system involvement, and lower frequencies of anemia and elevation of sedimentation rate than ACA-negative patients, but these differences were not statistically significant. They also had fewer manifestations of CREST syndrome. All 55 patients were studied for the Gm and Km allotypic markers. No association was found between Gm or Km allotypic markers and scleroderma or between the allotypic markers and the presence of ACA.

Immunofluorescence in skin specimens from three different biopsy sites in patients with scleroderma.

Immunofluorescence (IF) data from three different biopsy sites (nailfold, forearm, buttock) were studied in 18 patients with scleroderma (SD, systemic sclerosis) and the results compared with those obtained from 10 normal controls (NC) and 7 patients with systemic lupus erythematosus (SLE). Immunoglobulin (Ig) deposits were detected by direct IF technique at the dermo-epidermal junction (DEJ) in 8/14 nailfolds, 6/15 forearms and in none of the buttock specimens of SD patients. Epidermal nuclear staining was present in 6/14 nailfolds, and in 6/15 forearms and buttocks. The most prominent finding was the observation of multiple Ig deposits in the cuticle of 9/14 patients with SD. NC group was negative in all sites for epidermal nuclear staining and the only DEJ deposit occurred in the forearm of one subject. In conclusion, this study demonstrates that Ig deposits in SD, both at the DEJ and in the epidermal nuclei, occur more often than previously reported and are especially frequent in the nailfold & cuticle area.

Brief note. Induction of experimental allergic encephalomyelitis in sheep with purified bovine basic protein and adjuvant.

A large animal model for the induction of experimental allergic encephalomyelitis to study the immunological disease parameters through closed-circuit extracorporeal thoracic duct filtration was successfully produced in two different strains of sheep. This study demonstrates the advantages of using a well-defined antigen to obtain both rapid onset of disease and a higher percentage of animals succumbing to the illness.

Treatment of acute serum sickness by plasma ultrafiltration.

The clinical value of a scaled-down prototype of an extracorporeal plasma ultrafiltration system for the treatment of acute serum sickness in rabbits was examined. The system uses two filters: the primary separates red cells from plasma, and the secondary filter excludes high molecular weight proteins from plasma. In vitro and in vivo experiments showed that the secondary filters rejected substantially more IgM (80-90%) than IgG (10-30%) or albumin (10%) and totally rejected immune complexes (IC) prepared in vitro. Two groups of rabbits were submitted to either sham filtration with the primary filter only (n = 5), receiving back the remixed components of their blood, or to the complete ultrafiltration protocol (n = 7), with removal of high molecular weight proteins and IC. Several parameters were studied longitudinally, such as circulating IC, which appeared to rise more slowly in animals whose blood was ultrafiltered, and total proteinuria, which appeared to remain at lower levels in the same animals. Histologic examination of the kidneys, collected after killing, showed evidence of glomerular IC deposition in three of five sham-treated animals (a similar frequency to that observed in a separate group of five rabbits with acute serum sickness), while one of six treated animals had evidence of glomerular deposition of IC. These observations are tentative because of the small number of animals in each group, but are encouraging. Further studies with larger groups of animals are needed to determine whether the observed effects are reproducible and to better characterize the factors directly related to the removal of circulating IC.

Procainamide-induced systemic lupus erythematosus. Renal involvement with deposition of immune complexes.

A 54-year-old man sustained an acute anterior myocardial infarction and was subsequently treated with procainamide hydrochloride for 5 1/2 months, resulting in development of a drug-induced systemic lupus erythematosus (SLE) syndrome with renal involvement. In over 60 reported cases of procainamide-induced SLE, clinical renal evaluation has led to the concept that renal involvement is a rarity in this disease. Direct immunofluorescence examination of necropsy kidney tissue revealed mesangial deposition of immunoproteins and C3 in a granular pattern characteristically seen in idiopathic SLE. Ultrastructural studies confirm mesangial deposition of immune complexes. The immunological and ultrastructural pattern of immune complex deposition in glomeruli suggests similar pathological mechanisms for glomerular injury in procainamide as seen in idiopathic SLE.

Gold nephropathy. Ultrastructural, fluorescent, and energy-dispersive x-ray microanalysis study.

The nephrotic syndrome developed in a patient receiving therapy with gold for rheumatoid arthritis. The results of a histopathological examination of the renal biopsy specimen were unremarkable. Immunofluorescent studies showed deposits of immunoglobulins and C3 in a granular pattern in the glomerular basement membranes. Ultrastructurally, the discrete osmiophilic immune complexes were epimembranous. By x-ray microanalysis, gold that was complexed with sulfur was present in proximal tubular cytoplasmic vacuoles and nuclei. Gold and sulfur could not be demonstrated in glomerular epimembranous deposits. The results of these studies suggest that immune complex deposition does not involve gold and sulfur acting as haptens. Gold-salt therapy may result in damage to proximal tubules that leak renal tubular antigens, which in turn complex with autoantibody and produce an autoimmune membranous nephropathy. The evidence for this mechanism is not convincing. Although the data indicate an immune-complex cause for gold-salt nephropathy, the incident antigen (or antigens) and mechanism of action remain unidentified.

Byssinosis: thromboxane release from human platelets by cotton dust and bract extracts.

Extracts of cotton dust and bract induced in vitro release of arachidonic acid metabolite thromboxane A2 (TxA2), a potent bronchoconstricting agent, from human platelets. TxA2 release, determined by radioimmunoassay of the stabile metabolite TxB2, was both time- and dose-dependent. Cotton green bract extract was significantly more potent in releasing TxA2 from human platelets than cotton dust extract or cotton brown bract (field-dried bract) extract, while brown bract extract was least active, which suggests that the etiological agent(s) is of plant origin and not microbial in nature. TxA2 release was shown to be enzyme-dependent by inhibition of the reaction by indomethacin (cyclooxygenase inhibitor) and 7-(1-imidazolyl) heptanoic acid (thromboxane synthetase inhibitor). This study is the first to show in vitro platelet TxA2 release by cotton extracts and may provide an explanation for increased concentrations of TxB2, a metabolite of TxA2, in the bronchoalveolar spaces of rabbits exposed to aerosolized cotton dust extract. Platelet aggregation and production of TxA2 may prove to be an important mechanism in the pathogenesis of bronchoconstriction in byssinosis.

Byssinosis: serum immunoglobulin and complement concentrations in cotton mill workers.

In two separate studies (labeled A and B), concentrations of IgC, IgM, IgA, C3, and C4 were determined in the sera of byssinotic and non-byssinotic cotton mill workers. In study A, sera were collected on Monday morning and Monday afternoon and Friday afternoon in order to ascertain if the waning byssinotic response from Monday to Friday correlates with changes in serum immunoglobulin and complement concentrations. In study B, sera was collected on Monday morning and afternoon and was performed primarily to evaluate history of atopy and smoking as complicating factors in byssinosis. In study A, the concentrations of all immunoproteins were found to decrease from Monday morning to to Friday afternoon in all textile workers, and in study B only C3 concentration decreased morning to afternoon. There was, however, no difference between the immunoprotein changes for byssinotic workers when compared to non-byssinotic workers in either study. Due to the fact that hypersensitivity pneumonitis causes a decrease in immunoprotein concentrations in affected workers while no decreases are noticed in normal workers, our data do not support an immune complex etiology for byssinosis. In addition, decreased serum C3 concentrations observed in this study could be caused by complement activation; however, there was no difference in complement levels between byssinotic and non-byssinotic textile workers. Therefore, these data neither support nor eliminate the involvement of antibody-independent complement activation in the pathogenesis of byssinosis.