Carmen Gelpí

Enzyme‐linked immunosorbent assay (ELISA) and indirect immunofluorescence testing in a bullous pemphigoid and pemphigoid gestationis

Enzyme‐linked immunosorbent assay (ELISA) is an excellent tool for detection of circulating antibodies against the NC16A portion of BP180 antigen. We compared the sensitivity and specificity of a commercially available BP180‐NC16a domain ELISA with that of an indirect immunofluorescence (IIF) testing in the evaluation of bullous pemphigoid (BP) and pemphigoid gestationis (PG), and analyzed the relationship between ELISA results and the presence of IgG deposition, in an epidermal or combined pattern, on direct immunofluorescence (DIF) testing of salt‐split skin. ELISA was performed on serum from 28 patients (24 BP, 4 PG) and 50 controls. IIF testing was performed on serum from 27 patients and 98 controls. For the group of 28 patients with BP or PG, ELISA had a sensitivity of 93% and specificity of 96% (P < 0.001), while sensitivity was 74% and specificity 96% (P < 0.001) for IIF testing. In these patients, ELISA has a higher sensitivity than IIF testing, but similar specificity. Evaluation of controls who had IgG deposition on the dermal side of salt‐split skin on DIF testing showed specificity for the ELISA of 100% (all four cases negative) and 80% for IIF testing (one of five positive). Positive ELISA correlated with a diagnosis of BP or PG only in patients who had IgG at the basement membrane zone (BMZ) by DIF testing. Overall, ELISA appears to have greater sensitivity and specificity for BP or PG than does IIF testing.

Bullous pemphigoid in a patient with psoriasis during the course of PUVA therapy: study by ELISA test

A 65‐year‐old woman had a history of deep vein thrombosis and depression. Psoriasis was diagnosed in 1986 and various topical and systemic therapies, singly or in combination, were prescribed: tar, topical corticosteroids, cyclosporine, etretinate, and methotrexate. Two courses of oral and one course of bath psoralen plus UVA (PUVA) therapy (cumulative dose, 467 J/cm2) and UVB (2.96 J/cm2) had been given.

Paraneoplastic pemphigus with negative direct immunofluorescence in epidermis or mucosa but positive findings in adnexal structures

Introduction:  Paraneoplastic pemphigus (PNP) is considered an autoimmune, multiorgan disease caused by antiplakin antibodies. We present three PNP patients who had negative epithelial direct immunofluorescence (DIF) findings in one or more biopsies.

Usefulness of specific anti-desmoglein 1 and 3 enzyme-linked immunoassay and indirect immunofluorescence in the evaluation of pemphigus activity

The objective was to assess the relationship between enzyme‐linked immunoassay (ELISA) values of desmoglein (Dsg) 1 and Dsg3 antibodies and indirect immunofluorescence (IIF) values of anti‐epithelial antibodies with disease activity in patients with pemphigus.

Acquired bullous dermatosis associated with IgA multiple myeloma: a case report.

We present the case of a patient with IgA paraprotein who developed hemorrhagic subepidermal vesicles and bullae with numerous neutrophils. Direct immunofluorescence test (DIF) showed weak deposits of IgA lambda paraprotein at the dermal–epidermal junction and at the intercellular level in the basal layer of the epidermis, and stronger deposits in a perivascular and diffuse pattern in the dermis. Indirect immunofluorescence (IIF) test revealed the presence of circulating IgA lambda antibodies reacting with the intercellular space of monkey and guinea pig esophagus and human skin. A blood test revealed an IgA lambda paraprotein and multiple myeloma stage I(0) was diagnosed in a later hematological study. Dapsone was prescribed and cutaneous lesions improved. This is the second report of subepidermal vesicles and bullae with dermal deposits of IgA paraprotein appearing prior to diagnosis of an IgA multiple myeloma, and it is a unique case with circulating IgA lambda antibodies reacting with the intercellular space of epithelia.

The IL-12 Role in Donor Cell Engraftment in a Murine Model of Semiallogenic GVH Disease with Signs of Autoimmune Disease

We analyzed the IL-12 effect in an autoimmune disease induced in a semiallogenic murine model of graft-vs-host disease (GVHD) Balb/c semiallogenic lymphoid cells i.v. infected in hybrid mice (Balb/c×A/J) F1 (CAF). IL-12 was administered 1 h before cell transplantation following two different protocols: (a) injecting 2 μg of mrIL-12 (murine recombinant IL-12) per mouse before the first semiallogenic cell injection; or (b) injecting the 2 μg of mrIL-12 fractionated in 5 days. A Th1 response was produced but an acute GVHD did not appear although differences in class I and II major histocompatibility complex (MHC) antigens were present. Four days after the semiallogenic cell transfer, IL-12 treated mice showed a marked reduction in the percent of spleen B cells compared with CAF1 control and CAF1+Balb/c GVHD mice. After 5–6 months of follow-up, the donor cell chimerism increased significantly in spleen (70±31 vs. 43±31%) and in thymus. Flow cytometry of spleen lymphocytes demonstrated that donor chimerism was made up of TCD4, TCD8 and B lymphocytes and was higher in animals injected with IL-12. Moreover, CD8 T lymphocytes were 100% donor origin in the IL-12-injected group of GVHD animals and 50% origin in the IL-12-non-injected CAF1+Balb/c group of animals. This paper shows that: (1) IL-12 may play a role in the mechanisms of donor cell engraftment, probably produced by a CTL donor anti-host mechanism; (2) no acute GVHD was induced in spite of class I and II MHC differences; (3) IL-12 did not show any effect on the AR-like clinical signs of disease developed in this model of GVHD although histological subclinical signs were less frequent, and no glomerulonephritis was detected in the IL-12-treated GVHD mice.

Effect of enzymatic treatments on RNP and Sm antigenic reactivities—I. Loss of RNP but increase of Sm antigenic reactivity after RNase treatment of nuclear extract

Changes of RNP and Sm antigenic reactivities of a nuclear extract after enzymatic treatments were studied and quantified by the ELISA test. After RNase treatment of the nuclear extract, about a 300% increase of the Sm antigenic reactivity and more than a 95% decrease of RNP antigenic reactivity was found. Data from RNP-depleted nuclear extracts and column fractionation show that the increase in Sm antigenic reactivity after RNase treatment mainly comes from the RNP-Sm complex.