Systemic Lupus Erythematosus: A.Studies on an ldiotype Specific Dendritic Cell Vaccine. B.Association of Lupus Calcinosis with Calcifying Nanoparticles.

Systemic lupus erythrematosus (SLE) is an autoimmune disease characterized
by the production of anti-DNA antibodies. The primary goal of this study was to
activate T cells with specificity toward lupus B cells presenting anti-DNA antibody
idiotopes on their surface. Monocyte derived dendritic cells were obtained from
peripheral blood of healthy donors and lupus patients. Affinity purified anti-DNA
antibodies were obtained from lupus patients' plasmas. The efficacy of different
carrier proteins, conjugated to lgG, was evaluated, and KLH found to be the most
efficient for antigen uptake. The cognate dendritic cells were evaluated for their
capacity to activate autologous T cells, and generate a Th1 mediated response which was evaluated by proliferation assays and interferony secretion. During the
vaccine study a patient presented with panniculitis, CREST syndrome and a
calcinotic exudate. A secondary goal of my study was to analyze this exudate.
Calcifying nanoparticles were isolated in lymphocyte culture medium. They were
characterized by Von Kassa staining for hydroxyapatite, solubilization by the
calcium chelating agent EDT A, and light and scanning electron microscopy. A
novel method was developed, using a specific monoclonal antibody, to analyze
the calcifying nanoparticles. This method allowed for an approximate
quantification of the particles. These particles increased in numbers when
incubated for different time periods, and their numbers were decreased when
incubated in the presence of minocyclin. Concomitantly, the panniculi in the
patient underwent remission with long term antibiotic therapy. CNPs were also
obtained from fetal bovine serum and human plasma samples from both lupus
patients and healthy donors. Peripheral blood mononuclear cells from healthy
donors and lupus patients were analyzed in vitro for their reactivity when
incubated in the presence of a biofilm, generated by the calcifying nanoparticles.
Viability, proliferation, and co-stimulatory marker up-regulation were determined
in the presence or absence of the particles. Osteopontin was found highly
expressed in the supernatants of the cells grown with CNPs. Microarray of the
mononuclear cells of a healthy donor and a lupus patient incubated in the
presence or absence of CNPs was performed, and the results coincided with
those determined in vitro.