Toxicity following protein A treatment of metastatic breast adenocarcinoma.

The toxic effects of protein A (Prosorba, IMRE Corporation, Seattle, WA) treatments given as part of an on-line plasmapheresis or off-line procedure were determined in a Phase I Study. Patients were randomized and treated 12 times either once per week or three times per week with a Prosorba column containing 50 or 200 mg protein A. Treated plasma volumes varied from 150 ml off-line to 2000 ml on-line. Seven patients having advanced metastatic breast adenocarcinoma patients were evaluated. All had advanced progressive disease that was resistant to chemotherapy and/or radiation therapy. Greater than 50% regression of measurable tumor volume occurred in four of seven patients; an additional patient responded with 33.5% regression. Two patients with only bony metastases demonstrated stable disease for a 60-day period. Side effects resulting from protein A treatments included transient fever, chills, rigors, and infrequently nausea, vomiting, diarrhea, episodic hyper and/or hypotension, bronchospasm, venospasm, headache, joint and tumor pain. Mild to moderate reactions were seen in all patients regardless of clinical response, but abated spontaneously or were controlled with pretreatment and/or post treatment with antipyretics and/or antihistaminics. The side effects decreased notably during the course of the week with the more intense reaction occurring during the first treatment of the week. Side effects occurred regardless of column size or volume of plasma treated. In the course of 12 treatments, anemia requiring transfusion developed in two of seven patients. Significant tumor regression was obtained in this group of patients with advanced disease. In light of the mild to moderate side effects and tumor regression in five of seven of the patients treated, protein A treatment merits further evaluation to determine the effectiveness of this treatment in breast adenocarcinoma.

Altered villus vessel fibronectin in preeclampsia.

Fibronectin is a high molecular-weight glycoprotein found in most tissues and body fluids. Maternal plasma fibronectin levels have been shown to be elevated in preeclampsia, but little is known about placental fibronectin in preeclampsia. Fibronectin tissue distribution in placental villi was "blindly" graded by two examiners using placentas from eight nonpreeclamptic and six preeclamptic pregnancies. Selected frozen sections of normal-appearing areas from each placenta were incubated with rabbit antihuman fibronectin antiserum and then stained with fluorescein isothiocyanate-conjugated goat antirabbit immunoglobulin G and examined by fluorescent microscopy. We found less intensity of fetal vessel fibronectin staining in villi from placentas from preeclamptic pregnancies than in those of normal pregnancies (p less than 0.02, Mann-Whitney U test). It is unclear why fetal villous vessels in preeclampsia have decreased tissue fibronectin, but this may reflect an additional vascular abnormality associated with the preeclampsia syndrome.

Histopathologic evaluation of lupus patients with transient renal failure.

Patients with lupus nephritis and severe renal failure progress to end-stage renal disease despite aggressive therapy to suppress immunologic function. Within this group is a small subset presenting with rapid progression of renal failure and requiring dialytic support. We reviewed the clinicopathologic data of four such patients who were able to terminate dialysis after acute renal failure due to lupus nephritis. Three of these patients have remained independent of dialysis up to 4 years, and one patient returned to dialysis 1 month following discontinuation. Although glomerular pathology was variable in the four patients, a lesion common to all at presentation was acute tubular necrosis. It is suggested that tubular necrosis may cause reversible renal failure when part of the nephropathy of disseminated lupus treated with corticosteroids.

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.

Glomerular anionic site distribution in nonproteinuric rats. A computer-assisted morphometric analysis.

The cationic ultrastructural tracer polyethyleneimine (PEI: pI approximately equal to 11.0), binds electrophysically to uniformly spaced discrete electron-dense anionic sites present in the laminae rarae of the rat glomerular basement membrane (GBM), mesangial reflections of the GBM, Bowman's capsule, and tubular basement membranes when administered intravenously. Computer-assisted morphometric analysis of glomerular anionic sites reveals that the maximum concentration of stainable lamina rara externa (lre) sites (21/10,000 A GBM) occurs 60 minutes after PEI injection with a site-site interspacing of 460 A. Lamina rara interna (lri) sites similarly demonstrate a maximum concentration (20/10,000 A GBM) at 60 minutes with a periodicity of 497 A. The concentration and distribution of anionic sites within the lri was irregular in pattern and markedly decreased in number, while the lre possesses an electrical field that is highly regular at all time intervals analyzed (15, 30, 60, 120, 180, 240, and 300 minutes). Immersion and perfusion of renal tissue with PEI reveals additional heavy staining of the epithelial and endothelial cell sialoprotein coatings. PEI appears to bind to glomerular anionic sites reversibly: ie, between 60 and 180 minutes the concentration of stained sites decreases. At 300 minutes, the interspacing once again approaches the 60-minute concentration. This suggests a dynamic turnover or dissociation followed by a reassociation of glomerular negatively charged PEI binding sites. In contrast, morphometric analysis of anionic sites stained with lysozyme and protamine sulfate reveals interspacings of 642 A and 585 A, respectively; in addition, these tracers produce major glomerular ultrastructural alterations and induce transient proteinuria. PEI does not induce proteinuria in rats, nor does it produce glomerular morphologic alterations when ten times the tracer dosage is administered intravenously. These findings indicate that the choice of ultrastructural charge tracer, the method of administering the tracer, and the time selected for analysis of tissue after administration of tracer significantly influences results. Morphometric analysis of the distribution of glomerular anionic sites in nonproteinuric rats provides a method of evaluating quantitative alterations of the glomerular charge barrier in renal disease models.

