Robert E. Jordon

The late phase of the immediate wheal and flare skin reaction. Its dependence upon IgE antibodies.

IgE antibodies are usually thought to induce only immediate skin reactions. We have shown that the intradermal injection of a number of different allergens can produce a prolonged inflammatory reaction after the immediate wheal and flare in most sensitive subjects. This late inflammatory response occurs 6-12 h after challenge and is characterized by diffuse edema, erythema, pruritus, and heat. Both immediate and late responses can also be seen after passive sensitization of skin sites in nonatopic subjects. That IgE is involved in inducing the reaction was shown by the abolition of both immediate and late responses by passive transfer tests in the following experiments: (a) heating atopic serum at 56degreesC for 4 h, (b) removing IgE from the atopic serum by a solid phase anti-IgE immunoabsorbent, and (c) competitively inhibiting the binding of IgE antibodies to cells by an IgE myeloma protein. In addition, both responses were induced by affinity chromatography-purified IgE antibody, followed by antigenic challenge. Very similar lesions could also be induced by intradermal injection of Compound 48/80, thus suggesting a central role in the reaction for the mast cell or basophil. Histologically, the late phase is characterized by edema and a mixed cellular infiltration, predominantly lymphocytic but also containing eosinophils, neutrophils and basophils. Direct immunofluorescent staining did not show deposition of immunoglobulins or complement components, except IgM in 2 of 15 and C3 in 1 of 15 patients. This finding indicates that the late phase does not depend on the deposition of immune complexes. The results of the study suggest that IgE-allergen interaction on the surfaces of mast cells or on infiltrating basophils causes both immediate and late cutaneous responses.

The complement system in bullous pemphigoid. II. Immunofluorescent evidence for both classical and alternate-pathway activation

Abstract Skin lesions of 15 bullous pemphigoid patients were examined for the presence of Clq, C3, C3 proactivator (C3PA), and properdin in addition to immunoglobulins (IgG, IgA, and IgM) and fibrin using immunofluorescent (IF) techniques. Properdin and C3 were present along the skin basement-membrane zone in all of the lesions studied in addition to immunoglobulins. Deposition of C3PA was found in only 5 of 15 skin speciments with considerably less staining intensity than that observed for C3 and properdin. Deposition of Clq was present in 12 of 15 skin lesions while fibrin deposition was present in 11 of 15 skin lesions. Absorption of anti-C3PA and antiproperdin with C3PA and properdin, respectively, blocked the specific staining reactions. These studies, therefore, suggest local activation of complement, by both the classical and alternate pathways, in bullous pemphigoid skin lesions.

The Complement System in Bullous Pemphigoid. I. COMPLEMENT AND COMPONENT LEVELS IN SERA AND BLISTER FLUIDS

Compared with other serum and blister fluid proteins, total hemolytic complement was reduced in the blister fluid of six serologically positive bullous pemphigold patients while four serologically negative cases had blister fluid complement levels closely approaching the serum levels. Except for pemphigus vulgaris blisters. blister fluids from most patients with other bullous diseases and experimentally induced blisters had blister fluid complement levels more closely approaching the serum levels. With the exception of the two terminal components. C8 and C9, individual components of the complement sequence were also reduced in the blister fluids of the six bullous pemphigold patients with circulating basement membrane zone antibodies. On the other hand, transferrin and IgG levels of these same six serologically positive blister fluids closely approached the corresponding serum levels. Conversion of C3 proactivator was also demonstrable in the serologically positive bullous pemphigoid blister fluids, but not in the corresponding sera. Our studies, therefore, are suggestive of local activation of the complement sequence, by both the classical and alternate pathways, in blisters of serologically positive bullous pemphigold patients.

Cutaneous vascular immunofluorescence in rheumatoid arthritis. Correlation with circulating immune complexes and vasculitis

The presence of immunoglobulin and complement in the cutaneous blood vessels of clinically uninvolved forearm skin was studied in 70 patients with rheumatoid arthritis, using immunofluorescent techniques. Patients with evidence of these immune deposits had a greater prevalence of circulating immune complexes, vasculitic skin lesions, subcutaneous nodules, high titer rheumatoid factor and other findings suggestive of active vasculitis. Biopsy of uninvolved forearm skin may be a useful tool in assessing those patients with rheumatoid arthritis suspected of having a systemic vasculitis.

Circulating immune complexes in cutaneous vasculitis. Detection with C1q and monoclonal rheumatoid factor.

