Detection of circulating immune comlexes in some skin diseases by platelet aggregation test

Abstract

Since the observation of immune complexes in the circulation of patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis, much attention has been drawn to immune complexes as pathogenic factors. We here report the presence of immune complexes in the sera of patients with several kinds of skin diseases using the platelet aggregation (PA) test (Penttinen, 1977).The 128 fresh or stored (-8o°C) sera were from patients with the active stage of several skin diseases. Detection of immune complexes was performed by the PA test (Penttinen, 1977) with some modifications. Platelet suspensions were prepared as follows: 20 ml of venous blood were collected into 5 ml of anticoagulant solution. The platelet rich plasma was obtained by centrifugation at 200 g for 15 min. Platelets were then sedimented by centrifugation at 1000 g for 15 min and the pellet was washed with PBS. Finally, the platelets were suspended i-5-2-ox io'/mm^ in PBS (pH 7-8). The PA test was performed by using disposable U-microplates. PBS (pH 7-8, 100 /il) and twofold serial dilutions of tested sera (100 /il) were mixed in a U-microplate, and then the platelet suspensions (50 fil) were added to each well. After pipettings, the plates were incubated at 4°C for 18-20 h in a moisture chamber and aggregation patterns were read.The sensitivity of the PA test was shown by its ability to detect i /fg/ml of BSA, 2-5 /ig/ml of anti-BSA antibody and 6-3 ;Ug/ml of aggregated human IgG.PA titres of the tested sera are shown in Fig. i. All of the twenty-three sera from healthy volunteers showed PA titres of below 8. PA titres above 8 were noted in a considerable number of sera from the patients with erythema nodosum (78%), progressive systemic sclerosis (PSS) (71%), SLE (69%), pemphigus (56%), cutaneous angiitis (50%), generalized drug eruption (40%) and erythema multiforme (36%).The PA test is based on the phenomenon of interaction of immune complexes with the Fc receptors on the surface of platelets and has been proven to be one of the most sensitive assays for the detection of circulating large (19 s) immune complexes, although factors other than immune complexes (e.g. aggregated IgG, platelet antibodies and enzymes such as trypsin and thrombin) are also potent platelet agglutinators (Penttinen, 1977).In the present study, we have found the high PA titres in a considerable number of sera from patients with erythema nodosum, PSS, SLE, pemphigus, cutaneous angiitis, generalized drug eruption and erythema multiforme.With regard to collagen diseases such as SLE and PSS, the presence of circulating immune complexes has been mentioned by many authors. Jordon et al. (1976) suggested that pemphigus was an immune complex disease where local activation of complement was noted in blister fluids. Tappeiner et al. (1977) showed immune complexes in pemphigus sera by Clq-binding assay. Our results confirmed this.There have been several reports suggestive of immune complex formation in the sera of patients with erythema multiforme (Kazmierowski & Wuepper, 1977) and c...

Demonstration of circulating and tissue-fixed immune complexes in cutaneous necrotizing vasculitis

“…(2) described the presence of circulating immune complexes in all 6 patients with allergic vasculitis and 2 of 3 patients with livedo vasculitis, using the m1c I q deviation test. U sing the platelet aggrega tion test, we reported the presence of immune com plexes in the sera of 4 of 8 patients with cutaneous allergic vasculitis ( 13). These reports are consistent with our present results.…”

Treatment of Raynaud's phenomenon by intravenous infusion of prostacyclin (PGI2)

“…Modulation of immunological phenomena associated with systemic sclerosis and Raynaud's phenomenon (Van der Meulen et al, 1979;Cormane, Hamcrhorick & Nurizi, 1979;Yanase & Imamura, 1979) and alteration of neutrophil function may be the basis of the therapeutic response to prostacyclin.…”

Impaired Histamine Metabolism in Human Erythematous Dermatoses

“…Although accumulating evidence suggests that immune complex-mediated mechanisms are involved in the development of erythema multiforme and erythema nodosum (1)(2)(3)(4), the details are still obscure. As we have already demonstrated circulating and tissue-deposited immune complexes in patients with erythema multiforme (I), further studies to reveal the pathomechanisms for the development of erythematous dermatoses were attempted.…”

“…In spite of the multiplicity of etiologies, striking similarities in the clinical figures from each disease suggest that some common pathomechanisms exist. Although immunologic processes, especially immune complex-mediated ones (1)(2)(3)(4), are now considered to play important roles in the development of erythema multiforme and erythema nodosum, detailed mechanisms of how they act dynamically in skin tissue to produce lesions are still obscure. We previously reported impaired histamine metabolism in the Arthus reaction, a representative model of immune complex-mediated inflammation in guinea pig skin (5).…”