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.

Immunoglobulin E in psoriasis evaluated by paper radioimmunosorbent and paper enzyme-immunosorbent tests.

Serum IgE concentrations were determined by the paper radioimmunosorbent test in 56 patients with psoriasis and 50 normal controls, and by the paper enzyme-immunosorbent test in 32 of these patients and 50 controls. Elevated IgE levels were found in 26 (46%) of 56 patients with psoriasis and in 1 normal control (2%). The mean value (208 U/ml) in 56 patients was significantly higher than in normal controls (31 U/ml). Thirteen of 19 patients (68%) with extensive involvement (greater than 20% body surface) had an increased IgE level; the mean value (365 U/ml) was 4 times greater than in 17 patients with limited lesions (89 U/ml) and 12 times higher than in 50 normal controls (31 U/ml). No correlation was found between serum IgE levels and the presence of psoriatic arthritis. Both paper radioimmunosorbent and paper enzyme-immunosorbent testing produced similar results.

Fibronectin distribution in nailfold biopsies of scleroderma (systemic sclerosis) patients.

The distribution of fibronectin (FN) was studied in skin biopsies of 13 patients with scleroderma (SD), 7 patients with dermatomyositis (D), and 10 normal controls (NC) by direct immunofluorescence. In normal tissues, continuous or segmental linear staining of the dermal-epidermal junction (DEJ) was seen. Papillary, subpapillary dermis, and papillary capillary loops showed a reticular pattern of deposition with fibronectin. Scleroderma patients revealed similar staining in the dermis and DEJ. The reticular distribution of FN appeared to stain more intensely in the dermis than in controls, especially in deeper layers. The amount of FN in walls of blood vessels from SD patients was markedly increased; all dermal vessels stained with FN and revealed considerably thicker walls and larger lumens. FN distribution in DM patients was similar to that seen in SD with an increased amount of FN staining in capillary walls.

Complement-independent nephrotoxic serum nephritis in Munich Wistar rats. Immunologic and ultrastructural studies.

Immunologic mechanisms of proteinuria and ultrastructural alterations of the slit pore complex and glomerular charge barrier were investigated in Munich Wistar (MW) rats with nephrotoxic serum nephritis. Prior to disease induction, normal MW sera demonstrated 50% of the complement hemolytic activity compared with sera obtained from Sprague-Dawley rats. MW rats were sacrificed prior to, at onset (5 to 6 hours), and during maximal proteinuria (heterologous phase). Immunofluorescence revealed binding of rabbit antirat IgG antibodies in a linear pattern to the glomerular basement membrane (GBM) within 15 minutes postinjection. Complement deposition was not demonstrable in vivo in this model. Immediately after injection of nephrotoxic serum a decreased penetration of the GBM occurred, restricting ferritin to the level of the endothelium in in situ fixed glomeruli. GBM permeability to native ferritin did not increase despite areas of epithelial cell detachment, endothelial cell sloughing, and proteinuria between 2 and 24 hours postnephrotoxic serum injection. Colloidal iron initially decreased staining intensity between 6 and 8 hours, with a major decrease at 24 hours, indicating a loss in glomerular sialoprotein coincident with the onset of proteinuria. Polyethyleneimine (PEI) localization revealed an initial loss of anionic binding sites by 2 hours postinjection. At 6 hours peripheral capillary loops demonstrated only scattered, random polyethyleneimine-binding sites. Splitting of the lamina densa occurred at 24 hours with the exposure of previously undetected anionic binding sites within the lamina densa. Ultrastructurally, as early as 2 hours postnephrotoxic serum injection tissue perfused with tannic acid-glutaraldehyde showed epithelial membranes forming numerous pinocytotic vesicles. Blunting and retraction of foot processes caused displacement and stacking of slit diaphragms prior to the onset of proteinuria. Between 6 and 24 hours postinjection, slit diaphragms appeared to stretch and contract to compensate for epithelial cell retraction. Tangential sections showed neither alterations nor condensation products disrupting the isoporous substructure of the slit diaphragm 24 hours postnephrotoxic serum injection. Polymorphonuclear leukocytes were not found within capillary loops during the heterologous phase of nephrotoxic serum nephritis in MW rats. The absence of complement and polymorphonuclear leukocytes accompanying anti-GBM antibody deposition suggests that early epithelial cell injury and GBM charge alterations in MW rats are mediated by heterologous antibody via a complement-independent mechanism. The lower complement hemolytic activity in normal MW sera may explain the lack of complement involvement in renal lesions in this model of nephrotoxic serum nephritis. Loss of characteristic staining for both glomerular sialoprotein and discrete anionic sites in the GBM coincided with early epithelial cell alterations and occurred prior to the onset of measurable proteinuria.

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.