To investigate the pathogeneic significance of immune complexes in cutaneous vasculitis, 107 patients with various forms of cutaneous vasculitis, including 59 patients with necrotizing (leukocytoclastic) vasculitis (group 1), and 48 patients with lymphocytic vasculitis, or a predominately lymphocytic perivascular infiltrate (group 2), were studied. Immunoglobulins or complement components in cutaneous blood vessels were detected by direct immunofluorescence in high frequency in both groups (91 and 88%, respectively). Using two radioassays for circulating immune complexes, Clq or monoclonal rheumatoid factor (mRF) reactive material was detected in 68% of the patients with necrotizing vasculitis but only 44% of the patients in the lymphocytic-perivascular group. The mRF radioassay was elevated in 58% of the first group of patients and 41% of the patients in group 2, although Clq binding activity was increased in 54% of the patients with necrotizing vasculitis but only in 9% of the patients with a lymphocytic vasculitis or lymphocytic perivascular infiltrate. By using both sucrose density gradient ultracentrifugation and Sepharose 6B gel filtration, the Clq and mRF reactive material detected in some patients with necrotizing vasculitis eluted in high molecular weight fractions that were also anticomplementary. In one patient with necrotizing vasculitis and hepatitis B antigenemia, these heavy molecular weight Clq and mRF reactive fractions contained a two- to three-fold increase in hepatitis B surface antigen when compared with lighter molecular weight fractions. Heavy and light molecular weight mRF reactive material could be detected in selected patients in the lymphocytic-perivascular group as well as in the necrotizing vasculitis group. These studies suggest that cutaneous vasculitis, including acute necrotizing (leukocytoclastic) vasculitis and some forms of lymphocytic vasculitis, and perhaps some diseases characterized by a lymphocytic perivascular infiltrate, may represent cutaneous expressions of immune complex disease.

Dermal‐epidermal deposition of complement components and properdin in systemic lupus erythematosus

Skin lesions and clinically normal skin of thirteen patients with systemic lupus erythematosus (SLE) were examined, by the use of immunofluorescent techniques, to determine the presence of Ciq, C3, C3 proactivator (C3PA), properdin, immunoglobulins (IgG, IgA, and IgM), and fibrin. IgM deposition was present in all thirteen skin lesions and in all eleven normal areas tested, whereas IgG deposition occurred in eleven of the lesions but in only six normal areas. IgA was the least frequently encountered immunoglobulin.

The complement system in bullous pemphigoid: IV. Chemotactic activity in blister fluid

Abstract By a modified Boyden technique, chemotactic activity was present in bullous pemphigoid blister fluids but was also present in the corresponding sera. Heat inactivation (56°C for 30 min) only partially reduced the chemotactic activity of the blister fluid but almost completely inhibited the activity in pemphigoid sera. Both suction-induced and cantharidin-induced blister fluids exhibited some chemotactic activity but, in contrast to pemphigoid blister fluids, this activity was almost entirely abolished by heat inactivation. Blister fluids from some patients with other blistering skin diseases, however, did exhibit both heat-labile and heat-stable chemotactic activity. The chemotactic activity remaining in heat-inactivated pemphigoid blister fluid was inhibited by N-CBZ-α-glutamyl- l -tyrosine and by antiserum to C5 but not with antiserum to C3. Our studies suggest that complement-dependent chemotactic activity is present in bullous pemphigoid blister fluids, a finding that further implicates complement activation in the pathogenesis of this disease.

Direct immunofluorescence in the diagnosis of scleroderma syndromes.

Immunofluorescent study of the skin of nine patients with mesenchymal, inflammatory scleroderma (mixed connective tissue disease) revealed immunoglobulin and complement deposition at the basement membrane or within blood vessel walls. The skin specimens of ten patients with systemic scleroderma were negative for immunofluorescence. It is proposed that basement membrane or vascular (or both) immunofluorescence is an excellent means of identifying the infrequent patient who has scleroderma and myositis or lupus erythematosus in whom a corticosteroid response may occur.

Serum and Blister Fluid Anticomplementary Activity in Pemphigus and Bullous Pemphigoid. Sucrose Density Gradient Studies

Summary By sucrose density gradient ul-tracentrifugation and standard hemolytic complement assays, complement fixing activity of about 19 S and greater was found in blister fluids of one patient with pemphigus and three of five patients with bullous pem-phigoid. Control blister fluids, including experimentally induced blisters, lacked such activity. Complement fixing activity was apparent, however, in 6 of 10 pemphigus sera and in two of five bullous pemphigoid sera tested. The material present in both pemphigus and bullous pemphigoid appears to activate the classical complement pathway.

Systemic lupus erythematosus diagnosed during interferon alfa therapy

We describe a patient who had clinical manifestations of several autoimmune disorders: Sjögrens syndrome, benign hypergammaglobulinemic purpura of Waldenström, and systemic lupus erythematosus (SLE). The SLE was diagnosed during therapy with interferon alfa. Testing for anti-Ro and anti-La antibodies was negative until the serum was diluted to eliminate a possible prozone phenomenon of antibody excess.